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2018 Book PrinciplesOfFoodChemistry PDF

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Diana Uscanga
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Food Science Text Series

John M. deMan
John W. Finley
W. Jeffrey Hurst
Chang Yong Lee

Principles of
Food Chemistry
Fourth Edition
Food Science Text Series

Series Editor
Dennis R.  Heldman, Professor, Department of Food, Agricultural, and
Biological Engineering, The Ohio State University

Editorial Board
John Coupland, Professor of Food Science, Department of Food Science,
Penn State University
Mario Ferruzzi, Professor of Food Science and Nutrition, Department of
Food Bioprocessing and Nutrition, North Carolina State.
Richard W. Hartel, Professor of Food Engineering, Department of Food
Science, University of Wisconsin
Rubén Morawicki, Assistant Professor of Food Science, Department of Food
Science, Universisty of Arkansas
S. Suzanne Nielsen, Professor and Chair, Department of Food Science, Purdue
University
Juan L. Silva, Professor, Department of Food Science, Nutrition and Health
Promotion, Mississippi State University
The Food Science Text Series provides faculty with the leading teaching tools.
The Editorial Board has outlined the most appropriate and complete content
for each food science course in a typical food science program and has
identified textbooks of the highest quality, written by the leading food science
educators.

More information about this series at http://www.springer.com/series/5999


John M. deMan  •  John W. Finley
W. Jeffrey Hurst  •  Chang Yong Lee

Principles of Food
Chemistry

Fourth Edition
John M. deMan (Deceased) John W. Finley
Department of Food Science Louisiana State University
University of Guelph Lakewood Ranch, FL, USA
Guelph, ON, Canada
Chang Yong Lee
W. Jeffrey Hurst Department of Food Science
The Hershey Company Technical Center Cornell University
Hershey, PA, USA Ithaca, NY, USA

ISSN 1572-0330     ISSN 2214-7799 (electronic)


Food Science Text Series
ISBN 978-3-319-63605-4    ISBN 978-3-319-63607-8 (eBook)
https://doi.org/10.1007/978-3-319-63607-8

Library of Congress Control Number: 2017956195

1st edition: © AVI 1980


2nd edition: © AVI 1989
© Springer International Publishing AG 1999, 2018
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or
part of the material is concerned, specifically the rights of translation, reprinting, reuse of
illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way,
and transmission or information storage and retrieval, electronic adaptation, computer software,
or by similar or dissimilar methodology now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publication does not imply, even in the absence of a specific statement, that such names are
exempt from the relevant protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in
this book are believed to be true and accurate at the date of publication. Neither the publisher nor
the authors or the editors give a warranty, express or implied, with respect to the material
contained herein or for any errors or omissions that may have been made. The publisher remains
neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Printed on acid-free paper

This Springer imprint is published by Springer Nature


The registered company is Springer International Publishing AG
The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland
Preface

This book was designed to serve as an introductory text for courses in food
chemistry as part of food science programs meeting the Institute of Food
Technologists standards. The original concept for the preparation of this book
was to present basic information on the composition of foods and the chemical
and physical characteristics they undergo during processing, storage, and han-
dling. The basic principles of food chemistry remain the same, but much addi-
tional research carried out in recent years has expanded and in some cases
refined our knowledge. As with the third edition we have refined and expanded
the material in all chapters. Because of the rapidly growing interest we have
added chapters on transgenic crops as well as a chapter on beer and wine pro-
duction. We felt the transgenic crop chapter was important so that students
have a basic understanding of the technology and how it has evolved over the
last 10,000 years. The chapter on beer and wine production is included to help
the students appreciate the science behind fermented beverages. This knowl-
edge will be valuable because the opportunities for food scientists in those
areas are growing exponentially. In the area of water as a food component, the
issue of the glass transition has received much attention. This demonstrates the
important role of water in food properties. Carbohydrates and lipids are of
major sources of food energy and are of major interest for their functional and
nutritional properties in obesity and diabetes. Understanding how to the che­
mistry of these ingredients will help food scientists better formulate new nutri-
tionally superior foods in the future. Our understanding of the functionality of
proteins expands with increasing knowledge about their composition and
structure. Carbohydrates serve many functions in foods, and the non-caloric
dietary fiber has assumed an important role.
Color, flavor, and texture are important attributes of food quality, and in
these areas, especially those of flavor and texture, great advances have been
made in recent years. There is concern among consumers about the safety of
additives including colors and flavors. We have also included a section on
natural toxicants as well as ingredients that can cause adverse effects. It is
important to realize that many components in foods can be harmful or safe
depending on the concentrations in the foods. Enzymes are playing an ever
increasing part in the production and transformation of foods. Modern meth-
ods of biotechnology have produced a gamut of enzymes with new and
improved properties.
In the literature, information is found using different systems of units:
metric, SI, and the English system. Quotations from the literature are

v
vi Preface

p­ resented in their original form. It would be difficult to change all these units
in the book to one system. To assist the reader in converting these units, an
appendix is provided with conversion factors for all units found in the text.
It is hoped that this fourth edition will continue to fulfill the need for a
concise and relevant text for the teaching of food chemistry. We hope that this
edition will serve as a memorial to the enormous contributions of John deMan
and continue to provide teaching and reference material of value.

Guelph, ON John M. deMan


Lakewood Ranch, FL  John W. Finley
Hershey, PA  W. Jeffrey Hurst
Ithaca, NY  Chang Yong Lee
Contents

1 Water��������������������������������������������������������������������������������������������    1
Yrjo H. Roos, John W. Finley, and John M. deMan
Water in Foods ������������������������������������������������������������������������������    1
Physical Properties of Water and Ice ��������������������������������������������    1
Structure of the Water Molecule����������������������������������������������������    4
Structure of Ice������������������������������������������������������������������������������    6
Growth of Ice Crystals ������������������������������������������������������������������    7
Latent Heat of Fusion��������������������������������������������������������������������    7
Solidification Without Crystallization��������������������������������������������    8
Surface Tension of Water ��������������������������������������������������������������   10
Examples of Surface Tension����������������������������������������������������   11
Colligative Properties of Aqueous Solutions ��������������������������������   11
Freezing Point Depression ��������������������������������������������������������   11
Boiling Point Elevation��������������������������������������������������������������   12
Osmotic Pressure ����������������������������������������������������������������������   12
Vapor Pressure Lowering ����������������������������������������������������������   12
Ionic Interactions Are Attractions Between Oppositely
Charged Ions������������������������������������������������������������������������������   13
Properties of Hydrogen Bonds������������������������������������������������������   15
Hydrophobic Interactions��������������������������������������������������������������   16
Water Activity and Sorption Phenomena ��������������������������������������   17
Types of Water ������������������������������������������������������������������������������   24
Phase Diagram ������������������������������������������������������������������������������   26
The Glass Transition����������������������������������������������������������������������   27
Water Activity and Reaction Rate��������������������������������������������������   30
Water Activity and Food Spoilage ������������������������������������������������   30
Water Activity and Packaging��������������������������������������������������������   33
Water Activity and Food Processing����������������������������������������������   34
References��������������������������������������������������������������������������������������   35
2 Lipids��������������������������������������������������������������������������������������������   39
John W. Finley and John M. deMan
Shorthand Description of Fatty Acids and Glycerides������������������   40
Fatty Acids ������������������������������������������������������������������������������������   43
Lipid Nomenclature ������������������������������������������������������������������   43
Cis and trans Fatty Acids ����������������������������������������������������������   44

vii
viii Contents

Triglycerides����������������������������������������������������������������������������������   48
Component Glycerides������������������������������������������������������������������   53
Waxes ����������������������������������������������������������������������������������������   58
Phospholipids����������������������������������������������������������������������������   60
Unsaponifiables������������������������������������������������������������������������������   63
Terpenes ������������������������������������������������������������������������������������   63
Steroids������������������������������������������������������������������������������������������   63
Phytosterols������������������������������������������������������������������������������������   66
Lipid Reactions������������������������������������������������������������������������������   68
Fatty Acid Salts��������������������������������������������������������������������������   68
Hydrolysis����������������������������������������������������������������������������������   68
Interesterification ����������������������������������������������������������������������   68
Hydrogenation��������������������������������������������������������������������������������   72
Lipid Oxidation������������������������������������������������������������������������������   78
Initiation������������������������������������������������������������������������������������   79
Propagation��������������������������������������������������������������������������������   79
Termination��������������������������������������������������������������������������������   79
Photooxidation ������������������������������������������������������������������������������   88
Heated Fats: Frying������������������������������������������������������������������������   90
Flavor Reversion����������������������������������������������������������������������������   93
Physical Properties������������������������������������������������������������������������   95
Fractionation����������������������������������������������������������������������������������  105
Emulsions and Emulsifiers������������������������������������������������������������  106
Fat Replacers����������������������������������������������������������������������������������  110
References��������������������������������������������������������������������������������������  113
3 Amino Acids and Proteins ����������������������������������������������������������  117
Michael Appell, W. Jeffrey Hurst, John W. Finley,
and John M. deMan
Introduction������������������������������������������������������������������������������������  117
Amino Acids����������������������������������������������������������������������������������  117
Peptides and Proteins ��������������������������������������������������������������������  118
Protein Classification ��������������������������������������������������������������������  120
Simple Proteins��������������������������������������������������������������������������  120
Conjugated Proteins ������������������������������������������������������������������  123
Derived Proteins������������������������������������������������������������������������  123
Protein Structure����������������������������������������������������������������������������  123
Denaturation����������������������������������������������������������������������������������  125
Non-enzymic Browning����������������������������������������������������������������  127
Chemical Changes ������������������������������������������������������������������������  136
Functional Properties ��������������������������������������������������������������������  139
Surface Activity of Proteins ������������������������������������������������������  140
Gel Formation����������������������������������������������������������������������������  142
Animal Proteins ����������������������������������������������������������������������������  142
Milk Proteins������������������������������������������������������������������������������  142
Meat Proteins ��������������������������������������������������������������������������������  147
Meromysin ��������������������������������������������������������������������������������  147
Contents ix

Collagen ����������������������������������������������������������������������������������������  149


Fish Proteins������������������������������������������������������������������������������  152
Egg Proteins ����������������������������������������������������������������������������������  153
Plant Proteins ��������������������������������������������������������������������������������  154
Wheat Proteins ��������������������������������������������������������������������������  154
Maize Proteins ������������������������������������������������������������������������������  156
Rice Proteins����������������������������������������������������������������������������������  157
Soybean Proteins����������������������������������������������������������������������������  158
Gluten Sensitivity��������������������������������������������������������������������������  161
Test Methods������������������������������������������������������������������������������  161
References��������������������������������������������������������������������������������������  162
4 Carbohydrates������������������������������������������������������������������������������  165
Gillian Eggleston, John W. Finley, and John M. deMan
Introduction������������������������������������������������������������������������������������  165
Monosaccharides����������������������������������������������������������������������������  166
Related Compounds to Monosaccharides��������������������������������������  171
Amino Sugars����������������������������������������������������������������������������  171
Glycosides����������������������������������������������������������������������������������  172
Sugar Alcohols ��������������������������������������������������������������������������  172
Oligosaccharides����������������������������������������������������������������������������  175
Disaccharides ����������������������������������������������������������������������������  175
Chemical Reactions of Sugars ��������������������������������������������������  179
Compounds Related to Sugars ��������������������������������������������������  191
Polysaccharides������������������������������������������������������������������������������  191
Starch ����������������������������������������������������������������������������������������  192
Starch Degrading Enzymes��������������������������������������������������������  200
High Fructose Corn Syrup (HFCS)��������������������������������������������  201
Starch Hydrolyzates: Corn Sweeteners��������������������������������������  202
Modified Starches����������������������������������������������������������������������  202
Glycogen������������������������������������������������������������������������������������  207
Cellulose������������������������������������������������������������������������������������  209
Pentosans/Hemicelluloses����������������������������������������������������������  210
Lignin����������������������������������������������������������������������������������������  211
Cyclodextrins ����������������������������������������������������������������������������  212
Polydextrose������������������������������������������������������������������������������  214
Pectins����������������������������������������������������������������������������������������  215
Gums������������������������������������������������������������������������������������������  216
Dietary Fiber������������������������������������������������������������������������������  222
References��������������������������������������������������������������������������������������  226
5 Minerals����������������������������������������������������������������������������������������  231
John W. Finley and John M. deMan
Introduction������������������������������������������������������������������������������������  231
Interactions with Other Food Components��������������������������������  236
Major Minerals������������������������������������������������������������������������������  237
Sodium ��������������������������������������������������������������������������������������  237
Potassium ����������������������������������������������������������������������������������  237
Magnesium��������������������������������������������������������������������������������  238
x Contents

Calcium��������������������������������������������������������������������������������������  238
Phosphates����������������������������������������������������������������������������������  238
Minerals in Milk������������������������������������������������������������������������  239
Minerals in Meat������������������������������������������������������������������������  241
Minerals in Plant Products ��������������������������������������������������������  242
Chloride��������������������������������������������������������������������������������������  243
Trace Elements������������������������������������������������������������������������������  244
Iron��������������������������������������������������������������������������������������������  244
Cobalt����������������������������������������������������������������������������������������  245
Copper����������������������������������������������������������������������������������������  245
Zinc��������������������������������������������������������������������������������������������  245
Manganese ��������������������������������������������������������������������������������  245
Molybdenum������������������������������������������������������������������������������  246
Selenium������������������������������������������������������������������������������������  246
Fluorine��������������������������������������������������������������������������������������  246
Iodine ����������������������������������������������������������������������������������������  246
Chromium����������������������������������������������������������������������������������  246
Additional Information on Trace Elements��������������������������������  247
Metal Uptake in Canned Foods������������������������������������������������������  247
References��������������������������������������������������������������������������������������  250
6 Color and Food Colorants����������������������������������������������������������  253
John W. Finley, John M. deMan, and Chang Yong Lee
Introduction������������������������������������������������������������������������������������  253
CIE System������������������������������������������������������������������������������������  253
Munsell System������������������������������������������������������������������������������  258
Hunter System��������������������������������������������������������������������������������  260
Lovibond System ��������������������������������������������������������������������������  261
Gloss����������������������������������������������������������������������������������������������  262
Food Colorants������������������������������������������������������������������������������  262
Certified (Synthetic) Color��������������������������������������������������������  263
Colors Exempt from Certification (Natural)������������������������������  263
Tetrapyrrole Pigments��������������������������������������������������������������������  266
Myoglobins��������������������������������������������������������������������������������  266
Chlorophylls������������������������������������������������������������������������������  268
Isoprenoid Derivative Pigments ����������������������������������������������������  270
Carotenoids��������������������������������������������������������������������������������  270
Benzopyran Derivative Pigments��������������������������������������������������  276
Anthocyanins and Flavonoids����������������������������������������������������  276
Other Pigments������������������������������������������������������������������������������  281
Betalains������������������������������������������������������������������������������������  281
References��������������������������������������������������������������������������������������  283
7 Flavor��������������������������������������������������������������������������������������������  285
Han-Seok Seo, John W. Finley, and John M. deMan
Taste Sensations ����������������������������������������������������������������������������  290
Chemical Structure and Taste��������������������������������������������������������  290
Sweet Taste������������������������������������������������������������������������������������  293
Sour Taste��������������������������������������������������������������������������������������  294
Contents xi

Salty Taste��������������������������������������������������������������������������������������  295


Bitter Taste ������������������������������������������������������������������������������������  298
Other Aspects of Taste ������������������������������������������������������������������  301
Taste Inhibition and Modification��������������������������������������������������  303
Flavor Enhancement—Umami������������������������������������������������������  304
Odor ����������������������������������������������������������������������������������������������  306
Odor and Molecular Structure ������������������������������������������������������  310
Description of Food and Beverage Flavors������������������������������������  314
Astringency������������������������������������������������������������������������������������  316
Flavor and Off-Flavor��������������������������������������������������������������������  317
Flavor of Some Foods��������������������������������������������������������������������  319
Bread������������������������������������������������������������������������������������������  319
Meat ������������������������������������������������������������������������������������������  320
Fish��������������������������������������������������������������������������������������������  320
Milk��������������������������������������������������������������������������������������������  321
Cheese����������������������������������������������������������������������������������������  321
Fruits������������������������������������������������������������������������������������������  322
Vegetables����������������������������������������������������������������������������������  322
Tea����������������������������������������������������������������������������������������������  322
Coffee����������������������������������������������������������������������������������������  323
Alcoholic Beverages������������������������������������������������������������������  324
References��������������������������������������������������������������������������������������  325
8 Texture������������������������������������������������������������������������������������������  329
Harry Levine and John W. Finley
Introduction������������������������������������������������������������������������������������  329
Fluids���������������������������������������������������������������������������������������������  330
Texture of Solids����������������������������������������������������������������������������  333
Texture Profile��������������������������������������������������������������������������������  334
Measurement of Texture����������������������������������������������������������������  335
Force and Stress ������������������������������������������������������������������������  336
Deformation and Strain��������������������������������������������������������������  336
Principles of Measurement��������������������������������������������������������  337
Different Types of Bodies��������������������������������������������������������������  337
The Elastic Body������������������������������������������������������������������������  337
The Retarded Elastic Body��������������������������������������������������������  338
The Viscous Body����������������������������������������������������������������������  338
The Viscoelastic Body����������������������������������������������������������������  338
The Plastic Body������������������������������������������������������������������������  339
The Thixotropic Body����������������������������������������������������������������  341
Dynamic Behavior ��������������������������������������������������������������������  341
Rheology Applications in Foods����������������������������������������������������  342
Textural Properties of some Foods������������������������������������������������  348
Meat Texture������������������������������������������������������������������������������  348
Wheat Flour Dough��������������������������������������������������������������������  349
Fats��������������������������������������������������������������������������������������������  350
Fruits and Vegetables ����������������������������������������������������������������  353
Starch ����������������������������������������������������������������������������������������  353
xii Contents

Microstructure��������������������������������������������������������������������������������  355
Water Activity and Texture������������������������������������������������������������  359
References��������������������������������������������������������������������������������������  361
9 Vitamins����������������������������������������������������������������������������������������  365
John W. Finley and John M. deMan
Introduction������������������������������������������������������������������������������������  365
Fat-Soluble Vitamins����������������������������������������������������������������������  367
Vitamin A (Retinol)��������������������������������������������������������������������  367
Vitamin D����������������������������������������������������������������������������������  372
Tocopherols (Vitamin E)������������������������������������������������������������  372
Vitamin K����������������������������������������������������������������������������������  378
Water-Soluble Vitamins ����������������������������������������������������������������  378
Vitamin C (L-Ascorbic Acid)����������������������������������������������������  378
Vitamin B1 (Thiamin)����������������������������������������������������������������  382
Vitamin B2 (Riboflavin) ������������������������������������������������������������  383
Vitamin B6 (Pyridoxine)������������������������������������������������������������  385
Niacin����������������������������������������������������������������������������������������  386
Vitamin B12 (Cyanocobalamine)������������������������������������������������  388
Folic Acid (Folacin) ������������������������������������������������������������������  389
Pantothenic Acid������������������������������������������������������������������������  391
Biotin�����������������������������������������������������������������������������������������  391
Vitamins as Food Ingredients��������������������������������������������������������  392
References��������������������������������������������������������������������������������������  394
10 Enzymes����������������������������������������������������������������������������������������  397
John M. deMan and Chang Yong Lee
Introduction������������������������������������������������������������������������������������  397
Nature and Kinetics of Enzymes����������������������������������������������������  400
Nature of Enzymes��������������������������������������������������������������������  400
Kinetics of Enzymes������������������������������������������������������������������  400
Specificity����������������������������������������������������������������������������������  404
Classification����������������������������������������������������������������������������������  405
Enzyme Production������������������������������������������������������������������������  405
Hydrolases��������������������������������������������������������������������������������������  405
Esterases������������������������������������������������������������������������������������  405
Amylases������������������������������������������������������������������������������������  411
Pectic Enzymes��������������������������������������������������������������������������  413
Proteases������������������������������������������������������������������������������������  415
Protein Hydrolysates������������������������������������������������������������������  419
Oxidoreductases����������������������������������������������������������������������������  421
Phenolases����������������������������������������������������������������������������������  421
Glucose Oxidase (β-d-Glucose: Oxygen Oxidoreductase)��������  423
Catalase (Hydrogen Peroxide: Hydrogen Peroxide
Oxidoreductase) ��������������������������������������������������������������������  423
Peroxidase (Donor: Hydrogen Peroxide Oxidoreductase)��������  424
Contents xiii

Lipoxygenase (Linoleate: Oxygen Oxidoreductase) ����������������  426


Xanthine Oxidase (Xanthine: Oxygen Oxidoreductase)������������  427
Immobilized Enzymes ������������������������������������������������������������������  429
References��������������������������������������������������������������������������������������  432
11 Fruits and  Vegetables������������������������������������������������������������������  435
Chang Yong Lee
Major and Minor Components������������������������������������������������������  435
Water����������������������������������������������������������������������������������������������  435
Carbohydrates��������������������������������������������������������������������������������  436
Proteins and Nitrogenous Compounds������������������������������������������  438
Lipids ��������������������������������������������������������������������������������������������  438
Minor Composition������������������������������������������������������������������������  438
Organic Acids����������������������������������������������������������������������������  438
Phenolic Compounds ����������������������������������������������������������������  441
Minerals ������������������������������������������������������������������������������������  443
Pigments������������������������������������������������������������������������������������  444
Postharvest Deterioration��������������������������������������������������������������  445
Cellular Components and Physiology of Fruits
and Vegetables��������������������������������������������������������������������������������  446
Respiration ��������������������������������������������������������������������������������  447
Environmental Factors that Influence Deterioration������������������  448
Effects of Processing on Fruits and Vegetables ����������������������������  448
Carbohydrate Reactions ������������������������������������������������������������  449
Protein Reactions ����������������������������������������������������������������������  449
Lipid Reactions��������������������������������������������������������������������������  450
Color Change ����������������������������������������������������������������������������  450
Flavor Change����������������������������������������������������������������������������  451
Texture Change��������������������������������������������������������������������������  452
Nutrient Loss������������������������������������������������������������������������������  452
Dehydration of Fruits and Vegetables��������������������������������������������  453
Canning of Fruits and Vegetables��������������������������������������������������  454
Freezing of Fruits and Vegetables��������������������������������������������������  454
Lactic Acid Fermentation��������������������������������������������������������������  454
References��������������������������������������������������������������������������������������  455
12 Herbs and  Spices��������������������������������������������������������������������������  457
Zhuohong Xie and John W. Finley
Black Pepper����������������������������������������������������������������������������������  463
Vanilla��������������������������������������������������������������������������������������������  464
Cardamom��������������������������������������������������������������������������������������  468
Ginger��������������������������������������������������������������������������������������������  469
Turmeric����������������������������������������������������������������������������������������  471
Cinnamon��������������������������������������������������������������������������������������  473
Ginseng������������������������������������������������������������������������������������������  477
Ginkgo��������������������������������������������������������������������������������������������  478
Food Fraud Risks ��������������������������������������������������������������������������  478
References��������������������������������������������������������������������������������������  478
xiv Contents

13 Beer and  Wine������������������������������������������������������������������������������  483


John W. Finley
Alcoholic Fermentation ����������������������������������������������������������������  484
Beer������������������������������������������������������������������������������������������������  486
Raw Materials����������������������������������������������������������������������������  488
Hops ����������������������������������������������������������������������������������������������  489
Yeast����������������������������������������������������������������������������������������������  493
Wine ����������������������������������������������������������������������������������������������  495
Wine Grape Production��������������������������������������������������������������  496
Wine Production������������������������������������������������������������������������  498
References��������������������������������������������������������������������������������������  506
14 Genetically Modified Crops��������������������������������������������������������  511
W. Jeffrey Hurst and John W. Finley
Applications of Genetically Modified Crops ��������������������������������  518
Testing��������������������������������������������������������������������������������������������  521
Regulation��������������������������������������������������������������������������������������  522
Future Challenges��������������������������������������������������������������������������  522
GMO Dictionary������������������������������������������������������������������������  523
Flour������������������������������������������������������������������������������������������  525
References��������������������������������������������������������������������������������������  526
15 Additives and  Contaminants������������������������������������������������������  527
W. Jeffrey Hurst, John W. Finley and John M. deMan
Introduction������������������������������������������������������������������������������������  527
Food Toxins������������������������������������������������������������������������������������  527
Food Additives ������������������������������������������������������������������������������  531
Benzoic Acid������������������������������������������������������������������������������  533
Parabens ������������������������������������������������������������������������������������  533
Sorbic Acid��������������������������������������������������������������������������������  536
Sulfites���������������������������������������������������������������������������������������  536
Nitrates and Nitrites ������������������������������������������������������������������  538
Hydrogen Peroxide��������������������������������������������������������������������  539
Sodium Chloride������������������������������������������������������������������������  539
Bacteriocins��������������������������������������������������������������������������������  540
Antioxidants������������������������������������������������������������������������������  540
Emulsifiers ��������������������������������������������������������������������������������  542
Bread Improvers������������������������������������������������������������������������  543
Incidental Additives or Contaminants��������������������������������������������  549
Pesticides������������������������������������������������������������������������������������  549
Dioxin����������������������������������������������������������������������������������������  551
Polychlorinated Biphenyls (PCBs)��������������������������������������������  552
Antibiotics����������������������������������������������������������������������������������  555
Trace Metals������������������������������������������������������������������������������  556
Mercury��������������������������������������������������������������������������������������  556
Lead and Tin������������������������������������������������������������������������������  559
Contents xv

Cadmium������������������������������������������������������������������������������������  560
Arsenic ��������������������������������������������������������������������������������������  560
Polycyclic Aromatic Hydrocarbons (PAHs)������������������������������  560
Caffeine��������������������������������������������������������������������������������������  562
References��������������������������������������������������������������������������������������  563
Appendix A: Moisture Analysis����������������������������������������������������������  567
Appendix B: Units and Conversion Factors��������������������������������������  573
Appendix C: Greek Alphabet��������������������������������������������������������������  575
Index������������������������������������������������������������������������������������������������������  577
About the Editors

John M. deMan  (1926–2010) was a University Professor Emeritus in the


Department of Food Science at the University of Guelph, Ontario, Canada.
He was the Chairman of the Department and Past President of the Canadian
Institute of Food Science and Technology. He published over 250 papers and
book chapters on multiple aspects of food science and technology. He
received many professional awards, including the Dairy Research Award of
the American Dairy Science Association, the Institute Award of the Canadian
Institute of Food Science and Technology, the Alton E. Bailey Award of the
American Oil Chemist Society, the Stephen S. Chang Award of the Institute
of Food Technologists, and the Kaufmann Memorial Award of the International
Society of Fat Research. He was a Fellow of the Institute of Food Technologists,
the Canadian Institute of Food Science and Technology, and the Malaysian
Oil Science and Technology Association.
John W. Finley  received a B.S. in Chemistry from LeMoyne College and a
Ph.D. from Cornell University. Dr. Finley retired from Louisiana State
University where he was head of the Food Science program from 2007 to
2014. His laboratory studied low calorie ingredients, anti-inflammatory com-
pounds in the diet, modified nutritional lipids, and edible fiber. Previously he
headed Fundamental Science at Nabisco, was a Fellow at Kraft Food, served
as chief technology officer of A.M. Todd Co. and the leader of the Food
Science program at Monsanto, and Research Scientist with the USDA
Regional Research Center.
Dr. Finley is a Fellow of the American Chemical Society, Fellow of
Agricultural and Food Division of the American Chemical Society, Fellow of
the Royal Society of Chemistry, Fellow of the Institute of Food Technologists,
and Certified Food Technologist by Fellow Institute of Food Technologists. He
was recognized as an Outstanding Alumnus of Michigan State University.
Other awards include Harris Distinguished lecturer at the Ohio State University
and a Leadership Award at Nabisco, and his memberships include Sigma Xi at
Michigan State University and Phi Kappa Phi at Cornell University.
Dr. Finley has edited 8 books, holds 70 patents, and 135 publications.
W. Jeffrey Hurst  retired from the Hershey Company as Principal Scientist
after being with the corporation for over 39 years. His research focused on
monitoring new developments in measurement technology as they apply to
food systems and the review of new and emerging compounds important to

xvii
xviii About the Editors

the food industry. He is a member of the American Chemical Society, the


Institute of Food Technologist. He is a member of the American Society of
Mass Spectrometry and Fellow of the American Institute of Chemists (FAIC).
Furthermore, he was named a Fellow of the AOAC, a Pioneer in Laboratory
Robotics, and is a Diplomate of the American Association Integrated
Medicine. Dr. Hurst was a member of the US Air Force and retired as a Major.
He also serves as a member of the External Advisory Board of the University
of Illinois at Chicago NIH bv Botanical Center. This book will be the tenth
one that he has edited or written. He was founding editor of Lab Robotics
Automation and Seminars in Food Analysis. He has numerous patents with
over 300 papers and presentations.
Chang Yong Lee  received a B.S. in Chemistry from Chung-Ang University
in Seoul, Korea, and a Ph.D. from Utah State University. He has been work-
ing as a faculty member at Cornell University since 1969. Professor Lee has
been teaching food chemistry for a number of years in the Department of
Food Science. His research interests have been on biochemical aspects of
plant foods. Recently his laboratory has been working on the bioactivity of
phytochemicals that is related to health benefits. He served as Chair of the
Department of Food Science and Technology and Co-director of Cornell
Institute of Food Science (2002–2008). Dr. Lee has held visiting professor
appointments at several institutions, including Korea Institute of Science and
Technology; Inter-American Institute of Agricultural Science at EMBRAPA,
Brazil; Institut National de la Recherche Agronomique, Avignon, France;
Beijing Vegetable Research Center, China; Ecole Nationale Superieure
des Industries Agricoles et Alimentaire, France; Graduate School of
Biotechnology, Korea University; and Kyung Hee University, Korea.
Professor Lee has authored more than 300 research articles. He was a
recipient of Platinum Award on his edited books on polyphenols from the
American Chemical Society’s Division of Agricultural and Food Chemistry.
Journal of Agricultural and Food Chemistry and the Institute for Scientific
Information (ISI) acknowledged Professor Lee as one of the Highly Cited
Researchers (HCR) in 2004. Thomson Reuters selected him as one of 112
scientists in the world in the field of Agricultural Science during 2002–2012
who published the greatest number of highly cited papers ranked in the top
1% by citations. Again in 2015, Thomson Reuters listed Lee as one of the
World’s Most Influential Scientific Minds in Agricultural Science. Professor
Lee was awarded USDA Secretary’s Honor Award for Excellence in Research
in 2001 and 2004, and Babcock-Hart Award from the International Life
Science Institute and the Institute of Food Technologists in 2003. He is
elected Fellow of the American Chemical Society’s Division of Agriculture
and Food Chemistry (1991), the Institute of Food Technologists (1996), the
Korean Academy of Science and Technology (1998), and International
Academy of Food Science and Technology (2006). He was appointed as
International Scholar (2011–2014) at Kyung Hee University in Korea and
recently (2014–present) he has been serving as Adjunct Distinguished
Professor at King Abdulaziz University, Saudi Arabia.
Water
1
Yrjo H. Roos, John W. Finley, and John M. deMan

carbohydrate and fiber contents of a range of


Water in Foods foods.
Water determines the physical, chemical and
Water has a chemical formula of H2O which rep- microbiological stability of foods. When water
resents two hydrogen atoms covalently bound to freezes and thaws the nature of the food can
one oxygen atom. Water is an odorless, tasteless change dramatically. Many food processes
and transparent liquid at room temperature. It involve the addition or removal of water which
appears colorless in small quantities although in changes the stability or nature of the food.
larger bodies there is an inherent blue hue. Ice Frequently the process used to remove water has
and water vapor are also colorless, although ice a significant effect on the physical nature of the
under pressure as in glaciers exerts a range of food and the ability to rehydrate. For example,
blue colors. drum dried milk powder is much denser and
Water is the most abundant molecule in food more difficult to rehydrate than spray dried milk
and is an essential ingredient to support life and powder.
since all foods come from living organisms,
water is an essential component of foods. In
many foods both the intracellular water and inter- Physical Properties of Water and Ice
stitial water are essential. The ability of water to
dissolve a wide variety of materials makes it a Some of the physical properties of water and ice
nearly universal solvent. Water functions to which are considerations in foods are presented
determine the physical attributes of meat, vegeta- in Tables 1.2, 1.3, and 1.4. Much of this informa-
ble and fruit products. For food polymers, water tion was obtained from Landolt et al. (1923) and
serves as a structural component and a plasticizer Perry (1963). The physicochemical properties of
contributing to the attributes of proteins, starch water are important considerations in understand-
and food fibers. Water also serves as solvent or ing and showing how water contributes to food
dispersing medium in wide variety of foods processing. The exceptionally high values of the
including milk, juices and other beverages. Water thermodynamic parameters (energy to thaw ice
can be dispersed in emulsions in products like and convert water to steam) of water are of impor-
butter or margarine or be the continuous phase of tance for food processes and operations such as
emulsions such as mayonnaise. The water con- freezing and drying. The considerable expansion
tent of foods is extremely variable. Table 1.1 of water during freezing may contribute to struc-
­contains the water, energy, protein, lipid, ash, tural damages to foods when they are frozen.

© Springer International Publishing AG 2018 1


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_1
2 1 Water

Table 1.1  Typical water, energy and macronutrient content in 100 g of selected foods
Energy Carbo-­ Fiber
Food Water (g) (kcal) Protein (g) Lipids (g) Ash (g) hydrates (g) (g)
Butter, with salt 15.9 717 0.9 81.1 2.1 0.1 0
Cheese, mozzarella 50.0 300 22.2 22.4 3.3 2.2 0
Cheese, parmesan, hard 29.2 392 35.8 25.8 6.0 3.2 0
Milk, whole, 3.25% milkfat 88.1 61 3.2 3.3 0.7 4.8 0
Yogurt, plain, whole milk 87.9 61 3.5 3.3 0.7 4.7 0
Egg, WHL, RAW, FRSH 76.2 143 12.6 9.5 1.1 0.7 0
Yogurt, greek, plain, nonfat 85.1 59 10.2 0.4 0.7 3.6 0
Oil, soybean, salad or cooking 0 884 0 100 0 0 0
shortening 0 884 0 100 0.0 0 0
Chicken, broilers or fryers 65.4 237 16.7 18.3 0.8 0 0
Turkey, whole, meat only, raw 75.4 112 22.6 1.9 1.0 0.1 0
Salami, cooked, beef&pork 45.2 336 21.9 25.9 4.7 2.4 0
Frankfurter 53.9 302 10.7 26.4 3.8 5.2 0
Oatmeal 83.6 71 2.5 1.5 0.3 12.0 1.7
Cheerios 5.1 376 12.1 6.7 2.8 73.2 9.4
Apples, raw, with skin 85.6 52 0.3 0.2 0.2 13.8 2.4
Bananas, raw 74.9 89 1.1 0.3 0.8 22.8 2.6
Blueberries, raw 84.2 57 0.7 0.3 0.2 14.5 2.4
Orange juice, raw 88.3 45 0.7 0.2 0.4 10.4 0.2
Raw ham 62.5 245 17.43 18.87 0.88 0 0
Pork chops, bone-in, LN, raw 73.62 127 21.99 3.71 1.01 0 0
Cured, ham, raw 66.09 173 22.45 9.17 2.7 0.21 0
Frozen pinton beans 55.8 170 9.8 0.5 1.4 32.5 5.7
Broccoli, raw 89.3 34 2.82 0.37 0.87 6.64 2.6
Carrots, raw 88.29 41 0.93 0.24 0.97 9.58 2.8
Onions, raw 89.11 40 1.1 0.1 0.35 9.34 1.7
Potatoes, Flesh & SKN, raw 79.34 77 2.02 0.09 1.08 17.47 2.2
Squash, winter, acorn, raw 87.78 40 0.8 0.1 0.9 10.42 1.5
Tomatoes, red, ripe, raw 94.52 18 0.88 0.2 0.5 3.89 1.2
Almonds, dry RSTD, W/salt 2.41 598 20.96 52.54 3.07 21.01 10.9
Beef brisket 70.29 155 20.72 7.37 1.02 0 0
Sirloin, steak, broiled 60.33 212 29.33 9.67 1.26 0 0
Beef, ground, 70% lean 54.3 332 14.35 30 0.7 0 0
Cod, Atlantic, raw 81.22 82 17.81 0.67 1.16 0 0
Salmon, Atlantic, wild, raw 68.5 142 19.84 6.34 2.54 0 0
Tuna, fresh, Yellowfin, raw 74.03 109 24.4 0.49 1.64 0 0
Peanut butter 1.47 591 25.72 50.81 3.25 18.75 5.6
Beer 92.77 41 0.36 0 0.11 2.97 0
Red table wine 86.49 85 0.07 0 0.28 2.61 0
USDA Nutrient Composition Files: http://www.ars.usda.gov/Services/docs.htm

Freezing of water in foods occurs over a wide important quality attribute of frozen foods. An
temperature range, resulting in stresses in frozen example is that frozen strawberries loose much of
foods. Structural damage may result from fluctu- their texture after a freezing and thawing cycle
ating temperatures, even if the fluctuations remain because of loss of cellular integrity. Table 1.2
below the freezing point as ice crystals grow and contains the basic physico-­chemical properties of
causes stresses to cellular structures. This is an water.
Physical Properties of Water and Ice 3

Table 1.2  Physical–chemical properties of water This allows calculation of the pressure of the
Property Value vapor phase at a given temperature when the tem-
Molar mass (g/mol) 18.015 perature and pressure at another point, and the
Molar volume (mol/L) 55.5 enthalpy of vaporization are known. The boiling
Boiling point (BP, °C at 1 bar) 100 °C point is the temperature at which the vapor pres-
Freezing point (FP, °C at 1 bar) 0 °C sure at equilibrium exceeds atmospheric pres-
Triple point 0 °C, 6.1 mbar sure. In other words, Knowing the boiling point
Surface tension (20 °C) 73 dynes at 20 °C of a substance at 1 bar (the normal boiling point)
Vapor pressure (20 °C) 0.0212 atm at 20 °C with the heat of evaporation, ΔHvap for that sub-
∆H of vaporization 40.63 kJ/mol stance, the boiling point, T2 at another pressure
∆H of fusion 6.013 kJ/mol with values for P1 and T1 (the normal boiling
Heat capacity (Cp) 4.22 kJ/kg
point), P2, and ΔHvap, (also called the latent heat)
Viscosity (Pas, 20 °C) 1.002 centipoise at 20 °C
can be solved. The liquid–vapor transition fol-
Density (kg/m3) 1 g/mL
lows a very specific curve on the pressure–tem-
Maximum density (kg/m3) 4 °C
perature plane of a PVT diagram. This is given by
the Clausius–Clapeyron relation, which at tem-
The physical properties of water and ice are peratures and pressures that are not close to the
presented in Tables 1.3 and 1.4 according to data critical point, can be approximated as:
of Landolt et al. (1923) and Perry and Green Table 1.3 provides typical physical properties
9:2007. There is a significant variation in the prop- of water which are important to foods and food
erties of liquid water and ice with temperature. processing. The vapor pressure of water is defined
The influence of temperature is presented in Tables as the pressure at which air over the water is satu-
1.3 and 1.4. In comparison to other liquids higher rated. If the pressure is increased the water will
amounts of heat are required to convert water from condense, if the pressure decreases more water
solid ice to liquid water and to water vapor. will evaporate into the atmosphere. It is important
Within the three physical states of water (ice, to recognize the changes in vapor pressure with
liquid and steam) the physical properties have changes in temperature. This is an equilibrium
major impacts on food processing. The effects of where there is no net change although individual
heat on water drive processing conditions particu- atoms migrate between liquid and vapor phase.
larly freezing products, or water removal such as Density refers to the mass of a material divided by
concentration or drying. Figure 1.1 illustrates its volume. It can be seen that as the temperature
vapor pressure on the surface of water. As exter- of water increases the increased molecular motion
nal pressure increases it becomes increasingly from the heat results in a decline in density.
difficult for water molecules to escape from a liq- Table 1.4 illustrates vapor pressure of water in
uid, thus raising the boiling point of the water. ice as a function of temperature. This becomes
The vapor pressure of water is affected by tem- important in frozen foods as protecting the sur-
perature and therefore, if external pressure is face from water loss is required and conversely
lower water boils at lower temperatures. Such maximising the rate of water removal during
phenomenon occurs for example at higher alti- freeze drying. The changes in the coefficient of
tudes (Fig. 1.2). It is well known that water boils expansion are important in maintaining quality of
at 100 °C at sea level but at 95 °C at 1500 m above frozen foods. If the temperature of a frozen food
sea level. Therefore, cooking times of foods must stored at −30 °C increased to −5 °C there would
be extended at higher altitudes to achieve equiva- be a large increase in the size of the ice crystals
lent results such as making a hardboiled egg or potentially damaging product quality and texture.
hydrating pasta. At very high elevations cooking When ice and water are mixed together, then
times must be adjusted to obtain desired physical the temperature of the solution will be 0 °C as
changes such as gelation of starch, and to assure long as both liquid and solid phases coexist.
sufficient heat and time for microbial safety. Thus 0 °C is the freezing point for water or the
4 1 Water

Table 1.3  Physical properties of water at various temperatures


Temperature (°C)
Water 0 20 40 60 80 100
Vapor pressure (mbar) 6.11 23.37 73.75 199.18 473.56 1013.25
Density (kg/m3) 0.9998 0.9982 0.9922 0.9832 0.9718 0.9583
Specific heat (kJ/kg °C) 4.215 4.179 4.176 4.181 4.194 4.213
Heat of vaporization (kJ/kg) 2499 2452 2405 2357 2306 2255
Thermal conductance (kcal/m2 h °C) 0.565 0.599 0.628 0.652 0.670 0.680
Surface tension (mN/m) 75.62 72.75 69.55 66.17 62.60 58.84
Viscosity (mPa s) 1.792 1.002 0.653 0.466 0.355 0.282
Refractive index 1.3338 1.3330 1.3306 1.3272 1.3230 1.3180
Dielectric constant 88.0 80.4 73.3 66.7 60.8 55.3
Coefficient of thermal expansion × 10−4 – 2.07 3.87 5.38 6.57 –

Table 1.4  Physical properties of ice at various temperatures


Temperature (°C)
Ice 0 −5 −10 −15 −20 −25 −30
Vapor pressure (mbar) 6.11 4.01 2.60 1.65 1.03 0.63 0.37
Heat of fusion (kJ/kg) 334 – – – – – –
Heat of sublimation (kJ/kg) 2836 – 2813 – 2789 – 2771
Density (kg/m3) 0.9168 0.9171 0.9175 0.9178 0.9182 0.9185 0.9188
Specific heat (kJ/kg °C) 2.039 – 1.996 – 1944 – 1.884
Coefficient of thermal expansion × 10−5 9.2 7.1 5.5 4.4 3.9 3.6 –

The difference is due to the need for nucleation


to occur before an ice crystal growth is initiated.
Nucleation is the process by which a minimum
number of water molecules form an embryo
that can grow to form a crystal, after which ice
crystal growth results in continued expansion of
the crystals.

Structure of the Water Molecule


Fig. 1.1  At the boiling temperature of a liquid: vapor
The unique properties of water are a result of the
pressure = atmospheric pressure
structure of the water molecule (Fig. 1.3) and its
dielectric properties which allow it to form
melting point for ice. This is called the equilib- hydrogen bonds. Water molecules consist of two
rium point. If water is cooled to 0 °C it does not hydrogen atoms joined to an oxygen atom by
freeze; it must be cooled to below 0 °C before covalent bonds. Oxygen is more electronegative
freezing can occur. Likewise, ice has to be heated than hydrogen. That is, the high electronegativity
slightly above 0 °C before melting occurs. Unlike causes the oxygen atom to pull the shared pairs of
freezing, however, melting will begin as soon as electrons more towards the oxygen atom. As a
the temperature rises to above 0 °C. To initiate result, the O–H bond acquires polarity.
ice formation water must be cooled to tempera- Oxygen atoms have six electrons (1s2 2s2 2p4)
tures substantially below the freezing point. in its outermost shell. The ‘s’ and ‘p’ orbitals of
Structure of the Water Molecule 5

Fig. 1.2  Boiling point


of water at various
altitudes

Fig. 1.3  Angles of H


atoms, dipoles of lone
pairs of electrons
leading to the tetrahedral
arrangement of water

the valence shell are sp3 hybridized to form four form the angles of a regular tetrahedron. Because
sp3 hybrid orbitals oriented tetrahedrally around of the separation of charges in a water molecule,
the oxygen atom. Two of the hybrid orbitals are the attraction between neighboring molecules is
singly occupied with the half-filled orbital of the higher than is normal with van der Waals’ forces.
hydrogen atoms. Lone pairs of electrons occupy In the frozen state each water molecule accepts
the other two. Therefore, oxygen is bonded to the two hydrogen bonds from two other water mole-
two hydrogen atoms by two O–H covalent bonds, cules and donates two hydrogen atoms to form
and there are two lone-pairs of electrons on the hydrogen bonds with two more water molecules,
oxygen atom. The H–O–H bond angle is 104.5°, producing an open, cage like structure (Fig. 1.4).
which is slightly less than the tetrahedral angle of The structure of liquid water is very similar, but
109°28′. Therefore, the structure of water mole- in the liquid, the hydrogen bonds are continually
cule is an angular or bent structure. Figure 1.3 broken and formed because of rapid molecular
illustrates the tetrahedral structure of water. In motion.
the water molecule the atoms are arranged at an In the liquid state water molecules are held
angle of 104.5°, and the distance between the together by intermolecular hydrogen bonds. Each
nuclei of hydrogen and oxygen is 0.0957 nm. The oxygen atom can form two hydrogen bonds uti-
water molecule can be considered a spherical lizing both the lone pairs of electrons the oxygen
quadrupole with a diameter of 0.276 nm, where atom. Liquid water contains aggregates of vary-
the oxygen nucleus forms the center of the quad- ing number of water molecules held together by
rupole. The two negative and two positive charges hydrogen bonds and ‘free’ water molecules in
6 1 Water

Table 1.5  Physical properties of some hydrides at nor-


mal atmospheric conditions
Melting Boiling Molar heat of
Compound point (°C) point (°C) vaporization (kJ)
CH4 −184 −161 8.2
NH3 −78 −33 23.3
HF −92 +19 7.49
H 2O 0 +100 40.7

the properties of water with those of the hydrides


of elements near oxygen in the Periodic Table
(CH4, NH3, HF, DH3, H2S, HCl) indicates that
water has unusually high values for various phys-
ical constants, such as melting point, boiling
point, heat capacity, latent heat of fusion, latent
Fig. 1.4  Hydrogen bonds in water heat of vaporization, surface tension, and dielec-
tric constant. Some of these values are listed in
Table 1.5.
equilibrium. These intermolecular aggregates are
continually forming, collapsing at various rate
depending on temperature. Structure of Ice
Hydrogen bonding has very major influence
on the properties of water. In ice an individual water molecule connects to
In ice crystals a hexagonal matrix is formed four others in a tetrahedral arrangement. This
with tetrahedral structure of water molecules sur- arrangement results in the hexagonal crystal lat-
rounding each oxygen atom. One hydrogen atom tice in regular ice, as shown in Fig. 1.5. The lat-
exists between each pair of oxygen atoms. Thus, tice is loosely built and has relatively large hollow
each and every hydrogen atom is covalently spaces. These hollow spaces in typical ice struc-
bonded to one oxygen atom and hydrogen bonded ture result in expansion during freezing and cause
to another oxygen atom. This packing results in the high specific volume of ice. This is why ice
large open spaces between water molecules in the floats on the surface of water. In the hydrogen
ice resulting in the lower density of ice compared bonds, the hydrogen atom is 0.1 nm from one
to liquid water. oxygen atom and 0.176 nm from another hydro-
When ice melts some of the hydrogen bonds gen atom. When ice melts, the hydrogen bonds
are broken and the water molecules become more are broken and the water molecules pack together
closely packed. This results in an increase in the more compactly in a liquid state (http://www1.
density of water above its melting points lsbu.ac.uk/water/hexagonal_ice.html; http://
0 °C. Density of water attains a maximum value www.uwgb.edu/dutchs/petrology/Ice%20
of 1 g/mL at 4 °C; above 4 °C, the density Structure.HTM; Franks 2000).
decreases due to the normal temperature effects. Each oxygen atom inside the ice lattice is sur-
In ice, every H2O molecule is bound by four rounded by four other oxygen atoms in a tetrahe-
bridges to each neighbor. The binding energy of dral arrangement. The distance between oxygen
the hydrogen bond in ice amounts to 20.9/mol atoms is approximately 2.75 Å. The hydrogen
(Meryman and Pauling 1960). Similar strong atoms in ice are arranged following the Bernal-­
interactions occur between OH and NH and Fowler rules: (1) two protons are close (about
between small, strongly electronegative atoms 0.98 Å) to each oxygen atom, much like in a
such as O and N. This is the reason for the strong free water molecule; (2) each H2O molecule is
association in alcohols, fatty acids, and amines oriented so that the two protons point toward
and their great affinity to water. A comparison of two adjacent oxygen atoms; (3) there is only one
Latent Heat of Fusion 7

Fig. 1.5 Hydrogen
bonding among water
molecules leading to
hexagonal arrangement
of water molecules in
ice

proton between two adjacent oxygen atoms; (4) present in the liquid will be excluded from this
under ordinary conditions any of the large num- growing ice front. If the rate of crystal growth is
ber of possible configurations is equally probable faster than the rate at which diffusion of the par-
(http://www.its.caltech.edu/~atomic/snowcrys- ticular solutes can carry them away a concentra-
tals/ice/ice.htm). tion gradient will form in the liquid which
When there is a change of state from ice to surrounds the ice crystal. The concentrated solute
water, rigidity is lost, but water still retains a will lower the freezing point of the solution. The
large number of ice-like clusters. The term ice-­ solution at the interface will have a freezing point
like cluster does not imply an arrangement identi- equal to the temperature of the interface; at this
cal to that of crystalline ice. The HOH bond angle point, ice growth will be limited by diffusion of
of water is several degrees less than that of ice, the solute away from the crystal. When this
and the average distance between oxygen atoms occurs solution away from the ice crystal is
is 0.31 nm in water and 0.276 nm in ice. Research supercooled (a temperature below the melting
has not yet determined whether the ice-like clus- point). Eventually diffusion will ensure that the
ters of water exist in a tetrahedral arrangement, as system goes to equilibrium, however a situation
they do in ice. Since the average intermolecular of instability is created when this occurs.
distance is greater than in ice, it follows that the
greater density of water must be achieved by each
molecule having some neighbors. A cubic struc- Latent Heat of Fusion
ture with each HOH molecule surrounded by six
others has been suggested. When water freezes, heat is liberated by the pro-
At 0 °C, water contains ice-like clusters aver- cess. This latent heat of fusion is due to the
aging 90 molecules per cluster. With increasing energy released from hydrogen bonds in the crys-
temperature, clusters become smaller and more tal. In ice a water molecule is hydrogen bonded to
numerous. At 0 °C, approximately half of the four other neighboring water molecules, each
hydrogen bonds present remain unbroken, and bond having an energy between 10 and 40 kJ/
even at 100 °C approximately one-third are still mol. This equals an energy of 80 cal/g (335 J/g).
present. All hydrogen bonds are broken when The energy released in the transformation of 1 g
water changes into vapor at 100 °C. This explains of water at 0 °C to ice at the same temperature is
the large heat of vaporization of water. enough energy to raise the temperature of 1 g of
the water from 0 to 80 °C!
The speed of crystallization—that is, the prog-
Growth of Ice Crystals ress of the ice front in centimeters per second—is
determined by the removal of the heat of fusion
The crystal structure of ice is such that it does not from the area of crystallization. The speed
allow the inclusion of impurities, except within of crystallization is low at a high degree of
defects in the crystal structure. When an ice super-­cooling (Meryman 1966). This affects the
nucleus begins to grow, any solutes which are size of crystals in the ice. When large water
8 1 Water

masses are cooled slowly, there is sufficient time an amorphous solid, generally referred to as
for heterogeneous nucleation in the area of the glass. The liquid remains in a supercooled liquid
ice crystal growth. At that point the crystalliza- state until temperature is lowered to below the
tion speed is very rapid so that a few nuclei grow glass transition temperature (Tg) which is indi-
to a large size, resulting in a large crystalline cated by a decrease in heat capacity when mea-
structure. At greater cooling speed, high super- sured in differential scanning calorimetry. Once a
cooling occurs; this results in more nucleation supercooled liquid reaches temperatures below
and growth of smaller ice crystals. Tg, the system is not a viscous liquid, but is a
Upon freezing, H–O–H molecules associate in solid in a metastable state. Achieving vitrification
an orderly manner to form a rigid structure that is with pure water requires in laboratory very small
more open (less dense) than the liquid form. amounts of water and allowing an extremely fast
There still remains considerable movement of cooling. When high concentrations of solutes are
individual atoms and molecules in ice, particu- present, solutions can be vitrified more easily.
larly just below the freezing point. At 10 °C an (http://people.ucalgary.ca/~kmuldrew/cryo_
H–O–H molecule vibrates with an amplitude of course/cryo_chap6_2.html).
approximately 0.044 nm, nearly one-sixth the Crystal growth, in contrast to nucleation,
distance between adjacent HOH molecules. occurs readily at temperatures close to the freez-
Hydrogen atoms may wander from one oxygen ing point. It is more difficult to initiate crystalli-
atom to another. zation than to continue it. The rate of ice crystal
The production of heat from crystallization growth decreases with decreasing temperature.
also interferes with crystal growth. The heat is This is important in protecting quality of frozen
created at the crystal surface and must either be foods where excessive crystal growth can damage
diffused in the crystal or throughout the liquid. the structure of the food. A schematic graphical
The removal of this heat occurs by conduction representation of nucleation and crystal growth
and can only occur through the liquid if it is rates is given in Fig. 1.6. Solutes of many types at
supercooled when nucleation occurs. If the latent low concentrations will greatly slow ice crystal
heat of fusion is conducted away through the ice growth. The mechanism of this action is not
the growing crystal will remain essentially known. The effect of membranes on ice crystal
smooth the part of the interface which grows propagation was studied by Lusena and Cook
beyond the planar front. The system will not be (1953), who found that membranes can be either
able to lose its heat as quickly as the ice on either permeable, partly permeable, or impermeable to
side of it. If the heat is conducted away through growing ice crystals. Permeability to ice crystal
the liquid, growth occurs preferentially along the growth increases with porosity and is also affected
a-axes compared with growth along the c-axis.
This occurs because of the rise in temperature of
the liquid surrounding the crystal. As the mole-
cules become more energetic, they are less likely
to join a planar surface where they can only
hydrogen bond with a single neighbor. This is the
phenomenon responsible for the hexagonal sym-
metry that we see in growing ice crystals.

Solidification
Without Crystallization

If a liquid is cooled so quickly so that nucleation


cannot occur, it is possible to avoid ice formation. Fig. 1.6  Schematic representation of the rate of nucle-
This process is called vitrification and results in ation and crystal growth
Solidification Without Crystallization 9

by rate of cooling, membrane composition and controlling the pattern of propagation of the ice
membrane properties, and concentration of the front. Lusena and Cook (1955) also found that
solute(s) present in the aqueous phase. When ice solutes depressed the nucleation temperature to
crystal growth is retarded by solutes, the ice phase the same extent that they depressed the freezing
may become discontinuous either by the presence point. Solutes retard ice growth at 10 °C super-
of a membrane or spontaneously. cooling, with organic compounds having a
Ice crystal size at the completion of freezing is greater effect than inorganic ones. At low con-
related directly to the number of ice nuclei. The centrations, some proteins are as effective as
greater the number of nuclei, the smaller the size alcohols and sugars in retarding crystal growth.
of the crystals. This is the reason that the cryo-­ Once formed, crystals have a tendency to
freezing in food processing causes less damage to enlarge. Recrystallization is particularly evident
the cell structure and maintains better textural when storage temperatures are allowed to fluctu-
quality when the frozen products are thawed. In ate. This is evident in foods stored in freezers
liquid systems nuclei can be added by a process where temperatures fluctuate. The larger crystals
called seeding. Practical applications of seeding alter texture and can damage cellular structure in
include adding finely ground lactose to evapo- foods.
rated milk in the evaporator, and recirculating Slow freezing results in large ice crystals
some portion of crystallized fat in a heat exchanger located exclusively in extracellular areas. Rapid
during manufacture of margarine. If the system is freezing results in tiny ice crystals located both
maintained at a temperature close to the freezing extra- and intracellularly. During the freezing of
point (FP), where crystallization starts (Fig. 1.6), food, water is transformed to ice with a high
only a few nuclei form and each crystal grows degree of purity, and solute concentration in the
extensively. The slow removal of heat energy pro- unfrozen liquid is gradually increased. This is
duces an analogous situation, since the heat of accompanied by changes in pH, ionic strength,
crystallization released by the few growing crys- viscosity, osmotic pressure, vapor pressure, and
tals causes the temperature to remain near the other properties.
melting point, where nucleation is unlikely. In tis- When water freezes, it expands nearly 9%.
sue or unagitated fluid systems, slow removal of The volume change of a food that is frozen will
heat results in a continuous ice phase that slowly be determined by its water content and by solute
moves inward, with little if any nucleation. The concentration. Highly concentrated sucrose solu-
effect of temperature on the linear crystallization tions do not show significant expansion
velocity of water is given in Table 1.4. (Table 1.6). Air spaces may partially accommo-
When the temperature is lowered to below the date expanding ice crystals. Volume changes in
FP (Fig. 1.6), increasing rate of supercooling is some fruit products upon freezing are shown
required before nucleation becomes the predomi- in Table 1.7. The effect of air space is obvious.
nant factor and crystal growth appears thereafter.
At low supercooling large crystals are formed; as
supercooling increases, smaller crystals are Table 1.6  Volume change of water and sucrose solutions
formed. Control of crystal size is more difficult in on freezing
tissues than in agitated liquids. Agitation pro- Volume increase during temperature
motes nucleation and, therefore, reduces crystal Sucrose (%) change from 21 to −18 °C
size. Lusena and Cook (1954) suggested that 0 8.6
large ice crystals are formed when freezing takes 10 8.7
place above the critical nucleation temperature 20 8.2
(close to FP in Fig. 1.6). When freezing occurs at 30 6.2
the critical nucleation temperature, small ice 40 5.1
crystals form. The effect of solutes on nucleation 50 3.9
and rate of ice crystal growth is a major factor 60 None
10 1 Water

Table 1.7  Expansion of fruit products during freezing


H2O H2O H2O H2O
Percent volume increase
during temperature change
Product from 21 to −18 °C
Apple juice 8.3
H 2O H2O H2O H2O
Orange juice 8.0
Whole raspberries 4.0
Crushed raspberries 6.3
Whole strawberries 3.0 H 2O H2O H2O H2O
Crushed strawberries 8.2
Fig. 1.7 Interaction of water molecules at surface
increase surface tension
The expansion of water on freezing results
in local stresses that undoubtedly produce
mechanical damage in cellular materials. slightly negative charge. This polarity causes
Freezing may cause changes in frozen foods that water to be more cohesive and sticky.
make the product unacceptable. Such changes Water has greater molecular interaction at
may include destabilization of emulsions, floccu- surface:
lation of proteins, increase in toughness of fish Water molecules tend to be attracted to one
flesh, loss of textural integrity, and increase in another. At the surface, however, there are no
drip loss of meat. Ice formation can be influenced water molecules in the air above the liquid air
by the presence of carbohydrates. The effect of above which results in stronger bonds between
sucrose on the ice formation process has been those molecules at the surface as shown in
described by Roos and Karel (1991a, b, c). Fig.  1.7. This surface layer results in surface
tension which creates a barrier between the
atmosphere and the water. The hydrogen bond-
Surface Tension of  Water ing binding between water molecules results in
close alignment of water molecules at the
Water has a high surface tension which makes it water air interface. This interaction drives
sticky and as a result it tends to form droplets droplet formation and higher surface tension
rather than spread out as a film. This surface ten- in liquid water.
sion in water accounts for water’s ability to move In liquid, a water molecule will show net
in capillaries such as the roots of plants or in ves- force because the forces by the neighboring mol-
sels in living bodies. ecules all cancel out (Fig. 1.7). At the surface
When we look at a drop of water it almost there will be a net inward force because there are
appears to have skin around it making it appear no forces acting from above. This inward net
like a flattened sphere. This surface tension is force causes the molecules on the surface to con-
because water molecules are attracted to one tract and to resist being stretched or broken. The
another. Conversely non-polar compounds like surface tension of water is why small objects will
hexane do not form droplets because there is little “float” on the surface of water as long as the
intermolecular attraction. In water this attraction object cannot break through and separate the top
is because the two hydrogens line up one side of layer of water molecules. The surface of the
the oxygen atom and the hydrogens have a fluid, the surface under tension will behave like
slightly positive charge and the oxygen has a an elastic membrane.
Colligative Properties of Aqueous Solutions 11

Examples of Surface Tension depression, boiling point elevation, osmotic


pressure and vapor pressure.

Freezing Point Depression

When solutes are present in solution the freezing


point (defined as the highest temperature where
last ice crystals dissolve in heating) is lowered
relative to the freezing point of the pure solvent.
The depression of freezing point of an aqueous
solution is determined by the formula:
∆T f = iK f m

Insect on surface of water held by surface tension
where:
ΔTf = The depression in freezing point of the
solution
Water has a high surface tension against air; there- i = the number of ions or particles per molecule
fore, small insects such as the water strider can (van ’t Hoff factor)
walk on water because their weight is not enough Kf = freezing point depression constant for the
to penetrate the surface of the water. When soaps solvent (1.86 °C kg/mol for water)
and detergents are added to the water the surface m = Molar concentration of the solution
tension is reduced which makes the water better
able to wet other surfaces. Many food emulsifiers For example:
act like detergents to facilitate water contact with 1. A solution of sodium chloride containing 75 g
proteins or lipids in food systems. NaCl/500 mL.
When gases are formed in water the surface The molecular weight of NaCl is 58.44
tension of the water provides the necessary ten- m (molal) = 75 NaCl g/58.44/0.5 mL (500 mL
sion at the surface that tends to force the bubbles /1000) = 100/58.55/0.5 = 2.57
to become spherical. When water droplets are i = 2 (1 Na+ and 1 Cl−)
formed the surface tension phenomenon causes Kf = Constant for water is 1.86 °C kg/mol
the droplets to take on a spherical shape.
Therefore:
U.S. Department of the Interior | U.S.
Geological Survey ΔTf = iKfm
URL: http://water.usgs.gov/edu/surface-ten- ΔTf = 2 × 1.86 × 2.57 = 9.56
sion.html The sodium chloride solution will start to
freeze at −9.56 °C
2. A solution of 75 g glycerol/500 mL
 olligative Properties of Aqueous
C The molecular weight of glycerol is 92.09
Solutions m = 75 g/92.09/0.5 L (500 mL/1000) = 75/92.09
/0.5 = 1.62
The colligative properties of the solution are those i = 1 (glycerol is one molecule)
properties that are determined by the number of Kf = freezing constant for water is 1.86 °C kg/mol
particles dissolved in the solution. The freezing
point of a solution of water is lowered as a func- Therefore:
tion of the dissolved particles in the solution and ΔTf = iKfm
conversely, the boiling point is raised as a func- ΔTf = 1 × 1.86 × 1.62 = 4.82
tion of particles in solution. The colligative prop- The glycerol solution will show initial freezing at
erties important in food chemistry are freezing −4.82 °C
12 1 Water

Boiling Point Elevation Osmotic Pressure

When solutes are dissolved in a solution the boil- Osmosis is the process whereby a solvent passes
ing point will be raised. Like freezing point through a semipermeable membrane from one
depression, the boiling point elevation is depen- solution to another. Examples of semipermeable
dent on the number of particles in solution. The membranes are the cell membranes in cells of liv-
calculation for increase in boiling point is similar ing things (plants and animals) or synthetic mem-
to the equation for calculating the depression in branes for dialysis. Osmotic pressure drives
freezing point. The equation is: solvent molecules through the semipermeable
membrane from the low solute concentrations to
∆Tb = iK b m the high solute concentrations. When equilibrium

is reached the solute concentrations are equal on
both sides of the membrane. Osmotic pressure is
ΔTb = the increase in boiling point of the defined as the pressure applied to on the high
solution concentration side to stop osmosis.
i = the number of ions or particles per mole- Osmotic pressure is expressed by the formula:
cule (van ’t Hoff factor) Π = iMRT

Kb = boiling point increase constant for the
where
solvent (0.52 °C kg/mol for water)
Π is the osmotic pressure in atm
m = Molar concentration of the solution
i = van ’t Hoff factor of the solute.
M = molar concentration in mol/L
Using the same two examples as above:
R = universal gas constant = 0.08206 L atm/
mol K
1. A solution of sodium chloride containing 75 g
T = absolute temperature in K
NaCl/500 mL.
The molecular weight of NaCl is 58.44
An example of benefits in food from controlling
m (molal) = 75 NaCl g/58.44/0.5 mL (500 mL
osmotic pressure is dried fruit with added sugar. The
/1000) = 100/58.55/0.5 = 2.57
fruit cells are dehydrated preserving the fruit. When
i = 2 (1 Na+ and 1 Cl−)
the fruit is exposed to potential spoilage organisms
Kb = Boiling constant for water is 0.52 °C kg/
the moisture migrates from the spoilage organism to
mol
the fruit stopping growth of the microbe.
Therefore:
ΔTb = iKfm
ΔTb = 2 × 0.52 × 2.57 = 2.67 °C
The sodium chloride solution will boil at Vapor Pressure Lowering
102.67 °C
2. A solution of 75 g glycerol/500 mL Vapor pressure for a liquid is defined as the equi-
The molecular weight of glycerol is 92.09 librium pressure of gas molecules from a liquid
m = 75 g/92.09/0.5 L (500 mL/1000) = 75/92. that exists above the liquid in a closed system. In
09/0.5 = 1.62 a closed system, the vapor pressure above the liq-
i = 1 (glycerol is one molecule) uid is dependent on the temperature. The vapor
Kb = Boiling Constant for water is 0.52 °C kg/ pressure of pure water at room temperature
mol (20 °C) is about 2666 Pa, (which is about one
Therefore: forteith of the total atmospheric pressure at sea
ΔTb = iKbm level). When an aqueous solution is compared to
ΔTb = 1 × 0.52 × 1.62 = 0.84 pure water the vapor pressure will be lowered
The glycerol solution will boil at 100.84 °C proportional to the solute in solution. Raoult’s
law states that when a substance is added to a
Ionic Interactions Are Attractions Between Oppositely Charged Ions 13

solution, the vapor pressure of the solution will the chlorine atom. Even in solid crystals of NaCl,
decrease. This change depends on the mole frac- the sodium and chlorine atoms are ionized, so it
tion of the dissolved solute in solution and the is more accurate to write the formula for the com-
original vapor pressure of the solvent. pound as Na+Cl−.
These bonds are called ionic bonds (or inter-
Psolvent = X solvent P o solvent
actions) that result from the attraction of a posi-
tively charged ion—a cation—for a negatively
Psolvent = Vapor pressure of a solvent
charged ion—an anion. Unlike covalent or hydro-
Xsolvent = Mole fraction of solvent
gen bonds, ionic bonds do not have fixed or spe-
Posolvent = the vapor pressure of the pure solvent
cific geometric orientations. The electrostatic
at a particular temperature.
field around an ion—its attraction for an opposite
At any temperature there is a pressure at which
charge—is uniform in all directions. Crystals of
the vapor above the substance is in dynamic equi-
salts such as Na+Cl− do have very regular struc-
librium with its liquid or solid form. This is defined
tures because the specific crystalline structure is
as the vapor pressure of the substance at that
the energetically most favorable way of packing
temperature. At equilibrium, the rate at which the
together positive and negative ions. The force
solid or liquid evaporates is equal to the rate that
that stabilizes ionic crystals is called the lattice
the gas is condensing back to the solid or liquid
energy.
form. This pressure is constant regardless of how
In aqueous solutions, simple ions, such as
much of the substance is p­ resent. The law only
Na+, K+, Ca2+, Mg2+, and Cl−, do not exist as free,
works for ideal mixtures. In mixtures each compo-
isolated entities. Instead, each is surrounded by a
nent contributes to the total vapor pressure.
tightly held shell of water molecules (Fig. 1.8).
PA = X A × P o A Ionic interaction occurs between the ion and the
PB = X B × P o B oppositely charged end of the water dipole, as
Total Vapor Pressure = PA + PB shown below for the K+ ion:

where: Hd +
d−
XA = Mole fraction of Component A (moles A/ K+ ........ O
(moles A + moles B)
XB = Mole fraction of Component B Hd +
PoA = Vapor pressure of pure component A at a Sodium chloride is an example of an ionic sol-
specific temperature ute that dissolves in water. In water sodium chlo-
PoB = Vapor pressure of pure component B at a ride separates into positively charged sodium
specific temperature ions and negatively charged chloride ions. Each
PA = Partial pressure for Component A of these ions is surrounded by water molecules
PB = Partial Pressure for Component B breaking up the crystalline lattice of the salt and
pulling the ions into solution. The negative side

I onic Interactions Are Attractions


Between Oppositely Charged Ions

In compounds with ionic bonds, the bonded


atoms are sufficiently different in electronegativ-
ity that the bonding electrons are never shared:
these electrons are always found around the more
electronegative atom. For example, in sodium
chloride (NaCl), the bonding electron contributed
Fig. 1.8  Sodium and chloride ions surrounded by water
by the sodium atom is completely transferred to molecules with polarity of water stabilizing the solution
14 1 Water

of the water molecules is attracted to the positively intermolecular hydrogen bonding is why liquid
charged sodium ion and the positively charged water has a higher specific heat capacity.
sides of the water molecule are attracted to the Inorganic salts are considered to be soluble if
negatively charged chloride ion. the solubility is at least 0.1 mol/L at room tem-
With non-ionic molecules that have small perature, and not soluble if the solubility is less
dipole moments such as sugars, carboxylic acids than 0.001 M/L at room temperature. Table 1.8
and amines the water molecules surround the summarizes the water solubility’s of some repre-
­partially charged portion of the molecule and pull sentative inorganic salts.
it into solution. Water influences the conformation of macro-
Simple sugars like sucrose are dissolved in molecules because it exerts effects on many of
similar ways. The water breaks the weak bonds the non-covalent bonds that stabilize the confor-
holding the sugar crystals together and the water mation of the large molecules such as proteins,
molecules line up with the –OH groups on the starches and fibers (Klotz 1965). Three types of
sugar like they do with each other in solution. non-covalent bonds exert forces on macromole-
The solvation process requires energy because cules: hydrogen bonds, ionic bonds, or hydro-
the hydrogen bonding in water and in the sugar phobic interactions (apolar bonds). In proteins,
crystals must be broken. Some energy is given off water competes with the intermolecular amide
when the sugar molecules form intermolecular hydrogen bonds forming water–amide hydrogen
bonds with the water. bonds. The greater the hydrogen bonding ability
Molecular solids such as sucrose are dissolved of the solvent, the weaker the C=O⋯H–N bond.
when they are surrounded by water molecules. In aqueous solvents the heat of formation or dis-
Ionic solids such as sodium chloride are dis- ruption of this bond is zero. This means that a
solved as hydrates of sodium ions and chloride
ions in solution. Table 1.8  Examples of solubility of inorganic salts
When concentrations of ions become large
1. The Na+, K+, and NH4+ ions form soluble salts. Thus,
enough a reverse reaction occurs and the sodium NaCl, KNO3, (NH4)2SO4, Na2S, and (NH4)2CO3 are
and chloride ions re-associate forming new sol- soluble
ids. When the rate of precipitation and the rate of 2. The nitrate (NO3−) ion forms soluble salts. Thus,
solution are in equilibrium the solution defined as Cu(NO3)2 and Fe(NO3)3 are soluble
saturated. 3. The chloride (Cl−), bromide (Br−), and iodide (I−)
ions generally form soluble salts. Exceptions to this
The dipole charge differences in water allow
rule include salts of the Pb2+, Hg22+, Ag+, and Cu+
water molecules to be attracted to each other and ions. ZnCl2 is soluble, but CuBr is not
to other polar molecules through hydrogen bond- 4. The sulfate (SO42−) ion generally forms soluble salts.
ing. The hydrogen bond is an intermolecular Exceptions include BaSO4, SrSO4, and PbSO4, which
force that exists when two partial electric charges are insoluble, and Ag2SO4, CaSO4, and Hg2SO4,
which are slightly soluble
of opposite polarity interact with one another.
Insoluble salts
Chemically these bonds are much weaker than
1. Sulfides (S2−) are usually insoluble. Exceptions
covalent or ionic bonds. Hydrogen bonds can be include Na2S, K2S, (NH4)2S, MgS, CaS, SrS, and BaS
between molecules such as with water in solution 2. Oxides (O2−) are usually insoluble. Exceptions
or within a large macromolecule such as a protein include Na2O, K2O, SrO, and BaO, which are
where they help determine the three dimensional soluble, and CaO, which is slightly soluble
shape of the molecule. Hydrogen bonding in 3. Hydroxides (OH−) are usually insoluble. Exceptions
water determines the physical properties of water include NaOH, KOH, Sr(OH)2, and Ba(OH)2, which
are soluble, and Ca(OH)2, which is slightly soluble
such as relatively high boiling point and when
4. Chromates (CrO42−) are usually insoluble. Exceptions
water freezes the structure is of lower density. include Na2CrO4, K2CrO4, (NH4)2CrO4, and MgCrO4
Water like most other materials becomes denser 5. Phosphates (PO43−) and carbonates (CO32−) are
before freezing but unlike other liquids becomes usually insoluble. Exceptions include salts of the
less dense when ice crystals are formed. The Na+, K+, and NH4+ ions
Properties of Hydrogen Bonds 15

C=O⋯H–N hydrogen bond cannot provide stabi- d− d−


Dd − Hd + + : A Dd − Hd + ........: A
lization in aqueous solutions. The competitive
hydrogen bonding by H2O lessens the thermody- Hydrogen bond

namic stability of interamide hydrogen bonds.


For a hydrogen bond to form, the donor atom
must be electronegative, so that the covalent D–H
bond is polar. The acceptor atom also must be
Properties of Hydrogen Bonds
electronegative, and its outer shell must have at
least one nonbonding pair of electrons that
Hydrogen bonding of water molecules is of cru-
attracts the δ+ charge of the hydrogen atom. In
cial importance because all life forms exist in an
biological systems, both donors and acceptors are
aqueous environment constituted about 70–80%
usually nitrogen or oxygen atoms, especially
intracellular water. Although the exact structure
those atoms in amino (–NH 2) and hydroxyl
of liquid water is unknown, it appears that liquid
(–OH) groups. Because all covalent N–H and
water contains transient hydrogen-bonded net-
O–H bonds are polar, their H atoms can partici-
works. Water molecules are in rapid motion,
pate in hydrogen bonds. By contrast, C–H bonds
­constantly making and breaking hydrogen bonds
are nonpolar, so these H atoms are almost never
with adjacent molecules. As the temperature of
involved in a hydrogen bond.
water increases toward 100 °C at atmospheric
Liquid water molecules provide an excellent
pressure, the kinetic energy of its molecules
example of hydrogen bonding. The hydrogen
becomes greater than the energy of the hydrogen
atom in one water molecule is attracted to a pair
bonds connecting them, and the gaseous form of
of electrons in the outer shell of an oxygen atom
water appears.
in an adjacent molecule. Not only do water mol-
Normally, a hydrogen atom forms a covalent
ecules hydrogen-bond with one another, they also
bond with only one other atom as typically seen
form hydrogen bonds with other kinds of mole-
in hydrocarbon side chains. A hydrogen atom
cules as shown in Fig. 1.9. The presence of polar
covalently bonded to a donor atom, D, may form
groups such as hydroxyl (–OH) or amino (–NH2)
an additional weak association, the hydrogen
groups enhances the solubility of molecules in
bond, with an acceptor atom, A:

Fig. 1.9  Water readily


forms hydrogen bonds
with simple organic
compounds
16 1 Water

water. For instance, the hydroxyl group in metha- concept of a hydrophobic interaction has been
nol (CH3OH) and the amino group in methyl- schematically represented by Sa et al. (2013), as
amine (CH3NH2) can form several hydrogen shown in Fig. 1.10. Under appropriate conditions
bonds with water, enabling the molecules to dis- apolar molecules can form crystalline hydrates,
solve in water to high concentrations. in which the compound is enclosed within the
Most hydrogen bonds are 0.26 – 0.31 nm space formed by a polyhedron made up of water
long, about twice the length of covalent bonds molecules. Such polyhedrons can form a large
between the same atoms. The distance between lattice, as indicated in Fig. 1.10. The polyhedrons
the nuclei of the hydrogen and oxygen atoms of may enclose apolar guest molecules to form apo-
adjacent hydrogen-bonded molecules in water is lar hydrates (Speedy 1984; Sa et al. 2013). These
approximately 0.27 nm, about twice the length of pentagonal polyhedra of water molecules are
the covalent O–H bonds in water. The hydrogen unstable and normally change to liquid water
atom is closer to the donor atom, D, to which it above 0 °C and to normal hexagonal ice below
remains covalently bonded, than it is to the accep- 0 °C. In some cases, the hydrates melt well above
tor. The length of the covalent D–H bond is a bit 30 °C. There is a remarkable similarity between
longer than it would be if there were no hydrogen the small apolar molecules that form these
bonds, because the acceptor “pulls” the hydrogen clathrate-­like hydrates and the apolar side chains
away from the donor. The strength of a hydrogen of proteins. Some of these are shown in Fig. 1.11.
bond in water (≈5 kcal/mol) is much weaker than Because small molecules such as the ones shown
a covalent O–H bond (≈110 kcal/mol) (Lodish in Fig. 1.11 can form stable water cages, it may
et al. 2000) (http://www.ncbi.nlm.nih.gov/books/ be assumed that some of the apolar amino acid
NBK21726/). side chains in a polypeptide can do the same. The
concentration of such side chains in proteins is
high, and the combined effect of all these groups
Hydrophobic Interactions can be expected to result in the formation of a
stabilized and ordered water region around the
Water molecules surrounding an apolar or protein molecule. The stabilized water structure
non-­polar solutes are more ordered, leading to a around a protein molecule are illustrated in
loss in entropy. As a result, apolar groups in an Fig. 1.12.
aqueous environment tend to associate with each The hydrophobic effect is responsible for the
other rather than with the water molecules. This portioning in a mixture of oil and water, resulting

Fig. 1.10 Hydrophobic amino acids form different Han K, Lee K-H. (2013) Hydrophobic amino acids as a
hydrates in water. http://www.nature.com/articles/ new class of kinetic inhibitors for gas hydrate formation.
srep02428#f5, Sa J-H, Kwak G-H, Lee BR, Park D-H, Scientific Reports 3: 2428
Water Activity and Sorption Phenomena 17

Fig. 1.11 Comparison Crystal Hydrate formers Amino Acid


of hydrate-forming
molecules and amino
acid apolar side chains CH

Methane Methylmercaptan

Propane
Dimethyl sulfide

Isobutane
Benzene

Charton and Charton 1982; Breiten et al. 2013;


Lockett et al. 2013).

 ater Activity and Sorption


W
Phenomena

Water activity (aw) is the partial vapor pressure of


water in a substance divided by the partial vapor
pressure of water at the same temperature.
Essentially it is an expression of relative humid-
ity surrounding the food system. The standard
state vapor pressure is most often defined as the
partial vapor pressure of pure water at the same
temperature. The vapor pressure over pure dis-
tilled water has a water activity of exactly one. As
temperature increases, aw typically increases,
Fig. 1.12  Protein hydrated with water molecules stabiliz- except in some products with crystalline salt or
ing the three-dimensional structure of the protein sugar. Water activity is a physical chemistry
derived quantity. It applies to a system at equi-
in separation into two distinct phases of oil and librium and it should always be referred to
water. The hydrophobic effect is also seen in cell using a lower case a with subscript w, i.e., aw.
membranes providing the stability in the mem- Controlling the water activity in foods is more
brane network. It is also seen on the surface of critical to maintain quality and microbiological
milk fat globule membranes. Hydrophobic bond- stability of the food. Low water activity inhibits
ing in proteins facilitates protein folding as well microbial growth, provides textural characteristics
as the insertion of membrane proteins into the such as crispness and crunchiness in products like
nonpolar lipid environments (Kauzmann 1959; snack foods and ready to eat breakfast cereals.
18 1 Water

The sound produced by ‘crunching’ a breakfast mately gain enough moisture for mold growth on
cereal or a tortillas chip is lost if the aw exceeds the surface. Another example is that crackers
0.65. Rates of chemical reactions increase hydro- which have an aw of 0.3 are exposed to air with
phobic regions but reduced in hydrophilic regions. 80% relative humidity, the crackers will lose crisp-
The water activity must be carefully balanced to ness and snap when eaten. If salami (aw ≈ 0.87) is
obtain optimum quality of dehydrated foods and exposed to dry air (aww ≈ 0.5), the salami will lose
intermediate moisture foods (Leake 2006). Foods moisture and could adversely impact texture or
with an aw > 0.95 (equivalent to about 43% w/w oxidative stability of the product.
sucrose) tend to be highly perishable and prone to Water activity does not correlate directly with
microbial growth. Most bacteria growth is inhib- water content of the food. It is related to the
ited below about aw = 0.91 (equivalent to about amount of soluble solids and their molecular size
57% w/w sucrose); similarly, most yeasts cease in the material thus sweetened condensed milk
growing below aw = 0.87 (equivalent to about 65% contains over 30% water and small size sugars
w/w sucrose) and most molds cease growing but has the same aw as flour or rice which contain
below aw > 0.80 (equivalent to about 73% w/w around 12% moisture and starch and proteins as
sucrose). The lower limit of microbial growth is shown in Fig. 1.13.
about aw = 0.6. Water activity influences many characteristics
Products with higher aw tend to support more in foods that impact quality:
microbial growth. Bacteria usually require at least
0.91, and fungi at least 0.7. Water will migrate • aw ~ 0.2–0.3 below this value reactions requir-
from areas of high aw to areas of low aw. For exam- ing water phase do not occur
ple, if a sugar syrup or honey has an aw ≈ 0.6 and • aw ~ 0.2–0.3 below this rate of lipid oxidation
the relative humidity of air above it is 0.85, the increases
sugar solution will absorb moisture from the air. A • aw ~ 0.35–0.4 stickiness and caking are
consequence of this is that the surface could ulti- observed

Fig. 1.13  Water activity in foods at different moisture contents


Water Activity and Sorption Phenomena 19

• aw ~ 0.4–0.5 loss of crispness on gain of


aqueous solution in equilibrium with ice ( awi ) is
moisture
equal to the water vapor pressure over ice, to the
• aw ~ 0.5 onset of hardening (e.g. raisins) on
water pressure over pure liquid water and does
loss of moisture
not depend on the solute’s nature or concentra-
• aw ~ 0.6 onset of microbial growth
tion (Koop et al. 2000). Solutions with the same
• aw ~ 0.65–0.85 reaction rate in amorphous
ice melting point therefore have the same water
systems enzyme activity, lipid oxidation
activity.
• aw ~ 0.85 onset of growth of bacterial
Figure 1.14 is a water activity isotherm dis-
pathogens.
playing the hysteresis often encountered depend-
Water activity (aw) describes the effective ing on whether the water is being added to the dry
water concentration for hydration of materials. material or removed (drying) from the wet mate-
Water activity of 1.0 indicates pure water and rial. This hysteresis is due to non-reversible
zero is the complete absence of water molecules. structural changes and non-equilibrium effects.
Mixing of solute lowers the water activity of the There are many empirical equations but none
system when the system reaches equilibrium. predict with sufficient accuracy, therefore, water
Water activity has been reviewed for a variety of activity isotherm should be experimentally deter-
aqueous systems (Blandamer et al. 2005) and mined for each material.
biological systems (Schiraldi et al. 2012). Water Water activity is a property of aqueous solu-
activity can be determined from the dew-point tions and materials that contain water from the
temperature of the atmosphere in equilibrium liquid state to dry powders, aw is defined as the
with the material (Mathlouthi 2001; Blandamer ratio of the vapor pressures of pure water and a
et al. 2005). A high aw (>0.8) indicates a moist solution:
system and a low aw (<0.7) indicates a ‘dry’ sys- p
tem. The nature of a hydrocolloid or protein aw =
po
polymer network can influence the water activity
(Trombetta et al. 2005). The water activity of an

Fig. 1.14 Adsorption
desorption isotherms as
a function of water
activity. http://www1.
lsbu.ac.uk/water/water_
activity.html
20 1 Water

where aw = water activity cated in Fig. 1.13. Below moisture content of


p = partial pressure of water in a food about 50% the water activity decreases rapidly
po = vapor pressure of water at the same and the relationship between water content and
temperature relative humidity is represented by the sorption
According to Raoult’s law, the lowering of the isotherms. The adsorption and desorption pro-
vapor pressure of a solution is proportional to the cesses are not fully reversible; therefore, a dis-
mole fraction of the solute: aw can then be related tinction can be made between the adsorption
to the molar concentrations of soute (n1) and sol- and desorption isotherms by determining
vent (n2): whether a dry product’s moisture levels are
increasing, or whether the product’s moisture is
p n1 gradually lowering to reach equilibrium with its
aw = =
po n1 + n2 surroundings, implying that the product is being

dried (Fig. 1.15). Generally, the adsorption iso-
The extent to which a solute reduces aw is a therms are required for the observation of
function of the chemical nature of the solute. The hygroscopic products, and the desorption iso-
equilibrium relative humidity (ERH) in percent- therms are useful for investigation of the pro-
age is ERH/100. ERH is defined as: cess of drying. A steeply sloping curve indicates
p equ that the material is hygroscopic (curve A,
ERH = Fig. 1.15); a flat curve indicates a product that is
p sat
not very sensitive to moisture (curve B,
where Fig. 1.15). Many foods show the type of curves
pequ = partial pressure of water vapor in equi- given in Fig. 1.15C, where the first part of the
librium with the food at temperature T and 1 curve is quite flat, indicating a low hygroscopic-
atmosphere total pressure ity, and the end of the curve is quite steep, indi-
psat = the saturation partial pressure of water in cating highly hygroscopic conditions. Such
air at the same temperature and pressure curves are typical for foods with high sugar or
At high moisture contents, when the amount salt contents and low capillary adsorption. Such
of moisture in the product exceeds the soluble foods are hygroscopic. The reverse of this type
solids, the water activity is close to or equal to of curve is rarely encountered. These curves
1.0. When the moisture content is lower than the show that a hygroscopic product or hygroscopic
solids, water activity is lower than 1.0, as indi- conditions can be defined as the case where a

Fig. 1.15 Sorption
isotherms of
hygroscopic product (A)
and non-hygroscopic
product (B), high
crystalline sugar or salt
with low capillary
absorption (C)
Water Activity and Sorption Phenomena 21

small increase in relative humidity causes a When a/V is plotted versus a, the result is a
large increase in product moisture content. straight line with a slope equal to 1/Vm and the
Sorption isotherms usually have a sigmoid monolayer value can be calculated. In this form,
shape and can be divided into three areas that the equation has not been satisfactory for foods,
correspond to different conditions of the water because the heat of adsorption that enters into the
present in the food (Fig. 1.16). The first part (A) constant b is not constant over the whole surface,
of the isotherm, which is usually steep, corre- because of interaction between adsorbed mole-
sponds to the adsorption of a monomolecular cules, and because maximum adsorption is
layer of water; the second, flatter part (B) corre- greater than only a monolayer.
sponds to adsorption of additional layers of A form of isotherm widely used for foods is
water; and the third part (C) relates to condensa- the one described by Brunauer et al. (1938) and
tion of water in capillaries and pores of the mate- known as the BET isotherm or equation. A form
rial. There are no sharp divisions between these of the BET equation given by Labuza (1968) is
three regions, and no definite values of relative
a 1  a (C − 1) 
humidity exist to delineate these parts. Labuza = + 
(1968) has reviewed the various ways in which
(1 − a )V VmC  VmC 
the isotherms can be explained. The kinetic where
approach is based on the Langmuir equation, C = constant related to the heat of adsorption
which was initially developed for adsorption of A plot of a/(1 – a)V versus a gives a straight
gases and solids. This can be expressed in the fol- line, as indicated in Fig. 1.17 The monolayer cov-
lowing form: erage value can be calculated from the slope and
a  K  a the intercept of the line. The BET isotherm is
= + only applicable for values of a from 0.1 to 0.5. In
V  bVm  Vm
addition to monolayer coverage, the water sur-
where face area can be calculated by means of the fol-
a = water activity lowing equation:
b = a constant
1
K = 1/po and po = vapor pressure of water at To S o = Vm ⋅ ⋅ N o ⋅ AH2 O
V = volume adsorbed M H2 O
Vm = monolayer value = 3.5 × 103Vm

Fig. 1.16 Adsorption
and desorption
isotherms
22 1 Water

The Langmuir isotherm assumes that there are a


fixed number of sites where the sorption process
can occur. The process continues until all the
sites are filled. The BET theory assumes multiple
layers will continue to form after the first mono-
layer is complete.
Another method to analyze the sorption iso-
therms is the GAB sorption model described by
van den Berg and Bruin (1981) and used by Roos
(1993) and Jouppila and Roos (1994). The use of
the GAB model is described in Roos and Drusch
(2015).
The best explanation for syneresis phenome-
Fig. 1.17  BET monolayer plot. Adapted from Labuza non appears to be the so-called ink bottle theory
(1968). A = aw, V = Volume absorbed, C = heat of adsorp-
tion constant, Vm = monolayer value (Labuza 1968). It is assumed that the capillaries
have narrow necks and large bodies, as repre-
sented schematically in Fig. 1.18. During adsorp-
tion the capillary does not fill completely until
where an activity is reached that corresponds to the
large radius R. During desorption, the unfilling is
S = surface area,m 2 / g solid
controlled by the smaller radius r, thus lowering
M H2 O = molecular weight of water,18 the water activity. Several other theories have
N o = Avogadro ′s number, 6 × 10 23 been advanced to account for the hysteresis in
A H2 O = area of water molecule, 10.6 × 10 20 m 2 sorption. These have been summarized by
Kapsalis (1987).
The BET equation has been used in many The position of the sorption isotherms depends
cases to describe the sorption behavior of foods. on temperature: the higher the temperature, the
For example, note the work of Saravacos (1967) lower the position on the graph. This decrease in
on the sorption of dehydrated apple and potato. the amount adsorbed at higher temperatures fol-
The form of BET equation used for calculation of lows the Clausius Clapeyron relationship,
the monolayer value was
d ( ln a ) Qs
p 1 C − 1 Po =−
= + ⋅ d (1 / T ) R
W ( po − p ) W1C W1C P
where
where Qs = heat of adsorption
W = water content (in percent) R = gas constant
p = vapor pressure of sample T = absolute temperature
po = vapor pressure of water at same
temperature By plotting the natural logarithm of activity
C = heat of adsorption constant versus the reciprocal of absolute temperature at
W1 = moisture consent corresponding to constant moisture values, straight lines are
monolayer obtained with a slope of –Qs/R (Fig. 1.19). The
Labuza (1968) described several approaches values of Qs obtained in this way for foods having
to analyze the sorption isotherms. The Langmuir less than full monolayer coverage are between
isotherm as modified by Brunauer et al. (1938) about 2000 and 10,000 cal/mol, demonstrating
has been most widely used with food products. the strong binding of this water.
Water Activity and Sorption Phenomena 23

Fig. 1.18  Ink bottle


theory of hysteresis in
sorption. Adapted from:
Labuza (1968)

Fig. 1.19  Method for determination of heat of adsorp-


tion. Moisture content increases from M1 to M5. Adapted
from: Labuza (1968) Fig. 1.20  Relationship of heat of sorption and moisture
content as actually observed and according to BET
Theory. Adapted from Labuza (1968)
According to the principle of BET isotherm,
the heat of sorption Qs should be constant up to
monolayer coverage and then should suddenly
decrease. Labuza (1968) has pointed out that the where
latent heat of vaporization ΔΗv, about 10.4 kcal/ Qs = heat of adsorption
mol, should be added to obtain the total heat R = gas constant
value. The plot representing BET conditions as T = absolute temperature
well as actual findings are given in Fig. 1.20. The
observed heat of sorption at low moisture According to the principle of BET isotherm,
­contents is higher than theory indicates and falls the heat of sorption Qs should be constant up to
off gradually, indicating the gradual change from monolayer coverage and then should suddenly
Langmuir to capillary water. decrease. Labuza (1968) has pointed out that the
The position of the sorption isotherms depends latent heat of vaporization ΔΗv, about 10.4 kcal/
on temperature: the higher the temperature, the mol, should be added to obtain the total heat
lower the position on the graph. This decrease in value. The plot representing BET conditions as
the amount adsorbed at higher temperatures fol- well as actual findings are given in Fig. 1.20. The
lows the Clausius Clapeyron relationship, observed heat of sorption at low moisture con-
d ( ln a ) tents is higher than theory indicates and falls off
Qs
=− gradually, indicating the gradual change from
d (1 / T ) R Langmuir to capillary water.

24 1 Water

Types of  Water ods permits a distinct division between bulk and
strongly hydrogen bonding water, and results
The sorption isotherm indicates that differently obtained are not necessarily identical between
structured forms of water may be present in methods. This is not surprising since the adsorp-
foods. It is convenient to divide the water into tion isotherm indicates that the division between
three types: Langmuir or monolayer water, capil- the different forms of water is gradual rather than
lary water, and loosely bound water. The strongly sharp. A promising new method is the use of
interacting water can be attracted strongly and nuclear magnetic resonance, which can be
held in a rigid and orderly state. In this state the expected to give results based on the freedom of
water molecules have reduced solvent capacity movement of the hydrogen nuclei.
and may not form ice. It is difficult to provide a The main reason for the increased water con-
rigid definition for water strongly interacting tent at high values of water activity must be capil-
with food solids because much depends on the lary condensation. A liquid with surface tension
technique used for its measurement. There are σ in a capillary with radius r is subject to a pres-
two commonly used but not agreeable definitions sure loss, the capillary pressure po = 2σ/r, as evi-
for bound water as follows: denced by the rising of the liquid in the capillary.
As a result, there is a reduction in vapor pressure
1. Bound water is the water that remains unfro- in the capillary, which can be expressed by the
zen below −20 °C (note that all water mole- Thomson equation,
cules can form ice but they may have strong
p 2σ V
thermodynamic interactions with solids and ln =− ⋅
po r RT
not show ice formation).
2. Bound water is the amount of water in a sys- where
tem that is unavailable as a solvent (note that p = vapor pressure of liquid
all water molecules can act as solvent but their po = capillary vapor pressure
thermodynamic state may not favor solvent σ = surface tension
capacity). V = mole volume of liquid
R = gas constant
The amount of unfrozen water, based on pro- T = absolute temperature
tein content, appears to vary only slightly from
one food to another. About 8–10% of the total This permits the calculation of water activity
water in animal tissue may remain unfrozen in capillaries of different radii, as indicated in
(Meryman 1966). Egg white, egg yolk, meat, and Table 1.9. In water-rich foods, such as meat and
fish all contain approximately 0.4 g of unfrozen potatoes, the water is present in part in capillar-
water per gram of dry protein. This corresponds ies with a radius of 1 μm or more. The pressure
to 11.4% of total water in lean meat. Most fruits
and vegetables contain less than 6% unfrozen
water; whole grain corn, 34%. However, the pres- Table 1.9  Capillary radius and water activity
ent knowledge is that in most food materials an Radius (nm) Activity (a)
unfrozen maximally freeze-concentrated water-­ 0.51 0.116
solids phase forms at a low but composition spe- 1 0.340
cific temperature. Such unfrozen phase typically 2 0.583
contains 20% unfrozen water and 80% solids 5 0.806
(Levine and Slade 1986; Roos and Drusch 2015). 10 0.898
The “free” or bulk water is sometimes deter- 20 0.948
mined by pressing a food sample between filter 50 0.979
paper, by diluting with an added colored sub- 100 0.989
stance, or by centrifugation. None of these meth- 1000 0.999
Types of Water 25

Table 1.10  Pressure required to press of a hydrophilic surface freezes at a lower tem-
water from tissue at 20 °C perature than water. The closer to the surface, the
Radius Pressure in Pa lower the freezing temperature of the water. The
0.1 μm 140,003 water molecules closest to the surface, are very
1 μm 145,531 difficult to freeze or to remove. This is because
10 μm 14,553 the equilibrium hydration effect. The water near
0.1 mm 1455 a surface is affected by that surface. The surface
1 mm 145.5 of biomolecules usually affects the energy
and orientation of nearby water molecules.
necessary to remove this water is small. Calculated Some surfaces are charged or dipolar, and their
values of this pressure are given in Table 1.10 for electric field affects the orientation and energy of
water contained in capillaries ranging from nearby water molecules. The molecules compris-
0.1  μm to 1 mm radius. It is evident that water ing the surface form hydrogen bonds with water
from capillaries of 0.1 μm or larger can easily disrupting the water near the surface. Most sur-
drip out. Structural damage caused, for instance, faces don’t form hydrogen bonds with water, but
by freezing can easily result in drip loss in these they disrupt the water hydrogen bonds of the
products. The fact that water serves as a solvent water near the surface (Wolfe et al. 2002).
for many solutes such as salts and sugars is an This disturbance of the attracted water extends
additional factor in reducing the vapor pressure. several molecular layers from a hydrophilic
At low temperatures in some food systems water surface.
can remain unfrozen at ordinary pressures. Freezing The increasing energy required to remove
point depression can be caused by: 1—equilibrium water molecules that are closer to the hydrophilic
effects such as freezing point depression due to surface similarly gives rise to the greater freezing
hydration or osmotic effects, 2—supercooling point depression. Undisturbed, bulk water freezes
which is the delay or absence of formation of an ice below its equilibrium freezing temperature
crystal in the absence of efficient ice nucleators, (which depends on the solute concentration). The
3—the high viscosity produced by high concentra- more strongly the water near a surface is held (i.e.
tions of solutes (Wolfe et al. 2002). the closer it is to the surface), the lower the tem-
The freezing point of water can be lowered as perature at which it freezes (Wolfe et al. 2002).
a result of the osmotic effects of dissolved solutes Freezing point depression also occurs in non-­
such as salt, the hydration effects of macromole- equilibrium systems such as super cooling. Small
cules or ultrastructures, such as membranes. For volumes of water, or large volumes of very pure
each mole of soluble molecules (i.e. sugar) or ions water, can be cooled below 0 °C before they
(i.e. Na+ or Cl−) in kilogram of water, one lowers freeze. The water in biological systems is found
the equilibrium freezing point by (approximately) in very small volumes, such as the interiors of
2°. At equilibrium, freezing is a balance of mini- cells and organelles, where supercooling occurs
mizing energy and maximizing entropy. Molecules in processing such as freezing foods. Before
in ice have lower energy than liquid water, because freezing nuclei are formed and these nuclei must
of interaction with neighboring molecules, but reach a minimum size for crystallization to occur.
they also have lower entropy because they cannot The size of an ice crystal decreases with sub-­
move about. Solutes are minimally soluble in ice, freezing temperature, but just below freezing it is
so the entropy of ice is unaffected by their pres- large (Wolfe and Bryant 1999). The likelihood of
ence. However, dissolved solutes in water increase a large number of molecules spontaneously
the entropy of the water molecules which results forming ice crystals is small. Consequently,
­
in lowering energy and maximizing entropy at pure water can be cooled to well below zero
low temperatures (Wolfe et al. 2002). (sometimes to −40 °C) before ice forms. Usually
The depression of freezing point by hydration water will freeze at only a few degrees below
occurs when water within 1 nm (0.000001 mm) zero. This is because a range of substances act as
26 1 Water

ice nucleators: they act as surfaces on which an meat, 0.40 g/g protein. The unfrozen and
ice crystal can form an embryo for their growth. Langmuir water are probably not exactly the
The more efficient the nucleator, and the more of same. Wierbicki and Deatherage (1958) used a
them there are, the easier it is to freeze. pressure method to determine free water in meat.
Conversely, small volumes of very pure water The amount of free water in beef, pork, veal, and
can be supercooled most easily. lamb varies from 30 to 50% of total moisture,
When some of the water in a system freezes, depending on the kind of meat and the period of
the solutes are concentrated in the remaining aging. A sharp drop in such water occurs during
unfrozen water, producing more concentrated the first day after slaughter, and is followed by a
solutions. The increased concentration of solutes gradual, slight increase. Hamm and Deatherage
results in increased viscosity of the unfrozen (1960b) determined the changes in hydration
water. In the more viscous solution water mole- during the heating of meat. At the normal pH of
cules diffuse and rotate more slowly making it meat there is a considerable reduction of bound
less likely for ice nuclei to form and increased water. It is important to note that “bound” water
likelihood for supercooling to occur. At the same and “free” water may refer to water that can be
time water cannot quickly diffuse from the region mechanically removed from food and such defi-
near a hydrophilic surface to a region where ice nitions are not justified for quantification of water
has already started to form. Thus the amount of freezing. A new method to determine unfrozen
unfrozen water is higher than one would expect water contents in multicomponent food materials
at equilibrium. was presented by Roos and Potes (2015).
High viscosity and low temperature can pro-
duce a glass. A glass is a non-crystalline solid.
Window glass is an example. Glasses are formed Phase Diagram
when the viscosity of a liquid becomes so high
that it can resist shear stresses (resist changes in A phase diagram illustrates what phases of a sub-
shape) for extremely long periods. When the stance are present at any given temperature and
water in cells forms a glass, diffusion, freezing pressure. The phases are simply the solid, liquid
and biochemistry are virtually stopped. or vapor states of a pure substance. Figure 1.21
For a solute to cause significant freezing point demonstrates a phase diagram of a pure substance
depression it must be sufficiently soluble to
achieve high concentrations in water.
Supersaturation often occurs at freezing tempera-
tures. For osmotic freezing point depression, the
direct effect of small solutes is similar at low con-
centration. Provided that one counts dissociating
solutes (MgCl2 in solution is three solute ions),
the freezing point depression is approximately
proportional to concentration. At high concentra-
tion, the osmotic effects of salts may be less than
proportional to concentration. Conversely, the
osmotic effect of many solutes such as sugars
increases at high concentration by more than
simple proportionality (Wolfe and Bryant 2001).
The thermal behavior of water has been stud-
ied by Riedel (1959), who found that water in
bread did not freeze at all when moisture content
was below 18%. With this method it was possible
to determine the unfrozen water. For bread, the Fig. 1.21  Phase diagram of water. More detailed discussion:
value was 0.30 g/g dry matter, and for fish and http://www1.lsbu.ac.uk/water/water_phase_diagram.html
The Glass Transition 27

such as water. The phase diagram indicates the


existence of three phases: solid, liquid, and gas.
The conditions under which they exist are sepa-
rated by three equilibrium lines known as phase
boundaries: the vapor pressure line TA, the melt-
ing pressure line TC, and the sublimation pres-
sure line BT. The three lines meet at point T
(triple point), where all three phases are in equi-
librium. The diagram also illustrates that when
ice is heated at pressures below 6.1 mbar, it
changes directly into the vapor form. This is the
basis of freeze drying.

The Glass Transition

In aqueous systems containing polymeric sub- Fig. 1.22  Relationships among crystal growth, nucle-
stances or some low molecular weight materials ation, and crystallization rate between melting tempera-
including sugars and other carbohydrates, lower- ture (Tm) and glass temperature (Tg)
ing of the temperature may result in formation of
a glass. A glass is an amorphous solid material
rather than a crystalline solid. A glass is an super-
cooled liquid of high viscosity that exists in a
metastable solid state (Levine and Slade 1999). A
glass is formed when a liquid or an aqueous solu-
tion is cooled to a temperature that is consider-
ably lower than its melting temperature. This is
usually achieved at high cooling rates. The nor-
mal process of crystallization involves the con-
version of a disordered liquid molecular structure
to a highly ordered arrangement in a crystal. In a
crystal, molecules, atoms or ions are arranged in
a regular, three-dimensional array. In the forma-
tion of a glass, the disordered liquid state is
immobilized into a disordered glassy solid, which
has solid-like rheological properties but no Fig. 1.23  Simplified state diagram showing the effect of
moisture content on melting temperature (Tm) and glass
ordered crystalline structure. transition temperature (Tg)
The relationships among melting point (Tm),
glass transition temperature (Tg), and crystalliza-
tion are schematically represented in Fig. 1.22. crystals are formed and a glassy solid results.
At low degree of supercooling (just below Tm), During the transition from the molten state to the
nucleation is at a minimum and crystal growth glassy state, the moisture content plays an impor-
predominates. As the degree of supercooling tant role. This is illustrated by the phase diagram
increases, nucleation becomes the dominating of Fig. 1.23. When the temperature is lowered at
effect. The maximum overall crystallization rate sufficiently high moisture content, the system
is at a point about halfway between Tm and Tg. goes through a rubbery state before becoming
At high cooling rates and a degree of supercool- glassy (Chirife and Buera 1996). The glass tran-
ing that moves the temperature to below Tg, no sition temperature is characterized by very high
28 1 Water

Fig. 1.24  Relationship between water activity (aw) and Glass Dynamics and the Control of Microbiological
glass transition temperature (Tg) of some plant materials Growth in Foods, Critical Review Food Sci. Nutr., Vol.
and biopolymers. Source: Reprinted with permission from 36, No. 5, p. 490, ©1996. Copyright CRC Press, Boca
J. Cherife and M. del Pinar Buera, Water Activity, Water Raton, Florida

apparent viscosities of more than 105 Pa s may help prevent this from occurring. Such
(Aguilera et al. 1990). The rate of diffusion lim- ­ stabilization of frozen products is known as
ited processes is more rapid in the rubbery state cryoprotection, and the agents are known as
than in the glassy state, and this may be important cryoprotectants.
in the storage stability of certain foods. The effect When water is removed from foods during
of water activity on the glass transition tempera- processes such as extrusion, drying, or freezing, a
ture of a number of plant products (carrots, straw- glassy state may be produced (Roos and Drusch
berries, and potatoes) as well as some biopolymers 2015). The Tg values of high molecular mass
(gelatin, wheat gluten, and wheat starch) is shown food polymers, proteins, and polysaccharides are
in Fig. 1.24 (Chirife and Buera 1996). In the rub- high and cannot be determined experimentally,
bery state the rates of chemical reactions appear because of thermal decomposition. An example
to be higher than in the glassy state (Roos and of measured Tg values for low molecular mass
Karel 1991d). carbohydrates is given in Fig. 1.25. The value of
When water-containing foods are cooled to Tg for starch is obtained by extrapolation.
below the freezing point of water, ice may be Roos and Drusch (2015) used a combined
formed and the remaining water is increasingly sorption isotherm and state diagram to obtain
high in dissolved solids. When the glass transi- critical water activity and water content values
tion temperature is reached, the freeze-­ that result in depressing Tg to below ambient tem-
concentrated solids with remaining unfrozen perature (Fig. 1.26). This type of plot can be used
water is transformed into a glass. Ice formation to evaluate the stability of low-moisture foods
during freezing may destabilize sensitive prod- under different storage conditions. When the Tg is
ucts by rupturing cell walls and breaking emul- decreased to below ambient temperature, mole-
sions. The presence of glass-forming substances cules are mobilized because of plasticization and
The Glass Transition 29

reaction rates increase because of increased dif- Carbohydrate polymers with different molec-
fusion, which in turn may lead to deterioration. ular mass have different glass transition tempera-
Roos and Himberg (1994) and Roos et al. (1996) tures. Figure 1.27 illustrates the glass transition
have described how glass transition temperatures temperature (Tg) of different starch hydrolysates
influence nonenzymatic browning in model sys- at different moisture contents. From the figure it
tems. This deteriorative reaction showed an can be seen that the Tg decreases with decreasing
increased reaction rate as water content increased. molecular weight. All of the starches demon-
strate the reduction of Tg with increasing water
content.
The water present in foods may act as a plasti-
cizer. Plasticizers increase plasticity and flexibil-
ity of food polymers as a result of weakening of
the intermolecular forces existing between mol-
ecules. Increasing water content decreases Tg.
Roos and Karel (1991a) studied the plasticizing
effect of water on thermal behavior and crystal-
lization of amorphous food models. They found
that dried foods containing sugars behave like
amorphous materials, and that small amounts of
water decrease Tg to room temperature with the
result of structural collapse and formation of
Fig. 1.25  Glass transition temperature (Tg) for maltose, stickiness. Roos and Karel (1991d) reported a
maltose polymers, and extrapolated value for starch. M linearity between water activity (aw) and Tg over
indicates molecular weight. Adopted from from
the aw range of 0.1–0.8. This allows prediction of
Y.H. Roos, Glass Transition-Related Physico-Chemical
Changes in Foods, Food Technology, Vol. 49, No. 10, Tg at the aw range typical of dehydrated and inter-
p. 98, © 1995, Institute of Food Technologists mediate moisture foods.

Fig. 1.26  Modified state diagram showing relationship with critical values for water activity and water content.
between glass transition temperature (Tg), water activity Adapted from: Y.H. Roos, Glass Transition-Related
(GAB isotherm), and water content for an extruded snack Physico-Chemical Changes in Foods, Food Technology,
food model. Crispness is lost as water plasticization Vol. 49, No. 10, p. 99, © 1995, Institute of Food
depresses Tg to below 24 °C. Plasticization is indicated Technologists
30 1 Water

Fig. 1.27 Glass
Native starch [Roos, 1993]
transition temperature of 230 Maltodextrine DE 6.6
starch, maltodextrin, Wheat starch
Dextrose syrup DE 21
dextrose syrup and
180 Dextrose [Roos & Karel, 1993]
dextrose as a function of

Glass transition temp. Tg / °C


the water content.
Palzer, S. (2009) Maltodextrine DE 6
Influence of material 130
properties pm the Dextrose syrup DE 21
agglomeration of water
80 Amorphous dextrose
soluble particles.
Powder Technology
189:318 30

-20

-70
0 5 10 15 20 25 30
water content (wb) / %

Table 1.11  Reaction rates in foods as determined by


Water Activity and Reaction Rate water activity
Loosely
Water activity has a profound effect on the rate of Monolayer Capillary bound
many chemical reactions in foods and on the rate Reaction water water water
of microbial growth (Labuza 1980). This infor- Enzyme activity Zero Low High
mation is summarized in Table 1.11. Enzyme Mold growth Zero Lowa High
activity is virtually nonexistent in the monolayer Yeast growth Zero Lowa High
water (aw between 0 and 0.2). Not surprisingly, Bacterial growth Zero Zero High
growth of microorganisms at this level of aw is Hydrolysis Zero Rapid High
increase
also virtually zero. Molds and yeasts start to
Nonenzymic Zero Rapid High
grow at aw between 0.7 and 0.8, the upper limit of browning increase
capillary water. Bacterial growth takes place Lipid oxidation High Rapid High
when aw reaches 0.8, the limit of loosely bound increase
water. Enzyme activity increases gradually a
Growth starts at aw of 0.7–0.8
between aw of 0.3 and 0.8, then increases rapidly
in the loosely bound water area (aw 0.8–1.0).
Hydrolytic reactions and non-enzymic browning
do not proceed in the monolayer water range of Water Activity and Food Spoilage
aw (0.0–0.25). However, lipid oxidation rates are
high in this area, passing from a minimum at aw The influence of water activity on food quality
0.3–0.4, to a maximum at aw 0.8. The influence and spoilage is well established and widely rec-
of aw on chemical reactivity has been reviewed ognized as an important factor (Rockland and
by Leung (1987). The relationship between Nishi 1980). Moisture content and water activity
water activity and rates of several reactions and affect the progress of chemical and microbiolog-
enzyme activity is presented graphically in ical spoilage reactions in foods. Dried or freeze-­
Fig. 1.28 (Bone 1987). dried foods, which have great storage stability,
Water Activity and Food Spoilage 31

Fig. 1.28  Relationship between water activity and a number of reaction rates. http://www.aqualab.com/education/
measurement-of-water-activity-for-product-quality/

usually have water contents in the range of about these can adversely affect product quality. Lipase
5–15%. The group of intermediate-moisture can retain activity as at aw values as low as 0.3 or
foods, such as dates and cakes, may have sometimes 0.1 (Loncin et al. 1968). Acker (1969)
­moisture contents in the range of about 20–40%. provided examples of the effect of water activity
The dried foods correspond to the lower part of on some enzymatic reactions. Practical observa-
the sorption isotherms. This includes water in the tions of lipase and lipoxygenase activity can be
monolayer and multilayer category. Intermediate-­ observed in soy flour, whole grain wheat flours,
moisture foods have water activities generally crackers and peanuts where rancid aroma is
above 0.5, including the capillary water. observed in otherwise sable products with aw of 0.3
Reduction of water activity can be obtained by or lower. Table 1.12 contains the minimum aw and
drying or by adding water-soluble substances, temperatures for enzyme activity. The low water
such as sugar to jams or salt to pickled preserves. activity for lipases is taken advantage of to produce
Bacterial growth is virtually impossible below a interesterified lipids in non-aqueous systems.
water activity of 0.90. Molds and yeasts are usu- Acker found that for reactions in which lipo-
ally inhibited between 0.88 and 0.80, although lytic enzyme activity was measured, the manner
some osmophile yeast strains grow at water in which components of the food system were put
activities down to 0.65. into contact significantly influenced the enzyme
Uncontrolled enzyme activity in intermediate activity. Separation of substrate and enzyme
moisture foods can have detrimental effects on could greatly retard the reaction. Also, the sub-
quality. strate has to be in liquid form; for example, liquid
Most enzymes are inactive when the water oil could be hydrolyzed at water activity as low
activity falls below 0.85. Such enzymes include as 0.15, but solid fat was only slightly hydro-
invertases, amylases, phenoloxidases, peroxidases, lyzed. Oxidizing enzymes were affected by water
lipoxygenases and lipases (see Table 1.12). All of activity in about the same way as hydrolytic
32 1 Water

Table 1.12 Minimum aw values for enzyme activity in selected foods and model systems
Product/substrate Enzyme T (°C) aw threshold
Grains Phytases 20 0.90
Wheat germ Glucoside-hydrolases 20 0.20
Rye flour Amylases 30 0.75
Proteases
Macaroni Phospholipases 25-30 0.45
Wheat flour dough Proteases 35 0.96
Bread Amylases 30 0.36
Proteases
Starch Amylases 37 0.40/0.75
Potato Polyphenoloxidase 25 0.25
Galactose Galactosidase 30 0.40/0.60
Olive oil Lipase 5-40 0.025
Triolein, trilaurin Phospholipases 30 0.45
Glucose Glucose oxidase 30 0.40
Linoleic acid Lipoxygenase 25 0.50/0.70
Casein Trypsin 30 0.50
Adapted from Drapon (1985). Potato data from Acker (1969)

Fig. 1.29  Color change of milk powder (blue color J.J. Bimbenet, and J. Lenges, Influence of the Activity of
dashed line) and loss of free lysine in milk powder (red Water on the Spoilage of Foodstuffs, J. Food Technol.,
color dashed line) kept at 40 °C for 10 days as a function Vol. 3, pp. 131–142, 1968
of water activity. Data adopted from: From M. Loncin,

enzymes, as was shown by the example of pheno- parallels the color change, as is shown in
loxidase from potato. Fig. 1.29.
Nonenzymatic browning or Maillard reac- Labuza et al. (1970) have shown that, even at
tions are one of the most important factors caus- low water activities, sucrose may be hydrolyzed
ing spoilage in foods. These reactions are to form reducing sugars that may take part in
strongly dependent on water activity and reach a browning reactions. Browning reactions are usu-
maximum rate at a values of 0.6–0.7 (Loncin ally slow at low humidities and increase to a
et al. 1968). This is illustrated by the browning maximum in the range of intermediate-moisture
of milk powder kept at 40 °C for 10 days as a foods. Beyond this range the rate again decreases.
function of water activity (Fig. 1.29). The loss This behavior can be explained by the fact that, in
in lysine resulting from the browning reaction the intermediate range, the reactants are all dis-
Water Activity and Packaging 33

Fig. 1.30  Peroxide production in freeze-dried salmon on Rate of Deterioration of Freeze-Dried Salmon, J. Food
stored at different relative humidities. Source: From Sci., Vol. 33, pp. 241–247, 1968
F. Martinez and T.P. Labuza, Effect of Moisture Content

solved, and that further increase in moisture con- optimal conditions for long storage life. Sorption
tent leads to dilution of the reactants. isotherms play an important role in the selection
The effect of water activity on oxidation of of packaging materials. Hygroscopic products
fats is complex. Storage of freeze-dried and always have a steep sorption isotherm and reach
dehydrated foods at moisture levels above those the critical area of moisture content before reach-
giving monolayer coverage appears to give maxi- ing external climatic conditions. Such foods have
mum protection against oxidation. This has been to be packaged in glass containers with moisture-­
demonstrated by Martinez and Labuza (1968) proof seals or in watertight plastic (thick
with the oxidation of lipids in freeze-dried Polyvinylchloride). For example, consider instant
salmon (Fig. 1.30). Oxidation of the lipids was coffee, where the critical area is at about 50%
reduced as water content increased. Thus, condi- RH. Under these conditions the product cakes
tions that are optimal for protection against oxi- and loses its flowability. Other products might
dation may be conducive to other spoilage not be hygroscopic and no unfavorable reactions
reactions, such as browning. occur at normal conditions of storage. Such prod-
Other reactions that may be influenced by ucts can be packaged in polyethylene containers.
water activity are hydrolysis of protopectin, split- There are some foods where the equilibrium
ting and demethylation of pectin, autocatalytic relative humidity is above that of the external cli-
hydrolysis of fats, and the transformation of chlo- matic conditions. The packaging material then
rophyll into pheophytin (Loncin et al. 1968). serves the purpose of protecting the product from
moisture loss. This is the case with processed
cheese and baked goods.
Water Activity and Packaging Different problems may arise in composite
foods, such as soup mixes, where several distinct
Because water activity is a major factor influenc- ingredients are packaged together. In Fig. 1.31,
ing the keeping quality of a number of foods, it is for example, substance B with the steep isotherm
obvious that packaging can do much to maintain is more sensitive to moisture, and is mixed in
34 1 Water

Water Activity and Food Processing

Water activity is one of the criteria for establishing


good manufacturing practice (GMP) regulations
governing processing requirements and classifica-
tion of foods (Johnston and Lin 1987). Process
requirements for foods are governed by aw and pH;
aw controlled foods are those with pH greater than
4.6 and aw less than 0.85. At pH less than 4.6 and
aw greater than 0.85, foods fall into the category of
low-acid foods; when packaged in hermetically
sealed containers, these foods must be processed
Fig. 1.31  Sorption isotherms of materials A and B to achieve commercially sterile conditions.
Intermediate moisture foods are in the aw
equal quantities with substance A in an imperme- range of 0.90–0.60. They can achieve stability by
able package. The initial moisture content of B is a combination of aw with other factors, such as
X1 and after equilibration with A, the moisture pH, heat, preservatives, and Eh (equilibrium rela-
content is X2. The substances A and B will reach tive humidity).
a mean relative humidity of about 40%, but not a The aw level of 0.85 is applied as a point for
mean moisture content. If this were a dry soup determining whether a low-acid canned food or an
mix and the sensitive component was a freeze-­ acidified food is covered by FDA regulations. Low-
dried vegetable with a moisture content of 2% acid canned foods can be preserved by controlling
and the other component, a starch or flour with a water activity at levels above 0.85. The minimum
moisture content of 13%, the vegetable would be aw level for the growth of C. botulinum is approxi-
moistened to up to 9%. This would result in rapid mately 0.93. The regulations (21 CFR 113.3(e) (1)
quality deterioration due to nonenzymatic brown- (ii)) state that “commercial sterility can be achieved
ing reactions. In this case, the starch would have by the control of water activity and the application
to be postdried. of heat. The heat is generally necessary at aw levels
Salwin and Slawson (1959) found that stabil- above 0.85 to destroy vegetative cells of microor-
ity in dehydrated foods was impaired if several ganisms of public health significance (e.g., staphy-
products were packaged together. A transfer of lococci) and spoilage microorganisms which can
water could take place from items of higher grow in a reduced aw environment”. (See the fol-
moisture-vapor pressure to those of lower lowing other sections of the regulations which deal
moisture-­ vapor pressure. These authors deter- with aw controlled products:
mined packaging compatibility by examining the
respective sorption isotherms. They suggested a 21 CFR 113.10—Attendance at an approved
formula for calculation of the final equilibrium school giving instruction appropriate to the
moisture content of each component from the preservation technology involved.
isotherms of the mixed food and its equilibrium 21 CFR 113.40(i)—Equipment and procedures
relative humidity: for thermal processing of foods where critical
The initial relative humidity of A is 65% and factors such as water activity are used.
of B, 15%. 21 CFR 113.81(f)—Additional factors to be con-
trolled to prevent the growth of microogan-
(W ⋅ S ⋅ a ′ ) + (W2 ⋅ S 2 ⋅ aw 2′ )
aw = 1 1 w 1 isms not destroyed by the thermal process.

(W1 ⋅ S1 ) + (W2 ⋅ S 2 )
21 CFR 113.100(a) (6)—Record keeping require-
where ments for aw determinations.
W1 = gram solids of ingredient 1
S1 = linear slope of ingredient 1 Low-acid canned foods (LACF) include foods
aw1′ = initial aw of ingredient 1 with a finished equilibrium pH greater than 4.6
References 35

and a water activity greater than 0.85. This References


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Levine, H., & Slade, L. (1986). A polymer physico-­ Rockland, L. B., & Nishi, S. K. (1980). Influence of water
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in systems of biological interest. I Effect of mem- and delayed ice formation in sucrose solutions.
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in systems of biological interest. II Effect of solutes amorphous sucrose and frozen sucrose solutions.
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Lusena, C. V., & Cook, W. H. (1955). Ice propagation tein hydration, glass transitions, and structural relax-
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Salwin, H., & Slawson, V. (1959). Moisture transfer in In L. B. Rockland & G. F. Steward (Eds.), Water
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Saravacos, G. D. (1967). Effect of the drying method on Wierbicki, E., & Deatherage, F. E. (1958). Determination
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activity in biological systems—A review. Polish cells at low temperatures. In International congress
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The Journal of Physical Chemistry, 88, 3364–3373. (pp. 1377–1390).
Trombetta, G., Di Bona, C., & Grazi, E. (2005). The tran- Wolfe, J., & Bryant, G. (2001). Cellular cryobiology:
sition of polymers into a network of polymers alters thermodynamic and mechanical effects. International
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van den Berg, C., & Bruin, S. (1981). Water activity and ‘unfreezable water’, how unfreezable is it and how
its estimation in food systems: Theoretical aspects. much is there? CryoLetters, 23, 157–166.
Lipids
2
John W. Finley and John M. deMan

It is difficult to provide a clear and comprehensive for making fabricated foods, such as margarine
definition for the class of substances called lipids. and shortening, are almost pure triglyceride mix-
Early definitions were mainly based on whether tures. Fats are sometimes divided into visible and
the substance is soluble in organic solvents like invisible fats. In the United States, about 60% of
ether, benzene, or chloroform and is not soluble in total fat and oil consumed consists of invisible
water. In addition, definitions usually emphasize fats—that is, those contained in dairy products
the presence of fatty acids. Every definition pro- (excluding butter), eggs, meat, poultry, fish,
posed so far has some limitations. For example, fruits, vegetables, and grain products. The visible
monoglycerides of the short-chain fatty acids are fats, including lard, butter, margarine, shorten-
undoubtedly lipids, but they would not fit the defi- ing, and cooking oils, account for 40% of total fat
nition on the basis of solubility because they are intake. The interrelationship of most of the lipids
more soluble in water than in organic solvents. is represented in Fig. 2.1. A number of minor
Instead of trying to find a definition that would components, such as hydrocarbons, fat-soluble
include all lipids, it is better to provide a scheme vitamins, and pigments are not included in this
describing the lipids and their components, as scheme.
illustrated in Fig. 2.1 shows. The basic compo- Fats and oils may differ considerably in com-
nents of lipids are listed in the central column with position, depending on their origin. Both fatty
the fatty acids occupying the prominent position. acid and glyceride composition may result in dif-
The left column lists the lipids known as phospho- ferent properties. Fats and oils can be classified
lipids. The right column of the diagram includes broadly as of animal or vegetable origin. Animal
the compounds most important from a quantitative fats can be further subdivided into mammal depot
standpoint in foods. These are mostly esters of fat (lard and tallow) and milk fat (ruminant) and
fatty acids and glycerol. Up to 99% of the lipids in marine oils (fish and whale oil). Vegetable oils
plant and animal material consist of such esters, and fats can be divided into seed oils (such as
known as fats and oils. soybean, canola), fruit coat fats (palm and olive
The fat content of foods can range from very oils), and kernel oils (coconut and palm kernel).
low to very high in both vegetable and animal The scientific names for esters of glycerol and
products, as indicated in Table 2.1. In non-­ fatty acids is acylglycerols. Triacylglycerols, dia-
modified foods, such as meat, milk, cereals, and cylglycerols, and monoacylglycerols which have
fish, the lipids are mixtures of many of the com- three, two, or one fatty acid ester linkages respec-
pounds listed in Fig. 2.1, with triglycerides com- tively. The common names for these compounds
prising the major portion. The fats and oils used are glycerides, e.g. triglycerides, diglycerides,

© Springer International Publishing AG 2018 39


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_2
40 2 Lipids

Fig. 2.1  Interrelationships of lipids

Table 2.1 Composition of Lipids in various foods There is no single common structure for lip-
expressed as % of total lipids (Belitz et al. 2009) ids, however, the most abundant group of lipids
Milk Soya Wheat Apple in foods are triglycerides, which are commonly
Total lipids 3.6 23 1.5 0.088 grouped as fats and oils. Glycerides have a glyc-
Triacyglycerols 94 88 41 5 erol backbone appended with from one to three
Mono-, Diacylglycerol 1.5 1 fatty acids through ester linkages. Triglycerides
Sterol <1 1 15 are found in most plant, animal and microbial tis-
Steroid Ester 1 2 sue. They serve as energy storage or energy
Phospholipids 1.5 10 20 47 sources and are included in membranes. In ani-
Glycolipids 1.5 29 17 mals triglycerides are found in all tissues to vary-
Sulfolipids 1 ing degrees.
Others 0.54 7 15 Table 2.2 includes the principal lipid fractions
found in some common foods. The data illustrate
and monoglycerides. The scientific and common that most foods contain both triglycerides and
names are used interchangeably in the literature, phospholipids. The compositions very widely,
and this practice is followed in this book. the contents of cholesterol, saturated fat and
Lipids are organic molecules generally formed trans-fat in foods have raised some health
by the esterification of an fatty acid to an alcohol. concerns.
The simplest definition is a lipid as any molecule
that is insoluble in water and soluble in organic
solvents. Most lipids are soluble to some extent  horthand Description of Fatty
S
in organic solvents such as hexane, ether, chloro- Acids and Glycerides
form or benzene. They constitute a complex col-
lection organic compounds that include fatty To describe the composition of fatty acids it is
acids, (A,D, E and K). Lipids have many impor- sometimes useful to use a shorthand designa-
tant functions in vivo and in foods. The diversity tion. In this convention the composition of a
of some of the most abundant lipid structures is fatty acid can be described by two numbers sep-
illustrated in Fig. 2.2. arated by a colon. The first number indicates the
Shorthand Description of Fatty Acids and Glycerides 41

Fig. 2.2  Major lipid structures

Table 2.2  Fat contents of some foods


Total fat Saturated Fat Mono- Poly- Trans Cholesterol
Food g/100 g g/100 g g/100 g g/100 g g/100 g mg/100 g
Cooked rice 0.370 0.074 0.074 0.091 0 0
Macaroni cooked 0.930 0.176 0.131 0.319 0 0
Instant Oat Meal 1.360 0.226 0.391 0.426 0 0
Miniwheats 2.000 0.500 0.300 1.100 0 0
Whole Wheat Bread 3.500 0.722 0.620 1.592 0 0
Cooked Asparagas 0.220 0.048 0.000 0.105 0 0
Pinto Beans 0.480 0.058 0.035 0.276 0 0

(continued)
42 2 Lipids

Table 2.2 (continued)
Total fat Saturated Fat Mono- Poly- Trans Cholesterol
Food g/100 g g/100 g g/100 g g/100 g g/100 g mg/100 g
Carrots 0.180 0.030 0.006 0.089 0 0
Sweet corn 1.500 0.197 0.374 0.603 0 0
Almonds 49.930 3.802 31.551 12.329 0 0
English Walnuts 65.210 6.126 8.933 55.194 0 0
Mayonnaise 74.850 11.703 16.843 44.690 0.187 42
Margarine (80%) 80.170 14.224 36.435 26.741 5.827 0
Butter (salted) 81.110 51.368 21.021 3.043 3.278 215
Cheddar Cheese 33.820 19.368 8.428 1.433 1.179 102
Whole Milk 3.250 1.865 0.812 0.195 0 10
Chicken 13.600 3.790 5.340 2.970 0 88
Beef Steak 11.360 4.407 5.034 0.569 0.565 85
Ground Beef 21.830 8.805 9.356 0.656 0 84

Fig. 2.3 The n − 3 Family Polyunsaturated Fatty Acids through desaturation (vertical arrows) and chain elonga-
Based on Linolenic Acid. The heavy arrows show the tion (horizontal arrows)
relationship between the most important n  − 3 acids

number of carbon atoms in the fatty acid chain, These have also been described as ω6 and ω3.
the second number indicates the number of dou- The reason for this type of description is that the
ble bonds. Thus, 4:0 is short for butyric acid, members of each group n − 6 or n − 3 are related
16:0 for palmitic acid, 18:1 for oleic acid, etc. biosynthetically through processes involving
The two numbers provide a complete description desaturation, chain elongation, and chain short-
of a saturated fatty acid. For unsaturated fatty ening (Gunstone 1986) (Fig. 2.3).
acids, information about the location of double Triglycerides can be abbreviated by using the
bonds and their stereo isomers can be given as first letters of the common names of the compo-
follows: oleic acid (the cis isomer) is 18:1c9; nent fatty acids. SSS indicates tristearin, PPP tri-
elaidic acid (the trans isomer) is 18:1t9. The palmitin, and SOS a triglyceride with two
numbering of carbon atoms in fatty acids starts palmitic acid residues in the 1 and 3 positions and
normally with the carboxyl carbon as number oleic acid in the 2 position. In some cases, glyc-
one. In some cases polyunsaturated fatty acids eride compositions are discussed in terms of sat-
are numbered starting at the methyl end; for urated and unsaturated component fatty acids. In
instance, linoleic acid is represented as 18:2n − 6 this case, S and U are used and glycerides would
and linolenic acid 18:3n  − 3. These symbols be indicated as SSS for trisaturated glyceride and
indicate straight-chain, 18-carbon fatty acids SUS for a glyceride with an unsaturated fatty
with two and three methylene interrupted cis acid in the 2 position. In other cases, the total
double bonds that start at the sixth and third number of carbon atoms in a glyceride is impor-
carbon from the methyl end, respectively.
­ tant, and this can be shortened to glycerides with
Fatty Acids 43

carbon numbers 54, 52, and so on. A glyceride higher the melting point. Short chain length and
with carbon number 54 could be made up of three unsaturation enhance the fluidity of lipids.
fatty acids with 18 carbons, most likely to happen The common feature of these lipids is that they
if the glyceride originated from one of the seed are all esters of moderate to long chain fatty acids.
oils. A glyceride with carbon number 52 could Acid or base-catalyzed hydrolysis yields the
have two component fatty acids with 18 carbons component fatty acid, some examples of which
and one with 16 carbons. The carbon number are given in the following table, together with the
does not give any information about saturation alcohol component of the lipid. These long-chain
and unsaturation. carboxylic acids are generally referred to by their
common names, which in most cases reflect their
sources. Natural fatty acids may be saturated or
Fatty Acids unsaturated, and as the following data indicate,
the saturated acids have higher melting points
Fatty acids are composed of hydrocarbon chains than unsaturated acids of corresponding size. The
with a carboxylic acid on one end of the mole- double bonds in the unsaturated compounds listed
cule. Generally in foods fatty acids exist as esters on the right are all cis (or Z).
of alcohols. When esterified to a long chain fatty
alcohol they are called waxes and when bound to
glycerol they are called glycerides. The variation Lipid Nomenclature
in chain length and saturation of the hydrocarbon
chain of fatty acids is important in determining In nature even numbered straight chain fatty acids
the physical and biological properties of lipids. constitute the majority of the fatty acids found
Fatty acids in biological systems are generally in triglycerides and phospholipids. Branched
composed of an even number of carbon atoms, chain fatty acids and hydroxyl fatty acids are
typically between 14 and 24, although the 16- found in nature but not commonly. One hydroxyl
and 18-carbon fatty acids are the most abundant. fatty acid of importance is ricinoleic acid which
Fatty acids typically contain an even number of is found in Castor plant (Ricinus communisL.,
carbon atoms because of the way in which fatty Euphorbiaceae) seeds or in sclerotium of ergot
acids are biosynthesized. The hydrocarbon side (Claviceps purpurea Tul., Clavicipitaceae). In
chains in animal fatty acids have hydrocarbon castor oil approximately 90% of the fatty acid
chains which are almost invariably un-branched. content in the triglyceride is ricinoleic acid.
The side chain may be saturated or it may contain Ricinoleic acid exerts analgesic and anti-inflam-
one or more double bonds. In unsaturated fatty matory effects. It is also frequently used in lubri-
acids, the double bonds are in the cis formation. cants including motor oil (Gunstone et al. 2007;
In polyunsaturated fatty acids the double bonds Vieira et al. 2000).
are generally separated by at least one methylene Fatty acids are also broken in groups by chain
group. The chain length and degree of saturation length. Short chain fatty acids include: Formic
control the properties that are found within the acid, Acetic acid, Propionic acid, Isobutyric acid
fatty acids and lipids. Unsaturated fatty acids (2-methylpropanoic acid), Butyric acid,
have lower melting points than saturated fatty Isovaleric acid (3-methylbutanoic acid), Valeric
acids of the same length. The cis-double bonds acid (pentanoic acid). Formic acid is the simplest
cause curvature in the hydrocarbon chain which carboxylic acid. Its chemical formula is HCOOH
prevents alignment with other side chains and or HCO2H. It is an important intermediate in
limits crystallization. The fatty acids cannot com- chemical synthesis and occurs naturally, most
pact tightly together, reducing the van der Waals notably in ant venom. Acetic acid is widely dis-
interaction between the fatty acids. The melting tributed in nature and is an important food addi-
point of fatty acids is also affected by chain tive contributing pH control, typical acid taste
length. The longer the hydrocarbon chain is, the and antibacterial properties. Propionic acid is an
44 2 Lipids

important aroma constituent in Swiss cheese, at 14 carbon atoms; and long-chain fatty acids, 16
higher concentrations it is a pungent unpleasant or more carbon atoms. Yet another division dif-
aroma. Butyric acid, the only short chain fatty ferentiates between essential and nonessential
acid commonly found in triglycerides is a major fatty acids.
component in butter fat. Free butyric acid exhib- Some of the more important saturated fatty
its a strong aroma which at low levels is an acids are listed with their systematic and com-
important note in dairy flavors ranging from very mon names in Table 2.3, and some of the unsatu-
small amounts in butter to high levels in lim- rated fatty acids are listed in Table 2.3. The
burger cheese. The other short chain fatty acids naturally occurring unsaturated fatty acids in fats
exhibit strong aromas and are found at low levels are almost exclusively in the cis-form (Table 2.3),
in food flavors. although trans-acids are present in ruminant milk
Medium chain fatty acids generally include fats and in catalytically hydrogenated fats.
saturated fatty acids with 6–12 carbons includ-
ing caproic, caprylic, capric and lauric acids.
The medium chain fatty acids are found in tri- Cis and trans Fatty Acids
glycerides which are frequently used in foods
because of their oxidative stability and low melt- In fats and oils fatty acids containing double
ing pint making them liquid at room tempera- bonds can occur in two forms cis and trans as
tures. They are also frequently used in cosmetic shown in Fig. 2.4. The fatty acids have identical
formulations. chemical composition the only differences is the
The long chain fatty acids are 14 or greater orientation around the double bond. As we will
straight chain fatty acids. Long chain fatty acids see late this can have a profound effect on the
occur as saturated fatty acids or with varying physical properties of the fat or oil.
degrees of unsaturation. Most fatty acids found in human body are cis
Even-numbered, straight-chain saturated and fatty acids except for Retinoic acid present in the
unsaturated fatty acids make up the greatest pro- eye. Vegetable oils are typically rich in mono-
portion of the fatty acids of natural fats. However, and polyunsaturated fatty acids. Saturated fatty
it is now known that many other fatty acids may acids can pack together more effectively result-
be present in small amounts. Some of these ing in a more solid fat whereas unsaturated fatty
include odd carbon number acids, branched-­ acids containing cis double bonds do not line up
chain acids, and hydroxyl-acids. These may effectively to form solid structures thus resisting
occur in natural fats (products that occur in solidification. Vegetable oils tend to be rich in
nature), as well as in processed fats. The latter mono and poly- unsaturated fatty acids. The
category may, in addition, contain a variety of unsaturated vegetable oils are converted from liq-
isomeric fatty acids not normally found in natural uids to solids by the hydrogenation reaction. The
fats. It is customary to divide the fatty acids into hydrogenation results in hardening of the fats
different groups, for example, into saturated and making them solid or semi-solids. Vegetable oils
unsaturated ones. This particular division is use- which are partially hydrogenated, are partially
ful in food technology because saturated fatty saturated so the melting point increases to the
acids have a much higher melting point than point where a solid is present at room tempera-
unsaturated ones, so the ratio of saturated fatty ture. The degree of hydrogenation of unsaturated
acids to unsaturated ones significantly affects the oils controls the final consistency of the product.
physical properties of a fat or oil. Another com- Partial hydrogenation results in the production
mon division is into short-chain, medium-chain, of trans fats. Trans fats are the result of a side
and long-chain fatty acids. Unfortunately, there is reaction with the catalyst of the hydrogenation
no generally accepted division of these groups. process. Unsaturated fat which is normally found
Generally, short-chain fatty acids have from 4 to as a cis isomer converts to a trans isomer of the
10 carbon atoms; medium-chain fatty acids, 12 or unsaturated fat. Isomers are molecules that have
Table 2.3  Structures and nomenclature of common fatty acids found in plants and animals
Name Formula Mass Melting
Lipid number Common Systematic Molecular Structural (g/mol) Point Diagram
Short and medium chain fatty acids
C4:0 Butyric acid Butanoic acid C4H8O2 CH3(CH2)2COOH 88.11 - 5.1 °C

C6:0 Caproic acid Hexanoic acid C6H12O2 CH3(CH2)4COOH 116.16 - 3.4 °C

C8:0 Caprylic acid Octanoic acid C8H16O2 CH3(CH2)6COOH 144.21 16.7 °C


C10:0 Capric acid Decanoic acid C10H20O2 CH3(CH2)8COOH 172.26 31.6 °C

C12:0 Lauric acid C12:0 C12H24O2 CH3(CH2)10COOH 200.32 43.8 °C


C14:0 Myristic Acid Tetradecanoic acid C12H24O2 CH3(CH2)10COOH 228.37 54.4 °C

Long chain saturated fatty acids


C16:0 Palmitic Acid Hexadecanoic acid C16H32O2 CH3(CH2)14CO2H 62.9 °C

C18:0 stearic acid Octadecanoic Acid C18H36O2 CH3(CH2)16CO2H 284.48 69.3 °C

C20:0 arachidic acid Eicassanoic C20H40O2 CH3(CH2)18CO2H 312.53 75.5 °C

Unsaturated fatty acids


C16:1 Palmitoleic acid 9-cis-Hexadecenoic acid C16H30O2 CH3(CH2)5CH = CH(CH2)7COOH 254.41 0.1 °C

C18:1(CH Oleic acid cis-9-Octadecenoic acid C18H34O2 CH3(CH2)7CH = CH(CH2)7COOH 282.46 13–14 °C

C18:1 t Vaccenic (E)-Octadec-11-enoic acid C18H34O2 CH3(CH2)5CH = CH(CH2)9COOH 282.46 44–45 °C

C18:1 t Elacdic Acid (E)-octadec-9-enoic acid C18H34O2 CH3(CH2)7CH = CH(CH2)7COOH 282.46 41–45 °C

C18:2ω6 Linoleic acid (9Z,12Z)-9,12-Octadecadienoic C18H32O2 CH3(CH2)4(CH = CHCH2)2(CH2)6COOH 280.45 −5 °C


acid

(continued)
Table 2.3 (continued)
Name Formula Mass Melting
Lipid number Common Systematic Molecular Structural (g/mol) Point Diagram
C18:3 ω3 α−Linolenic acid cis,cis,cis-9,12,15-Octadecatrienoic C18H30O2 CH3(CH2CH = CH)3(CH2)7COOH 278.43 −11 °C
acid; (9Z,12Z,15Z)-octadeca-
9,12,15-trienoic acid

C18:3 ω6 γ−Linolenic acid Cis-cis-cis-6,9,12-octadecatrienoic C18H30O2 CH3(CH2)4(CH2CH = CH)3(CH2)3COOH 278.43 - 11 °C


acid

C20:4 ω6 Arachidonic acid (5Z,8Z,11Z,14Z)-5,8,11,14- C20H32O2 CH3(CH2)4(CH2CH = CH)4(CH2)2COOH 304.47 −49 °C


Eicosatetraenoic acid

C18:4 ω3 Stearidonic (18:4) (6Z,9Z,12Z,15Z)-6,9,12,15- CH3CH2(CH = CH CH2)4(CH2)3COOH 276.40 N/A


octadecatetraenoic acid

C20:5 ω3 Eicosapentaenoic (5Z,8Z,11Z,14Z,17Z)-5,8,11,14,17-­ C20H30O2 CH3CH2(CH = CH CH2)5(CH2)2COOH 302.45


EPA icosapentaenoic acid

C22:6 ω3 Docosahexaenoic (4Z,7Z,10Z,13Z,16Z,19Z)-docosa-­ C22H32O2 CH3CH2(CH = CHCH2)5(CH2)2COOH 328.49


DHA 4,7,10,13,16,19-hexaenoic acid
Fatty Acids 47

Fig. 2.4 Hydrogenation OH
of linoleic acid yields O
monounsaturated fatty
acids and ultimately
reaches saturated stearic LinoleicAcid
H2 H2
acid. Eladic acid is the 18:2cis,cis-9,12.
preferred pathway for
monounsaturated OH OH
because trans forms are
O O
more thermodynamically
stable
Elaidic acid
Oleic Acid 18:1 trans-9
18:1 cis-9

H2 H2
OH

O
Stearic acid
18:0

Fig. 2.5  Isomers of linoleic acid

the same molecular formula but are bonded alkane (saturated). The hydrogenation reaction is
together differently. Focusing on the Sp2 double (Fig. 2.5):
bonded carbons, a cis isomer has the hydrogens The trans fatty acids are chemically “monoun-
on the same side. Due to the added energy from saturated” or “polyunsaturated” but they differ
the hydrogenation process, the activation energy from the cis monounsaturated or polyunsaturated
is reached to convert the cis isomers of the fatty acids. The trans fatty acids (although chem-
­unsaturated fat to a trans isomer of the unsatu- ically still unsaturated) produced by the partial
rated fat. The effect is putting one of the hydro- hydrogenation process act more like saturated fat
gens on the opposite side of one of the carbons. because they form similar linear structures that
This results in a trans configuration of the double result in higher melting points than a cis fatty
bonded carbons. acid of the same length.
Unsaturated fatty acids are converted to satu- The methods of hydrogenation of fats were
rated fatty acids by the relatively simple hydroge- developed in early 1900s for the purpose of devel-
nation reaction. The addition of hydrogen to an oping solid fats for making soaps. Later they were
alkene (unsaturated double bond) results in an used to hydrogenate dietary fatty acids such as
48 2 Lipids

soybean oils. Hydrogenated oils are more stable to with either one or two fatty acids are referred to as
oxidation and therefore are less prone to produc- mono- and di- glycerides. The mono- and di-­
ing rancid or oxidized off flavors in the products. glycerides are frequently used as emulsifiers in
The hydrogenation process allowed processors to food systems. Triglycerides (or triacylglycerols)
dial in melting ranges and physical properties of are found in both plants and animals, and compose
the hydrogenated fats for specific food applica- one of the major food groups of our diet. Figure 2.6
tions such as bakery shortening or frying oil. represents a typical glyceride structures. It should
While there are many modifications of the be noted that the fatty acid in monoglycerides can
process, the major process is to heat the unsatu- occur at any of the three positions on the glycerol
rated oils to above 200 °C, add powdered nickel and the two fatty acids on diglycerides can be on
(as catalyst) bubble hydrogen through it. The any two of the glycerol hydroxyl positions.
double bonds get saturated. However, all the dou- Triglycerides of animal origins such as lard or
ble bonds do not get saturated. The first hydrogen beef tallow tend to be solid or semisolid at room
addition is reversible. When the c-c bonding temperature are classified as fats. Those triglyc-
rotates to the trans form, which is more stable, erides that are liquid are called oils and originate
subtraction of the hydrogen results in the forma- chiefly in plants, although triglycerides from fish
tion of the trans-fat. The trans configuration is are also largely oils. Some examples of the com-
more stable than cis. In that process at the newer position of triglycerides from various sources are
position they become trans double bond. Cis given in Table 2.4. Animal fats exhibit large vari-
configuration has more strain in it than trans. As ations in fatty acid composition. Chicken and
it is seen on the packet ingredient list, they are turkey tend to have higher levels of polyunsatu-
written as “partially hydrogenated”. rates than beef, sheep and pig (Table 2.5).
Fatty acids can also form structures known as Natural fats can be defined as mixtures of
micelles in an aqueous solution. The structure is mixed triglycerides. Simple triglycerides are vir-
formed when the hydrocarbon tails form a hydro- tually absent in natural fats, and the distribution
phobic center, while the polar heads form a of fatty acids both between and within glycerides
hydrophilic shell outside the interior. The signifi- is selective rather than random. When asymmet-
cance of micelles is that they act as emulsifiers, ric substitution in a glycerol molecule occurs,
thus dissolving fat-soluble vitamins or other lip- enantiomorphic forms are produced (Kuksis
ids that need to be absorbed. 1972; Villeneuve and Foglia 1997). This is illus-
trated in Fig. 2.7. Glycerol has a plane of sym-
metry or mirror plane, because two of the four
Triglycerides substituents on the central carbon atom are iden-
tical. When one of the carbon atoms is esterified
The predominant form of lipids in fats and oils are with a fatty acid, a monoglyceride results and two
as the triesters of fatty acids with glycerol non-superimposable structures exist. These are
(1,2,3-trihydroxypropane). Glycerol appended called enantiomers and are also referred to as

Fig. 2.6  Structures of


glycerol lipids
Triglycerides

Table 2.4  Principal fatty acid composition and melting points of food fats and oils
% Total Fatty acids
Oil or Fat Saturated Monounsaturated Polyunsaturated Melting point
o
4:0 6:0 8:0 10:0 12:0 14:0 16:0 18:0 16:1 18:1 18:2 18:3 C
Beef tallow 3 24 19 4 43 3 1 41–42
Butterfat 4 2 1 3 3 11 27 12 2 29 2 1 −10
Canola oil 4 2 62 22 10 14
Cocoa butter 26 34 34 3 24–38
Coconut 1 8 6 47 18 9 3 6 2 25
Corn 11 2 28 58 1 11
Cottonseed 1 22 3 1 19 54 1 48
Lard 2 26 14 3 44 10 36–45
Olive 13 3 1 71 10 6
Palm 1 45 4 40 10 35
Palm Kernal 3 4 48 16 8 3 15 2 3
Peanut 11 2 48 32 3
Safflower 7 2 13 78 −17
Soybean 11 4 24 54 7 −16
Sunflower 7 5 19 68 1 −17
49
50 2 Lipids

Fig. 2.7 A typical gas–liquid chromatogram of total numbering of all the peaks in the region between 18:0 and
bovine milk fatty acid methyl ester prepared by NaOCH3/ 18:2n − 6 using this column. [22:6n − 3] is not present in
methanol (10 min at 50 °C) followed by HCl/methanol these milk samples but the position at which it should
(10 min at 80 °C), and separated on a 100-m fused silica emerge in the chromatogram is indicated. Kramer, J.K.G.,
capillary column (SP-2560; Supelco Inc., Bellefonte, PA). Fellner, V., Dugan, M.E.R. et al. Lipids (1997) 32: 1219.
i iso, a anteiso; numbers 1–20 are an arbitrary consecutive doi:10.1007/s11745-997-0156-3

Table 2.5  Component fatty acids of animal depot fats Table  2.4. The kind of feed consumed by the
Fatty Acids Wt % ­animals may greatly influence the composition of
Animal 14:0 16:0 16:1 18:0 18:1 18:2 18:3 the depot fats. Animal depot fats are character-
Pig 1 24 3 13 41 10 1 ized by the presence of 20–30% palmitic acid, a
Beef 4 25 5 19 36 4 Trace property shared by human depot fat. Many of the
Sheep 3 21 2 25 34 5 3 seed oils, in contrast, are very low in palmitic
Chicken 1 24 6 6 40 17 1 acid. The influence of food consumption applies
Turkey 1 20 6 6 38 24 2 equally for the depot fat of chicken and turkey
(Marion et al. 1970; Jen et al. 1971). The animal
chiral. A racemic mixture is a mixture of equal depot fats are generally low in polyunsaturated
amounts of enantiomers. Asymmetric or chiral fatty acids. The iodine value of beef fat is about
compounds are formed in 1-monoglycerides; all 50 and of lard about 60. Iodine value is generally
1, 2-diglycerides; 1, 3-diglycerides containing used in the food industry as a measure of total
unlike substituents; and all triglycerides in which unsaturation in a fat.
the 1- and 3-positions carry different acyl groups. Iodine value is a measure of the degree of
The depot fats of higher land animals, espe- unsaturation of an oil or wax. The oil is reacted
cially mammals, have relatively simple fatty acid with is reacted with iodine, which adds across the
composition. The fats of birds are somewhat double bond. The iodine unreacted iodine is then
more complex. The fatty acid compositions of measured and the differences is the iodine num-
some major food fats of this group are listed in ber. A saturate fat would have a value of zero.
Triglycerides 51

Cocoa butter typically has iodine values of 30–40 It is impossible to determine all of the constitu-
and highly unsaturated oil like soy has a value of ents of milk fatty acids by a normal chromato-
130–136. These values are used by the industry as graphic technique, because many of the minor
an index of unsaturation and vulnerability to oxi- component fatty acids are either not resolved or
dation. The iodine addition reaction is shown as: are covered by peaks of other major fatty acids. A
milk fat chromatogram of fatty acid composition
is shown in Fig. 2.6. Such fatty acid compositions
as reported are therefore only to be considered as
Ruminant milk fat is extremely complex in approximations of the major component fatty
fatty acid composition. By using gas chromatog- acids; these are listed in Table 2.6. This table
raphy in combination with fractional distillation reports results of the major component fatty acids
of the methyl esters and adsorption chromatogra- in bovine milk fat as well as their distribution
phy, Magidman et al. (1962) and Herb et al. among the sn-1, sn-2, and sn-3 positions in the
(1962) identified at least 60 fatty acids in cow’s triacylglycerols (Jensen and Newburg 1995). Milk
milk fat. Several additional minor fatty acid com- fatty acids can vary considerably. Seasonal varia-
ponents have been found in other recent studies. tions because of changes in feed are one of the
About 12 fatty acids occur in amounts greater major factors (Jensen 2000; Månsson 2008). The
than 1% (Jensen and Newburg 1995). Among composition of bovine milk lipids over a 5 year
these, the short-chain fatty acids from butyric to period was reviewed by Jensen (2002).
capric are characteristic of ruminant milk fat. In most natural fats the double bonds of unsat-
Data provided by Hilditch and Williams (1964) urated fatty acids occur in the cis configuration.
and Markiewicz-Kęszycka et al. 2013, on the In milk fat a considerable proportion is in the
component fatty acids of some milk fats are listed trans configuration. These trans bonds result
in Table 2.6. Fatty acid compositions are usually from microbial action in the rumen where poly-
reported in percentage by weight, but in the case unsaturated fatty acids of the feed are partially
of fats containing short-chain fatty acids (or very hydrogenated. Catalytic hydrogenation of oils
long-chain fatty acids) this method may not give a in the fat industry also results in trans isomer
good impression of the molecular proportions of
fatty acids present. Therefore, in many instances, Table 2.7  Major fatty acids of bovine milk fat and their
the fatty acid composition is reported in mole per- distribution in the triacylglycerols
cent, as is the case with the data in Table 2.7. Bovine milk fat
Fatty acids (mol%) TG sn-1 sn-2 sn-3
Table 2.6  The component fatty acids of some milk fats
4:0 11.8 – – 35.4
in g/100 g
6:0 4.6 – 0.9 12.9
Fatty acids in g/100 g fat Goat Sheep Cow 8:0 1.9 1.4 0.7 3.6
C4:0 Butyric 2.03 2.57 2.87 10:0 37 1.9 3.0 6.2
C6:0 Caproic 2.78 1.87 2.01 12:0 3.9 4.9 6.2 0.6
C8:0 Caprylic 2.92 1.87 1.39 14:0 11.2 9.7 17.5 6.4
C10:0 Capric 9.59 6.33 3.03 15:0 2.1 2.0 2.9 1.4
C12:0 Lauric 4.52 3.99 3.64 16:0 23.9 34.0 32.3 5.4
C14:0 Myristic 9.83 10.17 10.92 16:1 2.6 2.8 3.6 1.4
C16:0 Palmitic 24.64 25.10 28.70 17:0 0.8 1.3 1.0 0.1
C18:0 Stearic 8.87 8.85 11.23 18:0 7.0 10.3 9.5 1.2
C18:1 cis-9 Oleic 18.65 20.18 22.39 18:1 24.0 30.0 18.9 23.1
C18:2 cis-9, cis-12 Linoleic 2.25 2.32 2.57 18:2 2.5 1.7 3.5 2.3
C18:2 cis-9, trans-11 CLA 0.45 0.76 0.57 18:3 Trace – – –
C18:3 cis-9, cis-12, cis-15 0.77 0.92 0.50 Adapted from R.G. Jensen and D.S. Newburg, Milk
Linolenic Lipids, in Handbook of Milk Composition, R.G. Jensen,
Adapted from Markiewicz-Kęszycka et al. (2013) ed., p. 546, © 1995, Academic Press
52 2 Lipids

f­ ormation. The level of trans isomers in milk fat there contain one or more double bonds causing
has been reported as 2–4% (deMan and deMan curvature in the chain and results in lower the
1983). Since the total content of unsaturated fatty melting point of the fatty acid. In most fats and oils
acids in milk fat is about 34%, trans isomers may the natural form of the fatty acid \double bond is in
constitute about 10% of total unsaturation (Akoh the cis form. The cis-double bond(s) in the unsatu-
1997). The complexity of the mixture of different rated fatty acids introduces a kink in chain, which
isomers is demonstrated by the distribution of makes it more difficult to pack their molecules
positional and geometric isomers in the monoe- together in a stable repeating array or crystalline
noic fatty acids of milk fat (Table 2.8) and in the lattice. The trans-double bond isomer of oleic
unconjugated 18:2 fatty acids (Table 2.9). The acid, known as elaidic acid, has a linear shape and
iodine value of milk fat is in the range of 30–35, a melting point of 45 °C (32 °C higher than its cis
much lower than that of lard, shortening, or mar- isomer). The shapes of stearic and oleic acids are
garine, which have similar consistencies. displayed in the models below.
The higher melting points of the saturated fatty Marine oils have also been found to contain a
acids are because the uniform rod-like shape of large number of component fatty acids. Ackman
their molecules which allows the fatty acids to (1972) has reported as many as 50 or 60 compo-
align in crystal like forms. Unsaturated fatty acids nents. Only about 14 of these are of importance
in terms of weight percent of the total. These con-
Table 2.8  Positional and geometric isomers of bovine sist of relatively few saturated fatty acids (14:0,
milk lipid monoenoic fatty acids (Wt%) 16:0, and 18:0) and a larger number of unsatu-
cis Isomers trans Isomers rated fatty acids with 16–22 carbon atoms and up
Position of to 6 double bonds. This provides the possibility
double bond 14:1 16:1 17:1 18:1 16:1 18:1 for many positional isomers.
5 1.0 Tr 2.2 The complexity of the fatty acid composition
6 0.8 1.3 3.4 7.8 1.0
of marine oils is evident from the chromatogram
7 0.9 5.6 2.1 6.7 0.8
shown in Fig. 2.8. The end structure of the poly-
8 0.6 Tr 20.1 1.7 5.0 3.2
unsaturated fatty acids is of nutritional impor-
9 96.6 88.7 71.3 95.8 32.8 10.2
tance, especially eicosapentaenoic acid (EPA),
10 – Tr Tr Tr 1.7 10.5
11 – 2.6 2.9 2.5 10.6 35.7
20:5ω3 or 20:5 n-3, and docosahexaenoic acid
12 – Tr Tr – 12.9 4.1 (DHA), 22:6ω3 or 22:6 n-3. The double bonds in
13 – – – – 10.6 10.5 marine oils occur exclusively in the cis configu-
14 – – – – – 9.0 ration. EPA and DHA can be produced slowly
15 – – – – – 6.8 from linolenic acid by herbivore animals, but not
16 – – – – – 7.5 by humans. EPA and DHA occur in major
Source: From R.G. Jensen, Composition of Bovine Milk amounts in fish from cold, deep waters, such as
Lipids, J. Am. Oil Chem. Soc., Vol. 50, pp. 186–192, 1973 cod, mackerel, tuna, swordfish, sardines, and

Table 2.9  Location of double bonds in unconjugated 18:2 isomers of milk lipids
cis, cis cis, trans or trans, cis trans, trans
11, 15 11, 16 and/or 11, 15 12, 16
10,15 10, 16 and/or 10, 15 11, 16 and/or 11, 15
9, 15 9, 15 and/or 9, 16 10,16 and/or 10, 15
8, 15 and/or 8, 126 8, 16 and/or 8, 15 and/or 8, 12 9, 16 and/or 9, 15 and/or 9, 13
7, 15 and/or 7, 12
15 and/or 6, 12
Adopted from R.G. Jensen, Composition of Bovine Milk Lipids, J. Am. Oil Chem. Soc., Vol. 50, pp. 186–192, 1973;
Morrison, W.R., in “Topics in Lipid Chemistry,” Vol. 1, Ed. by F.D. Gunstone, Logos Press, London, 1970, p. 51.; Van
der Wel, H., and K. De Jong, Fette Seifen Anstrichm. 67:279 (1967)
Component Glycerides 53

C20:5 (EPA)
DB-23
Norm.

C20:1n9
C14:0

C16:1

C18:1. cis

C22:6 (DHA)
100

90

80

70

C18:2. cis
60
C16:0

C18:3 n3
50
C18:0 C18:1. trans
C17:0

C24:0
C18:2. trans

C22:1

C24:1
C22:0
40

C20:4 n6

C22:2
C17:1

C20:2

C23:0
C18:3 n6

C20:3 n3
C14:1

C20:3 n6
C15:0

30
C12:0

C20:0

C21:0
C15:1

20

8 10 12 14 16 18 20 22 24

Fig. 2.8  Analysis fatty acid methyl esters of a typical marine oil sample. www.agilent.com/cs/library/applications/5989-
3760EN.pdf

herring (Ackman 1988a; Simopoulos 1988). 3% flax, soy or canola oil. The flax oil which is
Arachidonic acid is the precursor in the human rich in omega-3 fatty acids resulted in higher lev-
system of prostanoids and leukotrienes. els of omega-3 fatty acids in the egg yolk. All three
Ackman (1988b) has drawn attention to the experimental diets resulted in higher levels of
view that the fatty acid compositions of marine mono and poly unsaturated fatty acids (Table 2.12).
oils are all much the same and vary only in the Commonly used food oils have a wide range of
proportions of fatty acids. The previously held fatty acid composition which impacts functional-
view was that marine oils were species-specific. ity in food systems, nutritional profile and oxida-
The major fatty acids of commonly consumed tive stability. Table 2.13 summarizes some of the
seafood are found in Table 2.10. variations seen between vegetable oils.
The fatty acid composition of egg yolk is
given in Table 2.11. The main fatty acids are pal-
mitic, oleic, and linoleic. The yolk constitutes Component Glycerides
about one-third of the weight of the edible egg
portion. The relative amounts of egg yolk and Natural fats can be defined as mixtures of mixed
white vary with the size of the egg. Small eggs triglycerides. Simple triglycerides are virtually
have relatively higher amounts of yolk. The egg absent in natural fats, and the distribution of fatty
white is virtually devoid of fat. acids both between and within glycerides is
Diet can exert a significant impact on the lipid selective rather than random. When asymmetric
profile of foods. Milinsk et al. fed chickens diets substitution in a glycerol molecule occurs, enan-
enriched with various lipids and observed that the tiomorphic forms are produced (Kuksis 1972;
fatty acid profiles in egg yolk are altered by the Villeneuve and Foglia 1997). This is illustrated in
diet. The control diet was based on soy and the Fig.  2.9. Glycerol has a plane of symmetry or
flax, soy and canola diets were all enriched with mirror plane, because two of the four substituents
54 2 Lipids

Table 2.10  Total fat content, fatty acid content of raw seafood
Atlantic
Anchovy Salmon Cod Raw Shrimp raw Tuna Raw, Bluefin Bluefish Raw
Nutrient g/100 g Edible portion
Water 73.37 68.5 81.22 78.45 68.09 70.86
Total lipid (fat) 4.84 6.34 0.67 0.51 4.9 4.24
Lipids
Fatty acids, total saturated 1.282 0.981 0.131 0.101 1.257 0.915
C14:0 0.302 0.137 0.009 0.001 0.139 0.179
C16:0 0.715 0.632 0.091 0.052 0.81 0.576
C18:0 0.252 0.21 0.03 0.037 0.307 0.16
Fatty acids, total 1.182 2.103 0.094 0.086 1.6 1.793
monounsaturated
C16:1 0.4 0.251 0.016 0.003 0.162 0.277
C18:1 0.624 1.351 0.061 0.038 0.924 0.684
C20:1 0 0.223 0.015 0.002 0.277 0.34
C22:1 0.115 0.279 0.003 0 0.237 0.492
Fatty acids, total 1.637 2.539 0.231 0.152 1.433 1.06
polyunsaturated
C18:2 0.097 0.172 0.005 0.032 0.053 0.06
C18:3 0 0.295 0.001 0.002 0 0
C18:4 0.055 0.083 0.001 0 0.039 0.167
C20:4 0.007 0.267 0.022 0.012 0.043 0
20:5 n-3 (EPA) 0.538 0.321 0.064 0.03 0.283 0.252
22:5 n-3 (DPA) 0.029 0.287 0.01 0.002 0.125 0.062
22:6 n-3 (DHA) 0.911 1.115 0.12 0.031 0.89 0.519
Cholesterol (mg) 60 55 43 161 38 59
National Nutrient Database for Standard Reference Release 28 slightly revised May, 2016 https://ndb.nal.usda.gov/ndb/
foods

Table 2.11  Fatty acid composition of egg yolk racemic mixture is a mixture of equal amounts of
Fatty Acid % enantiomers. Asymmetric or chiral compounds
Total saturated 36.2 are formed in 1-monoglycerides; all 1,
 14:0 0.3 2-­diglycerides; 1, 3-diglycerides containing
 16:0 26.6 unlike substituents; and all triglycerides in which
 18:0 9.3 the 1- and 3-positions carry different acyl groups.
Total monounsaturated 48.2 The glyceride molecule can be represented in
 16:1 4.0 the wedge and slash form (Fig. 2.10). In this spa-
 18:1 44.1 tial representation, the wedge indicates a substit-
Total polyunsaturated 14.7 uent coming out of the plane toward the observer,
 18:2 13.4 and the slash indicates a substituent going away
 18:3 0.3
from the observer. The three carbon atoms of the
 20:4 1.0
glycerol are then described by the stereospecific
numbering (sn) with the three carbon atoms des-
on the central carbon atom are identical. When ignated sn-1 from the top to sn-3 at the bottom.
one of the carbon atoms is esterified with a fatty When a fat or oil is characterized by determi-
acid, a monoglyceride results and two nonsuper- nation of its component fatty acids, there still
imposable structures exist. These are called enan- remains the question as to how these acids are
tiomers and are also referred to as chiral. A distributed among and within the glycerides.
Component Glycerides 55

Table 2.12  Influence of lipid supplementation on fatty formed. In strictly random distribution the
acid distribution in egg yolk fatty acids
amount of GA3 in a fat would be proportional to
Control Flax Soy Canola the cube of the percentage of A present. For
C14:0 1.27 0.29 0.29 0.3 example, at 30% A there would be 2.7% of GA3,
C16:0 25 22.5 23.2 22.9 which under rules of even distribution would
C:16:1n9 0.7 0.94 0.83 0.94 occur only at levels of A over 70% (Fig. 2.11).
C16:1n7 2.14 2.47 1.91 1.91 The theory of restricted random distribution
C17:0 0.17 0.16 0.18 0.18
was proposed by Kartha (1953). In this theory the
C17:1n10 0.12 0.16 0.13 0.14
fatty acids are distributed at random, but the con-
C18:0 12.4 8.87 9.46 8.83
tent of fully saturated glycerides is limited to the
C18:1n9 39.6 43.3 41.5 44
amount that can remain fluid in vivo. This theory
C18:2n6 14.7 13.7 17.8 15.8
C18:3n6 0.06 0.08 0.13 0.13
is followed by the 1,3 random, 2 random distribu-
C18:3n3 0.22 3.4 0.63 0.55
tion hypothesis of Vander Wal (1964). According
C18:4n3 Nd 0.06 0.05 0.08 to this theory, all acyl groups at the 2-positions of
C20:1n9 0.27 0.17 0.24 0.34 the glycerol moieties of a fat are distributed
C20:2n6 0.22 0.21 0.24 0.26 therein at random. Equally, all acyl groups at the
C20:3n6 0.2 0.18 0.23 0.3 1- and 3-positions are distributed at random and
C20:4n6 2.63 1.17 1.74 1.94 these positions are identical. Application of this
C20:5n3 Nd 0.18 Nd Nd theory to the results obtained with a number of
C22:4n6 0.52 0.09 0.2 0.18 fats gave good agreement (Vander Wal 1964), as
C22:5n6 Nd 0.08 0.38 0.4 Table 2.14 shows.
C22:5n3 Nd 0.29 0.1 0.1 In vegetable fats and oils, the saturated fatty
C22:6n3 0.64 1.55 0.65 0.65 acyl groups have a tendency to occupy the 1- and
Total Fat 4.12 4.44 5.22 5.70 3- positions in the glycerides and the unsaturated
0Saturated FA 38.84 31.82 33.13 32.21 acyl groups occupy the 2-position (Fig. 2.12).
Mono Unsaturated 42.83 47.04 44.61 47.33 Since these fats contain a limited number of fatty
Total PUFA 19.19 20.99 22.15 20.39 acids, it is customary to show the glyceride com-
n-3 FA 0.86 5.48 1.43 1.38
position in terms of saturated (S) and unsaturated
Adapted from Milinsk et al. (2003) (U) acids. The predominant glyceride types in
these fats and oils are S-U-S and S-U-U. Lard is
Originally theories of glyceride distribution were an exception—saturated acyl groups predomi-
attempts by means of mathematical schemes to nate in the 2-position. The glyceride distribution
explain the occurrence of particular kinds and of cocoa butter results in a fat with a sharp melt-
amounts of glycerides in natural fats. Subsequent ing point of about 30–34 °C. It is hard and brittle
theories have been refinements attempting to below the melting point, which makes the fat use-
relate to the biochemical mechanisms of glycer- ful for chocolate and confectionery manufacture.
ide synthesis. Hilditch proposed the concept of Other fats with similar fatty acid composition,
even distribution (Gunstone 1967). In the rule of such as sheep depot fat (see Table 2.4), have a
even (or widest) distribution, each fatty acid in a greater variety of glycerides, giving the fat a
fat is distributed as widely as possible among higher melting point (about 45 °C) and a wider
glyceride molecules. This means that when a melting range, and a greasy and soft appearance.
given fatty acid A constitutes about 35 mole per- Brockerhoff et al. (1966) studied the fatty acid
cent or more of the total fatty acids (A + X), it distribution in the 1-, 2-, and 3-positions of the tri-
will occur at least once in all triglyceride mole- glycerides of animal depot fats by stereospecific
cules, as represented by GAX2. If A occurs at lev- analysis. The distribution among the three posi-
els of 35–70 mole percent, it will occur twice in tions was nonrandom. The distribution of fatty
an increasing number of triglycerides GA2X. At acids seems to be governed by chain length and
levels over 70%, simple triglycerides GA3 are unsaturation. In most fats a short chain and unsatu-
56

Table 2.13  Typical component fatty acids of some vegetable oils (Wt %)
Unsat/ Saturated Mono unsaturated Poly unsaturated
Oil or Fat sat ratio Capric acid Lauric acid Myristic acid Palmitic acid Stearic acid Oleic acid Linoleic acid (ω−6) Alpha linolenic acid (ω−3)
C10:0 C12:0 C14:0 C16:0 C18:0 C18:1 C18:2 C18:3
Canola Oil 15.7 – – – 4 2 62 22 10
Cocoa Butter 0.6 – – – 25 38 32  3 –
Cocoanut Oil 0.1 6 47 18 9 3  6  2 –
Corn Oil 6.7 – – – 11 2 28 58 1
Cottonseed Oil 2.8 – – – 22 3 19 54 1
Flaxseed Oil 9 – – – 3 7 21 16 53
Olive Oil 4.6 – – – 13 3 71 10 1
Palm Oil 1 – – 1 45 4 40 10 –
Palm Olein 1.3 – – 1 37 4 46 11 –
Palm Kernal Oil 0.2 4 48 16 8 3 15  2 –
Peanut Oil 4 – – – 11 2 48 32 –
Safflower 10.1 – – – 7 2 13 78 –
Sesame Oil 6.6 – – – 9 4 41 45 –
Soybean Oil 5.7 – – – 11 4 24 54 7
http://www.scientificpsychic.com/fitness/fattyacids1.html
2 Lipids
Component Glycerides 57

Fig. 2.9  Plane of


symmetry of a glycerol
molecule (top) and
mirror image of two
enantiomers of a
mono-acylglycerol
(bottom)

1- and 3-positions, whereas in cocoa butter the


major portion of the unsaturation is located in the
2-position. This difference accounts for the dif-
ference in physical properties of the two fats
(deMan et al. 1987).
Milk fat, with its great variety of fatty acids,
also has a very large number of glycerides. It is
Fig. 2.10  Stereospecific numbering of the carbons in a
triacylglycerol
possible, by, for example, fractional crystalliza-
tion, to separate milk fat in a number of fractions
with different melting points (Chen and deMan
ration direct a fatty acid toward position 2. The 1966). Milk fat is peculiar in some respects. Its
depot fat of pigs is an exception, palmitic acid short-chain fatty acids are classified chemically
being predominant in position 2. In the fats of as saturated compounds but behave physically
marine animals, chain length is the directing factor, like unsaturated fatty acids. One of the unsatu-
with polyunsaturated and short-chain fatty acids rated fatty acids, the so-called oleic acid, is partly
accumulated in the 2-position and long chains in trans and has a much higher melting point than
the 1- and 3- positions. In the fats of birds, unsatu- the cis isomers. In the highest melting fraction
ration seems to be the only directing factor and from milk fat, there is very little short-chain fatty
these acids accumulate in the 2-position. acid and little unsaturation, mostly in the trans
The positional distribution of fatty acids in pig configuration (Woodrow and deMan 1968). The
fat (lard) and cocoa butter is shown in Table 2.15. low melting fractions are high in short-chain
Most of the unsaturation in lard is located in the fatty acids and unsaturation (cis). The general
58 2 Lipids

distribution of major fatty acids in whole milk fat


is as follows (Morrison 1970): 4:0 and 6:0 are
located largely sn1 and sn-3 positions; 18:0 and
18:1 are preferentially in primary positions; 10:0,
12:0, and 16:0 are distributed randomly or with a
slight preference for the secondary position; and
14:0 is predominantly in the secondary position.
Prosser et al. (Table 2.16) have shown that C14
and C16 are predominantly in the sn-2 position
of the triglyceride in both human and cow milk.

Waxes

Waxes are esters of fatty acids with long chain


monohydric alcohols (one hydroxyl group).
Fig. 2.11  Calculated values for glyceride types in ran- Natural waxes are often mixtures of such esters,
dom distribution (solid lines) and even distribution (dotted and may also contain hydrocarbons. The formu-
lines). Adapted from: F.D. Gunstone, An Introduction to las for three well known waxes are given below,
the Chemistry of Fats and Fatty Acids, 1967, Chapman
with the carboxylic acid moiety colored red and
and Hall
the alcohol colored blue.

wax ester O
O
Spermaceti Beeswax Carnuba wax
CH3(CH2)14CO2-(CH2)15CH3 CH3(CH2)24CO2-(CH2)29CH3 CH3(CH2)30CO2-(CH2)33CH3

Spermaceti Beeswax Carnuba wax


CH3(CH2)14CO2─(CH2)15CH3 CH3(CH2)24CO2─(CH2)29CH3 CH3(CH2)30CO2─(CH2)33CH3

Table 2.14  Comparison of the glyceride composition of some natural fats as determined experimentally and as calcu-
lated by 1,3 random, 2 random hypothesis
Molecular species
SSS SUS SSU USU UUS UUU
Fat Method (Mole %) (Mole %) (Mole %) (Mole %) (Mole %) (Mole %)
Lard Experiment 8 0 29 36 15 12
Lard Calculated 6 2 29 36 12 15
Chicken fat Experiment 3 10 9 12 38 28
Chicken fat Calculated 3 10 10 9 36 32
Cocoa butter Experiment 5 66 7 3 20 1
Cocoa butter Calculated 5 69 2 0 22 2
Adapted from R.J. Vander Wal, Triglyceride Structure, Adv. Lipid Res., Vol. 2, pp. 1–16, 1964
Component Glycerides 59

Fig. 2.12  Fatty acid


distribution in the
triacylglycerols of
vegetable oils

Table 2.15  Positional distribution fatty acids in pig fat


and cocoa butter
Waxes are widely distributed in nature on
leaves and fruits of many plants. The waxy coat-
Fatty acid (Mole %)
ings may protect the leaves and fruit from dehy-
Fat Position 14:0 16:0 16:1 18:0 18:1 18:2
dration and small predators. Bird feathers and the
Pig fat 1 0.9 9.5 2.4 29.5 51.3 6.4
fur of some animals have similar coatings which
2 4.1 72.3 4.8 2.1 13.4 3.3
serve as a water repellent.
3 0 0.4 1.5 7.4 72.7 18.2
Waxes occur primarily in plants and are gener-
Cocoa 1 – 34.0 0.6 50.4 12.3 1.3
butter 2 ally composed of long chain alcohols esterified to
– 1.7 0.2 2.1 87.4 8.6
3 –
fatty acids. The chain-length and degree of unsat-
36.5 0.3 52.8 8.6 0.4
uration and branching of the aliphatic constituents
Source: From W.C. Breckenridge, Stereospecific Analysis
of Triacylglycerols, in Fatty Acids and Glycerides, will vary with the origin of the wax. Some waxes
A. Kuksis, ed., 1978, Plenum Press of marine origin or from some higher animals, the

Table 2.16  Fatty acid contents of cow milk and breast milk and the fatty acid distribution in the sn-2 position of the
triglycerides Adapted from Prosser et al. 2010
Cow Milk Breast Milk
Total Fatty acids In sn-2 position Total Fatty acids In sn-2 position
mol/100 mol mole/100 mol mol/100 mol mole/100 mol
C4:0 8.4 ND
C6:0 4.5 0.9
C8:0 2.2 1.7 0.1 ND
C10:0 4.3 4.2 1.2 0.3
C12:0 4.3 5.9 5.5 4.2
C14:0 13.0 21.7 6.0 9.6
C15:0 1.4 2.0 0.2 0.5
C16:0 31.0 40.8 21.0 57.0
C17:0 0.8 0.7 0.4 0.4
C18:0 9.6 4.9 7.4 1.4
C18:1n9 14.0 10.2 42.0 16.0
C18:2n6 0.6 1.2 18.0 13.0
C18:3n3 0.7 1.6 0.8 0.7
C20:0 0.1 ND 0.2 0.1
60 2 Lipids

aliphatic moieties tend to be saturated or monoe- Waxes are widely distributed in nature on
noic. A number of waxes are produced commer- leaves and fruits of many plants. The waxy coat-
cially in large amounts for use in food coatings, ings, may protect the leaves and fruit from dehy-
cosmetics, lubricants, polishes, inks and many dration and small predators. Bird feathers and the
other applications. Bees secrete a wax, which fur of some animals have similar coatings which
they use to construct the honeycomb. The wax is serve as a water repellent.
recovered as a by-product when the honey is har-
vested and refined. It contains a high proportion
of wax esters (35–80%). The hydrocarbon con- Phospholipids
tent is highly variable. The wax esters consist of
C40 to C46 molecular species, based on 16:0 and After triglycerides, phospholipids are the second
18:0 fatty acids some with hydroxyl groups in the most abundant class of lipids found in nature.
ω-2 and ω-3 positions. In addition, some diesters Phospholipids are found in animal, plant and
with up to 64 carbons may be present, together microbial cell membranes. Like triglycerides
with triesters, hydroxy-polyesters and free acids phospholipids contain a glycerol backbone
(which are different in composition and nature appended with two fatty acids, plus phosphoric
from the esterified acids). The jojoba plant acid and a low-molecular-weight alcohol on one
(Simmondsia chinensis), which grows in the of the hydroxyl groups of the glycerol backbone.
semi-arid regions of Mexico and the U.S.A., pro- Common phospholipids include lecithins and
duces wax esters rather than triacylglycerols in its cephalins.
seeds. The wax consists mainly of 18:1 (6%), Phospholipids are the primary constituents of
20:1 (35%) and 22:1 (7%) fatty acids linked to cell membranes. They resemble the triglycerides
20:1 (22%), 22:1 (21%) and 24:1 (4%) fatty alco- in being ester derivatives of glycerol appended
hols. Therefore, it contains C38 to C44 esters with with fatty acids and phosphoric acid. The phos-
one double bond in each alkyl moiety. As methy- phate moiety of the resulting phosphatidic acid is
lene-interrupted double bonds are absent, the wax further esterified with ethanolamine, choline or
is relatively resistant to oxidation. serine. Figure 2.13 illustrates some of the struc-
The leaves of the carnauba palm, Copernicia tural components of fatty acids and Fig. 2.14
cerifera which grows in Brazil, have a thick coat- illustrates the structures of the most abundant
ing of wax, which is harvested from the dried phospholipids. Note that the fatty acid compo-
leaves. It contains mainly wax esters (85%), nents (R & R’) may be saturated or unsaturated.
accompanied by small amounts of free acids and Phospholipids are an important class of bio-
alcohols, hydrocarbons and resins. The wax molecules. Phospholipids are the fundamental
esters constitute C:16 to C:20 fatty acids linked building blocks of cellular membranes. These
to C30 to C34 alcohols, resulting in C:46 to C:54 molecules have a glycerol backbone, a polar or
molecular species. charged head group and a pair of nonpolar fatty

Fig. 2.13  Components of phospholipids


Component Glycerides 61

Fig. 2.14  Structure of


the major phospholipids

acid tails esterified to the glycerol backbone. The (PE). Phosphatidyl choline commercially called
combination of polar and nonpolar segments is lecithin is important in foods as an emulsifier. A
termed amphiphilic, and the word describes the typical phospholipid contains a saturated fatty
tendency of these molecules to assemble at inter- acid, such as palmitic or stearic acid, at the sn1
faces between polar and nonpolar phases. position, and an unsaturated or polyunsaturated
Two fatty acid chains, each typically having fatty acid, such as oleic or arachodonic acid.
an even number of carbon atoms between 14 and Phosphatidylcholine (lecithin) is usually the
20, attach via esterification to the first and second most abundant phospholipid in plants, often
carbons of the glycerol molecule, denoted as the amounting to almost 50% of the total phospho-
sn1 and sn2 positions, respectively. The third lipid in the system. Phosphotidyl choline is a key
hydroxyl group of glycerol, at position sn3, is building block of membrane bilayers. It makes
appended with phosphoric acid to form phospha- up a very high proportion of the outer leaflet of
tidate bond. Phospholipids are widely distributed the plasma membrane. Phosphatidylcholine is
in nature, generally one of the groups are bound also the principal phospholipid circulating in
to the phosphatidic acid moiety such as serine, plasma, where it is an integral component of the
ethanolamine, choline, glyercol, or inositol. The lipoproteins, especially the HDL.
resulting phospholipids may be charged, for In food systems where it is more commonly
example, phosphatidyl serine (PS), phosphatidyl referred to as lecithin it is a valuable emulsifier.
inositol (PI), and phosphatidyl glyercol (PG); or Phospholipids are amphoteric (mixed ionic
dipolar (having separate positively and nega- charges), thus, they can aggregate or self-­
tively charged regions), for example, phosphati- assemble when mixed with water. This aggrega-
dyl choline (PC), and phosphatidyl ethanolamine tion differs from the surface activity of soaps and
62 2 Lipids

detergents. Soaps and detergents tend to form Liposomes are microscopic vesicles consisting of
micelles. The two pendant alkyl chains present in an aqueous core enclosed in one or more phos-
phospholipids and the unusual mixed charges in pholipid layers. They are formed when phospho-
their head groups, micelle formation is unfavor- lipids are vigorously mixed with water. Unlike
able relative to a bilayer structure. As shown in micelles, liposomes have both aqueous interiors
Fig. 2.15, the polar head groups on the faces of and exteriors. Liposomes therefore are extremely
the bilayer contact water, and the hydrophobic valuable for encapsulation of aqueous food, fla-
alkyl chains form a nonpolar interior. The phos- vor components or lipid soluble components. The
pholipid molecules can move about in their half liposome protects the guest material from oxida-
the bilayer, but there is a significant energy bar- tion or enzymatic attack.
rier preventing migration to the other side of the Soy beans are a rich source of phospholipids
bilayer. The bilayer membrane structure is also which are extracted and refined for use as emulsi-
found in aggregate structures called liposomes. fiers n foods. Table 2.17 illustrates the range and

water

30 Å

O water
H O P O C C N(CH3)3 planar bilayer
H2 H2
H2C C C O
H2
O O O O

aggregation
in water

aqueous
interior

liposome
phospholipid
aqueous exterior

Fig. 2.15  Phospholipids form lipid bilayers and lipid micelles

Table 2.17  Range of phospholipids in soy


Component Shorthand abbreviation Low Intermediate High
Phosphatidylcholine PC 12.0–21.0 29.0–39.0 41.0–46.0
Phosphatoidylethanolamine PE 8.5–9.5 20.0–26.3 31.0–34.0
Phosphatidylinositol PI 1.7–7.0 13.0–17.5 19.0–21.0
Phosphatoidic Acid PA 0.2–1.5 5.0–9.0 14
Phosphatidyl Serine PS 0.2 5.9–6.6 –
Lysophosphatidylcholine LPC 1.5 8.5 –
Lysophosphatidylinositol PLI 0.4–1.8 – –
Lysophosphatidylserine LPS 1 – –
Lysophosphatidic acid LPA 1 – –
Phytoglycolipids – – 14.3–15.4 29.6
Szuhaj (1989)
Steroids 63

Table 2.18  Total and principal phospholipids of different foods


Food Total lipid Total phospholipid PC PE PS PI
g/100 g product mg/100 g product
Beef (L. dorsi) Fattened 12.4 690 340a 124 96b 32
Beef (L. dorsi) Lean 1.7 597 260a 106 48b 44
Pork (L. dorsi) 2.58 596 304 167 57c
Chicken breast 1.12 782 391 187 100
Chicken thigh 3.26 1386 662 352 186 tr
Turkey breast 0.73 418 231 92 33b
Turkey thigh 2.48 418 231 92 34b
Cod 0.59 520 359 99 26
Tuna (Dorsal) 3.79 617 166 132 93
Cow milk 3.66 34 12 10 1 2
Egg Yolk 31.8 10,306 6771 1917 64
Peanut 48.5 620 270 50 150
Soybean 20.8 2038 917 536 287
Corn 3.7 213 139 15 26
Hard wheat (whole grain) 2.5 1060 164 56 69
a
PC + LPC
b
PS + PA + CL
c
PS + PI
PC phosphatidyl choline, PE phosphatidyl ethanolamine, PS phosphatidyl Serine, PI phosphatidyl Inositol, LPC lyso-
phosphatidyl choline
Adapted from Weihrauch and Son (1983)

distribution of soy phospholipids. Table 2.18 plants, but some of the larger and more complex
contains distribution of phospholipids found in a terpenes (e.g. squalene & lanosterol) occur in
variety of foods. Frequently the phosphatidyl animals. Are classified by the number and struc-
choline is refined and sold as lecithin which can tural organization. Terpenes are primarily made
be used as an emulsifier in foods and cosmetics. of isoprene (more accurately isopentane) units,
an empirical feature known as the isoprene rule.
Because of this, terpenes usually have 5n carbon
Unsaponifiables atoms (n is an integer), and are subdivided as fol-
lows (Table 2.20):
The unsaponifiable fraction of fats contains ste- Isoprene itself, a C5H8 gaseous hydrocarbon,
rols, terpenic alcohols, aliphatic alcohols, squa- is emitted by the leaves of various plants as a
lene, and hydrocarbons. The distribution of the natural byproduct of plant metabolism. Next to
various components of the unsaponifiable frac- methane it is the most common volatile organic
tion in some fats and oils is given in Table 2.19. compound found in the atmosphere. Examples of
In most fats the major components of the unsa- C10 and higher terpenes, representing the four
ponifiable fraction are sterols. most common classes are shown in Fig. 2.16.

Terpenes Steroids

Compounds classified as terpenes constitute a The class of lipids called steroids is recognized by
large and diverse group of natural compounds. A their tetracyclic skeleton, consisting of three fused
majority of these compounds are found only in six-membered and one five-membered ring.
64 2 Lipids

Table 2.19  Composition of the unsaponifiable fraction of some fats and oils
Oils Hydrocarbons Squalene Aliphatic alcohols Terpenic alcohols Sterols
Olive 2.8–3.5 32–50 0.5 20–26 20–30
Linseed 3.7–14.0 1.0–3.9 2.5–5.9 29–30 34.5–52
Tea seed 3.4 2.6 – – 22.7
Soybean 3.8 2.5 4.9 23.2 58.4
Rapeseed 8.7 4.3 7.2 9.2 63.6
Corn 1.4 2.2 5.0 6.7 81.3
Lard 23.8 4.6 2.1 7.1 47.0
Tallow 11.8 1.2 2.4 5.5 64.0
Source: From G. Jacini, E. Fedeli, and A. Lanzani, Research in the Nonglyceride Substances of Vegetable Oils, J. Assoc.
Off. Anal. Chem., Vol. 50, pp. 84–90, 1967

Table 2.20 Terpene
Classification Isoprene units Carbon atoms are excellent sources of phytosterols. Sterols con-
monoterpenes 2 C10 tain the perhydrocyclopenteno-­ phenanthrene
sesquiterpenes 3 C15 nucleus, which is shared in common with many
diterpenes 4 C20 other natural compounds, including bile acids,
sesterterpenes 5 C25 hormones, and vitamin D. The nucleus and the
triterpenes 6 C30 description of the four rings, as well as the sys-
tem of numbering of the carbon atoms, are shown
in Fig. 2.18a. The sterols generally have high
Steroids structures are not based a glycerol back- melting points and therefore are solid at room or
bone or ester linkages. The four rings in sterols body temperatures. Stereochemically they are
structures are designated A, B, C & D shown in relatively flat molecules, usually with all trans
Fig. 2.17. The numbers in red represent the num- linkages, as shown in Fig. 2.18. The ring junction
bering of the ring carbon atoms in the sterol struc- between rings A and B is trans in some steroids,
ture. The substituents designated by R are often cis in others. The junctions between B and C and
alkyl groups. The R group at the A:B ring fusion is C and D are normally trans. Substituents that lie
most commonly methyl or hydrogen, the R group above the plane, as drawn in Fig. 2.18c, are
at the C:D fusion is generally a methyl group. The named β, those below the plane, α. The 3-OH
R- substituent at C-17 varies considerably, and is group in cholesterol (Fig. 2.18c) is the
usually larger than methyl if it is not a functional β-configuration, and it is this group that may form
group. The most common locations of functional ester linkages. The structuresof the major plant
groups are C-3, C-4, C-7, C-11, C-12 & C-17. sterols is given in Fig. 2.15. Part of the sterols in
Animal fats contain cholesterol and, in some natural fats are present as esters of fatty acids; for
cases, minor amounts of other sterols such as example, in milk fat, about 10% of the choles-
lanosterol. Plant fats and oils contain phytoster- terol occurs in the form of cholesterol esters.
ols, usually at least three, and sometimes four Cholesterol has 256 stereoisomers, although
(Fedeli and Jacini 1971). Plants can contain trace only two of them are of biochemical significance
amounts or cholesterol. The predominant phytos- (nat-cholesterol and ent-cholesterol,[and only
terol found in plants is β-sitosterol; others include one of them occurs naturally (nat-cholesterol). In
campesterol and stigmasterol. In rapeseed oil, animal tissues, cholesterol (cholest-5-en-3β-ol)
brassicasterol takes the place of stigmasterol is by far the most abundant member of a family
(Table 2.21). Table 2.22 contains the typical phy- of polycyclic compounds known as sterols.
tosterol content found in servings of plant foods. In animals cholesterol has an essential struc-
Plant sterols are beneficial in helping control tural role in membranes and in lipid metabolism.
cholesterol re-absorption thus possibly educing Cholesterol is the biosynthetic precursor of bile
serum cholesterol. Wheat germ and rice bran oil acids, vitamin D and steroid hormones (glucocor-
Steroids 65

Fig. 2.16  Structures of monoterpenes

Fig. 2.17  Sterol structure and major plant sterols


66 2 Lipids

Table 2.21  Sterol content of fats and oils


Fat Sterol (%)
Lard 0.12
Beef tallow 0.08
Milk fat 0.3
Herring 0.2–0.6
Cottonseed 1.4
Soybean 0.7
Corn 1.0
Rapeseed 0.4
Coconut 0.08
Cocoa butter 0.2

Table 2.22  Phytosterol content of selected food oils


Total Phytosterol per serving of various food oils
Food Serving Phytosterols (mg)
Wheat germ ½ cup (57 g) 197
Rice bran oil 1 tablespoon (14 g) 162
Sesame oil 1 tablespoon (14 g) 118
Corn oil 1 tablespoon (14 g) 102
Canola oil 1 tablespoon (14 g) 92
Peanuts 1 ounce (28 g) 62
Wheat bran ½ cup (29 g) 58
Almonds 1 ounce (28 g) 39 Fig. 2.18  Sterols. (a) Structure of the steroid nucleus, (b)
Brussels ½ cup (78 g) 34 stereochemical representation, and (c) Cholesterol
sprouts
Rye bread 2 slices (64 g) 33
Macadamia 1 ounce (28 g) 33
of the cholesterol in the cell). Cholesterol levels
nuts in mitochondria and the endoplasmic reticulum
Olive oil 1 tablespoon (14 g) 22 are very low and the Golgi contains intermediate
Benecol® 1 tablespoon (14 g) 850 mg plant stanol levels. The brain contains more cholesterol than
spread esters (500 mg free any other organ, where it comprises roughly a
stanols)
quarter of the total free cholesterol in the human
http://lpi.oregonstate.edu/mic/dietary-factors/phytochem- body. Cholesterol can occur in the free form,
icals/phytosterols; USDA Nutrient Database for Standard
Reference, Release 20. 2007. Available at: http://ndb.nal. esterified to long-chain fatty acids (cholesterol
usda.gov//. Accessed 7/24/08.; (Norman et al. 2002; esters), and in other covalent and non-covalent
Normen et al. 1999; Phillips et al. 2002) linkages in animal tissues, including the plasma
lipoproteins. In plants, it tends to be a minor
ticoids, estrogens, progesterone’s, androgens and component only of a complex mixture of struc-
aldosterone), the central nervous system, and it turally related phytosterols, although there are
has major functions in signal transduction and exceptions, but it is nevertheless importance as a
males,m it is required for sperm development. precursor of some plant hormones. http://lipidli-
Plasma cholesterol levels are also risk factors for brary.aocs.org/Lipids/simple.html.
heart disease because they can be a major con-
tributory factor to atherogenesis. Cholesterol is
ubiquitous in in the membranes all animal tissues Phytosterols
(and of some fungi, although it is not evenly dis-
tributed. The highest levels of unesterified cho- The sterols provide a method of distinguishing
lesterol are found in plasma membrane (roughly between animal and vegetable fats by means of
30–50% of the lipid in the membrane or 60–80% their acetates. Cholesterol acetate has a melting
Phytosterols 67

The substances are insoluble in water, but soluble


in non-polar solvents, such as hexane, iso-octane
and 2-propanol. The esters are also soluble in
vegetable fats and oils. Phytostanol and phytos-
terol esters are added to margarine and promoted
for reduction of serum cholesterol.
Edible vegetable oils, extracted from oil seeds,
are typically refined to remove minor oil compo-
nents such as phosphatides, free fatty acids,
­oxidized fatty acids, pigments and odors, while
minimizing damage to the glycerides. The most
common processes are referred to as either physi-
cal or chemical. Chemical or alkaline refining an
alkali is used to neutralize the free fatty acids
which are removed as soapstock. The chemical
refining process consists of water degumming
which where the gums a dried and used to pro-
duce lecithin, aalkal neutralization resulting in
sproduction of soap stock, dewaxing, bleaching,
Fig. 2.19  Isomeric forms of cholesterol
and deodorization yielding edible oils. In physi-
cal refining acid degumming yields gums, dewax-
ing bleaching, are followed by deacidification
point of 114 °C, whereas phytosterol acetates yielding fatty acids and deodorization procedure
melt in the range of 126–137 °C. This provides a comprises degumming, neutralization, bleaching
way to detect adulteration of animal fats with and deodorization. In physical refining the neu-
vegetable fats. tralization step is omitted and the residual free
The most common phytosterols and phytosta- fatty acids are removed in the final deodorization
nols (examples of structures are shown in step. Deodorization is the last step in the edible
Fig.  2.19 are sitosterol (3β-stigmast-5-en-3ol; oil refining process in which volatiles are
CAS number 83-46-5), sitostanol (3β,5α-­ removed that can adversely affect the stability,
stigmastan-­3-ol; CAS Number 83-45-4), campes- flavor, and odor of the oil.
terol (3β-Ergost-5-en-3-ol; CAS Number This process relies on the large volatility dif-
474-62-4), campestanol (3β,5α-ergostan-3-ol; ferences between the oil itself (triglycerides) and
CAS Number 474-60-2), stigmasterol the volatile compounds to be removed and is car-
(3β-stigmasta-5,22-dien-3-ol; CAS Number ried out under reduced pressure, an elevated tem-
83-48-7) and brassicasterol (3β-ergosta-5,22-­ perature in the presence of a stripping gas. The
dien-­3-ol; CAS Number 474-67-9). Each com- volatiles are recovered in a vapor condenser. This
mercial source has its typical phytosterols distillate mainly contains free fatty acids, but also
composition. Commercially, phytosterols are iso- significant levels of tocopherols (5–15%) and
lated from vegetable oils, such as soybean oil, phytosterols (8–20%).
rapeseed (canola) oil, sunflower oil or corn oil, or In a transesterification (methanolysis) step,
from so-called “tall oil”, a by-product of the the glycerides are converted into fatty acid methyl
manufacture of wood pulp. Phytosterols can be esters and glycerol and the phytosterol-esters into
hydrogenated to obtain phytostanols. Phytosterols free phytosterols and fatty acid methyl esters.
and phytostanols are high melting powders. After removal of the methanol/glycerol phase,
Phytostanol and phytosterol esters are chemically the methyl esters are removed and the free phy-
stable materials, having comparable chemical tosterols and tocopherols removed by distillation.
and physical properties to edible fats and oils. The phytosterols are separated from the tocoph-
68 2 Lipids

Table 2.23  Hydrocarbon composition of some vegetable oils


Oils n-Paraffins iso-and/or ante-iso Paraffins Unidentified Total Hydrocarbons
Corn C11–31 C11–21 8 40
Peanut C11–30 C11–23 7 40
Rapeseed C11–31 C11–17, C19–21 6 36
Linseed C11–35 C11–21 7 43–45
Olive C11, C13–30 – 6 29
Source: From G. Jacini, E. Fedeli, and A. Lanzani, Research in the Nonglyceride Substances of Vegetable Oils, J. Assoc.
Off. Anal. Chem., Vol. 50, pp. 84–90, 1967

erols by solvent crystallization and filtration Hydrolysis


using food grade solvent. The phytosterols are
further purified by re-crystallisation, mainly to
remove wax-esters (Table 2.23). Interesterification
Plants also produce low levels of paraffin
hydrocarbons. Traces of hydrocarbons remain in Interesterification can be defined as a redistribu-
vegetable oils, however most are removed during tion of the fatty acid moieties present in triglycer-
refining of the oil. Table 2.23 the contents of ides. In the presence of certain catalysts, the fatty
hydrocarbons in some common vegetable oils. acid radicals can be made to move between
hydroxyl positions so that an essentially random
fatty acid distribution results, according to the
Lipid Reactions following reaction pattem (Formo 1954):

Lipids contain two main functions which become


involved in chemical reactions. Lipids that are
unsaturated can undergo oxidation reactions both Interesterification is used in industry to modify
in food and in vivo. Lipids with ester bonds such the crystallization behavior and the physical prop-
as triglycerides, phospholipids and waxes can erties of fats. It can also be used to produce solid
undergo hydrolysis or interesterification. The fats for margarine and shortening that are low in
interesterification reactions are commercially trans fatty acids. An additional advantage is that
important for formation of modified triglycerides polyunsaturated fatty acids, which are destroyed
and for analytical purposes forming esters of during hydrogenation, are not affected. Several
fatty acids for analysis. Interesterifcation reac- types of interesterification are possible. A fat can be
tions are important for forming new triglycerides randomized by carrying out the reaction at tem-
with desired functionalities. peratures above its melting point, several raw
The acidity of the carboxylic acids function on materials may be interesterified together so that a
fatty acids results in react with bases to form new product with desired physical properties
ionic salts, as shown below (Fig. 2.20). In the results. Fat can be interesterified at a t­emperature
case of alkali metal hydroxides and simple below its melting point so that only the liquid frac-
amines (or ammonia) the resulting salts have pro- tion reacts (this is known as directed interesterifica-
nounced ionic character and are usually soluble tion). The effect of randomization can be
in water. demonstrated with the case of a mixture of equal
amounts of two simple glycerides, such as triolein
and tristearin. Interesterification of two equal glyc-
Fatty Acid Salts erides (Fig. 2.21 OOO SSS) such as triolein and
tristearin results in the formation of six possible
RCO2H + NaHCO3 → RCO2(−) Na(+) + CO2 + H2O triglycerides. When the blend of the two glycerides
RCO2H + (CH3)3N: → RCO2(−) (CH3)3NH(+) is other than in equal quantities the results can be
Lipid Reactions 69

Fig. 2.20 Salt
formation of fatty acids
and hydrolysis of
triglycrides

Fig. 2.21 The
interesterification of
equamolar quantities of
triolein and tristearin

Fig. 2.22 
Triacylglycerol
distribution in a
randomized binary
mixture. Adapted from:
A. Rozenaal,
Interesterification of
Oils and Fats, Inform, 3,
pp. 1233–1235, © 1992,
AOCS Press

derived from a graph such as the one in Fig. 2.22. Table 2.24 Theoretical Triacylglycerol Composition
The graph indicates that the maximum levels of the after Interesterification for n Fatty Acids (A, B, C, D) with
Molar Fractions a, b, c, d
intermediate glycerides A2B and AB2 are formed at
molar fractions of one-third A or one-third B. Type Number Amount
The theoretical number of glycerides formed Mono acid n a3, b3, c3
(AAA, BBB)
by interesterification of mixtures containing dif-
Diacid (AAB, AAC) n(n–1) 3a2b, 3ab2, 3a2c
ferent fatty acids has been described by Rozenaal
Triacid (ABC, BCD) 1/6 n(n–1) (n–2) 6abc, 6acd
(1992) and is shown in Table 2.24. The table also
Total n3/6 + n2/2 + n/3
gives the formula for calculating the total number
70 2 Lipids

of glycerides formed. For example, for n = 4 the acid methyl ester into free fatty acid. The reaction
number of glycerides formed is 20 and for n = 6 is intramolecular as well as intermolecular. An
the number is 56. Thus, interesterification results active catalyst for interesterification can be devel-
in increased complexity of the oil. oped with sodium hydroxide and glycerol which
This is results in a randomized distribution of form an active catalyst as illustrated in Fig. 2.24.
the fatty acids on the glycerol moiety when the Freeman (1968) has reported that the intramolecu-
reaction is carried out in a single, liquid phase. lar rearrangement occurs at a faster rate than the
In directed interesterification, one of the reac- general randomization.
tion components is removed from the reaction Rozenaal determined the reaction rate for the
mixture. This can be achieved by selecting a randomization of palm oil (Fig. 2.25). The reac-
reaction temperature at which the trisaturated tion rate, which was measured by determination
glycerides become insoluble and precipitate. The of the solid fat content, increased with
equilibrium is then disturbed and more trisatu- ­temperature. There is evidence of an induction
rates are formed, which can then be precipitated. period at lower temperatures.
Because of the low temperature employed, the Random interesterification can result in either
reaction is up to 10–20 times slower than the ran- an increase or a decrease in melting point and
dom process. Another procedure of directed solid fat content, depending on the composition of
interesterification involves the continuous distill- the original fat or fat blend. When cocoa butter is
ing of low molecular weight fatty acids, such as interesterified, the unique melting properties are
those present in coconut oil with high free fatty completely changed (Fig. 2.26). Cocoa butter is a
acid content (Hustedt 1976). relatively expensive fat, used in confectionery,
The reaction mechanism of interesterification because of its sharp melting point between room
using sodium methoxide as a catalyst is a two-­step temperature and body temperature; chocolate lit-
process (Sreenivasan 1978; Rozenaal 1992). First, erally melts in the mouth. This is due to the fairly
the catalyst combines with the glyceride at one of small variation in the structure of the constituent
the carbonyl locations (Fig. 2.23). Then the anion triglycerides; 80% have palmitic acid or stearic
of the catalyst and the alkoxy group of the ester are acid in the 1 and 3 positions with oleic acid in the
exchanged. The catalyst has changed but remains central 2 position. Cocoa butter substitutes have
active. At the end of the reaction there remains an been produced from palm oil, the acid hydrolysis
amount of fatty acid methyl ester equivalent to the is accomplished by dissolving stearic acid in hex-
amount of sodium methoxide catalyst used. The ane containing enough water to activate the lipase.
randomization reaction continues until equilib- Olive oil may be similarly improved by exchang-
rium has been reached. The reaction is terminated ing its 1,3-oleic acid residues for palmityl groups.
by destroying the catalyst through addition of The products may be recovered by recrystalliza-
water or organic acid, which converts the fatty tion from aqueous acetone.

Fig. 2.23  Reaction mechanism of the interesterification process


Lipid Reactions 71

Fig. 2.24  Formation of interesterification catalyst from sodium hydroxide and glycerol in a two-step process. Adapted
from: A. Rozenaal, Interesterification of Oils and Fats, Inform, 3, pp. 1233–1235, © 1992, AOCS Press

Fig. 2.25  Randomization of palm oil at different tem-


peratures. Adapted from: A. Rozenaal, Interesterification
of Oils and Fats, Inform, 3, pp. 1233–1235, © 1992,
AOCS Press

Interesterification of lard has been used exten-


sively. Lard produces coarse crystals because it Fig. 2.26  Solid fat index (SFI) of cocoa butter before and
tends to crystallize in the β form. Palmitic acid is after interesterification
mostly located in the sn-2 position of the disatu-
rated glycerides (S2U). When lard is randomized, and makes a better shortening. Palm oil shows the
the level of palmitic acid in the sn-2 position phenomenon of post-hardening or post-­
drops from 64 to 24%. The result is a smooth-­ crystallization. This is a disadvantage in a num-
textured fat that crystallizes in the β′ form. ber of applications. Interesterification eliminates
Randomized lard has an improved plastic range this problem.
72 2 Lipids

In the formulation of margarines and shorten- carried out by using lipase enzymes as a cata-
ings, a hardstock is often combined with unmodi- lyst. This type of application is described in
fied liquid oil. A useful hardstock for the Chap. 10.
formulation of soft margarines is an interesteri-
fied blend of palm stearin and palm kernel oil or
fully hydrogenated palm kernel oil. Hydrogenation
Interesterification is used to produce trans free
fats for making margarines and shortenings. The Hydrogenation of fats is a chemical reaction con-
traditional method for transforming oils into fats sisting of addition of hydrogen at double bonds
involves hydrogenation and this results in high of unsaturated acyl groups. This reaction is of
trans levels. The physical and chemical proper- great importance to industry, because it permits
ties of trans free fats made by interesterfication the conversion of liquid oils into plastic fats for
have been described by Petrauskaite et al. (1998). the production of margarine and shortening. For
Ester interchange of fats with a large excess some oils, the process also results in a decreased
of glycerol, at high temperature, under vacuum, susceptibility to oxidative deterioration. In the
and in the presence of a catalyst, results in an hydrogenation reaction, gaseous hydrogen, liq-
equilibrium mixture of mono-, di-, and triglyc- uid oil, and solid catalyst participate under agita-
erides. After removal of excess glycerol, the tion in a closed vessel. Although most industrial
mixture is called technical monoglyceride and processes use solid nickel catalysts, interest in
contains about 40% of 1-monoglyceride. organometallic compounds that serve as homo-
Technical monoglycerides are used as emulsify- geneous catalysts has increased greatly. Frankel
ing agents in foods. Molecular distillation yields and Dutton (1970) have represented catalytic
products with well over 90% 1-monoglycerides; hydrogenation by the following scheme, in which
the distilled monoglycerides are also widely the reacting species are the olefinic substrate (S),
used in foods. Interesterification can also be the metal catalyst (M), and H2:

The intermediates 1, 2, and 3 are organometal- in a natural fat consisting of many component
lic species. If the reaction involves heterogeneous glycerides and different component unsaturated
catalysis, the olefins and hydrogen are bound to fatty acids, the result depends on many factors, if
the metal by chemisorption. If homogeneous the reaction is not carried to completion.
catalysis takes place, the intermediates are Generally, hydrogenation of fats is not carried to
organometallic complexes. The intermediates are completion and fats are hydrogenated only par-
labile and short-lived and cannot usually be iso- tially. Under these conditions, hydrogenation
lated. In heterogeneous catalysis, the surface of may be selective or nonselective. Selectivity
the metal performs the function of catalyst and means that hydrogen is added first to the most
the preparation of the catalyst is of major impor- unsaturated fatty acids. Selectivity is increased
tance. When hydrogen is added to double bonds by increasing hydrogenation temperature and
Hydrogenation 73

decreased by increasing pressure and agitation. intermediate. If a second hydrogen atom is then
Table 2.25 shows the effect of selectivity on the added to this intermediate, the original double
properties of soybean oil. The selectively hydro- bond has been saturated but because the first
genated oil is more resistant to oxidation because addition is reversible, the intermediate can also
of the preferential hydrogenation of the linolenic dissociate. http://lipidlibrary.aocs.org/process-
acid. The influence of selectivity conditions on ing/hydrog-mech/index.htm.
the fatty acids of hydrogenated cottonseed and Figure 2.27 shows how dienes (D for short)
peanut oil is demonstrated by the data presented are hydrogenated to from monoenes (M) and
in Table 2.26 The higher the selectivity, the lower finally stearic acid (D). So linoleic acid (9c,12c–
the level of polyunsaturated fatty acids will be octadecadienoic acid, c,c-D) is reversibly
and the higher the level of monounsaturates. adsorbed in reaction 3 and a hydrogen atom H* is
It is now commonly accepted that the nickel-­ reversibly added to the adsorbed linoleic acid
catalyzed hydrogenation of unsaturated fatty (c,c-D*) to form a half-hydrogenated intermedi-
acids follows the Horiuti-Polanyi mechanism. ate (c-DH*). This is still adsorbed as shown by
According to this mechanism, molecular hydro- the asterisk (*) but has only a single double bond
gen is adsorbed onto the nickel surface (reaction left that has retained its cis-configuration. This
1 in Fig. 2.27 where adsorbed species are indi- half-hydrogenated intermediate can do one of
cated by an asterisk) and dissociated into two several things. It can react irreversibly with a fur-
hydrogen atoms (reaction 2). Fatty acids are also ther hydrogen atom (H*) in reaction 10 or it can
adsorbed onto this nickel surface by their double dissociate.
bond or bonds and in a first step, a hydrogen atom The hydrogen atom leaving on this dissocia-
is added to this bond to form a half-hydrogenated tion can be the same as the one that has been
added as shown in reaction −4 (where the minus
sign indicates the reverse reaction), it can be a
Table 2.25  Differences in selective and nonselective different atom on the same carbon atom or it can
hydrogenation of soybean oil
be a hydrogen atom leaving from a different car-
Characteristic Selective Non-selective bon atom. Accordingly, the fatty acid resulting
Induction period AOM (h) 240 31 from this dissociation can have undergone geo-
Micropenetration 70 (more 30 metrical isomerisation so that the original cis-­
plastic)
configuration of the double bond has been
Capillary mp (°C) 39 55
changed into a trans-configuration as shown in
Condition
   Temp (°C) 177 121
reaction 5. It can also have undergone positional
  Pressure (psi) 5 50 isomerisation meaning that the double bond has
   Ni catalyst (%) 0.05 0.05 shifted one position along the fatty acid chain;
Source: From W.O. Lundberg, Autoxidation and
this type of isomerisation is not shown in the
Antioxidants, 1961, John Wiley & Sons figure. In methylene-interrupted polyunsatu-

Table 2.26  Fatty acid composition of cottonseed and peanut oil hydrogenated under different conditions of selectivity
to iodine value 65
Fatty acids
Oil Hydrogenation conditions Saturated (%) Oleic (%) Linoleic (%)
Cottonseed Moderately selective 31.5 64.5 4.0
Peanut Moderately selective 27.5 72.5 –
Cottonseed Nonselective 36.0 56.0 8.0
Peanut Nonselective 30.0 67.0 3.0
Cottonseed Very nonselective 39.5 48.5 12.0
Peanut Very nonselective 33.0 61.0 6.0
74 2 Lipids

Fig. 2.27  Horiuti-Polanyi mechanism for hydrogenation with nickel catalyst. http://lipidlibrary.aocs.org/processing/
hydrog-mech/index.htm

rated fatty acids this can lead to the formation of 16) leads to stearic acid. (http://lipidlibrary.aocs.
conjugated double bonds. Because the double org/processing/hydrog-mech/index.htm).
bond is no longer present in the half-hydroge- Another important factor in hydrogenation is
nated intermediate, this intermediate can rotate the formation of positional and geometrical iso-
around the original double bond and this can mers. Formation of trans isomers is rapid and
result in an isomerisation that is both geometri- extensive. The isomerization can be understood
cal and positional. http://lipidlibrary.aocs.org/ by the reversible character of chemisorption.
processing/hydrog-mech/index.htm. When the olefinic bond reacts, two carbon-metal
When a half-hydrogenated intermediate is bonds are formed as an intermediate stage (repre-
saturated by reacting with another hydrogen sented by an asterisk in Fig. 2.28). The intermedi-
atom, heat is liberated since the hydrogenation ate may react with an atom of adsorbed hydrogen
process is strongly exothermic. Consequently, to yield the “half-hydrogenated” compound,
the reaction product is so ‘hot’ that it immedi- which remains attached by only one bond.
ately leaves the catalyst surface. By sharing its Additional reaction with hydrogen results in for-
kinetic energy with its surroundings, it cools mation of the saturated compound. There is also
down so that it can be re-adsorbed at its remain- the possibility that the half-hydrogenated olefin
ing double bond (c-M*). This monoene can then may again attach itself to the catalyst surface at a
react with an adsorbed hydrogen to form the half-­ carbon on either side of the existing bond, with
hydrogenated monoene (MH*) that just as above simultaneous loss of hydrogen. Upon desorption
can react in several ways one of which (reaction of this species, a positional or geometrical isomer
Hydrogenation 75

Fig. 2.28  Hydrogenation of an olefinic compound

may result. The proportion of trans acids is high canola oil, and fish oils (Wijesundera et al. 1988).
because this is the more stable configuration. High-erucic rapeseed oil is very difficult to
Double bond migration occurs in both direc- hydrogenate unless it is deodorized (deMan et al.
tions and more extensively in the direction away 1995). Canola oils of the double-zero variety that
from the ester group. This is true not only for the are low in erucic acid and glucosinolates still
trans isomers that are formed but also for the cis contain traces of sulfur in the form of isothiocya-
isomers. The composition of the positional iso- nates (Abraham and deMan 1985).
mers in a partially hydrogenated margarine fat is When catalysts are poisoned by sulfur the
shown in Figs. 2.29 and 2.30 (Craig-Schmidt hydrogenation reaction is slowed down and the
1992). In a partially hydrogenated fat, the analy- formation of trans isomers is increased. U.S.
sis of component fatty acids by gas-liquid chro- stick margarines are reported to contain 24% of
matography is difficult because of the presence of trans fatty acids, and soft margarines contain
many isomeric fatty acids. This is shown in the 14–18%. Shortenings contain 22.5%, and fats in
chromatogram in Fig. 2.31 (Ratnayake 1994). snack foods contain up to 46% of trans fatty
Nickel catalysts are poisoned by sulfur and acids (Craig-Schmidt 1992).
phosphorous compounds, free fatty acids, and It is difficult to eliminate oxidation-sensitive
residual soaps. Oils are refined and sometimes polyunsaturated fatty acids by partial
bleached before hydrogenation. Sulfur com- ­hydrogenation of fish oils. This has been demon-
pounds are not easily removed from the oil. Oils strated by Ackman (1973) in the progressive
that contain sulfur compounds are rapeseed oil, hydrogenation of anchovetta oil. The original
76 2 Lipids

Fig. 2.29  Positional isomers of 18:1 cis formed in the Fatty Acids in Foods and Their Health Implications,
partial hydrogenation of a margarine fat. Adapted from C.K. Chow, ed., p. 369, 1992, by courtesy of Marcel
M.C. Craig-Schmidt, Fatty Acid Isomers in Foods, in Dekker, Inc.

Fig. 2.30  Positional isomers of 18:1 trans formed in the Fatty Acids in Foods and Their Health Implications,
partial hydrogenation of a margarine fat. Adapted from C.K. Chow, ed., p. 369, 1992, by courtesy of Marcel
M.C. Craig-Schmidt, Fatty Acid Isomers in Foods, in Dekker, Inc.

eicosapentaenoic acid (20:5 ω 3) is not com- In the nonselective hydrogenation of typical


pletely removed until an iodine value of 107.5 is seed oils, polyunsaturated fatty acids are rapidly
reached. Even at this point there are other poly- reduced and trans-isomer levels increase to high
unsaturated fatty acids present that may be sus- values. Figure 2.32 shows the hydrogenation of
ceptible to flavor reversion. canola oil (de El-Shattory et al. 1982).
Fig. 2.31  Gas chromatogram of the fatty acid methyl by Infrared Spectrophotometry and Deterinination of
esters from partially hydrogenated soybean oil, using a Fatty Acid Composition of Partially Hydrogenated
100-m fused silica capillary column coated with SP2560. Vegetable Oils and Animal Fats by Gas Chromatography/
Source: Reprinted with permission from Infrared Spectrophotometry: Collaborative Study,
W.M.N. Ratnayake, Determination of Trans Unsaturation J.A.O.A.C. Intern., Vol. 78, pp. 783–802, © 1994

Fig. 2.32  Change in


fatty acid composition
during hydrogenation of
canola oil
78 2 Lipids

Lipid Oxidation

The unsaturated bonds present in fats and oils


represent active centers that, among other things,
may react with oxygen. This reaction leads to the
formation of primary, secondary, and tertiary oxi-
dation products that may make the fat or fat-­
containing foods unsuitable for consumption.
The process of autoxidation and the resulting
deterioration in flavor of fats and fatty foods are
often described by the term rancidity. Usually
rancidity refers to oxidative deterioration but, in
the field of dairy science, rancidity refers usually
to hydrolytic changes resulting from enzyme
activity. Lundberg (1961) distinguishes several
types of rancidity. In fats such as lard, common
oxidative rancidity results from exposure to oxy-
gen; this is characterized by a sweet but undesir-
able odor and flavor that become progressively
more intense and unpleasant as oxidation pro- Fig. 2.33  Mechanism of lipid oxidation. http://www.
ncbi.nlm.nih.gov/pmc/articles/PMC2868362/
gresses. Flavor reversion is the term used for the
objectionable flavors that develop in oils contain-
ing linolenic acid. This type of oxidation is pro- During the propagation phase the chain reaction
duced with considerably less oxygen than with between fatty acid radicals and molecular oxygen
common oxidation. A type of oxidation similar to leads to the formation and accumulation of the
reversion may take place in dairy products, where primary hydroperoxide products. Reactions
a very small amount of oxygen may result in between radicals leading to non-radical products
intense oxidation off-flavors. It is interesting to dominate during the termination phase.
note that the linolenic acid content of milk fat is In the initiation part, hydrogen is abstracted
quite low. from an olefinic compound to yield a free radical.
Among the many factors that affect the rate of The removal of hydrogen takes place at the car-
oxidation are the following: bon atom next to the double bond and can be
­initiated by light or metal ions such as iron or
• amount of oxygen present copper. The dissociation energy of hydrogen in
• degree of unsaturation of the lipids various olefinic compounds has been listed by
• presence of antioxidants Ohloff (1973) and is shown in Table 2.27. Once a
• presence of prooxidants, especially copper, free radical has been formed, it will combine with
and some organic compounds such as heme-­ oxygen to form a peroxy-free radical, which can
containing molecules and lipoxidase in turn abstract hydrogen from another unsatu-
• nature of packaging material rated molecule to yield a peroxide and a new free
• light exposure radical, thus starting the propagation reaction.
• temperature of storage This reaction may be repeated up to several thou-
sand times and has the nature of a chain reaction.
The lipid oxidation process during lipid per- The hydroperoxides formed in the propagation
oxidation consists of three partially overlapping part of the reaction are the primary oxidation prod-
phases of radical reactions which can be distin- ucts. The hydroperoxide mechanism of autoxida-
guished: initiation, propagation, and termina- tion was first proposed by Farmer (1946). These
tion (Fig. 2.33). In the initiation phase reactions oxidation products are generally unstable and
prevail that form and expand the pool of radicals. decompose into the secondary oxidation products,
Lipid Oxidation 79

Table 2.27  Dissociation energy for the abstraction of used to measure the progress of oxidation.
hydrogen from olefinic compounds and peroxides
Organoleptic changes are more closely related to
Compound ΔΕ (kcal/mole) the secondary oxidation products, which can be
H—CH═CH2 103 measured by various procedures, including the
H—CH2—CH2—CH3 100 benzidine value, which is related to aldehyde
H—CH2—CH═CH2 85 decomposition products. As the aldehydes are
H — CH — CH = CH — CH 2 — 77 themselves oxidized, fatty acids are formed; these
| free fatty acids may be considered tertiary oxida-
CH 3 tion products. The length of the induction period,
65 therefore, depends on the method used to deter-
= =
— CH CH — CH — CH CH —
mine oxidation products.
|
H
H—OO—R 90 Initiation
Source: From G. Ohloff, Fats as Precursors, in Functional
Properties of Fats in Foods, J. Solms, ed., 1973, Forster The attack of a ROS able to abstract a hydrogen
Publishing
atom from a methylene group (─CH2─), generat-
ing free radicals from polyunsaturated fatty acids.
which include a variety of compounds, including OH is the most efficient ROS to do that attack.
carbonyls, which are the most important. The per-
oxides have no importance to flavor deterioration,
which is wholly caused by the secondary oxida-
tion products. The nature of the process can be
represented by the curves of Fig. 2.34 (Pokorny
1971). In the initial stages of the reaction, the
amount of hydroperoxides increases slowly; this
stage is termed the induction period. At the end of
the induction period, there is a sudden increase in
peroxide content. Because peroxides are easily Energy requirement for radical production by
determined in fats, the peroxide value is frequently rupture of a CH bond is about 80 kcal.

Propagation

Termination
80 2 Lipids

Fig. 2.34 Autoxidation
of Lard comparing (a)
peroxide value, (b)
benzidine value, (c) acid
value. Adapted from
J. Pokorny, Stabilization
of Fats by Phenolic
Antioxidants, Can. Inst.
Food Sci. Technol. J.,
Vol. 4, pp. 68–74, 1971

Fig. 2.35  Peroxide formation and decomposition as a function of time

The mechanism of the autoxidation of polyun- The propagation can be followed by termina-
saturated fatty acids as a radical chain reaction tion if the free radicals react with themselves to
was established in the middle of the twentieth yield non-active products, as shown here:
century is illustrated in Fig. 2.34. Soon after fol-
lowed the elucidation of the role of antioxidants R• + R• → R — R
as agents that break the radical chain (Inglold R • + RO•2 → RO2 R
1961), and the identification of secondary trans- nRO•2 → ( RO2 )n
formation products of the primary hydroperox-
ides through enzymatic and non-enzymatic The rate and course of autoxidation depend pri-
transformations, the latter an active area of marily on the composition of the fat—its degree of
research that continues to unravel novel enzymes unsaturation and the types of unsaturated fatty
and products (Gardner 1989; Gerwick 1996; acids present. The absence, or at least a low value,
Grechkin 1998; Tijet and Brash 2002; Schneider of peroxides does not necessarily indicate that an
et al. 2007). oil is not oxidized. As Fig. 2.35 indicates, perox-
ides are labile and may be transformed into sec-
Lipid Oxidation 81

ondary oxidation products. A combined index of tion period. At the end of the induction period,
primary and secondary oxidation products gives a there is a sudden increase in peroxide content.
better evaluation of the state of oxidation of an oil. Because peroxides are easily determined in fats,
This is expressed as Totox value: Totox value = 2 × the peroxide value is frequently used to measure
peroxide value + anisidine value. (Anisidine value the progress of oxidation. Organoleptic changes
is a measure of secondary oxidation products.) are more closely related to the secondary oxida-
Removal of oxygen from foods will prevent oxida- tion products, which can be measured by various
tion, but, in practice, this is not easy to accomplish procedures, including the benzidine value, which
in many cases. At high temperatures (100–140 °C) is related to aldehyde decomposition products.
such as those used in the accelerated tests for oil As the aldehydes are themselves oxidized, fatty
stability (active oxygen method), formic acid is acids are formed; these free fatty acids may be
produced, which can be used to indicate the end of considered tertiary oxidation products.
the induction period. The formation of formic acid Although even saturated fatty acids may be
results from aldehyde decomposition. Peroxidation oxidized, the rate of oxidation greatly depends on
of aldehydes establishes a resonance equilibrium the degree of unsaturation. In the series of
between two limiting forms. 18-­carbon-atom fatty acids 18:0, 18:1, 18:2, 18:3,
The hydroperoxides formed in the propaga- the relative rate of oxidation has been reported to
tion part of the reaction are the primary oxidation be in the ratio of 1:100:1200:2500. The reaction
products. The hydroperoxide mechanism of of unsaturated compounds proceeds by the
autoxidation was first proposed by Farmer abstraction of hydrogen from the α carbon, and
(1946). These oxidation products are generally the resulting free radical is stabilized by reso-
unstable and decompose into the secondary oxi- nance as follows:
dation products, which include a variety of com-
pounds, including carbonyls, which are the most
important. The peroxides have no importance to
flavor deterioration, which is wholly caused by
the secondary oxidation products. The nature of If oleic acid is taken as example of a mono-­
the process can be represented by the curves of ethenoid compound (cis-9-octadecenoic acid),
Fig. 2.36 (Labuza 1971). In the initial stages of the reaction will proceed by abstraction of hydro-
the reaction, the amount of hydroperoxides gen from carbons 8 or 11, resulting in two pairs
increases slowly; this stage is termed the induc- of resonance hybrids.

Fig. 2.36  The volatiles and non-volatiles contain the secondary oxidative by-products. Adapted from: Labuza, T. P.
1971. Kinetics of Lipid Oxidation in Foods, 2: 355–405. CRC Crit. Rev. Food Technol
82 2 Lipids

This leads to the formation of the following may be in the trans configuration (Lundberg
four isomeric hydroperoxides: 1961).
From linoleic acid (cis-cis-9,12-octadeca-­
dienoic acid), three isomeric hydroperoxides can
be formed as shown in the next formula. In this
mixture of 9, 11, and 13 hydroperoxides, the con-
jugated ones occur in greatest quantity because
they are the more stable forms. The hydroperox-
ides occur in the cis-trans and trans-trans con-
figurations, the content of the latter being greater
with higher temperature and greater extent of
oxidation. From the oxidation of linolenic acid
(cis, cis, cis-9,12,15-octadecatrienoic acid), six
isometric hydroperoxides can be expected
according to theory, as shown:

In addition to the changes in double bond


position, there is isomerization from cis to
trans, and 90% of the peroxides formed
Lipid Oxidation 83

Hydroperoxides of linolenate decompose are in great part responsible for the oxidized fla-
more readily than those of oleate and linoleate vor of fats.
because active methylene groups are present. The The alkoxy radical may also abstract a hydro-
active methylene groups are the ones located gen atom from another molecule to yield an alco-
between a single double bond and a conjugated hol and a new free radical, as shown:
diene group. The hydrogen at this methylene
group could readily be abstracted to form dihy-
droperoxides. The possibilities here for decom- R — CH — R + R1H → R — CH — R + R1•
position products are obviously more abundant | |
than with oleate oxidation. O• OH
The decomposition of hydroperoxides has
been outlined by Keeney (1962). The first step The new free radicals formed may participate
involves decomposition to the alkoxy and in propagation of the chain reaction. Some of the
hydroxy free radicals. free radicals may interact with themselves to ter-
minate the chain, and this could lead to the for-
mation of ketones as follows:

R — CH — R + R1· → R — C — R + R1H
The alkoxy radical can react to form | ||
aldehydes. O· O

As indicated, a variety of aldehydes have been
demonstrated in oxidized fats. Alcohols have also
been identified, but the presence of ketones is not
This reaction involves fission of the chain and as certain. Keeney (1962) has listed the alde-
can occur on either side of the free radical. The hydes that may be formed from breakdown of
aldehyde that is formed can be a short-chain vol- hydroperoxides of oxidized oleic, linoleic, linole-
atile compound, or it can be attached to the glyc- nic, and arachidonic acids (Table 2.28). The alde-
eride part of the molecule; in this case, the hydes are powerful flavor compounds and have
compound is nonvolatile. The volatile aldehydes very low flavor thresholds; for example,
84 2 Lipids

Table 2.28  Hydroperoxides and aldehydes (with single oxygen function) that may be formed in autoxidation of some
unsaturated fatty acids
Isomeric hydroperoxides formed from the
Methylene Group structures contributing to the intermediate Aldehydes formed by decomposition
Fatty acid Involveda free radical resonance hybrid of the hydroperoxides
Oleic 11 11-hydroperoxy-9-ene octanal
9-hydroperoxy-10-ene 2-decenal
8 8-hydroperoxy-9-ene 2-undecenal
10-hydroperoxy-8-ene nonanal
Linoleic 11 13-hydroperoxy-9,11-diene hexanal
11-hydroperoxy-9,12-diene 2-octenal
9-hydroperoxy-10,12-diene 2,4-decadienal
Linolenic 14 16-hydroperoxy-9,12,14-triene propanal
14-hydroperoxy-9,12,15-triene 2-pentenal
12-hydroperoxy-9,13,15-triene 2,4-heptadienal
11 13-hydroperoxy-9,11,15-triene 3-hexenal
11-hydroperoxy-9,12,15-triene 2,5-octadienal
9-hydroperoxy-10,12,15-triene 2,4,7-decatrienal
Arachidonic 13 15-hydroperoxy-5,8,11,13-tetraene hexanal
13-hydroperoxy-5,8,11,14-tetraene 2-octenal
11-hydroperoxy-5,8,12,14-tetraene 2,4-decadienal
10 12-hydroperoxy-5,8,10,14-tetraene 3-nonenal
10-hydroperoxy-5,8,11,14-tetraene 2,5-undecadienal
8-hydroperoxy-5,9,11,14-tetraene 2,4,7-tridecatrienal
7 9-hydroperoxy-5,7,11,14-tetraene 3,6-dodecadienal
7-hydroperoxy-5,8,11,14-tetraene 2,5,8-tetradecatrienal
5-hydroperoxy-6,8,11,14-tetraene 2,4,7,10-hexadecatetraenal
Source: From M. Keeney, Secondary Degradation Products, in Lipids and Their Oxidation, H.W. Schultz et al., eds.,
1962, AVI Publishing Co.
a
Only the most active methylene groups in each acid are considered

2,4-­decadienal has a flavor threshold of less than The rate and course of autoxidation depend
one part per billion. The presence of a double primarily on the composition of the fat—its
bond in an aldehyde generally lowers the flavor degree of unsaturation and the types of unsatu-
threshold considerably. The aldehydes can be rated fatty acids present. The absence, or at least
further oxidized to carboxylic acids or other ter- a low value, of peroxides does not necessarily
tiary oxidation products. indicate that an oil is not oxidized. As Fig. 2.16
When chain fission of the alkoxy radical indicates, peroxides are labile and may be trans-
occurs on the other side of the free radical group, formed into secondary oxidation products. A
the reaction will not yield volatile aldehydes but combined index of primary and secondary oxida-
will instead form nonvolatile aldehydo-­tion products gives a better evaluation of the state
glycerides. Volatile oxidation products can be of oxidation of an oil. This is expressed as Totox
removed in the refining process during deodor- value: Totox value = 2 × peroxide value + anisi-
ization, but the nonvolatile products remain; this dine value. (Anisidine value is a measure of sec-
can result in a lower oxidative stability of oils ondary oxidation products.) Removal of oxygen
that have already oxidized before refining. from foods will prevent oxidation, but, in prac-
tice, this is not easy to accomplish in many cases.
R – CH 2 – CO  R – CH – CHO
At high temperatures (100–140 °C) such as those
used in the accelerated tests for oil stability
Lipid Oxidation 85

(active oxygen method), formic acid is produced, AH + R· → RH + A·


which can be used to indicate the end of the AH + RO2· → RO2 H + A·
induction period. The formation of formic acid
results from aldehyde decomposition. The antioxidant free radical deactivated by
Peroxidation of aldehydes establishes a reso- further oxidation to quinones, thus terminating
nance equilibrium between two limiting forms. the chain. Only phenolic compounds that can
The second hybrid ties up oxygen at the α car- easily produce quinones are active as antioxi-
bon to yield the α-hydroperoxy aldehyde as dants (Pokorny 1971). At high concentrations
follows: antioxidants may have a prooxidant effect and
one of the reactions may be as follows:

| A· + RH → AH + R
O Tocopherols in natural fats are usually present
| at optimum levels. Addition of antioxidant
· beyond optimum amounts may result in increas-
R – C H – CHO + O2 → R – CH – CHO ing the extent of prooxidant action. Lard is an

example of a fat with very low natural antioxi-
Breakdown of oxygen and carbon bonds dant activity and antioxidant must be added to it,
yields formic acid and a new aldehyde. to provide protection. The effect of antioxidants
can be expressed in terms of protection factor, as
O· shown in Fig. 2.37 (Pokorny 1971). The highly
| active antioxidants that are used in the food
O industry are active at about 10–50 parts per mil-
| lion (ppm). Chemical structure of the antioxi-
dants is the most important factor affecting their
R – C – CH + CHO → HCOOH + RCHO
activity. The number of synthetic antioxidants
deMan et al. (1987) investigated this reaction permitted in foods is limited, and the structure of
with a variety of oils and found that although for- the most widely used compounds is shown in
mic acid was the main reaction product, other Fig. 2.38. Propyl gallate is more soluble in water
short-chain acids from acetic to caproic were also than in fats. The octyl and dodecyl esters are
formed. Trace metals, especially copper, and to a more fat soluble. They are heat resistant and non-
lesser extent iron, will catalyze fat oxidation; volatile with steam, making them useful for fry-
metal deactivators such as citric acid can be used ing oils and in baked products. These are
to reduce the effect. Lipoxygenase (lipoxidase) considered to have carry-through properties.
and heme compounds act as catalysts of lipid oxi- Butylated hydroxyanisole (BHA) has carry-­
dation. Antioxidants can be very effective in through properties but butylated hydroxy toluene
slowing down oxidation and increasing the (BHT) does not, because it is volatile with steam.
induction period. Many foods contain natural The compound tert-butyl hydroquinine (TBHQ)
antioxidants; the tocopherols are the most impor- is used for its effectiveness in increasing oxida-
tant of these. They are present in greater amounts tive stability of polyunsaturated oils and fats. It
in vegetable oils than in animal fats, which may also provides carry-through protection for fried
explain the former’s greater stability. foods. Antioxidants are frequently used in com-
Antioxidants such as tocopherols may be nat- bination or together with synergists. The latter
urally present; they may be induced by processes are frequently metal deactivators that have the
such as smoking or roasting, or added as syn- ability to chelate metal ions. An example of the
thetic antioxidants. Antioxidants act by reacting combined effect of antioxidants is shown in
with free radicals, thus terminating the chain. The Fig. 2.39. It has been pointed out (Zambiazi and
antioxidant AH may react with the fatty acid free Przybylski 1998) that fatty acid composition can
radical or with the peroxy free radical, explain only about half of the oxidative stability
86 2 Lipids

Fig. 2.37 
Determination of
protection factor. (a)
Lard, (b) lard +
antioxidant. Adapted
from J. Pokomy,
Stabilization of Fats by
Phenolic Antioxidants,
Can. Inst. Food Sci.
Technol. J., Vol. 4,
pp. 68–74, 1971

Fig. 2.38  Mechanism of oxidation, propagation and antioxidant protection

of a vegetable oil. The other half can be contrib- The formation of 9-, 11-, and 13-hydroperox-
uted to minor components including tocopherols, ides is expected based on the three mesomeric
metals, pigments, free fatty acids, phenols, phos- structures for the pentadienyl radical of linoleic
pholipids, and sterols. acid. That implicate localization (and therefore
Lipid Oxidation 87

Fig. 2.39  Structure of propyl gallate (PG), butylated hydroxyanisole (BHA), butylated hydroxy toluene (BHT), and
tert-butyl hydroquinone (TBHQ)

Fig. 2.40  Effect of


copper concentration on
protective effect of
antioxidants in lard.
(Adapted from
J. Pokorny, Stabilization
of Fats by Phenolic
Antioxidants, Can. Inst.
Food Sci. Technol. J.,
Vol. 4, pp. 68–74, 1971

reactivity with O2) of the radical at carbons 9, 11, as reversible and competing reactions help
and 13 (Fig. 2.38). The (9- and 13-)hydroperox- explain the chemistry. This will help explain how
ides are easily identified in autoxidation reac- the rate constants of different and competing
tions, the 11-hydroperoxide has proven elusive reactions are a determining factors explaining
for decades (Haslbeck et al. 1983). The instabil- products formed during lipid autoxidation
ity of the intermediate bis-allylic peroxyl radical (Fig.  2.40). http://www.ncbi.nlm.nih.gov/pmc/
of this hydroperoxide has made it difficult to articles/PMC2868362/.
identify and isolate. In order to explain the mech- There are a number of antioxidants that can
anistic basis for formation of the bis-allylic be applied to obtain certain effects in fats and
hydroperoxide, the concept of radical reactions oils.
88 2 Lipids

Antioxidants used to control oxidation fat or oil Singlet oxygen is short-lived and reverts back
Butylated Improves oxidative stability, to the ground state with the emission of light.
hydroxyanisole antioxidants This light is fluorescent, which means that the
(BHA)
wavelength of the emitted light is higher than that
Butylated Improves oxidative stability,
hydroxytoluene antioxidants
of the light that was absorbed for the excitation.
(BHT) The reactivity of singlet oxygen is 1500 greater
Carotene (Pro-Vitamin Enhances color of finished than that of ground-state oxygen. Compounds
A) foods; color additive that can act as sensitizers are widely occurring
Citric acid Inhibit metal-catalized food components, including chlorophyll, myo-
oxidation and production of globin, riboflavin, and heavy metals. Most of
dark colors; metal chelating
agents. these compounds promote type II oxidation reac-
Methyl silicone Inhibits oxidation; antifoam tions. In these reactions the sensitizer is trans-
agent formed into the activated state by light. The
Phosphoric acid Inhibit metal-catalyzed activated sensitizer then reacts with oxygen to
oxidation and production of produce singlet oxygen.
dark colors; metal chelating
agent hv

tertiary Improves oxidative stability,


sen → sen∗

butylhydroquinone antioxidants sen + O2 → sen +1 O2
(TBHQ)
Tocopherols Natural antioxidant, The singlet oxygen can react directly with
improves oxidative stability unsaturated fatty acids.


1
O2 + RH → ROOH
Photooxidation The singlet oxygen reacts directly with the
double bond by addition, and shifts the double
Oxidation of lipids, in addition to the free radical bond one carbon away. The singlet oxygen
process, can be brought about by at least two attack on linoleate produces four hydroperox-
other mechanisms—photooxidation and enzymic ides as shown in Fig. 2.41. Photooxidation has
oxidation by lipoxygenase. The latter is dealt no induction period, but the reaction can be
with in Chap. 10. Light-induced oxidation or quenched by carotenoids that effectively com-
photooxidation results from the reactivity of an pete for the singlet oxygen and bring it back to
excited state of oxygen, known as singlet oxygen the ground state.
(1O2). Ground-state or normal oxygen is triplet Phenolic antioxidants do not protect fats from
oxygen (3O2). The activation energy for the reac- oxidation by singlet oxidation (Yasaei et al.
tion of normal oxygen with an unsaturated fatty 1996). However, the antioxidant ascorbyl palmi-
acid is very high, of the order of 146–273 kJ/ tate is an effective singlet oxygen quencher (Lee
mole. When oxygen is converted from the ground et al. 1997). Carotenoids are widely used as
state to the singlet state, energy is taken up quenchers. Rahmani and Csallany (1998)
amounting to 92 kJ/mole, and in this state the reported that in the photooxidation of virgin olive
oxygen is much more reactive. Singlet-state oxy- oil, pheophytin A functioned as sensitizer, while
gen production requires the presence of a sensi- β-carotene acted as a quencher.
tizer. The sensitizer is activated by light, and can The combination of light and sensitizers is
then either react directly with the substrate (type present in many foods displayed in transparent
I sensitizer) or activate oxygen to the singlet state containers in brightly lit supermarkets. The light-­
(type II sensitizer). In both cases unsaturated induced deterioration of milk has been studied
fatty acid residues are converted into hydroper- extensively. Sattar et al. (1976) reported on the
oxides. The light can be from the visible or ultra- light-induced flavor deterioration of several oils
violet region of the spectrum. and fats. Of the five fats examined, milk fat and
Photooxidation 89

Fig. 2.41 
Photooxidation.
Singlet-oxygen attack on
oleate produces two
hydroperoxides;
linoleate yields four
hydroperoxides

Fig. 2.42  Effect of


temperature on rate of
oxidation of illuminated
com oil. Adapted from:
M.H. Chahine and J.M.
deMan, Autoxidation of
Com Oil under the
Influence of Fluorescent
Light, Can. Inst. Food
Sci. Technol. J., Vol. 4,
pp. 24–28,1971

soybean oil were most susceptible and corn oil deMan (1971) (Fig. 2.42). They found that tem-
least susceptible to singlet oxygen attack. The perature has an important effect on photooxida-
effect of temperature on the rate of oxidation of tion rates, but even freezing does not completely
illuminated corn oil was reported by Chahine and prevent oxidation.
90 2 Lipids

Heated Fats: Frying high levels of free fatty acids demonstrated that
the formation of monoacylglycerols is minimal
Fats and oils are heated during commercial pro- and also that the reaction is not selective and con-
cessing and during frying. Heating during pro- sequently, is independent of the fatty acid com-
cessing mainly involves hydrogenation, physical position of the frying oil. Hydrolysis is a relatively
refining, and deodorization. Temperature used in simple reaction but many consider it to be one of
these processes may range from 120 °C to the most important reactions during frying.
270 °C. The oil is not in contact with air, which Pokorny et al. (1998) reported that used frying
eliminates the possibility of oxidation. At the oils from fast food operations exhibited high con-
high temperatures used in physical refining and tents of both diacylglycerols and fatty acids. In
deodorization, several chemical changes may well-­controlled frying operations of potatoes
take place (Erickson and Frey 1994). These substrate had a very high content of water over a
include randomization of the glyceride structure, wide range of conditions, hydrolytic products were
dimer formation, cis-trans isomerization, and minor compounds within the total recovered deg-
formation of conjugated fatty acids (positional radation compounds (Dobarganes et al. 1993).
isomerization) of polyunsaturated fatty acids Lipid oxidation at high temperatures such as
(Hoffmann 1989). Heating oils during frying can those encountered in baking and frying is very
result in three types of chemical modification of complex because both oxidative and thermal
the oil. They are hydrolysis, oxidation and ther- reactions are proceeding simultaneously.
mal degradation. Table 2.29 summarizes the prin- Although oxygen is highly soluble in cooking
cipal degradation products form these reactions. oils, the oil solubility decreases at high tempera-
Hydrolysis is the splitting of the triacylglyc- ture, however, the high temperatures cause the
erol molecule, with formation of diacylglycerols oxidation reactions to increase (Velasco et al.
and fatty acids resulting in nonvolatile fatty acids 2008; Frankel 1997).
with molecular weight significantly lower than Figure 2.43 illustrates the oxidation process
that of the parent triacylglycerols. Analysis of that occurs during frying. The oxidation proceeds
glycerides and free fatty acids in frying oils with via a free radical mechanism with chain reactions,
where RH represents here the triacylglycerol
Table 2.29  Principal degradation products compounds molecule undergoing oxidation in one of its
formed during frying unsaturated fatty acyl groups. In the initiation
Reaction Products found stage, an alkyl radical is formed by abstraction of
Reaction Cause in oil a hydrogen radical from an allylic or bis allylic
Hydrolysis Moisture Fatty acids position of an unsaturated fatty acid. In the propa-
Diacylglycerols gation step, the alkyl radical reacts with oxygen at
Oxidation Air Oxidized monomeric rates controlled by diffusion to form peroxyl radi-
triacylglycerols cals that in turn react with new triacylglycerol
Oxidized dimeric and molecules giving rise to hydroperoxides as the
oligomeric
triacylglycerols
primary oxidation products and new alkyl radi-
Volatile compounds cals that propagate the reaction chain. Finally, in
(aldehydes, ketones, the termination stage, radicals react between them
alcohols, hydrocarbons, to yield relatively stable non-radical species.
etc.) (http://lipidlibrary.aocs.org/frying/frying.html).
Thermal High Cyclic monomeric
The oxygen content becomes depleted at fry-
degradation Temperature triacylglycerols
Isomeric monomeric
ing temperatures thus the alkylperoxyl radicals
triacylglycerols (ROO• are diminished and the concentration of
Nonpolar dimeric and alkyl radicals (R•) increases. These changes result
oligomeric in the formation of more polymeric products
triacylglycerols through reactions mainly involving alkyl (R•) and
Heated Fats: Frying 91

Fig. 2.43  Simplified scheme of thermal oxidation. http://lipidlibrary.aocs.org/frying/frying.html

alkoxyl (RO•) radicals. Polymerization com- range of 160–195 °C. At lower temperatures fry-
pounds become significant in the accelerated stage ing takes longer, and at higher temperatures dete-
of oxidation after the end of the induction period rioration of the oil is the limiting factor. Deep
(Márquez-Ruiz and Dobarganes 2005). At high frying is a complex process involving both the oil
temperatures, formation of new compounds is and the food to be fried. The reactions taking
very rapid, ROOH are practically absent above place are schematically presented in Fig. 2.44.
150 °C, indicating that the rate of ROOH decom- Steam is given off during the frying, which
position becomes higher than that of their forma- removes volatile antioxidants, free fatty acids,
tion, and polymeric compounds are formed from and other volátiles. Contact with the air leads to
the very early stages of heating. Also, the forma- autoxidation and the formation of a large number
tion of significant amounts of non-polar triacylg- of degradation products. The presence of steam
lycerol dimers (R-R), typical compounds formed results in hydrolysis, with the production of free
in the absence of oxygen through interaction of fatty acids and partial glycerides. At lower frying
alkyl radicals, is a clear indication of the low oxy- temperatures the food has to be fried longer to
gen concentration (Dobarganes and Pérez-­Camino reach the desirable color, and this results in
1987). Dimers and oligomers: are the major com- higher oil uptake. Oil absorption by fried foods
pounds in used frying fats and are formed through may range from 10 to 40%, depending on condi-
interaction between triglyceride radicals. tions of frying and the nature and size of the food.
Conditions prevailing during frying are less Oils used in deep frying must be of high qual-
favorable than those encountered in the above-­ ity because of the harsh conditions during deep
mentioned processes. Deep frying, where the frying and to provide satisfactory shelf life in
food is heated by immersion in hot oil, is prac- fried foods. The suitability of an oil for frying is
ticed in commercial frying as well as in food ser- directly related to its content of unsaturated fatty
vice operations. The temperatures used are in the acids, especially linolenic acid. This has been
92 2 Lipids

Fig. 2.44  Summary of


chemical reactions
occurring during deep
frying. Source:
Reprinted with
permission from
F.T. Orthoefer,
S. Gurkin, and K. Lui,
Dynamics of Frying in
Deep Frying, in
Chemistry, Nutrition and
Practical Applications,
E.G. Perkins and
M.D. Erickson, eds.,
p. 224. © 1996, AOCS
Press

described by Erickson (1996) as “inherent stabil- The stability of frying oils and fats is usually
ity” calculated from the level of each of the unsat- measured by an accelerated test known as the
urated fatty acids (oleic, linoleic, and linolenic) active oxygen method (AOM). In this test, air is
and their relative reaction rate with oxygen. The bubbled through an oil sample maintained at
inherent stability calculated for a number of oils 95 °C and the peroxide value is measured at
is given in Table 2.30. The higher the inherent intervals. At the end point the peroxide value
stability, the less suitable the oil is for frying. The shows a sharp increase, and this represents the
liquid seed oils, such as soybean and sunflower AOM value in hours. Typical AOM values for liq-
oil, are not suitable for deep frying and are usu- uid seed oils range from 10 to 30 h; heavy-duty
ally partially hydrogenated for this purpose. Such frying shortenings range from 200 to 300 h. AOM
hydrogenated oils can take the form of shorten- values of some oils and fats determined by mea-
ings, which may be plastic solids or pourable sus- suring the peroxide value and using an automatic
pensions. Through plant breeding and genetic recording of volatile acids produced during the
engineering, oils with higher inherent stability test are given in Table 2.31 (deMan et al. 1987).
can be obtained, such as high-oleic sunflower oil, As shown in Fig. 2.44, oil breakdown during
low-linolenic canola oil, and low-linolenic soy- frying can be caused by oxidation and thermal
bean oil. alteration. Oxidation can result in the formation
Flavor Reversion 93

Table 2.30  Inherent stability of oils for use in frying dimeric and oligomeric triglycerides. The polym-
Oil Iodine value Inherent stabilitya erization reaction may take place by conversion
Soybean 130 7.4 of part of the cis-cis-1,4 diene system of linole-
Sunflower 120 7.7 ates to the trans-trans conjugated diene. The 1,4
High-oleic 90 2.0 and dienes can combine in a Diels-Alder type
sunflower addition reaction to produce a dimer as shown in
Corn 110 6.2 Fig. 2.45. Other possible routes for dimer forma-
Cottonseed 98 5.2
tion are through free radical reactions. As shown
Canola 110 5.4
in Fig. 2.46, this may involve combination of
Peanut 92 4.5
radicals, intermolecular addition, and intramo-
Lard 60 1.4
lecular addition. From dimers, higher oligomers
Olive 88 1.8
Palm 55 1.4
can be produced; the structure of these is still
Palm olein 58 1.6
relatively unknown.
Palm stearin 35 1.0 Another class of compounds formed during
Tallow 50 0.7 frying is cyclic monomers of fatty acids. Linoleic
Palm kernel 17 0.5 acid can react at either the C9 or C12 double
Coconut 9 0.4 bonds to give rings between carbons 5 and 9, 5
a
Inherent stability calculated from decimal fraction of and 10, 8 and 12, 12 and 17, and 13 and 17.
fatty acids multiplied by relative reaction rates with oxy- Cyclic monomers with a cyclopentenyl ring have
gen, assuming rate for oleic acid = 1, linoleic acid = 10, been isolated from heated sunflower oil, and their
and linolenic acid = 25
structure is illustrated in Fig. 2.47 (Le Quéré and
Sébédio 1996).
Table 2.31  Active oxygen method (AOM) time of sev- Some countries such as France require that
eral oils and fats as determined by peroxide value and frying oils contain less than 2% linolenic acid.
conductivity measurements Several European countries have set maximum
AOM time limits for the level of polar compounds or for the
Oil AOM Time (POV)a (conductivity)b level of free fatty acids beyond which the fat is
Sunflower 6.2 7.1 considered unfit for human consumption. In con-
Canola 14.0 15.8 tinuous industrial frying, oil is constantly being
Olive 17.8 17.8 removed from the fryer with the fried food and
Corn 12.4 13.8 replenished with fresh oil so that the quality of
Peanut 21.1 21.5 the oil can remain satisfactory. This is more dif-
Soybean 11.0 10.4
ficult in intermittent frying operations.
Triolein 8.1 7.4
Lard 42.7 43.2
Butterfat 2.8 2.0
Flavor Reversion
Source: Reprinted with permission from J.M. deMan,
et al., Formation of Short Chain Volatile Organic Acids in
the Automated AOM Method, J.A.O.C.S., Vol. 64, p. 996, Soybean oil and other fats and oils containing
© 1987, American Oil Chemists’ Society linolenic acid show the reversion phenomenon
a
At peroxide value 100 when exposed to air. Reversion flavor is a partic-
b
At intercept of conductivity curve and time axis
ular type of oxidized flavor that develops at com-
paratively low levels of oxidation. The off-flavors
of oxidized monomeric, dimeric, and oligomeric may develop in oils that have a peroxide value of
triglycerides as well as volatile compounds as little as 1 or 2. Other oils may not become ran-
including aldehydes, ketones, alcohols, and cid until the peroxide value reaches 100.
hydrocarbons. In addition, oxidized sterols may Linolenic acid is generally recognized as the
be formed. Thermal degradation can result in determining factor of inversion flavors. These
cyclic monomeric triglycerides and nonpolar off-flavors are variously described as grassy,
94 2 Lipids

Fig. 2.45  Polymerization of Diene systems to form dimers

Fig. 2.46 Nonpolar
dimer formation through
free radical reactions

Fig. 2.47  Cyclic fatty acid monomers formed from linoleic acid in heated sunflower oil
Physical Properties 95

fishy, and painty (Cowan and Evans 1961). The Physical Properties
origin of these flavors appears to be the volatile
oxidation products resulting from the terminal Fats and oils are mixtures of mixed triglycerides.
pentene radical of linolenic acid, CH3–CH2– Fats are semisolid at room temperature; they are
CH=CH–CH2–. Hoffmann (1962) has listed the known as plastic fats. The solid character of fats
flavor descriptions of reverted soybean oil is the result of the presence of a certain propor-
(Table 2.32) and the volatile decomposition prod- tion of crystallized triglycerides. Most fats con-
ucts isolated from reverted or oxidized soybean tain a range of triglycerides of different melting
oil (Table 2.33). points from very high to very low. When a fat is
The first perceptible reversion flavor was liquefied by heating, all glycerides are in the liq-
found to be caused by 3-cis-hexenal, which has a uid state; upon cooling, some of the higher melt-
pronounced green bean odor. Other flavorful ing fractions become insoluble and crystallize.
aldehydes isolated were 2-trans-hexenal (green, As the temperature is lowered, more glycerides
grassy), 2-trans-nonenal (rancid), and 2-trans-­6- become insoluble and crystallize, thereby
cis-nonadienal (cucumber flavor). These findings increasing the solid fat content.
illustrate the complexity of the reversion flavor. Crystallization of a fat is a slow process,
Similar problems occur with other polyunsatu- whereas melting is instantaneous. When a fat
rated oils such as fish oil and rapeseed oil. crystallizes, the latent heat of crystallization is
liberated and a volume contraction takes place.
When a fat melts, the heat effect is negative and
volume expands (Fig. 2.48). These changes have
Table 2.32  Flavor descriptions used for crude, pro- been used for the measurement of some fat prop-
cessed, and reverted soybean oil erties such as melting point, solidification tem-
State Flavor perature, and solid fat content. Because fats
Crude Grassy, beany contain a range of glycerides of different melting
Freshly Sweet, pleasant, nutty points, there is no distinct melting point but rather
processed a melting range. Nevertheless, melting points of
Reverted Grassy, bany, buttery, melony,
fats are often determined. These are not real melt-
tallowy, painty, fishy
ing points, but some arbitrary temperature at
Source: From G. Hoffmann, Vegetable Oils, in Lipids and
Their Oxidation, H.W. Schultz et al., eds., 1962, AVI which virtually all of the fat has become liquid.
Publishing Co. The value of these melting points depends on the

Table 2.33  Volatile compounds isolated from reverted or oxidized soybean oil
Aldehydes
Saturated Δ2 Unsaturated Δ2,4 Unsaturated Δ3 Unsaturated
C1C2C3* C4C5C6* C4C5C6C7*C8 C6C7C7* C8*C9 C6C6

C7C8C9 C9*C10C11 C10C10* C12

Ketones and Dicarb. Alcohols and Esters Acids


Methyl-pentyl ketone 1-octen-3-ol Saturated
Di-n-propyl ketone Ethanol C1–C9 or C10
Malondialdehyde Ethyl formate
Ethyl acetate
*Main products
Stereo-isomers

Source: From G. Hoffmann, Vegetable Oils, in Lipids and Their Oxidation, H.W. Schultz et al., eds., 1962, AVI
Publishing Co.
96 2 Lipids

Fig. 2.48  Melting and solidification of fats

Table 2.34  Melting points of some fatty acids


Fatty Acid MP(°C)
Oleic (cis) 13
Elaidic (trans) 44
Stearic 70
Linoleic (cis-cis) –5
Linelaidic (trans-trans) 28
Butyric –8

Table 2.35  Melting points of some triglycerides


Triglycerides MP (°C)
Trisaturated S-S-S 72
P-P-P 65
S-P-P 62
Fig. 2.49  Physical properties of tallow and cocoa butter
Disaturated P-0-P 37 as influenced by solid fat profile. Adapted from”:
O-P-P 34 U. Bracco, Effect of Triglyceride Structure on Fat
S-0-P 35 Absorption, American Journal of Clinical Nutrition, Vol.
S-P-O 39 60, (Suppl.) p. 1008S, © 1994, American Society for
Clinical Nutrition
P-S-O 36
Diunsaturated O-O-P 19
O-O-S 23 solid fat content. Because the solid fat content is
Triunsaturated O-O-O 5 dependent on temperature, it is common to deter-
E-E-E 42
mine the solid fat profile, which is a graph repre-
L-L-L −10
senting the relationship between solid fat content
S stearic, P palmitic, O oleic, E elaidic, L linoleic
and temperature. Earlier methods for measuring
solid fat were based on dilatometry, a technique
measurement technique employed (Mertens and using the melting expansion of fats on heating.
deMan 1972). The melting point of a fat is basi- Modem methods employ pulsed nuclear mag-
cally determined by the melting points of its con- netic resonance. The dilatometer technique gives
stituent fatty acids. As shown in Table 2.34, chain an approximation of the true solid fat content and
length and unsaturation of fatty acids affect melt- is reported as solid fat index (SFI). The nuclear
ing point. In addition, the configuration around magnetic resonance results are true solid fat mea-
the double bond is important. The arrangement of surements and reported as solid fat content (SFC).
the fatty acids in different glyceride types also The relationship between solid fat content and
affects the melting point, as shown in Table 2.35. physical properties is demonstrated in Fig. 2.49
One of the most important properties of fats is the (Bracco 1994), where the difference in properties
Physical Properties 97

of cocoa butter and tallow is demonstrated. The Control of the crystalline microstructure in
steep solid fat curve of cocoa butter provides lipid systems provides control of the physical
coolness and flavor release, and its high solids properties of the food. For example, to obtain the
content at room temperature ensures hardness desired polymorphic form of chocolate temper-
and heat resistance. Tallow has a less steep solids ing prior to molding or enrobing is applied to
curve with solids beyond 37 °C, which gives rise control crystallization of the cocoa butter into a
to a waxy mouthfeel. large number of very small crystals. The cocoa
The rate at which a liquid fat is cooled is butter crystals in chocolate contribute to the
important in establishing solid fat content and desired appearance (shine or gloss), snap, and fla-
crystal size. In contrast to the glassy state form- vor release, melting in the mouth, and stability
ing systems described in Chap. 1, fats do not during shelf life. Control of fat crystallization is
form a glassy state. The rate of supercooling also important in butter, margarine, whipped
determines whether nucleation or crystal growth cream, ice cream, shortening, peanut butter, and
will predominate. At low supercooling, nucle- other foods.
ation is at a minimum and large crystals are Fats crystallization is often used to modify the
formed. At high supercooling, nucleation is high properties of the fat. For example, winterization
and small crystals result. Another result of high of vegetable oils removes fractions that crystal-
supercooling is the formation of solid solutions lize at higher temperatures assuring that oil
or mixed crystals. When fats are cooled quickly remains a clear liquid even when stored at low
to well below their melting point (e.g., 0 °C), the temperatures for extended time periods. Many
high melting glycerides crystallize together with fats, including palm oil, palm-kernel oil, milk fat,
some of the lower melting ones into mixed crys- and tallow, are fractionated by crystallization to
tals. When the fat is subsequently tempered at a produce different functional fats.
temperature close to the melting point, the mixed Generally about 98% of the content of fats are
crystals recrystallize into crystals of more uni- present as triglycerides (TAGs), the remainder
form composition. This means that a rapidly lipids like diacylglycerols (DAGs), monoacylg-
cooled fat contains more solid fat than the same lycerols (MAGs), free fatty acids (FFAs), phos-
fat that has been tempered after the same cooling pholipids, glycolipids, sterols, and other trace
treatment (Table 2.36). components. These minor lipids are partially
In many food products and processing opera- removed resulting in lower concentrations than in
tions, control of lipid crystallization is important unrefined fats. Although the TAGs form the main
to obtain the desired number of fat crystals, their crystalline phase, the minor components, or
size distribution, polymorphic form, and impurities, can influence how crystallization
­dispersion of the crystalline phase. Crystallization occurs and as a result crystallization may be dif-
of triacylglycerols (TAG) is the most important ferent in refined oil than in the unrefined starting
factor, however crystallization of other lipid com- material.
ponents (i.e., monoacylglycerols, diacylglycerols, It is important to understand the nature of the
phospholipids, etc.) can impact product quality. liquid phase prior to crystallization to understand
how crystals form. Lipids retain some degree of
Table 2.36  Solid fat content of a soft margarine fat with order in the liquid phase, with temperatures well
and without tempering at °C above the meltin. When melting fats, this liquid
Solid fat content ordering is termed a crystalline memory effect,
Temperature (°C) Tempered Not tempered where subsequent recooling leads to formation of
10 22.3 30.7 a different (usually more stable) phase than
20 11.7 16.0 would occur if the fat was heated to higher tem-
30 4.1 6.3 peratures to destroy the liquid memory (Larsson
35 2.8 1.9 1972; Hernqvist 1984).
98 2 Lipids

Nucleation, refers to the organization of mol-


ecules leading to the formation of the crystalline
phase from the liquid. The inherent ordering of
the liquid phase in lipids leads to crystal forma-
tion. Rapid cooling of liquid lipids results in the
formation of a diffuse crystalline phase (low-­
energy polymorph) because of the ordering struc-
ture in the liquid phase. This effect differs from
rapid cooling of other systems, most notably sug-
ars and starches which results in the formation of
a glassy state where molecules that are randomly
organized together with no long-term ordering.
Slower cooling from the liquid, the lipid mol-
ecules have time to organize into lamellae (1) and
eventually can form coherent, three-dimensional
crystals (shown schematically in Fig. 2.50). The
arrangement of the molecules into the crystalline
state depends on such factors as the cooling rate,
the temperature at which crystallization occurs,
the agitation rate, and the composition of the
lipid phase.
Polymorphism is the ability of a molecule to
take more than one crystalline form depending
on its arrangement within the crystal lattice. The
polymorphic forms in lipids are determined by
the differences in hydrocarbon chain packing
and variations in the angle of tilt of the hydro-
carbon chain. The crystallization behavior of
TAG, including crystallization rate, crystal size,
morphology, and total crystallinity, are affected
by polymorphism. The molecular structure of Fig. 2.50  Proposed mechanism (highly schematic) for
the TAG and several external factors like tem- nucleation of triacylglycerols (TAGs). Straight chains
perature, pressure, rate of crystallization, impu- indicate crystallized TAGs, whereas bent chains indicate
fluid TAGs (Hartel 2001)
rities, and shear rate influence polymorphism
(Sato 2001).
Polymorphism is described a s the ability of a TAGs are oriented in a chair or tuning fork
chemical compound to exist in differing crystal- configuration in the crystalline lattice. The TAG
line or liquid crystalline forms. Table 2.37 can take either a double or triple chain-length
describes the basic physical properties of the structure as seen in Fig. 2.51 The fatty acids of
three main polymorphic forms found in fats. The TAG pairs overlap in a double chain-length struc-
three forms are α, β’ and β. Polymorph α is the ture whereas in triple chain packing, the fatty
least stable is readily transformed to either β or β’ acids do not overlap. The height of these chair
forms. Polymorph β’ is a meta-stable form which structures and the distance between the mole-
is found in margarine and shortening because the cules in the chair structures are found by using
fat networks provide the desired physical and the X-ray spectra as the long and short spacings,
rheological properties. The β form is the most respectively.
stable, forming large plate like structures which The polymorphic forms of fats are often sim-
result in poor margarine or shortening qualities. ply classified into three categories, α, β′, and β, in
Physical Properties 99

Table 2.37  Physical properties of the three principal polymorphic forms found in fat
Form Stability Density Melting point Morphology Unit cell
α Least Lowest Lowest Amorphous Hexagonal
β' Metastable Intermediate Intermediate Rectangular Orthorhombic
β Most Highest Highest Needle-shaped Triclinic
Adopted from Sato and Ueno 2014

Fig. 2.51 Packing
arrangements of
triacylglycerol
molecules in the crystal
lattice

increasing order of stability. The α form is the subcell (O⊥); and β polymorphs are triclinic–par-
least stable polymorph with the lowest melting allel subcell (T//) (Larsson 1966;) as illustrated in
point and latent heat of fusion. The β form is the Fig. 2.52a.
most stable, with the highest melting point and Figure 2.52b illustrates the chain-length struc-
latent heat. Each polymorphic form has distinct ture, applying the repetitive sequence of the acyl
short spacings (the distances between parallel chains involved in a unit cell lamella along the
acyl groups on the TAG) that are used to distin- long-chain axis (Larsson 1972). A double chain-­
guish the polymorphic forms based on their length structure is formed when the chemical
X-ray diffraction patterns, as summarized in properties of the three acid moieties are the same
Table 2.37 Based on the unique configuration of or very similar as in tripalmitin or a homologus
the molecules within the crystal lattice, each fat with high content of long chain fatty acids or
polymorph has a different crystallographic unit trans fatty acids. Conversely, when the chemical
cell, also shown in Table 2.37. properties of one or two fatty acid side chains are
Applying measuring X-ray diffraction (XRD) different, a triple chain-length (TCL) structure is
short spacing patterns of poly-crystalline samples formed because of chain sorting. TAGs with three
the subcellular structures of the three polymorphs saturated fatty acids crystallize in double chain-­
can be determined. The three fat polymorphs, α, length packing, whereas triple chain-length pack-
β’ and β of fats, exhibit differing subcell struc- ing is obtained if the TAG contains fatty acids
tures: α polymorphs are hexagonal sub-cell (H); with different chain lengths and unsaturation).
β’ polymorphs are orthorhombic–perpendicular Lutton (1950) stated that if the fatty acids of a
100 2 Lipids

Fig. 2.52  Proposed structure of the form and form of tri- methyl end-group regions are somewhat disordered, as in
glycerides with double chain length longitudinal organi- liquid crystals. (Lopez et al. 2006) (Permission requested)
zations as seen in the ca projection. For the form, the

TAG differ in length by more than four carbons, length structures can be determined solely by
it forms a triple chain-length structure. Triple measuring the XRD long spacing patterns of the
chain-length packing is also observed in TAG poly-crystalline samples. In food fats, transforma-
containing a cis-unsaturated fatty acid because tion from polymorph β’ to polymorph β can result
this causes a kink in the structure, as seen in in deterioration of the end product resulting from
Fig. 2.52. Because the fatty acids are not linear, changes in the crystal morphology and network,
cis-unsaturated fatty acids do not mix in one as shown in Table 2.37. The β-type polymorph is
layer with the more linear saturated fatty acids, found in confectionery fats made of cocoa butter
and triple chain-length crystals are formed. (Timms 2003). There are two β-type crystals: a
Trans-unsaturated fatty acids incorporate into a meta-stable β2 form is more useful than the more
crystal structure in the same way as the saturated stable β1 form (Sato and Koyano 2001; Van
fatty acids. This is why oils are hydrogenated to Mechelen et al. 2006a, b; Sato and Ueno 2014).
produce solid components in the fat for marga- Monotropic polymorphism occurs in lipids
rines and shortenings. The chain-length structure where unstable forms are the first to crystallize in
influences the mixing-phase behavior of different a subcooled fat, because of their lower energy
types of TAGs in solid phases (Sato 2001). The state. Subsequently these unstable forms trans-
triple chain-length structure has greater long form into more stable forms. The changes con-
spacings than does the double chain-length struc- tinue to occur until the most stable polymorph for
ture. (Metin and Hartel 1998) a given lipid is reached. The transformations
The relevance of the chain-length structure is occur at temperatures slightly above the melting
revealed in the mixing phase behavior of the dif- temperature of the less stable form. The increase
ferent types of the TAGs in the solid phase: when in temperature first causes the melting of the
the fatty acids with different chain lengths or unstable forms followed by solidification in a
degrees of unsaturation are mixed with the TCL more stable form. Transformation to a more sta-
fats, phase separation readily occurs. The chain ble form can also take place without melting. The
Physical Properties 101

Fig. 2.53  Illustration of fat crystallization in milk fat. from the melt. β-crystals (triclinic subcell) are formed via
α-crystals (hexagonal subcell structure), forms directly recrystallization from β’. Representation of the Packing of
from the melt, while β’-crystals (orthorhombic subcell) Triacylglycerols in the Three Main Polymorphic Forms.
forms either via recrystallization of α to β∋ or directly (Rønholt et al. 2014)

Fig. 2.54  Cross-sectional structures of long-chain compounds (Lutton 1972)

polymorphic state becomes more stable and typically crystallize at rates in order of their sta-
higher melting as the molecules become more bility (α < β′ < β). Thus, the least-stable polymor-
tightly arranged in the crystal lattice. Figure 2.53 phic form typically crystallizes first in a strongly
represents the differences in packing of the three subcooled molten fat because of the lower sur-
polymorphic forms. It is assumed that the chair face energy (Bailey 1950).
structure is maintained during polymorphic Assignment of the polymorphic form of a fat
transformations (Larsson 1966). The layer can be done unequivocally only by X-ray diffrac-
arrangement of the α polymorph does not change tion, sometimes supported by differential scan-
when it is transformed to the β′ polymorph, ning calorimetry. The characteristics of the three
although its lateral chain packing and angle of tilt polymorphs are as follows:
changes during polymorphic transformation.
The hydrocarbon chain packing of the β poly- • α form—one strong, short spacing in the X-ray
morph is denser than that of the α polymorph. diffraction pattem at 0.42 nm. The chains are
The denser chain packing in the β polymorph arranged in a hexagonal crystal structure (H) with
gives increased stability compared with the α no order of the zigzag chain planes. The chains
polymorph. The different polymorphic forms have no angle of tilt (Lutton 1972) (Fig. 2.54).
102 2 Lipids

• β′ form—two strong, short spacings in the Table 2.38  Polymorphic forms of cocoa butter melting
temperature (°C)
X-ray diffraction pattem at 0.38 and 0.42 nm.
The chains are arranged in an orthorhombic Form Wille and Lutton (13) Davis and Dimick (13)
crystal structure. The zigzag planes are arranged I γ 17.3 13.1
in a perpendicular crystal structure. The chains II α 23.3 17.7
have an angle of tilt between 50 and 70°. III β'2 25.5 22.4
• β form—one strong, short spacing at 0.46 nm IV β'1 27.5 26.4
and two spacings at 0.37 and 0.39 nm. This V β2 33.8 30.7
form is the most densely packed and is in a VI β 36.3 33.8
triclinic structure. The zigzag planes are in a
parallel arrangement, and the chains have an given the subscript 1, and other polymorphs
angle of tilt of 50–70°. within that form are ordered in decreasing stabil-
ity or melting temperature. For example, cocoa
The rate of polymorphic transformation butter has two β′ forms, with the β′1 form having
depends on the length of the fatty acid chain. The the highest melting point (most stable) (Mertin
rate of change is longest for TAGs with short-­ and Hartel 1998).
chain fatty acids such as butterfat (Bailey 1950). Chocolate is first melted to eliminate existing
Natural fats generally contain a large number of crystalline structures. The molten chocolate is
different TAGs. This heterogeneity results in then cooled and allowed for seeds to form or fre-
very slow transition form unstable to stable forms quently type V seeds are added to initiate crystal-
is often very slow. As mentioned previously, the line growth. The chocolate is then warmed
α- form is generally formed first in a rapidly slightly and maintained at that temperature to
cooled liquid fat, but it is usually very unstable allow conversion of most of the structure to Type
and rapidly transforms to the β′ form. The β′ V crystals. The chocolate is then cooled to room
form may remain for hours to days. In many fats temperature and allowed to solidify. This product
the β′ However, in many natural fats, the β′ poly- will then have appropriate shine, snap in biting
morph can exist for long periods of time because and cooling in the mouth. The temperature choc-
the complex mixtures result in the development olate is also resistant to bloom. Bloom is the
of a solid solution (Walstra et al. 1987). In some migration of fat to the surface of eh product leav-
mixed-acid TAGs, no β polymorph may form and ing a unsightly white residue on the surface.
β′ is the most stable. In some systems both β The polymorphs of chocolate exhibit differing
forms may be present (Sato 2001). characteristics as shown in Table 2.39 (Beckett
More than one subtype within the main poly- 2007).
morphic grouping has been identified in some
fats. For example, six different polymorphic Melting
Form temperature Typical properties
forms have been identified in cocoa butter,
I γ 17 °C Produced when liquid
although there is still some debate whether they chocolate is cooled rapidly.
are all truly unique polymorphs (Table 2.38). Unstable form results in soft
Two β′ and two β forms have been identified for crumbly product that will
bloom.
cocoa butter (Loisel et al. 1998). These poly-
II α 21 °C After storing rapidly cooled
morphs have slightly different melting points, but
form chocolate morphs to this
they have X-ray spectra that fit within the defini- form. Soft, crumbly and tends
tion of that polymorph (Metin and Hartel 1998; to boom
van Malssen et al. 1996a, b, c). III β'2 26 °C Form II crystals become form
Different nomenclatures for cocoa butter have III after storage at low
temperatures
been used to describe the polymorphic forms as
seen in Table 2.39. In the Greek nomenclature,
where polymorphs are given a Greek letter, the
most stable form within a polymorph type is
Physical Properties 103

Melting ides with melting points considerably higher than


Form temperature Typical properties those of the whole fat. These triglycerides are
IV β'1 28 °C Produced by cooling chocolate referred to as high melting glycerides (HMGs)
to room temperature with no
and can be obtained by fractionation from ace-
further tempering. Product will
be firm but not snap, Melts at tone (D’Souza et al. 1991). When the HMGs con-
too low a temperature sist of a high level of the same fatty acid such as
V β2 34 °C Produced by properly palmitic acid or stearic acid or a mixture of ­stearic
tempering chocolate. Very and any trans isomeric form 18:1, the fat crystals
stable, high glows, good snap,
resistant to bloom
quickly transform from β′ to β form. This hap-
VI β 36 °C Produced when well-­tempered pens when canola oil or sunflower oil is hydroge-
chocolate is stored at room nated and used as a sole hardstock in margarines
temperature for many months. and shortenings (Hernqvist 1984, 1988). The
Product will be hard and fatty acid composition of canola oil (see
difficult to melt
Table  2.13) consists mainly of 18-carbon fatty
The tendency of fats to remain stable in the β′ acids such as 18:0, 18:1, 18:2, and 18:3; it con-
form for a long time or to quickly convert to the β tains only 4% palmitic acid. When canola oil is
form is more difficult to predict than that of pure hydrogenated, the solids that are produced con-
triglycerides. In margarines and shortenings it is sist of triglycerides of 18:0 or 18:1 trans fatty
desirable that the fat crystals are in the β′ form. In acid, which results in a high level of 54 (3 × 18)
margarines this results in a smooth texture, shiny carbon triglycerides. These triglycerides are lia-
surface, and good meltdown properties. The β′ ble to undergo polymorphic transition to the β
crystals in shortenings give good aeration in form. Triglycerides consisting of only palmitic
cakes and creams. The solids or fat crystals in acid or tripalmitin (48-carbon triglycerides) are
margarines and shortenings consist of triglycer- equally likely to convert to the β form. This can

Table 2.39  Characteristics of various crystalline forms of cocoa butter


Melting
Polymorph of cocoa butter point oF Properties
Form I 61–67 Produced by rapid cooling such as placing liquid cocoa butter n freezer
Beta prime 2 Most unstable form
γ Soft, crumbly, blooms and melts at low temperature
Form II 70–73 Produced by rapid cooling placing chocolate in refrigerator of freezer
Alpha Alpha crystals form after short time storage at freezing temperatures
α Melts at low temperature, contains many polymorps, soft crumbly and
blooms heavily
Form III 77–78 Produced by too rapid cooling in freezer
Mixture of Alpha and Form after storage at cool temperatures above freezing
Beta-prime I
Form IV 81–84 Cooling at room temperature with no further tempering
Beta-prime Crystals are firm but do not have snap
β’ Product melts below body temperature and blooms
Form V 92–95 Produced by tempering. After tempering cool slowly at room
temperature
Beta 2 Highly desirable polymorph
β Very stable, smooth shiny surface, good snap, melts readily when eaten
Form VI 97 Formed on long term storage of tempered chocolate at room
temperature
Beta 1 Very hard, does not melt easily
β Can be reversed by retempering
104 2 Lipids

Table 2.40  Triacylglycerol (TAG) composition (%) by Table 2.41  Major triacylglycerols (TAGs) of palm oil
carbon number of common vegetable oils and their melting points
TAG (Carbon Number) Melting Point (°C)
Oil 48 50 52 54 56 TAG Carbon Number % β′ β
Canola – 1.1 13.0 76.8 5.6 a
PPP 48 6 56.7 66.2
Soybean – 3.3 27.6 66.7 1.7 a
PPS 50 1 59.9 62.9
Sunflower – 2.8 20.2 75.1 0.7 a
PSP 50 0.5 68.8 –
Sunflower high olein – 2.0 15.0 80.6 1.0 POP 50 26 30.5 35.3
Corn – 4.6 30.4 64.2 0.8 a
PPO 50 6 35.4 40.4
Olive – 4.7 27.7 66.7 0.9 PLP 50 7 18.6 NA
Peanut – 5.5 30.9 54.2 5.3 POO 52 19 14.2 19.2
Cottonseed 0.9 13.6 43.5 40.5 1.3 POS 52 3 33.2 38.2
Palm 8.0 42.5 40.5 9.0 – PLO 52 4 NA NA
OOO 54 3 −11.8 5.1
Likely to be in the palm stearin fraction
a

NA Not available
happen when palm stearin, which is obtained
from fractionated palm oil, is used as the sole
hardstock for a margarine or shortening (deMan PEP, which provides a high level of desirable β′
and deMan 1995). Soybean oil contains 11% pal- triglycerides. In addition, the solid 52-carbon tri-
mitic acid (see Table 2.13). When soybean oil is glycerides that are produced are also β′ tending.
hydrogenated, the solids are more diverse in fatty The β′ stability of hydrogenated canola oil can be
acid chain length than those of hydrogenated greatly enhanced by incorporation of hydroge-
canola or sunflower oil of similar consistency. nated palm oil at a level of 10%. For margarines
The triglyceride composition (carbon number) of where it is desirable to have low levels of trans
common vegetable oils is displayed in Table 2.40. fatty acids, palm mid-­fraction that contains very
Canola and sunflower oil contain high levels of high levels of POP and PLP (up to 60%) can be
54-carbon triacylglycerol (TAG), while palm oil hydrogenated and incorporated at lower levels to
has the lowest level. When these oils are hydro- secure a desirable level of PSP and PEP.
genated, the solids in the early stages of hydroge- Diversification of the fatty acid composition
nation contain less 54-carbon and more 50-carbon of the solids can also be achieved by including
triglycerides than the original oils, because the lauric oils such as palm kernel or coconut oil,
50-carbon triglycerides already contain two satu- preferably in the fully hydrogenated form (Nor
rated fatty acids and are likely to be included in Aini et al. 1996, 1997). A β′ stable hardstock can,
the solids at the early stage. The 50-carbon tri- upon dilution with liquid oil, crystallize in the β
glycerides of the solids consist mainly of PSP or form, as is done in soft margarines and to a cer-
PEP (E stands for elaidic acid or any form of iso- tain extent in stick margarines. Elevated storage
meric 18:1 trans). These triglycerides are very temperatures can also induce a polymorphic tran-
stable in the β′ form, as mentioned earlier, and sition. β crystals are initially small but upon stor-
preferably should be part of the solids. The solid age can grow into large, needle-like agglomerates
52-carbon triglycerides are mainly made up of that make the product grainy, make the texture
PSS or PEE. Palmitic acid in vegetable oils is brittle, and give the surface a dull appearance.
mainly located in the 1 or the 3 position. Palm oil Shortenings always contain about 10–12%
has a high level of 50- and 52-carbon triglycer- hydrogenated palm oil—usually in the fully
ides and a low level of 54-­ carbon TAG hydrogenated form, which extends the plastic
(Table 2.41). The 50-carbon TAG consist mainly range so that the product is still plastic at higher
of POP (26%) and PLP (8%). When palm oil is temperatures. The melting point of shortenings is
hydrogenated, POP and PLP change to PSP or in the low range of 40–45 °C.
Fractionation 105

Fractionation Table 2.42  Composition and slip melting point (SMP)


of palm oil and its fractions

Fats can be separated into fractions with different Fatty Acid (Wt %)
physical characteristics by fractional crystalliza- Product 16:0 18:0 18:1 18:2 SMP (°C)
tion from a solvent or by fractionation from the Palm oil 44.1 4.4 39.0 10.6 36.7
melt. The former process gives sharply defined Palm olein 40.9 4.2 41.5 11.6 21.5
fractions but is only used for production of high-­ Palm stearin 56.8 4.9 29.0 7.2 51.4
value fats; the latter process is much simpler and Source: Reprinted with permission from W.L. Siew, et al.,
Identity Characteristics of Malaysian Palm Oil Products:
more cost-effective. Fractionation from the melt Fatty Acid and Triglyceride Composition and Solid Fat
or dry fractionation is carried out on a very large Content, E.L.A.E.I.S., Vol. 5, No. 1, p. 40, © 1993, Palm
scale with palm oil and other fats including beef Oil Research Institute of Malaysia
tallow, lard, and milk fat.
There are several reasons for employing frac- Table 2.43  Composition of palm oil and palm olein
tional crystallization (Hamm 1995): Triglyceride Palm oil Palm olein
SSS (mainly PPP) 8 0.5
• To remove small quantities of high melting SUS (mainly POP) 50 48
components that might result in cloudiness of SUU (mainly POO) 37 44
an oil. This can be either a triacylglycerol UUU 5 7
fraction or non-triacylglycerol compound. Iodine value 51–53 57–58
The former happens when soybean oil is Source: Reprinted with permission from W.L. Siew, et al.,
lightly hydrogenated to convert it to a more Identity Characteristics of Malaysian Palm Oil Products:
stable oil. The resulting solid triglycerides Fatty Acid and Triglyceride Composition and Solid Fat
Content, E.L.A.E.I.S., Vol. 4, No. 1, p. 40, ©1993, Palm
have to be removed to yield a clear oil. The
Oil Research Institute of Malaysia
latter occurs when waxes crystallize from oils
such as sunflower oil. This type of fraction-
ation is known as winterization. The major glycerides present in palm oil are listed
• To change a fat or oil into two or more frac- in Table 2.42, together with their melting points.
tions with different melting characteristics. In When a liquid fat is cooled, a crystalline solid is
simple dry fractionation, a hard fraction (stea- formed; its composition and yield are determined
rin) and a liquid fraction (olein) are obtained. by the final temperature of the oil. From Table 2.43
This is by far the most common application of it is obvious that the glycerides most likely to crys-
fractionation. tallize are PPP, PPS, and PSP. It is theoretically
• To produce well-defined fractions with special impossible to separate these glycerides sharply
physical properties that can be used as spe- from the rest of the glycerides. There are two rea-
cialty fats or confectionery fats. This is often sons: (1) the formation of solid solutions and (2)
done by solvent fractionation. the problem of entrainment.
The formation of solid solutions can be
The process of fractionation involves the con- explained by the phase diagram of Fig. 2.55
trolled and limited crystallization of a melted fat. (Timms 1984, 1995). This relates to two triglycer-
By careful management of the rate of cooling and ides A and B, which form a solid solution. In
the intensity of agitation, it is possible to produce other words, they crystallize together into mixed
a slurry of relatively large crystals that can be sep- crystals. At temperature T1 the solid phase has the
arated from the remaining liquid oil by filtration. composition indicated by c and the liquid phase
The major application of fractionation is with composition is a. The fraction of the solid phase is
palm oil; many millions of tons of palm oil are equal to ab/ac. As the crystallization temperature
fractionated into palm stearin and palm olein every differs from T1, the composition of the solid and
year. Palm oil is unusual among vegetable oils. It liquid phases will also differ. Solid solutions are
has a high level of palmitic acid; contains a sub- formed when two or more solutes have melting
stantial amount of a trisaturated simple glyceride, points that are not very different. If the solute and
tripalmitin; and has a high level of SUS glycerides. solvent molecules have widely differing melting
106 2 Lipids

Fig. 2.55 Typical 50
temperature profiles to Melt to destroy
temper chocolate 45

40 Warm and maintain


Temperature
35
Until Type IV are
Cool to room
30 temperature
Seed and cool Allow to set
25
Allow type V
Crystals to grow
20
Start Melt Seed Destrov IV Hold Cool
crystals
Dark Chocolate Milk Chocolate White Chocolate

points, no solid solutions will be formed and the


solubility is dependent only on the solute. This is
known as ideal solubility. A plot of the solubility
of tripalmitin in 2-oleo-dipalmitin is presented in
Fig. 2.56. At all temperatures the actual solubility
is higher than the ideal solubility, and the result-
ing solid phase that crystallizes is composed not
only of tripalmitin but a solid solution of tripalmi-
tin and 2-oleo-dipalmitin (Fig. 2.57).
Entrainment results from mechanical entrap-
ment of liquid phase in the crystal cake obtained
by filtration. The composition of the filter cake is
highly dependent on the degree of pressure applied
in the filtration process. The composition of palm
oil and its main fractions are given in Table 2.42
(Siew et al. 1992, 1993). The triglycerides in palm
oil and palm olein are listed in Table 2.43
Depending on the process used, the stearin can be
obtained in yields of 20–40%, with iodine values
ranging from 29 to 47. This is important because Fig. 2.56  Phase diagram of mixture of triglycerides A
the olein is considered the more valuable com- and B, showing a continuous solid solution. Source:
modity. Palm olein is widely used as a liquid oil in Reprinted from R.E. Timms, Crystallization of Fats, in
Developments in Oils and Fats, R.J. Hamilton, ed., p. 206,
tropical climates. However, in moderate climates, © 1995, Aspen Publishers, Inc.
it crystallizes at lower temperatures, just as olive
oil and peanut oil do. Palm stearin is finding
increasing application as a nonhydrogenated hard Milk fat fractionation has been described by
fat and as a component in interesterification for Deffense (1993). By combining multistep fraction-
the production of no-trans margarines. ation and blending, it is possible to produce modi-
The fractionation of palm oil can be carried fied milk fats with improved functional properties.
out in a number of ways to yield a variety of
products as shown in Fig. 2.58. In this multistage
process, a palm midfraction is obtained that can Emulsions and Emulsifiers
be further fractionated to yield a cocoa butter
equivalent (Kellens et al. 1994). Palm oil can be A satisfactory definition of emulsions has been
double fractionated to yield a so-called super provided by Becher (1965), who states that an
olein with iodine value of 65. emulsion is a heterogeneous system, consisting
Fig. 2.57 Solubility
diagram of tripalmitin in
2-oleo-dipalmitin.
Source: Reprinted from
R.E. Timms,
Crystallization of Fats,
in Developments in Oils
and Fats, R.J. Hamilton,
ed., p. 209, © 1995,
Aspen Publishers, Inc.

Fig. 2.58  Products obtained by multistage fractionation of palm oil. Source: Reprinted with permission from
Developments in Fat Fractionation Technology, Paper No. 0042, Kellens et al. 1994, p. 29, © Mark Kellens, PhD
108 2 Lipids

of one immiscible liquid intimately dispersed in used (Friberg 1976). The HLB scale goes from 0
another one, in the form of droplets with a to 20, in theory at least, since at each end of the
­diameter generally over 0.1 μm. These systems scale the compounds would have little emulsify-
have a minimal stability, which can be enhanced ing activity. The HLB values of some commer-
by surface-­ active agents and some other sub- cial nonionic emulsifiers are given in Table 2.44
stances. In foods, emulsions usually contain the (Griffin 1965).
two phases, oil and water. If water is the continu- Foods contain many natural emulsifiers, of
ous phase and oil the disperse phase, the emul- which phospholipids are the most common. Crude
sion is of the oil in water (O/W) type. In the phospholipid mixtures obtained by degumming of
reverse case, the emulsion is of the water in oil soya oil are utilized extensively as food emulsifiers
(W/O) type. A third material or combination of and are known as soya-lecithin. This product con-
several materials is required to confer stability tains a variety of phospholipids, not just lecithin.
upon the emulsion. These are surface-active Emulsifiers can be tailor-made to serve in
agents called emulsifiers. The action of emulsifi- many food emulsion systems. Probably the most
ers can be enhanced by the presence of stabiliz- widely used is the group of monoglycerides
ers. Emulsifiers are surface-­ active compounds obtained by glycerolysis of fats. Reaction of an
that have the ability to reduce the interfacial ten- excess of glycerol with a fat under vacuum at high
sion between air-liquid and liquid-liquid inter- temperature and in the presence of a catalyst
faces. This ability is the result of an emulsifier’s results in the formation of so-called technical
molecular structure: the molecules contain two monoglycerides. These are mixtures of mono-,
distinct sections, one having polar or hydrophilic di-, and triglycerides. Only the 1-monoglycerides
character, the other having nonpolar or hydro- are active as emulsifiers. By molecular distillation
phobic properties. The extent of the lowering of under high vacuum, a product can be obtained in
interfacial tension by surface-active agents is which the 1-monoglyceride content exceeds 90%.
shown in Fig. 2.59. Most surface-active agents The ability of emulsifier molecules to orient
reduce the surface tension from about 50 dynes/ themselves at interfaces is exemplified in the
cm to less than 10 dynes/cm when used in con- phase behavior of monoglycerides. The emulsi-
centrations below 0.2%.
The relative size of the hydrophilic and hydro-
phobic sections of an emulsifier molecule mostly
determines its behavior in emulsification. To make
the selection of the proper emulsifier for a given
application, the so-called HLB (hydrophile-­
lipophile balance) system was developed. It is a
numerical expression for the relative simultaneous
attraction of an emulsifier for water and for oil.
The HLB of an emulsifier is an indication of how
it will behave but not how efficient it is. Emulsifiers
with low HLB tend to form W/O emulsions, those
with intermediate HLB form OAV emulsions, and
those with high HLB are solubilizing agents. HLB
values can be calculated from the saponification
number of the ester (S) and the acid value of the
fatty acid radical (Av) as follows:
 S
HLB = 20  1 − 
 Av 
Fig. 2.59  Lowering of the surface tension of water by
most surface-active compounds. Source: From R Becher,
In many cases the HLB value is determined Emulsions—Theory and Practice, Becher 1965, Van
experimentally and various methods have been Nostrand Reinhold Publishing Co.
Emulsions and Emulsifiers 109

Table 2.44  HLB values of some commercial nonionic emulsifiers


Trade name Chemical designation HLB
Span 85 Sorbitan trioleate 1.8
Span 65 Sorbitan tristearate 2.1
Atmos 150 Mono- and diglycerides from the glycerolysis of edible fats 3.2
Atmul 500 Mono- and diglycerides from the glycerolysis of edible fats 3.5
Atmul 84 Glycerol monostearate 3.8
Span 80 Sorbitan monooleate 4.3
Span 60 Sorbitan monostearate 4.7
Span 40 Sorbitan monopalmitate 6.7
Span 20 Sorbitan monolaurate 8.6
Tween 61 Polyoxyethylene sorbitan monostearate 9.6
Tween 81 Polyoxyethylene sorbitan monooleate 10.0
Tween 85 Polyoxyethylene sorbitan trioleate 11.0
Arlacel 165 Glycerol monostearate (acid stable, self-emulsifying) 11.0
Myrj 45 Polyoxyethylene monostearate 11.1
Atlas G-2127 Polyoxyethylene monolaurate 12.8
Myrj 49 Polyoxyethylene monostearate 15.0
Myrj 51 Polyoxyethylene monostearate 16.0
Source: From W.C. Griffin, Emulsions, in Kirk-Othmer Encyclopedia of Chemical Technology, 2nd ed., Vol. 8, pp. 117–
154, 1965, John Wiley & Sons

fier molecules in aqueous systems show meso- the food system (Larsson and Dejmek 1990). For
morphism (formation of liquid crystalline example, consider the amylose-­complexing effect
phases). In such systems, several mesophases of emulsifiers (Krog 1971). This effect is useful
may exist, as Krog and Larsson (1968) have for improving the shelf life of bread (antifirming
shown. When 1-monopalmitin crystals in effect) and modifying the physical characteristics
20–30% of water are heated to 60 °C, a meso- of potato products, pasta, and similar foods.
phase, called the neat phase, is formed The amount of emulsifier required to provide
(Fig. 2.60a). This structure consists of bimolecu- a monomolecular layer for an emulsion of any
lar lipid layers separated by water, with the chains particle size can be calculated from the relation-
in a disordered state. If this phase is cooled, a gel ship A/V = 6/D, where A, V, and D are area, vol-
is formed (Fig. 2.60b). The structure is lamellar ume, and diameter, respectively, of the spherical
with the hydrocarbon chains extended and tilted particles. The total area for 1 mL of oil dispersed
in a 54° angle toward the water layers. When the as uniform spheres thus calculated is given in
neat phase is heated, a stiff cubic phase is formed, Table 2.45. If sodium stearate is used as the emul-
called viscous isotropic (Fig. 2.60c). It consists sifier, the amount required can be calculated,
of small water spheres arranged in a face-­centered since this molecule has a surface area of about
lattice with the polar groups pointed toward the 0.2 nm2.
water. On further heating, an isotropic fluid is
obtained (Fig. 2.60d), in which polar groups and W=
( )
Total area nm 2 × 306
water form disk-like arrangements. With excess 0.2 × 6 × 10 23
water, a dispersion is formed (Fig. 2.60e), which
consists of concentric bimolecular shells of where
monoglyceride molecules alternating with water. W = weight of emulsifier
Emulsions are stabilized by a variety of com- 306 = molecular weight of sodium stearate
pounds, mostly macromolecules such as proteins 6 × 1023 = Avogadro’s number
and starches. The amount of emulsifier required increases
Emulsifiers have many additional functions in sharply with decreasing particle size of the emul-
foods. They form complexes with food compo- sion, as shown in Table 2.46 This constitutes a
nents, resulting in modified physical properties of practical limitation to the lower limit of particle
110 2 Lipids

Fig. 2.60  Structure of the mesophases of monoglyceride-­ Properties of Aqueous Systems of Industrial Distilled
water systems. (a) Neat phase, (b) gel, (c) viscous isotro- Monoglycerides, Chem. Phys. Lipids, Vol. 2, pp. 129–
pic, (d) fluid isotropic, and (e) dispersion. Source: From 143, 1968
N. Krog and K. Larsson, Phase Behavior and Rheological

Table 2.45  Total surface area of 1 mL of oil dispersed as size that can be obtained in emulsification. It also
uniform spheres in an emulsion indicates that a substance that consists of mole-
Diameter Total surface area cules with a large ratio of surface area to molecu-
μm cm m2 cm2 nm2 lar weight and that yields a strong film should be
10 10−3 0.6 6 × 103 6 × 1017 an efficient emulsifier.
1 10−4 6 6 × 104 6 × 1018
0.1 10−5 60 6 × 105 6 × 1019
0.01 10−6 600 6 × 106 6 × 1020 Fat Replacers

Fat replacers are substances that are meant to


replace the calories provided by fat (9 kcal/g)
Table 2.46 Amount of sodium stearate emulsifier
either partly or completely. There are three basic
required to provide a monomolecular layer in emulsions groupings of fat replacers; 1- protein based. 2- car-
of different particle size bohydrate based and 3- fat based. The fat-like sub-
Sodium stearate stances that are either not absorbed, partially
Particle diameter micrometer (as % of oil) absorbed or intrinsically lower in calories. The
10 0.15 proteinaceous or carbohydrate compounds that
1 1.5 mimic the gustatory qualities of fats. The latter will
0.1 15 be dealt with in Chaps. 3 and 4. Table 2.47 sum-
0.01 150 marizes the fat substitutes used in food products.
Table 2.47  Fat replacers
Product Technology Application
Protein-based fat replacers
Fat Replacers

Microparticulated Protein Simplesse® Reduced-calorie (1–2 calorie/gram) ingredient made from whey Ice cream, butter, sour cream, cheese, yogurt), salad dressing,
protein or milk and egg protein. Digested as a protein margarine- and mayonnaise-type products, as well as baked
goods, coffee creamer, soups and sauces
Modified Whey Protein Concentrate Controlled thermal denaturation results in a functional protein Cheese, yogurt, sour cream, ice cream), baked goods,
Dairy-Lo R® with fat-like properties frostings, as well as salad dressing and mayonnaise-­type
products
Other (K-Blazer®, ULTRA-­BAKE™, One example a fat substitute based on egg white and milk Some blends of protein and carbohydrate can be used in
ULTRA-FREEZETM, Lita®) proteins. Similar to microparticulated protein but made by a frozen desserts and baked goods
different process. Another example is a reduced-calorie fat
replacer derived from a corn protein
Carbohydrate-based fat replacers
Cellulose (Avicel® cellulose gel, Non-caloric purified form of cellulose ground to microparticles Dairy-type products, sauces, frozen desserts and salad
MethocelTM, Solka-Floc®) which, when dispersed, form a network of particles with dressings
mouthfeel and flow properties similar to fat
Dextrins (Amylum, N-Oil®) Four calorie/gram fat replacers replace all or some of the fat in a Salad dressings, puddings, spreads, dairy-type products and
variety of products. frozen desserts
Fiber (Opta™, Oat Fiber, Snowite, Fiber provides structural integrity, volume, moisture holding Baked goods, meats, spreads and extruded products
Ultracel™, Z-Trim) capacity, adhesiveness and shelf stability in reduced-fat foods
Gums (KELCOGEL®, KELTROL®, Hydrophilic colloids: guar gum, gum arabic, locust bean gum, Reduced-calorie, fat-free salad dressings, formulated foods,
SlendidTM) xanthan gum, carrageenan and pectin. Non-caloric; provide including desserts and processed meats
thickening, sometimes gelling effect; promote creamy texture
Inulin (Raftiline®, Fruitafit®, Fibruline Reduced-calorie (1 to 1.2 calorie/gram) fat and sugar replacer, Yogurt, cheese, frozen desserts, baked goods, icings, fillings,
fiber and bulking agent extracted from chicory root whipped cream, dairy products, fiber supplements and
processed meats
Maltodextrins (CrystaLean®, Lorelite, Four calorie/gram gel or powder from corn, potato, wheat and Baked goods, dairy products, salad dressings, spreads, sauces,
Lycadex®, MALTRIN®, Paselli®D- tapioca. Fat replacer, texture modifier or bulking agent frostings, fillings, processed meat, frozen desserts, extruded
LITE, Paselli®EXCEL, Paselli®SA2, products and beverages
STAR-DRI®)
Nu-Trim Extraction process removes coarse fiber components from oat or Baked goods, dairy products, salad dressings, spreads, sauces,
Beta-glucan from oat or barley barley frostings, fillings, processed meat, frozen desserts, extruded
products and beverages
(continued)
111
Table 2.47 (continued)
112

Product Technology Application


Oatrim [Hydrolyzed oat flour] (Beta- A water-soluble from enzyme treated oat flour containing Baked goods, fillings and frostings, frozen desserts, dairy
TrimTM, TrimChoice beta-glucan soluble fiber, used as a fat replacer, bodying and beverages, cheese, salad dressings, processed meats and
texturizing ingredient. 1–4 calories/gram) confections
Polydextrose (Litesse®, Sta-LiteTM One calorie/gram) fat replacer and bulking agent. Water-soluble Baked goods, chewing gums, confections, salad dressings,
polymer of dextrose containing minor amounts of sorbitol and frozen dairy desserts, gelatins and puddings
citric acid.
Starch and Modified Food Starch 1–4 calories/gram as used, fat replacers, bodying agents, texture Processed meats, salad dressings, baked goods, fillings and
(Amalean®I & II, FairnexTMVA15, & modifiers. Can be derived from potato, corn, oat, rice, wheat or frostings, sauces, condiments, frozen desserts and dairy
VA20, Instant StellarTM, N-Lite, tapioca starches. Used with emulsifiers, proteins, gums and products
OptaGrade®#, PerfectamylTMAC, AX-1, other modified food starches
& AX-2, PURE-GEL®, STA-SLIMTM)
Z-Trim Zero calorie fat replacer from insoluble fiber from oat, soybean, Baked goods, burgers, hot dogs, cheese, ice cream and yogurt
pea and rice hulls or corn or wheat bran. It is heat stable
Fat-based fat replacers
Emulsifiers (Dur-Lo®, ECT-25) Vegetable oil mono- and diglyceride emulsifiers used with water Cake mixes, cookies, icings, and numerous vegetable dairy
Same caloric value as fat (9 calories/gram) but less is required. products
Sucrose fatty acid esters also can be used for emulsification.
Emulsion systems using soybean oil or milk fat can replace fat
on a one-to-one basis
Salatrim Short and long-chain acid triglyceride molecules. A five Baked goods, confections, dairy and other applications
BenefatT calorie-per-gram family of fats
Lipid (Fat/Oil) Analogs Reduced-calorie fat replacer. Can partially or fully replace fats Formulated products, baking and frying
 –  Esterified Propoxylated Glycerol and oils in all typical consumer and commercial applications
 – Olestra (Olean®) Calorie-free ingredient made from sucrose and edible fats and Fried products, Chips, dairy, confections and other food
oils. Not metabolized and unabsorbed by the body. Heat stable products
Sorbestrin Low-calorie, heat stable, liquid fat substitute composed of fatty Fried foods, salad dressing, mayonnaise and baked goods
acid esters of sorbitol and sorbitol anhydrides. Approximately
1.5 calories per gram and is suitable for use in all vegetable oil
applications
Based on: http://www.caloriecontrol.org/articles-and-video/feature-articles/glossary-of-fat-replacers
2 Lipids
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Amino Acids and Proteins
3
Michael Appell, W. Jeffrey Hurst, John W. Finley,
and John M. deMan

Proteins occur in animal as well as vegetable


Introduction products in important quantities. In the developed
countries, people obtain much of their protein
Proteins are polymers of some 21 different from animal products. In other parts of the world,
amino acids joined together by peptide bonds. the major portion of dietary protein is derived
Because of the variety of side chains that occur from plant products. Many plant proteins are often
when these amino acids are linked together, the deficient in one or more of the essential amino
different proteins may have different chemical acids. Essential amino acids are defined as those
properties and widely different secondary and that cannot be synthesized by an organism and are
tertiary structures. In peptides and proteins, the only obtained from the diet. The protein content of
amino acids joined in a peptide chain can be some selected foods is listed in Table 3.1.
grouped on the basis of the chemical nature of
the side chains (Krull and Wall 1969). The side
chains may be polar or nonpolar. High levels of Amino Acids
polar amino acid residues in a protein increase
water solubility. The most polar side chains are Amino acids have a range of side chains but all
those of the basic and acidic amino acids. These have an available carboxylic acid and an ammo-
amino acids are present at high levels in the nia group in the α-position.
soluble albumins and globulins. In contrast, the
wheat proteins, gliadin and glutenin, have low
levels of polar side chains and are quite insolu-
ble in water. The acidic amino acids may also be
present in proteins in the form of their amides,
glutamine and asparagine. Hydroxyl groups in
the side chains may become involved in ester
linkages with phosphoric acid and phosphates.
Sulfur amino acids may form disulfide cross- The structures of the amino acids can be bro-
links between neighboring peptide chains or ken into categories based on the structure and
between different parts of the same chain. chemical characteristics of the side chains. In
Proline and hydroxyproline impose significant Fig. 3.1 the structures and groupings of the amino
structural limitations on the geometry of the acids are illustrated. For convenience, amino
peptide chain. acids are abbreviated with either a three letter

© Springer International Publishing AG 2018 117


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_3
118 3  Amino Acids and Proteins

Table 3.1  Protein content of some selected foods online at http://fnic.nal.usda.gov/dietary-guidance/


Product Protein (g/100 g) dietary-reference-intakes/dri-reports.)
Meat The amounts of these essential amino acids
 Beef 16.5 present in a protein and their availability deter-
 Pork 10.2 mine the nutritional quality of the protein. In gen-
Chicken (light meat) 23.4 eral, animal proteins are of higher quality than
Fish plant proteins. Plant proteins can be upgraded
 Haddock 18.3 nutritionally by judicious blending or by genetic
 Cod 17.6 modification through plant breeding. The amino
Milk 3.6 acid composition of some selected animal and
Egg 12.9 vegetable proteins is given in Table 3.3.
Wheat 13.3 Protein digestibility-corrected amino acid
Bread 8.7
score (PDCAAS) is a method of evaluating the
Soybeans
protein quality based on both the amino acid
 Dry 34.1
requirements of humans and their ability to digest
 Raw cooked 11.0
it. The PDCAAS rating was adopted by the US
Peas 6.3
Beans
Food and Drug Administration (FDA) and the
 Dry 22.3 Food and Agricultural Organization of the United
 Raw cooked 7.8 Nations/World Health Organization (FAO/WHO)
Rice in 1993 as “the preferred ‘best’” method to deter-
 White 6.7 mine protein quality.
 Raw cooked 2.0 Egg protein is one of the best quality proteins
Cassava 1.6 and is considered to have a biological value of
Potato 2.0 100. It is widely used as a standard, and protein
efficiency ratio (PER) values sometimes use egg
abbreviation or a single letter designation which white as a standard. Cereal proteins are gener-
is frequently used when describing longer chains ally deficient in lysine and threonine, as indi-
and sequences of amino acids. cated in Table 3.4. Soybean is a good source of
There are 21 amino acids found in most pro- lysine but is deficient in methionine. Cottonseed
teins. An essential amino acid is an amino acid protein is deficient in lysine and peanut protein
that cannot be synthesized de novo (from scratch) in methionine and lysine. The protein of potato
by the organism, and thus must be supplied in the although present in small quantity (Table 3.1) is
diet. The nine amino acids humans cannot syn- of excellent quality and is equivalent to that of
thesize are phenylalanine, valine, threonine, tryp- whole egg.
tophan, methionine, leucine, isoleucine, lysine,
and histidine (Table 3.2). Six other amino acids
are considered conditionally essential in the Peptides and Proteins
human diet, meaning their synthesis can be lim-
ited under certain conditions, such as premature Proteins and peptides are formed from amino
birth in the infants or individuals in severe cata- acids by peptide bonds. The reaction of peptide
bolic distress. The six conditionally essential bond formation produces one peptide bond and
amino acids are arginine, cysteine, glycine, gluta- one molecule of water. Peptides are oligomers of
mine, proline and tyrosine. (Dietary Reference amino acids joined by peptide bonds and proteins
Intakes: The Essential Guide to Nutrient Requi­ can range from a few amino acids joined together
rements, published by the Institute of Medicine’s to many thousands of amino acid residues in
Food and Nutrition Board, currently available large proteins.
Peptides and Proteins 119

Fig. 3.1  Common amino acids found in proteins with short hand abbreviations, molecular weight and pKa values of
ionizable side chains

Proteins which make up more than half the dry chains form specific folded three-­ dimensional
weight of most cells are the most important mac- shapes which enable the proteins to perform spe-
romolecules in all living organisms. More than cific tasks such as structure or enzyme activity.
half of the dry weight of a cell is made up of pro- These tasks include transporting small molecules
teins of various shapes and sizes (Voet and Voet (e.g., the hemoglobin transports oxygen in the
2004). Proteins are based on sequences of amino bloodstream), catalyzing biological functions, pro­
acids connected by peptide bonds. The protein viding structure to collagen and skin, controlling
120 3  Amino Acids and Proteins

Table 3.2  Essential and nonessential amino acids The structure is stabilized when the parts of a
Essential Nonessential protein domain are locked into place by tertiary
Histidine Alanine interactions, such as salt bridges, hydrogen
Isoleucine Arginine bonds, the packing of side chains and disulfide
Leucine Aspartic acid bridges. The Quaternary structure is a large
Lysine Cysteine assembly of several protein molecules or poly-
Methionine Glutamic acid peptide chains generally referred to as subunits.
Phenylalanine Glutamine The quaternary structure is stabilized by the same
Threonine Glycine non-covalent interactions and disulfide bonds as
Tryptophan Proline the tertiary structure.
Valine Serine The native structure of a protein is its natural
Tyrosine state in the cell, unaltered by heat, chemicals,
Asparagine
enzyme action. In their native state proteins are
folded into their 3D structures which provide
sense, and regulating hormones (Pietzsch 2003). maximum stability and are reached at lowest
In peptides and proteins, the carboxyl terminus energy (Pietzsch 2003). When proteins structures
(known as C-terminus) of one amino acid with a are forced to be deformed they are considered to
free carboxyl group, the amino terminus (known be denatured (Anfinsen 1973; Tanford 1968). The
as N-terminus) of another amino acid with a free forces that cause denaturation can be chemicals
amino group. The size of the protein is determined or the application of heat. An example of heat
by the number of amino acids in the protein denaturation is the heating of egg white where
sequence. A protein sequence having n amino the protein is denatured resulting in a conversion
acids has n − 1 peptide bonds and n − 1 amide form liquid to solid form.
planes which are fairly rigid because of the struc-
ture of the peptide bond. The bond angle between
N of amine group and alpha carbon is denoted by Protein Classification
φ (phi) and the bond angle between C α and C of
carboxyl group is denoted by ψ (psi). Proteins are complex molecules, and classifica-
There are four different levels of protein struc- tion has been based mostly on solubility in dif-
ture. These are: Primary Structure, Secondary ferent solvents. Increasingly, however, as more
Structure, Tertiary Structure, and Quaternary knowledge about molecular composition and
Structure as depicted in Fig. 3.2. Primary struc- structure is obtained, other criteria are being
ture: The Primary structure is defined by the used for classification. These include behavior in
amino acid sequence of the polypeptide chain. the ultracentrifuge and electrophoretic pro­
The primary structure is held together by peptide perties. Proteins are divided into the following
bonds, which are formed during the process main groups: simple, conjugated, and derived
of protein synthesis. Secondary structure: The proteins.
Secondary Structure as shown in Fig. 3.2 refers
to highly regular local sub-structures such as the
alpha helix in Fig. 3.2. In the secondary structure Simple Proteins
includes the alpha helix, the beta sheet, and
the random coils. These secondary structures are Simple proteins yield only amino acids on hydro-
controlled by hydrogen bonds between the main-­ lysis and include the following classes:
chain peptide groups. The Tertiary structure as
shown in Fig. 3.2 represents the three-­dimensional • Albumins. Soluble in neutral, salt-free water.
structure of a single protein molecule where the Usually these are proteins of relatively low
alpha-helices and beta-sheets are folded into a molecular weight. Examples are egg albumin,
compact globule structure. lactalbumin, and serum albumin in the whey
Table 3.3  Composition and amino acid content of some selected foodsa
Whole egg Cows milk Ground beef 15% fat Hard red winter wheat Soy flour (defatted)
Energy 143 61 215 327 327
(kcal/100 g)
Composition in g/100 g
Water 76.15 88.13 55.81 13.1 7.25
Protein Classification

Carbohydrates  0.72  5.26 0 71.18 33.97


Fat  9.51  3.25 15  1.54 1.22
Protein 12.56  3.22 18.59 12.61 51.46
Amino acid content foods and protein g/100 g
AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g AA/100 g
food protein food protein food protein food protein food protein
Tryptophan 0.17 1.33 0.04 1.24 0.09 0.51 0.16 1.27 0.68 1.33
Threonine 0.56 4.43 0.13 4.16 0.72 3.87 0.37 2.89 2.04 3.97
Isoleucine 0.67 5.34 0.16 5.06 0.82 4.42 0.46 3.63 2.28 4.43
Leucine 1.09 8.65 0.30 9.29 1.45 7.80 0.85 6.77 3.83 7.44
Lysine 0.91 7.26 0.26 8.20 1.54 8.28 0.34 2.66 3.13 6.08
Methionine 0.38 3.03 0.08 2.58 0.48 2.57 0.20 1.59 0.63 1.23
Cystine 0.27 2.17 0.02 0.59 0.19 1.03 0.32 2.55 0.76 1.47
Phenylalanine 0.68 5.41 0.16 5.06 0.75 4.05 0.59 4.69 2.45 4.77
Tyrosine 0.50 3.97 0.16 4.94 0.57 3.08 0.39 3.07 1.78 3.46
Valine 0.86 6.83 0.21 6.40 0.91 4.92 0.56 4.41 2.35 4.56
Arginine 0.74 5.85 0.09 2.80 1.21 6.51 0.60 4.72 3.65 7.09
Histidine 0.31 2.46 0.10 2.95 0.60 3.25 0.29 2.26 1.27 2.46
Alanine 0.74 5.85 0.11 3.32 1.07 5.73 0.45 3.57 2.22 4.30
Aspartic acid 1.33 10.56 0.27 8.39 1.67 9.00 0.64 5.08 5.91 11.49
Glutamic acid 1.67 13.32 0.71 21.99 2.79 15.00 4.00 31.70 9.11 17.70
Glycine 0.43 3.44 0.06 1.93 1.26 6.79 0.53 4.19 2.17 4.22
Proline 0.51 4.08 0.31 9.66 0.95 5.10 1.29 10.22 2.75 5.34
Serine 0.97 7.73 0.19 5.90 0.74 4.00 0.59 4.65 2.73 5.30
Hyroxy-proline 0.27 1.46
PDCAASb 1 1 0.92 0.42 0.91
a
From USDA Nutrient data base
121

b
Schaafsma (2000)
122 3  Amino Acids and Proteins

proteins of milk, leucosin of cereals, and • Prolamins. Soluble in 50–90% ethanol and
­legumelin in legume seeds. insoluble in water. These proteins have large
• Globulins. Soluble in neutral salt solutions amounts of proline and glutamic acid and
and almost insoluble in water. Examples are occur in cereals. Examples are zein in corn,
serum globulins and β-lactoglobulin in milk, gliadin in wheat, and hordein in barley.
myosin and actin in meat, and glycinin in • Scleroproteins. Insoluble in water and neu-
soybeans. tral solvents and resistant to enzymic
• Glutelins. Soluble in very dilute acid or base ­hydrolysis. These are fibrous proteins serv-
and insoluble in neutral solvents. These pro- ing structural and binding purposes. Collagen
teins occur in cereals, such as glutenin in of muscle tissue is included in this group, as
wheat and oryzenin in rice. is gelatin, which is derived from it. Other
examples include elastin, a component of
Table 3.4  Limiting essential amino acids of some grain tendons, and keratin, a component of hair
proteins and hoofs.
First limiting Second limiting • Histones. Basic proteins, as defined by their
Grain amino acid amino acid high content of lysine and arginine. Soluble in
Wheat Lysine Threonine water and precipitated by ammonia.
Corn Lysine Tryptophan • Protamines. Strongly basic proteins of low
Rice Lysine Threonine molecular weight (4000–8000). They are rich
Sorghum Lysine Threonine in arginine. Examples are clupein from her-
Millet Lysine Threonine ring and scombrin from mackerel.

Fig. 3.2  The four levels of protein structure (Rashid et al. 2015)
Protein Structure 123

Conjugated Proteins solution. Peptides contain two or more amino


acid residues. These breakdown products are
Conjugated proteins contain an amino acid part formed during the processing of many foods, for
combined with a nonprotein material such as a example, during ripening of cheese.
lipid, nucleic acid, or carbohydrate. Some of the
major conjugated proteins are as follows:
Protein Structure
• Phosphoproteins. An important group that
includes many major food proteins. Phosphate Proteins are macromolecules with different levels
groups are linked to the hydroxyl groups of of structural organization. The primary structure
serine and threonine. This group includes of proteins relates to the peptide bonds between
casein of milk and the phosphoproteins of egg component amino acids and also to the amino
yolk. acid sequence in the molecule. Researchers have
• Lipoproteins. These are combinations of lipids elucidated the amino acid sequence in many pro-
with protein and have excellent emulsifying teins. For example, the amino acid composition
capacity. Lipoproteins occur in milk and egg and sequence for several milk proteins is now
yolk. well established (Swaisgood 1982).
• Nucleoproteins. These are combinations of Some proteolytic enzymes have quite specific
nucleic acids with protein. These compounds actions; they attack only a limited number of
are found in cell nuclei. bonds, involving only particular amino acid resi-
• Glycoproteins. These are combinations of car- dues in a particular sequence. This may lead to
bohydrates with protein. Usually the amount the accumulation of well-defined peptides during
of carbohydrate is small, but some glycopro- some enzymic proteolytic reactions in foods.
teins have carbohydrate contents of 8–20%. The secondary structure of proteins involves
An example of such a mucoprotein is ovomu- folding the primary structure. Hydrogen bonds
cin of egg white. between amide nitrogen and carbonyl oxygen are
• Chromoproteins. These are proteins with a the major stabilizing force. These bonds may be
colored prosthetic group. There are many formed between different areas of the same poly-
compounds of this type, including hemoglo- peptide chain or between adjacent chains. In
bin and myoglobin, chlorophyll, and aqueous media, the hydrogen bonds may be less
flavoproteins. significant, and van der Waals forces and hydro-
phobic interaction between apolar side chains
may contribute to the stability of the secondary
Derived Proteins structure. The secondary structure may be either
the α-helix or the sheet structure, as shown in
These are compounds obtained by chemical or Fig. 3.2. The helical structures are stabilized by
enzymatic methods and are divided into primary intramolecular hydrogen bonds, the sheet struc-
and secondary derivatives, depending on the tures by intermolecular hydrogen bonds. The
extent of change that has taken place. Primary requirements for maximum stability of the helix
derivatives are slightly modified and are insolu- structure were established by Pauling et al.
ble in water; rennet-coagulated casein is an (1951). The helix model involves a translation of
example of a primary derivative. Secondary 0.54 nm per turn along the central axis. A com-
derivatives are more extensively changed and plete turn is made for every 3.6 amino acid resi-
include proteoses, peptones, and peptides. The dues. Proteins do not necessarily have to occur in
difference between these breakdown products is a complete α-helix configuration; rather, only
in size and solubility. All are soluble in water and parts of the peptide chains may be helical, with
not coagulated by heat, but proteoses can be other areas of the chain in a more or less ­unordered
precipitated with saturated ammonium sulfate
­ configuration. Proteins with α-helix structure
124 3  Amino Acids and Proteins

Table 3.5  Bond and interaction energies of the interac- Table 3.6  Oligomeric food proteins
tions involved in protein structure
Molecular
Bond Bond energya (kcal/mol) Protein weight (Daltons) Subunits
Covalent C–C 83 Lactoglobulin 35,000 2
Covalent S–S 50 Hemoglobin 64,500 4
Hydrogen bond 3–7 Avidin 68,300 4
Electrostatic interactions 3–7 Lipoxygenase 108,000 2
Hydrophobic interactions 3–5 Tyrosinase 128,000 4
Van der Waals forces 1–2 Lactate dehydrogenase 140,000 4
a
These refer to free energy required to break the bonds: in 7S soy protein 200,000 9
the case of a hydrophobic bond, the free energy required Invertase 210,000 4
to unfold a nonpolar side chain from the interior of the Catalase 232,000 4
molecule into the aqueous medium
Collagen 300,000 3
11S soy protein 350,000 12
Legumin 360,000 6
may be either globular or fibrous. In the parallel Myosin 475,000 6
sheet structure, the polypeptide chains are almost
Source: Reprinted with permission from D.W. Stanley
fully extended and can form hydrogen bonds and R.Y. Yada, Thermal Reactions in Food Protein
between adjacent chains. Such structures are gen- Systems, Physical Chemistry of Foods, H.G. Schwartzberg
erally insoluble in aqueous solvents and are and R.H. Hartel, eds., p. 676, 1992, by courtesy of Marcel
Dekker, Inc.
fibrous in nature.
The tertiary structure of proteins involves a
pattern of folding of the chains into a compact similar subunits through noncovalent forces or
unit that is stabilized by hydrogen bonds, van der disulfide bonds to form an oligomeric macromol-
Waals forces, disulfide bridges, and hydrophobic ecule (Stanley and Yada 1992). Many food pro-
interactions. The tertiary structure results in the teins are oligomeric and consist of a number of
formation of a tightly packed unit with most of subunits, usually 2 or 4, but occasionally as many
the polar amino acid residues located on the out- as 24. A listing of some oligomeric food proteins
side and hydrated. This leaves the internal part is given in Table 3.6. The subunits of proteins are
with most of the apolar side chains and virtually held together by various types of bonds: electro-
no hydration. Certain amino acids, such as pro- static bonds involving carboxyl, amino, imidaz-
line, disrupt the α-helix, and this causes fold ole, and guanido groups; hydrogen bonds
regions with random structure (Kinsella 1982). involving hydroxyl, amide, and phenol groups;
The nature of the tertiary structure varies among hydrophobic bonds involving long-chain ali-
proteins as does the ratio of α-helix and random phatic residues or aromatic groups; and covalent
coil. Insulin is loosely folded, and its tertiary disulfide bonds involving cystine residues.
structure is stabilized by disulfide bridges. Hydrophobic bonds are not true bonds but have
Lysozyme and glycinin have disulfide bridges but been described as interactions of nonpolar
are compactly folded. groups. These nonpolar groups or areas have a
Large molecules of molecular weights above tendency to orient themselves to the interior of
about 50,000 may form quaternary structures by the protein molecule. This tendency depends on
association of subunits. These structures may be the relative number of nonpolar amino acid resi-
stabilized by hydrogen bonds, disulfide bridges, dues and their location in the peptide chain. Many
and hydrophobic interactions. The bond energies food proteins, especially plant storage proteins,
involved in forming these structures are listed in are highly hydrophobic—so much so that not all
Table 3.5. of the hydrophobic areas can be oriented toward
The term subunit denotes a protein chain pos- the inside and have to be located on the surface.
sessing an internal covalent and noncovalent This is a possible factor in subunits association
structure that is capable of joining with other and in some cases may result in aggregation.
Denaturation 125

The hydrophobicity values of some food proteins p­ roteins and affects different proteins to different
as reported by Stanley and Yada (1992) are listed degrees, depending on the structure of a protein.
in Table 3.7. Denaturation can be brought about by a variety of
The well-defined secondary, tertiary, and qua- agents, of which the most important are heat, pH,
ternary structures are thought to arise directly salts, and surface effects. Considering the com-
from the primary structure. This means that a plexity of many food systems, it is not surprising
given combination of amino acids will automati- that denaturation is a complex process that c­ annot
cally assume the type of structure that is most easily be described in simple terms. Denaturation
stable and possible given the considerations usually involves loss of biological activity and
described by Pauling et al. (1951). significant changes in some physical or func-
tional properties such as solubility. A simplified
view of protein denaturation and the renaturation
Denaturation are illustrated in Fig. 3.3. Protein denaturation
generally is not reversible. Thermal treatments of
Denaturation is a process that changes the molec- egg white protein are a good example of protein
ular structure without breaking any of the peptide denaturation.
bonds of a protein. The process is peculiar to The three-dimensional structure of proteins is
driven by a number of interactions in the side
chain of the protein. When proteins are exposed
Table 3.7  Hydrophobicity values of some food proteins to conditions such as extremes in pH or in the
Protein Hydrophobicity (cal/residue) heat all these are disrupted leaving the protein
Gliadin 1300 structure with reduced stabilizing forces which
Bovine serum albumin 1120–1000 results in the unraveling of the structure in rare
α-Lactalbumin 1050 instances an unraveled protein with its primary
β-Lactoglobulin 1050 structure intact can when under carefully con-
Actin 1000 trolled conditions the protein will spontaneously
Ovalbumin 980 fold and become renatured.
Collagen 880 The destruction of enzyme activity by heat is
Myosin 880 an important operation in food processing. In
Casein 725 most cases, denaturation is nonreversible; how-
Whey protein 387 ever, there are some exceptions, such as the
Gluten 349 recovery of some types of enzyme activity after
Source: Reprinted with permission from D.W. Stanley heating. Heat denaturation is sometimes desir-
and R.Y. Yada, Thermal Reactions in Food Protein
Systems, Physical Chemistry of Foods, H.G. Schwartzberg
able—for example, the denaturation of whey pro-
and R.H. Hartel, eds., p. 677, 1992, by courtesy of Marcel teins for the production of milk powder used in
Dekker, Inc. baking. The relationship among temperature,

Fig. 3.3 Thermal
aturat
Denloss of ion
denaturation of protein
and the renaturation
with loss and regain of biological
activity
biological activity

regains
activity
Normal protein Re Denatured protein
n a t u r a tio n
126 3  Amino Acids and Proteins

Fig. 3.4  Time-­temperature


relationships for the heat
denaturation of whey
proteins in skim milk.
Adapted from
H.A. Harland, S.T. Coulter,
and R. Jenness, The Effects
of Various Steps in the
Manufacture on the Extent
of Serum Protein
Denaturation in Nonfat
Dry Milk Solids. J. Dairy
Sci. 35: 363–368, 1952

heating time, and the extent of whey protein Table 3.8  Heat coagulation temperatures of some albu-
denaturation in skim milk is demonstrated in mins and globulins and casein
Fig. 3.4 (Harland et al. 1952). Protein Coagulation temperature (°C)
The proteins of egg white are readily dena- Egg albumin 56
tured by heat and by surface forces when egg Serum albumin (bovine) 67
white is whipped to a foam. Meat proteins are Milk albumin (bovine) 72
denatured in the temperature range 57–75 °C, Legumelin (pea) 60
which has a profound effect on texture, water Serum globulin (human) 75
holding capacity, and shrinkage. β-Lactoglobulin (bovine) 70–75
Fibrinogen (human) 56–64
Denaturation may sometimes result in the
Myosin (rabbit) 47–56
flocculation of globular proteins but may also
Casein (bovine) 160–200
lead to the formation of gels. Foods may be dena-
tured, and their proteins destabilized, during
freezing and frozen storage. Fish proteins are
particularly susceptible to destabilization. After p­ rotein. Denaturation can be defined as a major
freezing, fish may become tough and rubbery and change in the native structure that does not
lose moisture. The caseinate micelles of milk, involve alteration of the amino acid sequence.
which are quite stable to heat, may be destabi- The effect of heat usually involves a change in
lized by freezing. On frozen storage of milk, the the tertiary structure, leading to a less ordered
stability of the caseinate progressively decreases, arrangement of the polypeptide chains. The tem-
and this may lead to complete coagulation. perature range in which denaturation and coa­
Protein denaturation and coagulation are gulation of most proteins take place is about
aspects of heat stability that can be related to 55–75 °C, as indicated in Table 3.8. There are
the amino acid composition and sequence of the some notable exceptions to this general pattern.
Non-enzymic Browning 127

cysteine
C
C
CH2
CH2
SH
S
SH
S
Interchain
CH2
C CH2 disulfide
C oxidants C bonds
CH2 C
heat
CH2
SH
S Intrachain
SH
S disulfide
CH2 bonds
CH2
C
C

Fig. 3.5  Heat or oxidation of protein sulfhydryl groups from cysteine can form both intra- and intermolecular bonds

Table 3.9  Cysteine and cystine content of some proteins shown in Fig. 3.5. Casein, with its extremely low
(g amino acid/100 g protein) content of sulfur amino acids, exemplifies this
Protein Cysteine (%) Cystine (%) behavior. The heat stability of casein is also
Egg albumin 1.4 0.5 explained by the restraints against forming a
Serum albumin (bovine) 0.3 5.7 folded tertiary structure. These restraints are due
Milk albumin 6.4 – to the relatively high content of proline and
β-Lactoglobulin 1.1 2.3 hydroxyproline in the heat stable proteins
Fibrinogen 0.4 2.3 (Table  3.10). In a peptide chain free of proline,
Casein – 0.3
the possibility of forming inter- and intramolecu-
lar hydrogen bonds is better than in a chain con-
Cysteine in proteins can be involved in the taining many proline residues (Fig. 3.6). These
­denaturation process. As shown in Fig. 3.5 when considerations show how amino acid composi-
cysteine is oxidized to form new disulfide bridges tion directly relates to secondary and tertiary
they can be formed within the protein or in inter- structure of proteins; these structures are, in turn,
molecular bridges causing aggregation of the responsible for the functionality of the protein.
protein crosslinked protein. Casein and gelatin
are examples of proteins that can be boiled with-
out apparent change in stability. The exceptional Non-enzymic Browning
stability of casein makes it possible to boil, steril-
ize, and concentrate milk, without coagulation. The non-oxyenzymic browning or Maillard reac­
The reasons for this exceptional stability have tion is of great importance in food manufacturing
been discussed by Kirchmeier (1962). In the first and its results can be either desirable or undesir-
place, restricted formation of disulfide bonds due able. For example, the brown crust formation on
to low content of cystine and cysteine results in bread is desirable; the brown disco­loration of
increased stability. The relationship between evaporated and sterilized milk is undesirable. For
coagulation temperature as a measure of stability products in which the browning reaction is favor-
and sulfur amino acid content is shown in Tables able, the resulting color and flavor characteristics
3.8 and 3.9. Peptides, which are low in these par- are generally experienced as pleasant. In other
ticular amino acids, are less likely to become products, color and flavor may become quite
involved in the type of sulfhydryl agglomeration unpleasant.
128 3  Amino Acids and Proteins

The browning reaction can be defined as the of brown nitrogenous polymers or melanoidins
sequence of events that begins with the reaction (Ellis 1959).
of the amino group of amino acids, peptides, or The reaction velocity and pattern are influenced
proteins with a glycosidic hydroxyl group of sug- by the nature of the reacting amino acid or protein
ars; the sequence terminates with the formation and the carbohydrate. This means that each kind of
food may show a different browning pattern.
Generally, lysine is the most reactive amino acid
Table 3.10  Amino acid composition of serum albumin, because of the free ε-amino group. Since lysine is
casein, and gelatin (g amino acid/100 g protein) the limiting essential amino acid in many food
Serum proteins, its destruction can substantially reduce
Amino acid albumin Casein Gelatin the nutritional value of the protein. Foods that are
Glycine 1.8 1.9 27.5 rich in reducing sugars are very reactive, and this
Alanine 6.3 3.1 11.0
explains why lysine in milk is destroyed more eas-
Valine 5.9 6.8 2.6
ily than in other foods (Fig. 3.7). Other factors that
Leucine 12.3 9.2 3.3
influence the browning reaction are temperature,
Isoleucine 2.6 5.6 1.7
pH, moisture level, oxygen, metals, phosphates,
Serine 4.2 5.3 4.2
Threonine 5.8 4.4 2.2
sulfur dioxide, and other inhibitors.
Cystine ½ 6.0 0.3 0.0 The browning reaction involves a number of
Methionine 0.8 1.8 0.9 steps. An outline of the total pathway of melanoi-
Phenylalanine 6.6 5.3 2.2 din formation has been given by Hodge (1953)
Tyrosine 5.1 5.7 0.3 and is shown in Fig. 3.8. According to Hurst and
Proline 4.8 13.5 16.4 Ito (1972), the following five steps are involved
Hydroxyproline – – 14.1 in the process:
Aspartic acid 10.9 7.6 6.7
Glutamic acid 16.5 24.5 11.4 1. The production of an N-substituted glycosyl-
Lysine 12.8 8.9 4.5 amine from an aldose or ketose reacting with
Arginine 5.9 3.3 8.8 a primary amino group of an amino acid, pep-
Histidine 4.0 3.8 0.8 tide, or protein.

Fig. 3.6  Effect of


proline residues on
possible hydrogen bond
formation in peptide
chains. (a) Proline-free
chain; (b) proline-­
containing chain; (c)
hydrogen bond
formation in proline-free
and proline-containing
chains. Source: From
O. Kirchmeier, The
Physical-Chemical
Causes of the Heat
Stability of Milk
Proteins,
Milchwissenschaft
(German), Vol. 17,
pp. 408–412, 1962
Non-enzymic Browning 129

shown is thought to involve addition of the amine


to the carbonyl group of the open-chain form of
the sugar, elimination of a molecule of water, and
closure of the ring. The rate is high at low water
content; this explains the ease of browning in
dried and concentrated foods.
The Amadori rearrangement of the glycosyl-
amines involves the presence of an acid catalyst
and leads to the formation of ketoseamine or
1-amino-1-deoxyketose according to the scheme
shown in Fig. 3.10. In the reaction of d-glucose
with glycine, the amino acid reacts as the catalyst
and the compound produced is l-deoxy-1-­glycino-
β-d-fructose (Fig. 3.11). The ketoseamines are
relatively stable compounds, which are formed in
maximum yield in systems with 18% water con-
tent. A second type of rearrangement reaction is
the Heyns rearrangement, which is an alternative
to the Amadori rearrangement and leads to the
same type of transformation. The mechanism of
the Amadori rearrangement (Fig. 3.9) involves
protonation of the nitrogen atom at carbon 1. The
Heyns rearrangement (Fig. 3.12) involves proton-
Fig. 3.7  Loss of lysine occurring as a result of heating of ation of the oxygen at carbon 6.
several foods. Source: Adapted from: J. Adrian, The
Maillard Reaction. IV. Study on the Behavior of Some Secondary reactions lead to the formation of
Amino Acids During Roasting of Proteinaceous Foods, diketoseamines and diamino sugars. The formation
Ann. Nutr. Aliment. (French), Vol. 21, pp. 129–147, 1967 of these compounds involves complex reactions
and, in contrast to the formation of the primary
2. Rearrangement of the glycosylamine by an products, does not occur on a mole-for-­mole basis.
Amadori rearrangement type of reaction to In Step 4, the ketoseamines are decomposed
yield an aldoseamine or ketoseamine. by 1,2-enolization or 2,3-enolization. The former
3. A second rearrangement of the ketoseamine pathway appears to be the more important one for
with a second mole of aldose to form a dike- the formation of brown color, whereas the latter
toseamine, or the reaction of an aldoseamine results in the formation of flavor products.
with a second mole of amino acid to yield a According to Hurst and Ito (1972), the
diamino sugar. 1,2-­enolization pathway appears mainly to lead
4. Degradation of the amino sugars with loss of to browning but also contributes to formation of
one or more molecules of water to give amino off-flavors through hydroxymethylfurfural,
or nonamino compounds. which may be a factor in causing off-flavors in
5. Condensation of the compounds formed in stored, overheated, or dehydrated food products.
Step 4 with each other or with amino com- The mechanism of this reaction is shown in
pounds to form brown pigments and Fig. 3.13 (Hurst and Ito 1972). The ketoseamine
polymers. (1) is protonated in acid medium to yield (2).
This is changed in a reversible reaction into the
The formation of glycosylamines from the 1,2-enolamine (3) and this is assisted by the N
reaction of amino groups and sugars is reversible substituent on carbon 1. The following steps
(Fig.  3.9) and the equilibrium is highly depen- involve the β-elimination of the hydroxyl group
dent on the moisture level. The mechanism as on carbon 3. In (4) the enolamine is in the free
130 3  Amino Acids and Proteins

Fig. 3.8  Reaction pattern of the formation of melanoidins from aldose sugars and amino compounds. Source: From
J.E. Hodge, Chemistry of Browning Reactions in Model Systems, Agr. Food Chem., VoL 1, pp. 928–943, 1953

base form and converts to the Schiff base (5). The Following the production of 1,2-enol forms of
Schiff base may undergo hydrolysis and form the aldose and ketose amines, a series of degrada-
enolform (7) of 3-deoxyosulose (8). In another tions and condensations results in the formation
step the Schiff base (5) may lose a proton and of melanoidins. The α-β-dicarbonyl compounds
the hydroxyl from carbon 4 to yield a new enter into aldol type condensations, which lead to
Schiff base (6). Both this compound and the the formation of polymers, initially of small size,
3-­
deoxyosulose may be transformed into an highly hydrated, and in colloidal form. These ini-
unsaturated osulose (9), and by elimination of a tial products of condensation are fluorescent, and
proton and a hydroxyl group, hydroxymethylfur- continuation of the reaction results in the forma-
fural (10) is formed. tion of the brown melanoidins. These polymers
Non-enzymic Browning 131

Fig. 3.9  Reversible formation of glycosylamines in the Its Inhibition by Sulpher Dioxide, BFMIRA Scientific and
browning reaction. Source: From D.T. Hurst, Recent Technical Surveys, No. 75, Leatherhead, England, 1972
Development in the Study of Nonenzymic Browning and

Fig. 3.10  Amadori rearrangement. Source: From M.J. Kort, Reactions of Free Sugars with Aqueous Ammonia, Adv.
Carbohydrate Chem. Biochem., Vol. 25, pp. 311–349, 1970

are of nondistinct composition and contain r­eminiscent of caramelization. The Strecker


­varying levels of nitrogen. The composition var- ­degradation of α-amino acids is a reaction that
ies with the nature of the reaction partners, pH, also significantly contributes to the formation of
temperature, and other conditions. ­flavor compounds. The dicarbonyl compounds
The flavors produced by the Maillard reaction formed in the previously described schemes react
also vary widely. In some cases, the flavor is in the following manner with α-amino acids:
132 3  Amino Acids and Proteins

The amino acid is converted into an aldehyde are listed in Table 3.11. These are mostly vicinal
with one less carbon atom (Schönberg and polycarbonyl compounds and α,β-­unsaturated
Moubacher 1952). Some of the compounds of aldehydes. They condense rapidly to form mela-
browning flavor have been described by Hodge noidins. The Strecker degradation aldehydes con-
et al. (1972). Corny, nutty, bready, and crackery tribute to the aroma of bread, peanuts, cocoa, and
aroma compounds consist of planar unsaturated other roasted foods. Although acetic, phenyla­
heterocyclic compounds with one or two n­ itrogen cetic, isobutyric, and isovaleric aldehydes are
atoms in the ring. Other important members of prominent in the aromas of bread, malt, peanuts,
this group are partially saturated N-heterocyclics and cocoa, they are not really characteristic of
with alkyl or acetyl group substituents. Comp­ these foods (Hodge et al. 1972).
ounds that contribute to pungent, burnt aromas A somewhat different mechanism for the brown-
ing reaction has been proposed by Burton and
McWeeney (1964) and is shown in Fig. 3.14. After
formation of the aldosylamine, dehydration reac-
tions result in the production of 4- to 6-­membered
ring compounds. When the reaction proceeds under
conditions of moderate heating, fluorescent nitrog-
enous compounds are formed. These react rapidly
Fig. 3.11  Structure of 1-deoxy-l-glycino-β-d-fructose with glycine to yield melanoidins.

Fig. 3.12  Heyns rearrangement. Source: From M.J. Kort, Reactions of Free Sugars with Aqueous Ammonia, Adv.
Carbohydrate Chem. Biochem., Vol. 25, pp. 311–349, 1970
Non-enzymic Browning 133

Fig. 3.13  1,2-Enolization mechanism of the browning by Sulphur Dioxide, BFMIRA Scientific and Technical
reaction. Source: From D.T. Hurst, Recent Developments Surveys, No. 75, Leatherhead, England, 1972
in the Study of Nonenzymic Browning and Its Inhibition

The influence of reaction components and more rapid. Temperature also affects the compo-
reaction conditions results in a wide variety of sition of the pigment formed. At higher tempera-
reaction patterns. Many of these conditions are tures, the carbon content of the pigment increases
interdependent. Increasing temperature results in and more pigment is formed per mole of carbon
a rapidly increasing rate of browning; not only dioxide released. Color intensity of the pigment
reaction rate, but also the pattern of the reaction increases with increasing temperature. The effect
may change with temperature. In model systems, of temperature on the reaction rate of d-glucose
the rate of browning increases 2–3 times for each with dl-leucine is illustrated in Fig. 3.15.
10° rise in temperature. In foods containing fruc- In the Maillard reaction, the basic amino
tose, the increase may be 5–10 times for each 10° group disappears; therefore, the initial pH or the
rise. At high sugar contents, the rate may be even presence of a buffer has an important effect on
134 3  Amino Acids and Proteins

Table 3.11  Aroma and structure classification of browned flavor compounds


Aromas: Burnt (pungent, empyreumatic) Variable (aldehydic, ketonic)
Structure: Polycarbonyls(α,β-Unsat’d aldehydes Monocarbonyls (R–CHO,
–C:O–C:O–, =C–CHO R–C:O–CH3)
Examples of compounds: Glyoxal Strecker aldehydes
Pyruvaldehyde Isobutyric
Diacetyl Isovaleric
Mesoxalic dialdehyde Methional
Acrolein 2-Furaldehydes
Crotonaldehyde 2-Pyrrole aldehydes
C3–C6 Methyl ketones
Source: From J.E. Hodge, F.D. Mills, and B.E. Fisher, Compounds of Browned Flavor from Sugar-Amine Reactions,
Cereal Sci. Today, Vol. 17, pp. 34–40, 1972

Fig. 3.14  Browning pathway leading to melanoidin formation


Non-enzymic Browning 135

Fig. 3.15  Effect of


temperature on the
reaction rate of
d-glucose with
dl-leucine. Adapted
from: G. Haugard,
L. Tumerman, and
A. Sylvestri, A Study on
the Reaction of Aldoses
and Amino Acids, J. Am.
Chem. Soc., Vol. 73,
pp. 4594–4600, 1951

the reaction. The browning reaction is slowed series, glycine is the most reactive. Longer and
down by decreasing pH, and the browning reac- more complex substituent groups reduce the rate
tion can be said to be self-inhibitory since the pH of browning. In the ω-amino acid series, brown-
decreases with the loss of the basic amino group. ing rate increases with increasing chain length.
The effect of pH on the reaction rate of d-glucose Ornithine browns more rapidly than lysine. When
with dl-leucine is demonstrated in Fig. 3.16. The the reactant is a protein, particular sites in the
effect of pH on the browning reaction is highly molecule may react faster than others. In pro-
dependent on moisture content. When a large teins, the ε-amino group of lysine is particularly
amount of water is present, most of the browning vulnerable to attack by aldoses and ketoses.
is caused by caramelization, but at low water lev- Moisture content is an important factor in
els and at pH greater than 6, the Maillard reaction influencing the rate of the browning reaction.
is predominant. Browning occurs at low temperatures and inter-
The nature of the sugars in a nonenzymic mediate moisture content; the rate increases with
browning reaction determines their reactivity. increasing water content. The rate is extremely
Reactivity is related to their conformational sta- low below the glass transition temperature, prob-
bility or to the amount of open-chain structure ably because of limited diffusion (Roos and
present in solution. Pentoses are more reactive Himberg 1994; Roos et al. 1996a, b).
than hexoses, and hexoses more than reducing Methods of preventing browning could con-
disaccharides. Nonreducing disaccharides only sist of measures intended to slow reaction rates,
react after hydrolysis has taken place. The order such as control of moisture, temperature, or pH,
of reactivity of some of the aldohexoses is: man- or removal of an active intermediate. Generally, it
nose is more reactive than galactose, which is is easier to use an inhibitor. One of the most
more reactive than glucose. effective inhibitors of browning is sulfur dioxide
The effect of the type of amino acid can be or sodium bisulfite. The action of sulfur dioxide
summarized as follows. In the α-amino acid is unique and no other suitable inhibitor has been
136 3  Amino Acids and Proteins

Fig. 3.16  Effect of pH


on the reaction rate of
d-glucose with
dl-leucine. Adapted
from: G. Haugard,
L. Tumerman, and
A. Sylvestri, A Study on
the Reaction of Aldoses
and Amino Acids, J. Am.
Chem. Soc., Vol. 73,
pp. 4594–4600, 1951

found. It is known that sulfite can combine with others undesirable. Such chemical changes may
the carbonyl group of an aldose to give an addi- lead to compounds that are not hydrolyzable by
tion compound: intestinal enzymes or to modifications of the pep-
tide side chains that render certain amino acids
NaHSO3 + RCHO → RCHOHSO3 Na
unavailable. Mild heat treatments in the presence
However, this reaction cannot possibly of water can significantly improve the protein’s
account for the inhibitory effect of sulfite. It is nutritional value in some cases. Sulfur-containing
thought that sulfur dioxide reacts with the degra- amino acids may become more available and cer-
dation products of the amino sugars, thus pre- tain antinutritional factors such as the trypsin
venting these compounds from condensing into inhibitors of soybeans may be deactivated.
melanoidins. A serious drawback of the use of Excessive heat in the absence of water can be det-
sulfur dioxide is that it reacts with thiamine and rimental to protein quality; for example, in fish
proteins, thereby reducing the nutritional value of proteins, tryptophan, arginine, methionine, and
foods. Sulfur dioxide destroys thiamine and is lysine may be damaged. A number of chemical
therefore not permitted for use in foods contain- reactions may take place during heat treatment
ing this vitamin. including decomposition, dehydration of serine
and threonine, loss of sulfur from cysteine, oxida-
tion of cysteine and methionine, cyclization of
Chemical Changes glutamic and aspartic acids and threonine (Mauron
1970; Mauron and Antener 1983).
During processing and storage of foods, a number The nonenzymic browning, or Maillard,
of chemical changes involving proteins may occur reaction causes the decomposition of certain
­
(Hurrell 1984). Some of these may be desirable, amino acids. For this reaction, the presence of a
Chemical Changes 137

Fig. 3.17  Formation of


amide-type bonds from
the reaction between
ε-amino groups of lysine
and amide groups of
asparagine (n = 1)
glutamine (n = 2)

reducing sugar is required. Heat damage may conventional sterilization, and 20–50% in roller
also occur in the absence of sugars. Bjarnason drying (Hurrell 1984).
and Carpenter (1970) demonstrated that the heat- Some amino acids may be oxidized by react-
ing of bovine plasma albumin for 27 h at 115 °C ing with free radicals formed by lipid oxidation.
resulted in a 50% loss of cystine and a 4% loss of Methionine can react with a lipid peroxide to
lysine. These authors suggest that amide-type yield methionine sulfoxide. Cysteine can be
bonds are formed by reaction between the decomposed by a lipid free radical according to
ε-amino group of lysine and the amide groups of the following scheme:
asparagine or glutamine, with the reacting units
present either in the same peptide chain or in
neighboring ones (Fig. 3.17). The Maillard reac-
tion leads to the formation of brown pigments, or
melanoidins, which are not well defined and may
result in numerous flavor and odor compounds.
The browning reaction may also result in the The decomposition of unsaturated fatty acids
blocking of lysine. Lysine becomes unavailable produces reactive carbonyl compounds that may
when it is involved in the Amadori reaction, lead to reactions similar to those involved in non-
the first stage of browning. Blockage of lysine is enzymic browning. Methionine can be oxidized
nonexistent in pasteurization of milk products, under aerobic conditions in the presence of SO2
and is at 0–2% in UHT sterilization, 10–15% in as follows:

This reaction is catalyzed by manganese ions losses occurred of the amino acids lysine, trypto-
at pH values from 6 to 7.5. SO2 can also react phan, and histidine. Methionine was extensively
with cystine to yield a series of oxidation prod- oxidized to its sulfoxide. Increasing water activ-
ucts. Some of the possible reaction products ity increased losses of lysine and tryptophan but
resulting from the oxidation of sulfur amino had no effect on methionine oxidation.
acids are listed in Table 3.12. Nielsen et al. (1985) Alkali treatment of proteins is becoming more
studied the reactions between protein-bound common in the food industry and may result in
amino acids and oxidizing lipids. Significant several undesirable reactions. When cystine is
138 3  Amino Acids and Proteins

treated with calcium hydroxide, it is transformed free sulfur, and 2-methyl thiazolidine-2,
into amino-acrylic acid, hydrogen sulfide, 4-­dicarboxylic acid as follows:

This can also occur under alkaline conditions, Amino-acrylic acid (dehydroalanine) is very
when cystine is changed into amino acrylic acid reactive and can combine with the ε-amino
and thiocysteine by a β-elimination mechanism, group of lysine to yield lysinoalanine (Ziegler
as follows: 1964) as shown:

Table 3.12  Oxidation products of the sulfur-containing


amino acids
Name Formula
Methionine R–S–CH3
Sulfoxide R–SO–CH3
Sulfone R–SO2–CH3
Cystine R–S–S–R
Disulfoxide R–SO–
SO–R
Disulfone R–SO2–
SO2–R
Cysteine R–SH
Sulfenic R–SOH
Sulfinic R–SO2H
Sulfonic (or cysteic R–SO3H
acid)
Functional Properties 139

Lysinoalanine formation is not restricted to More severe treatment with alkali can decompose
alkaline conditions—it can also be formed by arginine into ornithine and urea. Ornithine can
prolonged heat treatment. Any factor favoring combine with dehydroalanine in a reaction simi-
lower pH and less drastic heat treatment will lar to the one giving lysinoalanine and, in this
reduce the formation of lysinoalanine. Hurrell case, omithinoalanine is formed.
(1984) found that dried whole milk and UHT Treatment of proteins with ammonia can
milk contained no lysinoalanine and that evapo- result in addition of ammonia to dehydroalanine
rated and sterilized milk contained 1000 ppm. to yield β-amino-alanine as follows:

Light-induced oxidation of proteins has been negatively charged planar carbanion. When a
shown to lead to off-flavors and destruction of proton is added back to this optically inactive
essential amino acids in milk. Patton (1954) dem- intermediate, either a d- or l-enantiomer may be
onstrated that sunlight attacks methionine and formed (Masters and Friedman 1980).
converts it into methional (β-­methylmercaptoprop Racemization leads to reduced digestibility and
ionaldehyde), which can cause a typical sunlight protein quality.
off-flavor at a level of 0.1 ppm. It was later dem-
onstrated by Finley and Shipe (1971) that the
source of the light-induced off-flavor in milk Functional Properties
resides in a low-density lipoprotein fraction.
Proteins react with polyphenols such as phe- Increasing emphasis is being placed on isolating
nolic acids, flavonoids, and tannins, which occur proteins from various sources and using them as
widely in plant products. These reactions may food ingredients. In many applications functional
result in the lowering of available lysine, protein properties are of great importance. Functional
digestibility, and biological value (Hurrell 1984). properties have been defined as those physical
Racemization is the result of heat and alkaline and chemical properties that affect the behavior
treatment of food proteins. The amino acids pres- of proteins in food systems during processing,
ent in proteins are of the L-series. The racemiza- storage, preparation, and consumption (Kinsella
tion reaction starts with the abstraction of an 1982). A summary of these properties is given in
α-proton from an amino acid residue to give a Table 3.13.
140 3  Amino Acids and Proteins

Table 3.13  Functional properties of food proteins


General property Functional criteria
Organoleptic Color, flavor, odor
Kinesthetic Texture, mouth feel, smoothness, grittiness, turbidity
Hydration Solubility, wettability, water absorption, swelling, thickening, gelling, syneresis, viscosity
Surface Emulsification, foaming (aeration, whipping), film formation
Binding Lipid-binding, flavor-binding
Structural Elasticity, cohesiveness, chewiness, adhesion, network cross-binding, aggregation, dough
formation, texturizability, fiber formation, extrudability
Rheological Viscosity, gelation
Enzymatic Coagulation (rennet), tenderization (papain), mellowing (“proteinases”)
“Blendability” Complementarity (wheat-soy, gluten-casein)
Antioxidant Off-flavor prevention (fluid emulsions)
Source: From J.E. Kinsella, Structure and Functional Properties of Food Proteins, in Food Proteins, P.F. Fox and
J.J. Condon, eds., 1982, Applied Science Publishers

Even when protein ingredients are added to Surface Activity of Proteins


food in relatively small amounts, they may sig-
nificantly influence some of the physical proper- Proteins can act as surfactants in stabilizing
ties of the food. Hermansson (1973) found that emulsions and foams. To perform this function
addition of 4% of a soybean protein isolate to proteins must be amphiphilic just of Food
processed meat significantly affected firmness, as Proteins like the emulsifiers discussed in Chap. 2.
measured by extrusion force, compression work, This is achieved when part of the protein struc-
and sensory evaluation. ture contains predominantly amino acids with
The emulsifying and foaming properties of hydrophobic side chains and another part con-
proteins relate to their adsorption at interfaces tains mostly hydrophilic side chains. The mole-
and to the structure of the protein film formed cule is then able to orient itself in the oil-water
there (Mitchell 1986). The emulsifying and emul- interface. Thus, the ability of proteins to serve as
sion stabilizing capacity of protein meat additives emulsifiers varies greatly among proteins. The
is important to the production of sausages. The emulsifying capacity of a protein depends not
emulsifying properties of proteins are also only on its overall hydrophobicity but, more
involved in the production of whipped toppings importantly, on the distribution of the hydropho-
and coffee whiteners. The whipping properties bic or charged groups along the polypeptide
of proteins are essential in the production of chain and the manner in which the chain is folded
whipped toppings. Paulsen and Horan (1965) (Dalgleish 1989). Hydrophobic side chains are
determined the functional characteristics of edi- likely to be folded into the inside of the molecule
ble soya flours, especially in relation to their use leaving the outside more hydrophilic. To be
in bakery products. They evaluated the measur- effective surfactants, proteins need to have flexi-
able parameters of functional properties such as ble polypeptide chains, so that they are able to
water dispersibility, wettability, solubility, and orient at the interface. Only proteins that have
foaming characteristics as those properties little secondary structure and are able to unfold at
affected the quality of baked products containing the interface are effective emulsifiers. Nakai and
added soya flour. Powrie (1981) have shown the relationship
Some typical functional properties of food between solubility, charge frequency, and hydro-
proteins are listed in Table 3.14. phobicity in graphical manner (Fig. 3.18) and in
Functional Properties 141

Table 3.14  Functional properties of proteins in food systems


Functional property Mode of action Food system
Solubility Protein solvation Beverages
Water absorption Hydrogen bonding of water; Meat, sausages, bread, cakes
and binding entrapment of water (no drip)
Viscosity Thickening; water binding Soups, gravies
Gelation Protein acts as adhesive material Meat, sausages, baked goods, pasta products
Elasticity Hydrophobic bonding in gluten; Meats, bakery products
disulfide links in gels
Emulsification Formation and stabilization of fat Sausages, bologna, soup, cakes
emulsion
Fat absorption Binding of free fat Meats, sausages, doughnuts
Flavor-binding Adsorption, entrapment, release Simulated meats, bakery products, etc.
Foaming Forms stable films to entrap gas Whipped toppings, chiffon, desserts, angel cakes
Source: From J.E. Kinsella, Structure and Functional Properties of Food Proteins, in Food Proteins, P.F. Fox and
J.J. Condon, eds., 1982, Applied Science Publishers

Fig. 3.18 Relationship
between solubility, charge
frequency, and
hydrophobicity of proteins.
Source: Reprinted with
permission from S. Nakai
and W.D. Powrie,
Modification of Proteins
for Functional and
Nutritional Improvements,
in Cereals—A Renewable
Resource, Theory and
Practice, Y. Pomeranz and
L. Munck, eds., p. 225, ©
1981, American
Association of Cereal
Chemists

the form of a table relating these parameters to can be stabilized by steric factors. This happens
the functional properties of proteins (Table 3.15). when disordered protein molecules at the interface
There are two important considerations in prevent emulsion droplets from approaching one
emulsion formation: the binding of the protein to another closely enough to permit coagulation as a
the oil-water interface and the stability of the result of attraction by van der Waals forces
emulsion. For an emulsion to possess stability, the (Dalgleish 1989).
proteins have to form a cohesive film. The cohe- The amount of protein required to form a sta-
siveness of such films is stabilized by ­intramolecular ble emulsion depends on the size of emulsion
disulfide bonds. In addition, emulsion particles droplets produced and the nature of the protein.
142 3  Amino Acids and Proteins

Table 3.15  Contribution of hydrophobicity, charge frequency, and structural parameters to functionality
of proteins
Hydrophobicity Charge frequency Structure
Solubility − + −
Emulsification +sur (−) +
Foaming +tot − +
Fat binding +sur (−) −
Water holding − + ?
Heat coagulation +tot − +
Dough making (+) − +
+: positive contribution; −: negative contribution; sur: surface hydrophobicity; tot: total hydrophobicity;
( ): contributes to a lesser extent
Source: Reprinted with permission from S. Nakai and W.D. Powrie, Modification of Proteins for
Functional and Nutritional Improvements, Cereals—Renewable Resource, Theory and Practice,
Y. Pomeranz and L. Munck, eds., p. 235, © 1981, American Association of Cereal Chemists

The size of the droplets, assuming the presence association and, therefore, the nature of the gel
of sufficient protein to cover the interfacial area, depends on a variety of covalent and noncovalent
is determined by the input of mechanical energy interactions involving disulfide bonds, hydrogen
by a device such as a homogenizer or colloid mill. bonds, ionic and hydrophobic interactions, or
According to Dalgleish (1989), the protein load combinations of these.
for monolayer coverage of the oil-water interface Protein gels can be divided into two types:
is in the order of a few mg/m2. β-casein is an aggregated gels and clear gels (Barbut 1994).
effective emulsifier protein and has been reported Aggregated gels are formed from casein and from
to give an interfacial loading of 2–3 mg/m2. egg white proteins and are opaque because of the
relatively large size of the protein aggregates. Clear
gels are formed from smaller particles, such as
those formed from whey protein isolate, and have
Gel Formation high water-holding capacity. The formation of such
gels from ovalbumin is illustrated in Fig. 3.19
Proteins can form gels by acid coagulation, action (Hatta and Koseki 1988), showing the influence of
of enzymes, heat, and storage. A gel is a protein protein concentration, pH, and ionic strength.
network that immobilizes a large amount of Many dairy products involve gel formation through
water. The network is formed by protein-protein the action of acid or by combined activity of acid
interactions. Gels are characterized by having and enzymes. Mayonnaise is an oil-in-water emul-
relatively high non-Newtonian viscosity, elastic- sion, in which egg yolk protein acts as the emulsi-
ity, and plasticity. Examples of protein gels are a fier. The presence of acetic acid in the form of
variety of dairy products including yogurt, soy- vinegar or citric acid from lemon juice leads to
bean curd (tofu), egg protein gels including may- interaction of the proteins covering the emulsion
onnaise, and meat and fish protein gels. Some droplets, resulting in a gel-type emulsion.
types of gel formation are reversible, especially
those produced by heat. Gelatin gels are pro-
duced when a heated solution of gelatin is cooled.
This sol-gel transformation is reversible. Most Animal Proteins
other types of gel formation are not reversible.
Gelation has been described as a two-stage pro- Milk Proteins
cess (Pomeranz 1991). The first stage is a dena-
turation of the native protein into unfolded The proteins of cow’s milk can be divided into
polypeptides, and the second stage is a gradual two groups: the caseins, which are phosphopro-
association to form the gel matrix. The type of teins and comprise 78% of the total weight, and
Animal Proteins 143

Fig. 3.19  Model for the formation of a gel network by of SH Groups to Functionality of Ovalbumin, in Food
heated ovalbumin, pi = isoelectric point. Source: Reprinted Proteins, J.E. Kinsella and W.G. Soucie, eds., p. 265, ©
with permission from H. Hatta and T. Koseki, Relationship 1988, American Oil Chemists’ Society

Table 3.16  Protein composition of mature bovine herd Table 3.17  Amino acid composition of milk proteins
milk
Amino acid Protein (g/kg)
Protein g/L Essential amino acids
Total protein 36  Threonine 46
Total casein 29.5  Valine 66
Whey protein 6.3  Methionine 26
αs1-Casein 11.9  Cystine 8
αs2-Casein 3.1  Isoleucine 59
β-Casein 9.8  Leucine 97
κ-Casein 3.5  Phenylalanine 49
γ-casein 1.2  Lysine 81
α-Lactalbumin 1.2  Histidine 27
β-Lactoglobulin 3.2  Arginine 35
Serum albumin 0.4  Tryptophan 17
Immunoglobulin 0.8 Nonessential amino acids
Proteose-peptones 1.0  Aspartic acid, asparagine 79
Source: Reprinted with permission from H.E. Swais-good,  Serine 56
Protein and Amino Acid Composition of Bovine Milk, in  Glutamic acid, glutamine 219
Handbook of Milk Composition, R.G. Jensen, ed., p. 465,
 Proline 99
© 1982, Academic Press
 Glycine 20
 Alanine 34
the milk serum proteins, which make up 17% of  Tyrosine 51
the total weight. The latter group includes
β-lactoglobulin (8.5%), a lactalbumin (5.1%),
immune globulins (1.7%), and serum albumins. and the amino acid composition of the milk
In addition, about 5% of milk’s total weight is ­proteins is shown in Table 3.17.
nonprotein nitrogen (NPN)-containing sub- Casein is defined as the heterogeneous group
stances, and these include peptides and amino of phosphoproteins precipitated from skim milk
acids. Milk also contains very small amounts of at pH 4.6 and 20 °C. The proteins remaining in
enzymes, including peroxidase, acid phospha- solution, the serum or whey proteins, can be sep-
tase, alkaline phosphatase, xanthine oxidase, and arated into the classic lactoglobulin and lactalbu-
amylase. The protein composition of bovine herd min fractions by half saturation with ammonium
milk is listed in Table 3.16 (Swaisgood 1995), sulfate or by full saturation with magnesium
144 3  Amino Acids and Proteins

Fig. 3.20  Separation of


milk proteins by
precipitation

casein can be separated from milk by rennet


action or by saturation with sodium chloride. The
composition of the casein depends on the method
of isolation. In the native state, the caseinate
­particles contain relatively large amounts of cal-
cium and phosphorus and smaller quantities of
­magnesium and citrate and are usually referred to
as calcium caseinatephosphate or calcium phos-
phocaseinate particles. When adding acid to
milk, the calcium and phosphorus are progres-
sively removed until, at the isoelectric point of
pH 4.6, the casein is completely free of salts.
Other methods of preparing casein yield other
products; for example, salt precipitation does not
remove the calcium and phosphorus, and rennet
Fig. 3.21  Electron photomicrograph of casein micelles action involves limited proteolysis. The rennet
in milk (http://www.usu.edu/westcent/microstructure_
food/Foods&bact.htm) casein is named paracasein.
Casein is a nonhomogeneous protein that con-
sists of four components, identified as αs1-, αs2-,
s­ulfate, as is shown in Fig. 3.20. However, this β-, and κ-casein. The γ-casein mentioned in the
separation is possible only with unheated milk. literature (see Table 3.16) has been identified as
After heating by, for example, boiling, about proteolytic fragments of β-casein (Wong et al.
80% of the whey proteins will precipitate with 1996). The four casein components occur as
the casein at pH 4.6; this property has been used genetic variants. Such genetically determined
to develop a method for measuring the degree of variants or polymorphs differ from one another
heat exposure of milk and milk products. by one or more amino acid substitutions and/or
Casein exists in milk as relatively large, nearly deletions. The complete amino acid sequence has
spherical particles of 30–300 nm in diameter been established, and the exact nature of the
(Fig.  3.21). In addition to acid precipitation, genetic variants has been determined. The genetic
Animal Proteins 145

Table 3.18  Genetic variants of caseins s­pecific hydrolysis of casein with proteolytic
Total amino acid Molecular enzymes has produced a number of large poly-
Variant residues weight peptides that resist further hydrolysis. An electro-
αs1-CN B-8P 199 23,614 phoretically homogeneous phosphopeptone has
αs2-CN A-11P 207 24,350 been isolated from a trypsin digest of β-casein by
β-CN A1-5P 209 23,982 Peterson et al. (1958). This phosphopeptone has a
κ-CN B-1P 169 19,023 molecular weight of about 3000 and consists of
24 amino acid residues of ten different amino
acids. The peptone contains five phosphate resi-
variants are described by the suffix CN (for dues linked to four serine and one threonine
casein) and a capital A, B, C, etc., as well as by groups. This constitutes essentially all of the
the number of phosphorylations. Genetic variants phosphorus of β-casein and it appears that the
of the four caseins are listed in Table 3.18 (Wong phosphate residues are localized in a relatively
et al. 1996). The caseins have sites of phosphory- small region of the casein molecule.
lation that have a unique amino acid sequence. In addition to ester phosphate, casein contains
The three–amino acid sequence is Ser-X-A, with calcium phosphate in the colloidal form. It
X being any amino acid and A either glutamic appears that the presence of this colloidal cal-
acid or serine-phosphate. One part of the α-casein cium phosphate helps maintain the structural
is precipitable by calcium ions and has been des- integrity of the casein micelle. Although the com-
ignated calcium-sensitive casein or αs. The non-­ position and structure of most of the casein frac-
calcium-­sensitive fraction, κ-casein, is the protein tions are now well established, the exact nature of
assumed to confer stability on the casein micelle; the arrangement of the caseins and calcium phos-
this has been found to be removed by the action phate into a micelle is not well known. Many
of rennin, thereby leaving the remaining casein models of micelle structure have been proposed
precipitable by calcium ions, κ-casein is the frac- (Farrell 1973; Farrell and Thompson 1974).
tion with the lowest phosphate content. The two These can be divided into three groups: coat-core
αs-caseins show strong association. The associa- models, internal structure models, and subunit
tion of β-casein is temperature dependent. At models. In the most popular, the coat-core model,
4 °C only monomers exist; at temperatures it is assumed that the core contains the calcium-­
greater than 8 °C association will occur. αsl-­ sensitive αs-caseins and that this core is covered
Casein has more acidic than basic amino acids by a layer of κ-casein. The function of the
and has a net negative charge of 22 at pH 6.5. The κ-casein coat is to protect the micelle from insol-
polypeptide chain contains 8.5% proline that is ubilization by calcium ions. The κ-casein is read-
distributed uniformly, resulting in no apparent ily attacked by the enzyme rennin, thus removing
secondary structure. αs2-Casein has the highest the coat and resulting in coagulation of the
number of phosphorylations and a low proline micelles. This model most readily accounts for
content, β-casein is a single polypeptide chain the action of rennin but does not explain the posi-
with a total of 209 amino acids, and has seven tion of the colloidal calcium phosphate.
genetic variants. The distribution of amino acids It appears that micelles are formed by cross-­
in the polypeptide chain is quite specific. The linking of some of the ester phosphate groups by
N-terminal segment has a high negative charge, calcium. Chelation of calcium results in dissocia-
giving it hydrophilic properties, the C-terminal tion and solubilization of the micelles, and the
portion is highly hydrophobic. This arrangement rate at which this happens corresponds to the
lends surfactant properties to this protein. ester phosphate content of the monomers (Aoki
Casein contains 0.86% phosphorus, and it is et al. 1987).
assumed that this is present exclusively in the The whey proteins of milk were originally
form of monophosphate esters with the hydroxyl thought to be composed of two main compo-
groups of serine and threonine. Limited and nents, lactalbumin and lactoglobulin, as indicated
146 3  Amino Acids and Proteins

Fig. 3.22  Production of protein products from skim milk

in Fig. 3.22. Then it was found that the lactalbumin heated milk. The cysteine group is also involved
contains a protein with the characteristics of a in thermal denaturation. At pH 6.7 and above
globulin. This protein, known as β-lactoglobulin, 67 °C, β-lactoglobulin denatures, followed by
is the most abundant of the whey proteins. It has aggregation. The first step in the denaturation is a
a molecular weight of 36,000. In addition to series of reversible conformational changes that
β-lactoglobulin, the classic lactalbumin fraction result in exposure of cysteine. The next step
contains α-lactalbumin, serum albumin, and at involves association through sulfhydryl-disulfide
least two minor components. exchange.
β-lactoglobulin is rich in lysine, leucine, The differences between genetic variants,
­glutamic acid, and aspartic acid. It is a globular although minor, may result in marked changes in
protein with five known genetic variants. Variants some properties (Aschaffenburg 1965). The two
A and B have 162 amino acids and molecular chains of β-lactoglobulin C differ from the chains
weights of 18,362 and 18,276, respectively. of the B variant in that a histidine residue has
β-lactoglobulin has a tightly packed structure and taken the place of a glutamic acid or glutamine
consists of eight strands of antiparallel β sheets. residue. The A chains differ in two places from
The interior of the molecule is hydrophobic. The the B chains: aspartic acid replaces glycine and
molecular structure also contains a certain amount valine replaces alanine. Because of these minute
of α helix, which plays a role in the formation of differences, A is less soluble and more stable
the usually occurring dimer. The association is pH when heated than B. Variant A has a tendency to
dependent, β-lactoglobulin A will form octamers form tetramers at pH 4.5, whereas this tendency
at low temperature and high concentration and at is absent in B. These differences are thought to be
pH values between 3.5 and 5.2. Below pH 3.5 the the result of differences in the three-dimensional
protein dissociates into monomers. This protein is folding or tertiary structure of the amino acid
the only milk protein c­ontaining cysteine and, chains.
therefore, contains free sulfhydryl groups, which α-Lactalbumin occurs as genetic variants A
play a role in the development of cooked flavor in and B, each with 123 amino acid residues.
Meat Proteins 147

The molecular weight of A is 14,147 and B is


14,175. The amino acid sequence of α-lactalbumin
is very similar to that of hen egg-white lysozyme.
α-Lactalbumin has a high binding capacity for
calcium and some other metals. It is insoluble at
the isoelectric range from pH 4 to 5. The calcium
in α-lactalbumin is bound very strongly and pro-
tects the stability of the molecule against thermal
denaturation.
The immune globulins were previously
divided into euglobulin and pseudoglobulin. The
level of these proteins in colostrum is very high
and they have been shown to be transferred to the
blood of the young calf, indicating that they are
absorbed unchanged. The three classes of immu-
noglobulins in milk are designated IgM(γM),
IgA(γA), and IgG(γG) (Gordon and Kalan 1974).
IgG is subdivided into IgG1 and IgG2. The serum Fig. 3.23  Processing of whey to produce whey protein
albumin has been shown to be identical to the concentrate by reverse osmosis (RO) and ultrafiltration
blood serum albumin. (UF). Source: From C. V. Morr, Production and Use of
Milk Proteins in Food, Food Technol., Vol. 38, No. 7,
Nonfat milk (skim milk) is the raw material pp. 39–18, 1984
from which a number of milk protein products
are manufactured. A schematic diagram of the
various products obtained by processing of skim
milk is given in Fig. 3.22 (Wong et al. 1996).
These products are used as raw materials in Meat Proteins
many manufactured foods and include caseins,
caseinates, and coprecipitates (Morr 1984). Acid Meromysin
casein results from isoelectric precipitation of
casein at pH 4.6–4.7. The curd is recovered by The proteins of muscle consist of about 70%
centrifugation, then washed and dried. Alkali structural or fibrillar proteins and about 30%
neutralization of the wet curd yields caseinate, water-soluble proteins. The fibrillar proteins con-
which is spray dried. Rennet casein is made by tain about 32–38% myosin, 13–17% actin, 7%
rennet coagulation followed by washing and dry- tropomyosin, and 6% stroma proteins. Meat and
ing of the curd. Coprecipitates include both fish proteins contribute to highly organized struc-
casein and whey proteins and are made from tures that lend particular properties to these prod-
heated skim milk. The heating denatures the ucts. Some of the other proteins discussed in this
whey proteins, which can then be precipitated chapter are more or less globular and consist of
with acid together with the casein. particles that are not normally involved in an
Whey protein concentrate is made from whey, extensive structural array. Examples are milk
the by-product of cheese making. Removal of proteins and the protein bodies in cereals and oil-
lactose and minerals requires reverse osmosis seeds. Extensive structure formation involving
end ultrafiltration processing (Fig. 3.23). these proteins may occur in various technological
An up-to-date coverage of our present knowl- processes such as making cheese from milk
edge of milk proteins is given by Wong et al. or texturized vegetable protein products from
(1996). oilseeds.
148 3  Amino Acids and Proteins

The fibrils are optically nonuniform, which


accounts for the striated appearance. Compounds
with different refractive indexes are arranged
along the fiber.
Meat contains three general types of proteins:
the soluble proteins, which can easily be removed
by extraction with weak salt solutions (ionic
strength ≤0.1); the contractile proteins; and the
stroma proteins of the connective tissue. The sol-
uble proteins are classed as myogens and myoal-
bumins. The myogens are a heterogeneous group
of metabolic enzymes. After extraction of the
soluble proteins, the fibril and stroma proteins
remain. They can be extracted with buffered
0.6 M potassium chloride to yield a viscous gel of
actomyosin.
Myosin is the most abundant of the muscle
proteins and makes up about 38% of the total.
Myosin is a highly asymmetric molecule with a
molecular weight of about 500,000 that contains
about 60–70% α-helix structure. The molecule
has a relatively high charge and contains large
amounts of glutamic and aspartic acids and diba-
sic amino acids. Myosin has enzyme activity and
can split ATP into ADP and monophosphate,
thereby liberating energy that is used in muscle
contraction. The myosin molecule is not a single
Fig. 3.24  Microscopic structure of striated muscle in entity. It can be separated into two subunits by
longitudinal section means of enzymes or action of the ultracentri-
fuge. The subunits with the higher molecular
Muscle is made up of fibers that are several weight, about 220,000, are called heavy mero-
centimeters long and measure 0.01–0.1 mm in myosin. Those with low molecular weight, about
diameter. The fibers are enclosed in membranes 20,000 are called light meromyosin. Only the
called sarcolemma and are arranged in bundles heavy meromyosin has ATP-ase activity.
that enclose fat and connective tissue. The fibers Actin makes up about 13% of the muscle pro-
are cross-striated, as indicated in Fig. 3.24, and tein, so the actin-myosin ratio is about 1:3. Actin
this is due to the presence of cross-striated myo- occurs in two forms: G-actin and F-actin (G and
fibrils. The myofibrils are embedded in the cell F denote globular and fibrous). G-actin is a
cytoplasm called sarcoplasm. The fibers contain monomer that has a molecular weight of about
peripherally distributed nuclei; a diagram of the 47,000 and is a molecule of almost spherical
arrangement of the various constituents of a mus- shape. Because of its relatively high proline con-
cle fiber is given in Fig. 3.25 (Cassens 1971). In tent, it has only about 30% of α-helix configura-
addition to the constituents mentioned, the mus- tion. F-actin is a large polymer and is formed
cle fibers contain other components including when ATP is split from G-actin. The units of actin
mitochondria, ribosomes, lysosomes, and glyco- combine to form a double helix of indefinite
gen granules. The fibers make up the largest part length, and molecular weights of actin have been
of the muscle volume, but there is from 12 to reported to be in the order of several millions.
18% of extracellular space. Bodwell and McClain (1971) indicate that actin
Collagen 149

Fig. 3.25 Diagrammatic Thin Filaments


representation of a Nucleus Thick Filaments
muscle fiber (http://
people.eku.edu/ Sarcoplasmic
reticulum
ritchisong/301notes3.
htm) Myofibril

T-tubules
Sarcolemma

Sarcolemma Sarcoplasm
Mitochondria

Z-line

I-band

H-band
M-line
A-band

One sarcomere

Copyright © 2006 Nature Publishing Group


Nature Reviews | Molecular Cell biology

polymers may have an apparent molecular weight


of over 14,000,000, with a length of 1160 nm and
Collagen
a diameter of 6 nm. The transformation of G-actin
The contractile meat proteins are separated and
into F-actin is schematically represented in
surrounded by layers of connective tissues. The
Fig. 3.26.
amount and nature of this connective tissue is an
Actomyosin is a complex of F-actin and myo-
important factor in the tenderness or toughness
sin and is responsible for muscle contraction and
and the resulting eating quality of meat. Collagens
relaxation. Contraction occurs when myosin
form the most widely occurring group of proteins
ATP-ase activity splits ATP to form phosphory-
in the animal body. They are part of the connec-
lated actin and ADP. For this reaction, the pres-
tive tissues in muscle and organs, skin, bone,
ence of K+ and Mg2+ is required. Relaxation of
teeth, and tendons. The collagens are a distinct
muscle depends on regeneration of ATP from
class of proteins as can be demonstrated by X-ray
ADP by phosphorylation from creatine phos-
diffraction analysis. This technique shows that
phate. The precise mechanism of contraction is
collagen fibrils have regular periodicity of 64 nm,
still unknown, although a working hypothesis is
which can be increased under tension to 400 nm.
available (Bailey 1982).
Collagen exists as a triple helix; the formation of
The composition of various cuts of meats and
collagen into the triple helix is shown in Fig. 3.27
their structural implications have been described
(Yamauchi and Sricholpech 2012). The biosyn-
by Ranken (1984).
thesis of collagen is a long complicated including,
150 3  Amino Acids and Proteins

Fig. 3.26  Transformation of G-actin into F-actin from dimers to trimers to polymeric F-actin

Fig. 3.27  Collagen peptides fold to form a right-handed length to form a fibril. Source: From Yamauchi M.,
superhelix. This is the tropocollagen molecule. Molecules Sricholpech (2012)
line up in a staggered fashion to overlap by one-quarter
Collagen 151

chain association and folding, secretion, The last of these is the only change essential
­procollagen processing, self-assembly and pro- for the conversion of collagen to gelatin. The
gressive cross-linking (Fig. 3.27). Type I collagen conditions employed during the production of
is a long (∼300 nm long, ∼1.5 nm thick) heterotri- gelatin determine its characteristics. If there is
meric molecule composed of two α1 chains and extensive breakdown of peptide bonds, many lat-
one α2 chain. An α1 homotrimeric form exists as eral bonds may remain intact and soluble frag-
a minor form. The molecule consists of three ments are produced. If many lateral bonds are
domains: the N-terminal non-triple helical domain destroyed, the gelatin molecules may have rela-
(N-telopeptide), the central triple helical domain and tively long chain lengths. Thus, there is a great
the C-terminal non-triple helical domain variety of gelatins. In normal production, the
(C-telopeptide). The single (uninterrupted) triple hides or bones are extracted first, under relatively
helical domain represents more than 95% of the mild conditions, followed by successive extrac-
molecule (Yamauchi and Sricholpech 2012). tions under more severe conditions. The first
The triple helix is the tropocollagen molecule; extraction yields the best-quality gelatin. The
these are lined up in a staggered array, overlap- term gelatin is used for products derived from
ping by one-quarter of their length to form a mammalian collagen that can be dispersed in
fibril. The fibrils are stacked in layers to form water and show a reversible sol-gel change with
connective tissue. Important in the formation of temperature. The gels formed by gelatin can be
these structures is the high content of hydroxy- considered as a partial return of the molecules to
proline and hydroxylysine. The content of diba- an ordered state. However, the return to the highly
sic and diacidic amino acids is also high, but ordered state of collagen is not possible. High-­
tryptophan and cystine are absent. As a result of quality gelatin has an average chain length of
this particular amino acid composition, there are 60,000–80,000, whereas the value for native col-
few interchain cross-bonds, and collagen swells lagen is infinite.
readily in acid or alkali. The process of gel formation is probably
Heating of collagen fibers in water to 60–70 °C closely associated with the presence of guanidino
shortens them by one-third or one-fourth of the groups of arginine. Hypobromite has the ability
original length. This temperature is characteristic to destroy guanidino groups and, when added to
of the type of collagen and is called shrink tem- gelatin, it inhibits gelation.
perature (Ts). The Ts of fish skin collagen is very There are three types of gelatin: alpha, with
low, 35 °C. When the temperature is increased to a molecular weight of 80,000–125,000; beta,
about 80 °C, mammalian collagen changes into with molecular weight of 160,000–250,000; and
gelatin. Certain amino acid sequences are common gamma, with molecular weight of 240,000–
in collagen, such as Gly-Pro-Hypro-Gly. In a triple 375,000 (Poppe 1992). The typical amino acid
helix, only certain sequences are permissible. The sequence in gelatin is Gly-X-Y, where X is
structural unit of the collagen fibrils is tropocolla- mostly proline and Y is mostly hydroxyproline
gen with a length of 280 nm, a diameter of 1.5 nm, (Fig. 3.28).
and a molecular weight of 360,000. Gelatin is a When gelatin is placed in cold water it will
soluble protein derived from insoluble collagen. absorb 5–10 times its own weight in water and
Although it can be made from different animal by- swell. When this material is heated to above the
products, hide is the common source of gelatin pro- melting point, between 27 and 34 °C, the swollen
duction. The process of transforming collagen into gelatin dissolves. This solgel transformation is
gelatin involves the following three changes: reversible. Poppe (1992) has described the mech-
anism of gel formation. It involves the reversion
1. rupture of a limited number of peptide bonds from a random coil to a helix structure. Upon
to reduce the length of the chains cooling, the imino acid-rich regions of different
2. rupture or disorganization of a number of the chains form a helical structure, which is s­ tabilized
lateral bonds between chains by hydrogen bonding. This then forms the three-
3. change in chain configuration dimensional gel matrix.
152 3  Amino Acids and Proteins

Fig. 3.28  Amino acid sequence in gelatin

Table 3.19  Division of the proteins of fish flesh according to solubility


Ionic strength at which soluble Name of group Location
Equal to or greater than 0 “Myogen” easily soluble Mainly sarcoplasm, muscle cell juice
Greater than about 0.3 “Structural” less soluble Mainly myofibrils, contractile elements
Insoluble “Stroma” Mainly connective tissues, cell walls, etc.

Commercial gelatin is available in two types, about three-quarters of the total muscle protein.
A and B. Type A gelatin has undergone an acid Fish actomyosin has been found to be quite labile
pretreatment and type B a lime pretreatment. and easily changed during processing and stor-
They differ in their viscosity and their ability to age. During frozen storage, the actomyosin
combine with negatively charged hydrocolloids becomes progressively less soluble, and the flesh
such as carrageenan. becomes increasingly tough. Connell (1962) has
described the changes that may occur in cod
myosin. When stored in dilute neutral solution,
Fish Proteins myosin rapidly denatures and forms aggregates
in a step-wise manner as follows:
The proteins of fish flesh can be divided into M → MD (3.1)

three groups on the basis of solubility, as indi-
cated in Table 3.19. The skeletal muscle of fish M D + M D → 2M D 
 (3.2)
consists of short fibers arranged between sheets 2 M D + M D → 3M D 
of connective tissue, although the amount of con-
nective tissue in fish muscle is less than that in Equation (3.1) represents the change from
mammalian tissue and the fibers are shorter. The native to denatured protein and follows first-order
myofibrils of fish muscle have a striated appear- reaction kinetics. In successive steps represented
ance similar to that of mammalian muscle and by Eq. (3.2), dimers, trimers, and higher poly-
contain the same major proteins, myosin, actin, mers are formed in a concentration-dependent
actomyosin, and tropomyosin. The soluble pro- reaction pattern. The aggregation is assumed to
teins include most of the muscle enzymes and be mostly in a lateral fashion with only little end-­
account for about 22% of the total protein. The to-­end aggregation. The instability of fish myosin
connective tissue of fish muscle is present in appears to be one of the major factors causing the
lower quantity than in mammalian muscle; the lability of fish muscle.
tissue has different physical properties, which The interest in using fish for the production of
result in a more tender texture of fish compared fish protein concentrate has waned because the
with meat. The structural proteins consist mainly product lacks satisfactory functional properties.
of actin and myosin, and actomyosin represents More promising ways of using fish resources
Egg Proteins 153

Table 3.20  Protein composition of egg white


Approximate Approximate isoelectric
Constituent amount (%) point (pH) Unique properties
Ovalbumin 54 4.6 Denatures easily, has sulfhydryls
Conalbumin 13 6.0 Complexes iron, antimicrobial
Ovomucoid 11 4.3 Inhibits enzyme trypsin
Lysozyme 3.5 10.7 Enzyme for polysaccharides
antimicrobial
Ovomucin 1.5 4.5 Viscous, high sialic acid, reacts
with viruses
Flavoprotein-apoprotein 0.8 4.1 Binds riboflavin
“Proteinase inhibitor” 0.1 5.2 Inhibits enzyme (bacterial
proteinase)
Avidin 0.05 9.5 Binds biotin, antimicrobial
Unidentified proteins 8 5.5, 7.5, 8.0, 9.0 Mainly globulins
Nonprotein 8 Primarily half glucose and salts
(poorly characterized)
Source: From R.R. Feeney and R.M. Hill, Protein Chemistry and Food Research, in Advances in Food Research,
Vol. 10, C.O. Chichester, E.M. Mrak, and G.F. Stewart, eds., 1960, Academic Press

involve fish protein gels (surimi), which can be phosphate groups and another component with
made into attractive consumer products (Mackie one phosphate group. Some of the diphosphate
1983). changes to monophosphate on storage. Oval­
bumin is readily denatured by heat.
Conalbumin has a molecular weight of 70,000
Egg Proteins and has iron-binding and antimicrobial pro­
perties. It can render iron unavailable to micro­
The proteins of eggs are characterized by their organisms; this property is lost after heat
high biological value and can be divided into the denaturation. Iron is bound in the ferric form by
egg white and egg yolk proteins. The egg white coordination. The groups involved in the binding
contains at least eight different proteins, which of iron are amino, carboxyl, guanidine, and
are listed in Table 3.20. Some of these proteins amides. When these groups are blocked, the iron-
have unusual properties, as indicated in the table; binding property is lost.
for example, lysozyme is an antibiotic, ovomu- Ovomucoid is a trypsin inhibitor and a gly­
coid is a trypsin inhibitor, ovomucin inhibits coprotein with a molecular weight of 27,000–
hemagglutination, avidin binds biotin, and conal- 29,000, containing mannose and glucosamine.
bumin binds iron. The antimicrobial properties This protein is highly resistant to denaturation.
help to protect the egg from bacterial invasion. Lysozyme is classed as a globulin and has
Liquid egg white contains 10–11% of protein, the ability to cause lysis of bacterial cells. There are
and the dried form contains about 83%. The most three fractions, designated G1, G2, and G3. The
abundant protein is ovalbumin, a phosphoprotein activity resides in the G1 fraction. The protein has a
with a molecular weight of 45,000 that contains a molecular weight of 14,000–17,000 and is a basic
small proportion of carbohydrate. The carbohy- protein with unusually high content of histidine,
drate is present as a polysaccharide composed of arginine, and lysine. It is stable to many agents that
two glucosamine and four mannose groups. denature other proteins, such as heat, cold, and
Ovalbumin can be separated by electrophoresis denaturing reagents. It is also quite resistant to pro-
into two components, one component with two teolysis by enzymes such as papain and trypsin.
154 3  Amino Acids and Proteins

Table 3.21  Protein components of egg yolk Plant Proteins


Approximate
Constituent amount (%) Particular properties As with animal proteins, plant proteins occur in
Livetin 5 Contains wide variety. Many plant proteins have until
enzymes—poorly
recently received much less study than the animal
characterized
Phosvitin 7 Contains 10%
proteins. This is gradually changing, and more
phosphorus information is now becoming available on many
Lipoproteins 21 Emulsifiers nontraditional food proteins. Proteins can be
(Total protein) (33) obtained from leaves, cereals, oilseeds, and nuts.
Source: From R.R. Feeney and R.M. Hill, Protein Leaf proteins have been extracted from macer-
Chemistry and Food Research, in Advances in Food ated leaves and are very labile. They are readily
Research, Vol. 10, C.O. Chichester, E.M. Mrak, and denatured at about 50 °C and undergo surface
G.F. Stewart, eds., 1960, Academic Press
denaturation in the pH range 4.5–6.0. Cereal seed
proteins are generally low in lysine. Peanut pro-
Ovomucin is an insoluble protein, which tein is poor in lysine, tryptophan, methionine,
p­recipitates from egg white on dilution with and threonine. Legume seeds are low in cyst(e)
water. It is not well known, has a high molecular ine and methionine. Great improvement in nutri-
weight (7,600,000), and is a mucoprotein. The tional value can sometimes be obtained by judi-
ability of certain viruses to agglutinate red blood cious mixing of different products.
cells, called hemagglutination, is inhibited by The proteins of cereal grains are very impor-
ovomucin. tant to their physical properties, even though the
Avidin is a protein characterized by its ability protein content of grains is not very high. Protein
to bind biotin and render it unavailable. Heat levels vary within wide limits, depending on spe-
denaturation destroys this property. cies, soil, fertilizer, and climate. The protein is
Egg yolk proteins precipitate when the yolk is nonuniformly distributed throughout the kernel,
diluted with water. The protein components of with the center having the lowest protein content.
egg yolk are listed in Table 3.21. The yolk con- Wheat has a protein content of 8–14%, rye about
tains a considerable amount of lipid, part of 12%, barley 10%, and rice 9%.
which occurs in bound form as lipoproteins.
Lipoproteins are excellent emulsifiers, and egg
yolk is widely used in foods for that reason. The Wheat Proteins
two lipoproteins are lipovitellin, which has
17–18% lipid, and lipovitellenin, which has Wheat proteins are unique among plant proteins
36–41% lipid. The protein portions of these and are responsible for bread-making properties
­compounds after removal of the lipid are named of wheat. The classic method of fractionation
vitellin and vitellenin. The former contains 1%% based on solubility characteristics indicates the
phosphorus, the latter 0.29%. presence of four main fractions (Fig. 3.29): albu-
The membranes of eggs consist of keratins min, which is water-soluble and coagulated by
and mucins. heat; globulin, soluble in neutral salt solution;
When fluid egg yolk is frozen, changes take gliadin, a prolamine soluble in 70% ethanol; and
place, causing the thawed yolk to form a gel glutenin, a glutelin insoluble in alcohol but solu-
(Powrie 1984). Gelation increases as the freezing ble in dilute acid or alkali.
temperature is lowered from −6 to −14 °C. The methods of gel electrophoresis and iso-
Gradual aggregation of lipoprotein is postulated electric focusing now provide highly efficient
as the cause of gelation. tools for separation of these proteins. By using
Plant Proteins 155

Fig. 3.29  Schematic representation of the main protein fractions of wheat flour. Source: From J. Holme, A Review of
Wheat Flour Proteins and Their Functional Properties, Bakers’ Dig., Vol. 40, No. 6, pp. 38–42, 78, 1966

these techniques, both gliadin and glutenin have Heat damage to gluten can result from
been shown to be complex mixtures. Gliadin and ­excessive air temperatures used in the drying of
glutenin are the storage, or gluten-forming, pro- wet grain. The gluten becomes tough and is more
teins of wheat. The formation of gluten takes difficult to extract. Heat-denatured wheat gives
place when flour is mixed with water. The gluten bread poor texture and loaf volume (Schofield
is a coherent elastic mass, which holds together and Booth 1983).
other bread components such as starch and gas Gliadin and glutenin are composed of many
bubbles, thus providing the basis for the crumb different molecular species. Gliadin proteins con-
structure of bread. The hydration of the gluten sist mostly of single-chain units and have molec-
proteins results in the formation of fibrils ular weights near 36,500 (Bietz and Wall 1972).
(Simmonds and Orth 1973), with gliadins form- Whole gliadin also contains polypeptides of
ing films and glutenins forming strands. molecular weight 11,400 that may be albumins, a
Gluten proteins have a high content of gluta- major polypeptide of molecular weight 44,200,
mine but are low in the essential amino acids and ω-gliadins of molecular weights 69,300 and
lysine, methionine, and tryptophan. The insolu- 78,100. The polypeptides of 44,200 and 36,500
bility of gluten proteins can be directly related to molecular weight are joined through disulfide
their amino acid composition. High levels of non- bonds into higher molecular weight proteins.
polar side chains result from the presence of glu- Glutenin consists of a series of at least 15 poly-
tamic and aspartic acids as the amides. Because peptides with molecular weights ranging from
these are not ionized, there is a high level of 11,600 to 133,000. These units are bound together
­apolar (hydrogen) bonding. This contributes to by disulfide bonds to form large molecules.
aggregation of the molecules and results in low The nongluten albumin and globulin proteins
solubility. represent from 13 to 35% of the total protein of
156 3  Amino Acids and Proteins

Fig. 3.30  Types of


disulfide bonds in cereal
proteins. Source: From
J.S. Wall, Disulfide
Bonds: Determination,
Location, and Influence
on Molecular Properties
of Proteins, Agr. Food
Chem., Vol. 19,
pp. 619–625, 1971

cereal flours. This protein fraction contains glutenin results in the unfolding of the peptide
­glycoproteins, nucleoproteins, lipoproteins, and a chains (Krull and Wall 1969), as indicated in
variety of enzymes. Fig.  3.31. This type of change has a profound
Chemical modification of gluten proteins effect on the rheological properties of dough
plays an important role in the industrial use of (Pomeranz 1968).
cereals. Especially reactions that lead to splitting
or formation of an SS bond can greatly influence
solubility and rheological properties such as Maize Proteins
extensibility and elasticity.
An example of the reduction of an SS bond by Maize (corn), wheat, and rice are the three main
means of an SH containing reagent is as follows: cereal crops of the world. The protein content of
maize varies widely depending on variety, cli-
mate, and other factors; it is generally in the
9–10% range. The main proteins of maize are the
storage proteins of the endosperm, namely zein
and glutelin. There are also minor amounts of
albumin and globulin. Maize proteins are low in
levels of the essential amino acids lysine and
tryptophan. To overcome this problem Mertz
et al. (1964) developed high-lysine corn,
Opaque-2. In this mutant the synthesis of zein,
the protein with the lowest lysine content, is
The disulfide bonds in wheat gluten play an suppressed.
important role in cross-linking polypeptide The storage proteins of maize can be divided
chains. Some of the bonds present in cereal into low molecular weight (zeins) and high
proteins are shown in Fig. 3.30 (Wall 1971).
­ molecular weight (glutelins). Zein is the protein
Reduction of the disulfide bonds in gliadin and group that is soluble in alcohols. The zeins can be
Rice Proteins 157

Fig. 3.31  Reduction of disulfide bonds in gliadin and glutenin. Source: From L.H. Krull and J.S. Wall, Relationship of
Amino Acid Composition and Wheat Protein Properties, Bakers’ Dig., Vol. 43, No. 4, pp. 30–39, 1969

separated into as many as 30 components, extensive processing such as milling and baking.
belonging to two major groups with molecular In contrast, rice is eaten mostly as the intact ker-
weights of about 22 and 24 kDa (Lasztity 1996). nel after the bran has been removed. In some
Zeins are asymmetric molecules containing about parts of the world, however, rice is also consumed
45% α-helix and 15% β-sheet; the rest is in the form of rice noodles. The protein content
aperiodic. of polished or white rice is lower than that of
The high molecular weight storage proteins are wheat. Values reported for protein content of rice
the glutelins. The glutelins are less well-­defined range from 6 to 9%. The nutritional value of rice
than the zeins. They have higher lysine, arginine, protein is high because of its relatively high con-
histidine, and tryptophan and lower glutamic acid tent of lysine, the first limiting essential amino
content than the zeins. Glutelin consists of several acid. Up to 18% of the protein in the rice kernel
subunits joined together by disulfide bonds. is lost in the bran and polish. In rice the main
The remaining proteins, present in amounts of storage protein is glutelin, in contrast to wheat,
about 8% each, are albumins and globulins. whose main storage protein is gliadin. The
These proteins are soluble in water and/or salt approximate protein distribution in rice proteins
solutions, and are characterized by higher levels is as follows: albumin 5%, globulin 10%, prola-
of essential amino acids and lower levels of glu- min 5%, and glutenin 80% (Lookhart 1991).
tamic acid. Albumins have higher contents of The protein in rice is present in the form of
aspartic acid, proline, glycine, and alanine, and encapsulated protein bodies, which are found
lower levels of glutamic acid and arginine than throughout the endosperm. The protein bodies
the globulins. may be small or large with the former containing
primarily glutelin and the latter containing prola-
min and glutelin (Hamaker 1994). The protein
Rice Proteins bodies are insoluble and remain intact during
cooking. Rice has little or no intergranular matrix
Rice and wheat are the two staple cereals for protein. This characteristic is different than in
much of the world population. The wheat kernel most other cereals, and it may have an effect in
is almost never eaten as such, but only after the process of noodle making.
158 3  Amino Acids and Proteins

Glutelin is the principal protein in the whole the unique proteins of wheat gluten. As a result,
grain as well as in milled rice and rice polish. The soy flour cannot be incorporated into bread with-
major proteins in rice bran are albumin and glob- out the use of special additives that improve loaf
ulin. This indicates that glutelin is concentrated volume. The soy proteins have a relatively high
in the milled rice and albumin and globulin are solubility in water or dilute salt solutions at pH
enriched in the bran and rice polish. These values below or above the isoelectric point. This
byproducts are mainly used as animal feed and means they are classified as globulins. There is as
would constitute a valuable food source if prop- yet no generally accepted nomenclature for the
erly processed. soy proteins, and only some of the common ter-
minology is used here. The complex character of
the mixture of proteins in soybeans is indicated
Soybean Proteins by the fact that starch gel electrophoresis of acid-
precipitated globulins in 5 M urea with alkaline
The proteins in soybeans are contained in protein buffer reveals 14 protein bands, and in acid buffer
bodies, or aleurone grains, which measure from 2 15 bands appear (Puski and Melnychyn 1968).
to 20 μm in diameter. The protein bodies can be Generally, soybean proteins are differentiated
visualized by electron microscopy (Fig. 3.32). on the basis of their behavior in the ultracentri-
Soy protein is a good source of all the essential fuge. Water-extractable proteins are separated
amino acids except methionine and tryptophan. into four fractions with approximate sedimenta-
The high lysine content makes it a good comple- tion rates of 2, 7, 11, and 15S. The percentage of
ment to cereal proteins, which are low in lysine. these fractions is indicated in Table 3.22, along
Soybean proteins have neither gliadin nor glutenin, with their components and their molecular

Fig. 3.32 Transmission
electron microscopy
observation of protein
bodies in transgenic
soybean seeds. Seeds
grown in presence of
0.5 mM magnesium
sulfate contain a few
endoplasmic reticulum-­
derived spherical protein
bodies (a, arrows) while
seeds grown in presence
of 2 mM magnesium
sulfate reveal numerous
dark staining spherical
protein bodies (b, c).
PSV, protein storage
vacuole; OB oil bodies,
PB protein body (http://
journal.frontiersin.org/
article/10.3389/
fpls.2014.00633/full)
Soybean Proteins 159

Table 3.22  Ultracentrifuge fractions of soybean proteins


Protein fraction Percentage of total Components Molecular weight
2S 22 Trypsin inhibitors 8000
21,500
Cytochrome c 12,000
2.3S Globulin 18,200
2.8S Globulin 32,000
Allantoinase 50,000
7S 37 Beta-amylase 61,700
Hemagglutinins 110,000
Lipoxygenases 108,000
7S Globulin 186,000–210,000
11S 31 11S Globulin 350,000
15S 11 – 600,000
Source: From W.J. Wolf, What Is Soy Protein, Food Technol., Vol. 26, No. 5, pp. 44–54, 1972b

weights. Several of the ultracentrifuge fractions Current information on soy protein fraction-
can be further separated into a number of compo- ation and nomenclature has been given by Brooks
nents. The 2S fraction contains trypsin inhibitors, and Moor (1985). The 7S globulins are classified
cytochrome c, allantoinase, and two globulins. into three major types. Type I is β-conglycinin
The 7S fraction contains β-amylase, hemagglu­ (B1–B6), type II is β-conglycinin (B0), and type
tinin, lipoxygenase, and 7S globulin. The 11S IIΙ is γ-conglycinin.
fraction consists mainly of 11S globulin. This Application of heat to soybeans or defatted
compound has been separated by electrophoresis soy meal makes the protein progressively more
into 18 bands in alkaline gels and 10 bands in insoluble. Hydrogen bonds and hydrophobicity
acid gels. The 11S protein is usually named gly- appear to be responsible for the decrease in solu-
cinin, and there are various proposals for naming bility of the proteins during heating.
other protein fractions conglycinin (Wolf 1969). Both 7S and 11S proteins show a complicated
The detailed subunit structures of 7S and 11S pattern of association and dissociation reactions.
globulins, the most important of the soy proteins, The 7S globulin at 0.5 ionic strength and pH 7.6
have been described by Wolf (1972b). The 11S is present as a monomer with molecular weight
globulin has a quaternary structure consisting of of 180,000–210,000. At 0.1 ionic strength, the
12 subunits. According to Catsimpoolas et al. molecule forms the 9S dimer. This reaction is
(1967), these have the following amino-terminal reversible. At pH 2 and low ionic strength, the 7S
residues: 8 glycine, 2 phenylalanine, and either 2 globulin forms 2S and 5S compounds; these are
leucine or 2 isoleucine. It appears that the 11S the result of dissociation into subunits. This reac-
protein is a dimer of two identical monomers, tion is reversed at higher ionic strengths.
each consisting of six subunits, three of which are Changes in the quaternary structure of the 11S
acidic and three basic. Interactions among these globulin have been summarized by Wolf (1972a).
subunits may be a factor in stabilizing the mole- Secondary and tertiary structures of this protein
cule. The 7S globulin consists of 9 subunits of involve no alpha helix structure but instead consist
single polypeptide chains. The protein is a glyco- of antiparallel beta-structure and disordered
protein and the polysaccharide is attached as a regions. The structure appears to be compact and is
single unit to one of the polypeptide chains. The sta­bilized by hydrophobic bonds. The changes in
carbohydrate consists of 38 mannose and 12 glu- ­quaternary structure that occur as a function of exp­
cosamine residues. erimental conditions are represented in Fig. 3.33.
160 3  Amino Acids and Proteins

Fig. 3.33  Schematic representation of the heat induced dissociated 7s subunits with the heat plus RSH dissociated
molecular complex between the subunits of 7s and 11s basic (B) subunits of 11s. The dissociated acidic subunits
globulins. α, α′, and β refer to the three subunits of 7s remain as monomers in solution. Further heating results in
globulin and A and B refer to the acidic and basic subunits gradual polymerization to a soluble macromolecular com-
of 11s globulin. These are generally linked by one inter- plex composed mostly of β subunits of 7s associated elec-
molecular disulfide (SS) as depicted by the black line trostatically with BS of 11s at contact points that are
linking the A and B subunits in the half 11s molecular relatively nonpolar. A few a and d subunitn and some
shown. The first stage of the reaction is a heat induced disulfide linked basic subunits occur in the complex.
dissociation at >80 “C in the presence of thiol (RSH) Modified from Utsumi et al. (1984)
reagent and the initial electrostatic association of the heat-­

At ionic strength of 0.1, the protein partially associ- For direct human consumption, soybeans are
ates into agglomerates with a higher sedimentation mainly used as soymilk and tofu or bean curd. In
velocity. Increasing the ionic strength reverses this the preparation of soymilk the extractability of
reaction. Various conditions promote dissociation proteins is related to the age and storage
of the 11S protein into half-molecules with a sedi- ­conditions of soybeans (Thomas and Robertson
mentation velocity of 7S. Further breakdown of the 1989). Adverse storage conditions result in low
half-molecules may occur and will result in the for- protein extraction and poor quality of tofu. Tofu
mation of unfolded subunits. It has been suggested is manufactured from soymilk with 10% of total
(Catsimpoolas 1969) that the 11S molecule is made solids by coagulation of the proteins with the
up of two annular-hexagonal structures, each con- following coagulants: CaSO4, MgSO4, CaCl2,
­
taining six alternating acidic and basic subunits. MgCl2, or GDL (glucono delta lactone) (deMan
This feature would result in a stabilization of the et al. 1986). When using the salts, the Ca and Mg
structure by ionic bonds (Utsumi et al. 1984). ions are the prime reactants. CaSO4 is only
Soybean whey proteins are obtained in the slightly soluble in water; the reaction is slow, but
solution left after acid precipitation of protein the resulting curd is cohesive and, when pressed,
from an aqueous extract. The solution contains an has a soft texture. With soluble salts such as
unknown number of albumins and globulins, in CaCl2 the reaction is fast and the resulting curd is
addition to water-soluble carbohydrates, nonpro- fragmented. When pressed, the tofu has a higher
tein nitrogen, salts, vitamins, and phytates. In the protein content and the texture is firm.
production of soy protein isolates, the whey pro- Coagulation with GDL is a different process. In
teins may create a disposal problem. freshly prepared tofu utilizing GDL the pH
Gluten Sensitivity 161

slowly decreases and coagulation occurs at the quantitative results. Lateral-flow devices generally
isoelectric point of the proteins. The texture of provide qualitative results, indicating the pres-
GDL tofu is very homogeneous. ence of gluten above a threshold level, but in
some instances can also provide semi-­quantitative
results. Other types of tests include polymerase
Gluten Sensitivity chain reaction (PCR), which detects DNA rather
than protein; adenosine triphosphate (ATP) swab
Gluten is a mixture of prolamin (gliadin in wheat) tests for assessing cleanliness of equipment sur-
and glutelin proteins naturally present in wheat, faces; and general protein swabs, which are not
rye, barley, and related grains, including those specific to gluten but detect all types of protein
wheat varieties known by such names as durum and can be used for assessing cleanliness.
(semolina), spelt, einkorn, emmer, khorasan The most common form of ELISA for gluten
(Kamut), club wheat, triticale, and farro. It is detection is the sandwich format. In sandwich
most commonly present in products made from ELISAs the antigen (gluten proteins in this case)
wheat flour and in certain other food products in binds to anti-gluten antibodies that are affixed to
which it is used as an ingredient, providing elas- a surface, generally a microwell plate. Then a
ticity in baked goods, for example, as well as tex- second gluten-specific antibody—this one linked
ture, moisture retention, and flavor. to an enzyme—is applied over the surface and
Celiac disease, also referred to as celiac sprue, also binds to any gluten that is now affixed to the
is a genetic disease that is said to affect about 1% surface. Finally, a substance is added that the
of the people in North America and Europe. The enzyme can convert into a detectable signal, such
immune system of people with the disease as a color change.
responds to the consumption of gluten by damag- Lateral flow tests, also known as immuno-
ing the lining of the small intestine, thus interfer- chromatographic assays, are usually available in
ing with absorption of nutrients. The disease has dipstick format, in which the test sample flows
no cure but can be managed by avoiding gluten in along a solid substrate by capillary action. When
the diet. the sample is applied to the strip, it mixes with a
The number of products marketed as gluten-­ colored reagent and moves with the substrate into
free is increasing worldwide, but even with the specific zones on the strip that contain the spe-
establishment of regulations allowing such label- cific antibodies. When liquid from a sample or
ing, it is possible that foods labeled as gluten-free wet equipment surface passes over this zone, the
may be contaminated during processing by gluten will bind to the antibody. Color also forms
equipment previously used for gluten-containing as a line in this zone on the strip. A control zone
foods. Because of the high prevalence of wheat in is also usually included that will form a color that
the food supply, even products that are formu- merely indicates that the strip has worked cor-
lated or processed to not contain it may still rectly. Thus, a negative test is the formation of
­contain enough trace amounts of gluten to pro- one line while a positive test is the formation
duce symptoms in gluten-sensitive individuals. of two lines.
Consequently, reliable tests are required for the Most commercially available kits for routine
detection of gluten in foods. food allergen analysis rely on immunological
methods such as enzyme-linked immunosorbent
assay (ELISA) or on polymerase chain reaction
 est Methods
T (PCR) approaches. ELISA has the advantage of
being relatively quick and simple to perform and
There are three types of methods in gluten testing can be used to detect many known allergens;
with the majority of tests for gluten in food prod- however, it can only detect one allergen at a time,
ucts are enzyme-linked immunosorbent assays is susceptible to cross-reactivity, and can lead to
(ELISAs). Microwell versions of ELISAs ­provide the generation of false positive and false negative
162 3  Amino Acids and Proteins

results. PCR methods are complementary to Bietz, J. A., & Wall, J. S. (1972). Wheat gluten subunits:
ELISA methods, and are used to amplify and Molecular weights determined by sodium dodecyl
sulfate-polyaerylamide gel electrophoresis. Cereal
detect the DNA of an allergen. PCR approaches Chemistry, 49, 416–430.
are highly specific, sensitive to very low levels of Bjarnason, J., & Carpenter, K. J. (1970). Mechanisms
allergen, and can be used to detect more than two of heat damage in proteins. 2. Chemical changes in
allergens at once. However, the PCR is limited pure proteins. The British Journal of Nutrition, 24(1),
313–329.
because some food processing methods can Bodwell, C. E., & McClain, P. E. (1971). Proteins. In J. F.
destroy detectable DNA, causing false negative Price & B. S. Schweigert (Eds.), The science of meat
results. PCR methods are also highly susceptible and meat products. San Francisco: W.H. Freeman and
to food matrix interferences. Liquid chromatog- Company.
Brooks, J. R., & Moor, C. V. (1985). Current aspects
raphy tandem mass spectrometry (LC–MS–MS) of soy protein fractionation and nomenclature.
is an alternative method for allergen detection Journal of the American Oil Chemists’ Society, 62,
that is highly specific, sensitive, and has the abil- 1347–1354.
ity to directly analyze multiple allergens in a Burton, H. S., & McWeeney, D. J. (1964). Non-enzymatic
browning: Routes to the production of melanoidins
single analysis. LC–MS–MS detection is also not from aldoses and amino compounds. Chemistry &
as strongly influenced by food processing, and Industry, 11, 462–463.
has the capability for accurate quantitation of the Cassens, R. G. (1971). Microscopic structure of animal
allergens. LC–MS–MS allows direct analysis of tissues. In J. F. Price & B. S. Schweigert (Eds.), The
science of meat and meat products. San Francisco:
multiple allergenic proteins in a single prepara- W.H. Freeman and Company.
tion; is more sensitive; and allows more accurate Catsimpoolas, N. (1969). Isolation of glycinin subunits by
quantitation than traditional approaches. isoelectric focusing in urea-mercaptoethanol. FEBS
LC–MS–MS has grown in popularity for Letters, 4(4), 259–261.
Catsimpoolas, N., et al. (1967). Purification and structural
allergen testing because it provides quantitative studies of the 11S component of soybean proteins.
capabilities for multiple allergens simultane- Cereal Chemistry, 44, 631–637.
ously. Allergenic proteins are extracted from Connell, J. J. (1962). Fish muscle proteins. In J. Hawthorn
samples and are subsequently digested into & J. M. Leitch (Eds.), Recent advances in food science
(Vol. 1). London: Butterworth.
peptide fragments that are directly analyzed
­ Dalgleish, D. G. (1989). Protein-stabilized emulsions and
using their characteristic molecular masses. The their properties. In T. M. Hardman (Ed.), Water and
analysis of multiple target peptides and their food quality. London: Elsevier.
unique masses and fragmentation patterns Ellis, G. P. (1959). The Maillard reaction. Advances in
Carbohydrate Chemistry, 14, 63–134.
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tion. Journal of Dairy Science, 56(9), 1195–1206.
Farrell, H. M., Jr., & Thompson, M. P. (1974). Physical
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Carbohydrates
4
Gillian Eggleston, John W. Finley,
and John M. deMan

such as cellulose and hemicellulose. Gums are a


Introduction varied group of polysaccharides obtained from
plants, seaweeds, and microorganisms. Because
Carbohydrates or saccharides (from the Greek of their useful physical properties, the gums have
word sakkharon meaning sugar) occur in plant found widespread application in food processing.
and animal tissues as well as in microorganisms; The main carbohydrates that occur in a number
as macronutrients they are the human body’s pre- of example food products are listed in Table 4.1.
ferred energy source, providing fuel for the cen- When consumed as part of the diet, carbohy-
tral nervous system and energy for working drates cause a range of physiological effects.
muscles. Carbohydrates also serve as (1) a short- Simple sugars are generally absorbed and rapidly
term energy source for all organisms, (2) struc- utilized for energy or converted to fat for storage.
tural molecules in plants, and (3) storage forms The more complex carbohydrates range from
of foods in plants and animals. Carbohydrates are those that are highly digestible, i.e., starch to those
technically hydrates of carbon with the empirical that are non-digestible, i.e., celluloses and chitins.
formula Cm(H2O)n (where m could be different Hemi-cellulose and some oligosaccharides are not
from n), but structurally they are more accurately digestible but are fermented in the colon resulting
viewed as polyhydroxy aldehydes and ketones. in production of beneficial short chain fatty acids.
Carbohydrates can be divided into three chemical In addition to the physiological effect in vivo the
groups: monosaccharides, oligosaccharides, and carbohydrates can serve as bulking agents. In
polysaccharides, with the first being small (lower foods the carbohydrates have a wide range of
molecular weight) and commonly referred to as functions. The simple sugars can add sweetness,
simple sugars. Carbohydrates in food can also be alter the boiling point and freezing point of foods,
classified as simple or complex, with the differ- and help stabilize proteins in solution.
ence between the two forms being the chemical Carbohydrates are generally based on simple sug-
structure and how quickly they are absorbed and ars and polymers of widely varying chain length.
digested. In animal organisms, the main sugar is Carbohydrates are generally and broadly classified
glucose and the storage carbohydrate is glyco- according to their size or degree of polymerization
gen; in milk, the main sugar is almost exclusively (DP). This is a rather broad definition and as you
the disaccharide lactose. In plant organisms, a will see in this chapter there are many exceptions.
wide variety of monosaccharides and oligosac- We will start with a discussion of the simple sugars
charides occur, as well as storage polysaccha- and advance through the larger and more complex
rides such as starch, and structural polysaccharides carbohydrates later in the chapter.

© Springer International Publishing AG 2018 165


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_4
166 4 Carbohydrates

Table 4.1  Carbohydrates in some example foods and food products


Product Total sugar (%) Mono- and disaccharides (%) Polysaccharides (%)
Fruits
Apple 14.5 Glucose 1.2; fructose 6.0; sucrose 3.8; mannose trace Starch 1.5; cellulose 1.0
Grape 17.3 Glucose 5.4; fructose 5.3; sucrose 1.3; mannose 2.2 Cellulose 0.6
Strawberry 8.4 Glucose 2.1; fructose 2.4; sucrose 1.0; mannose 0.1 Cellulose 1.3
Vegetables
Carrot 9.7 Glucose 0.9; fructose 0.9; sucrose 4.3 Starch 7.8; cellulose 1.0
Onion 8.7 Glucose 2.1; fructose 1.1; sucrose 0.9 Cellulose 0.7
Peanuts 18.6 Sucrose 4–12 Cellulose 2.4
Potato 17.1 Starch 14; cellulose 0.5
Sweet corn 22.1 Sucrose 12–17 Cellulose 0.7
Sweet potato 26.3 Glucose 0.9; sucrose 2–3 Starch 14.7; cellulose 0.7
Turnip 6.6 Glucose 1.5; fructose 1.2; sucrose 0.4 Cellulose 0.9
Others
Sugar beet juice 18–20 Glucose 0.01; fructose 0.01; sucrose 16–18
Sugar cane juice 14–28 Glucose + fructose 0.7–1.5; sucrose 11.2–22.4
Maple syrup 65.5 Sucrose 58.2–65.5; hexoses 0.0–7.9
Honey 82.3 Glucose 28–35; fructose 34–41; sucrose 1–5
Meat Glucose 0.01 Glycogen 0.1
Milk 4.9 Lactose 4.9

Monosaccharides

The simplest sugars or monomeric units are


called monosaccharides, which cannot be hydro-
lyzed into smaller carbohydrates. Chemically,
they are aldehydes or ketones with two or more
hydroxyl groups. The general chemical formula
of an unmodified monosaccharide is Cn(H2O)n. Fig. 4.1  Structures of the simplest carbohydrates: three
Monosaccharides range from three to eight car- carbon sugars (trioses) with an aldehyde (aldose) or
bon atoms which constitute the backbone of the ketone (ketose) group
sugar. The carbon backbone is then appended
with Hydrogen (H) or Hydroxy (−OH) groups. Carbohydrates have a chiral center around
Monosaccharides are classified according to which the OH group is either to the left (L) or to
three different characteristics: (1) the placement the right (D). Most natural sugars are members of
of the carbonyl group, (2) the number of carbon the D series (see Fig. 4.1). D and L structures are
atoms it contains, and (3) chirality. If the car- non-superimposable mirror images, named enan-
bonyl group is an aldehyde then the sugar is an tiomers. The simplest of the sugars are the three
aldose; if the carbohyl group is a ketone the sugar carbon sugars and, unfortunately, their most
is a ketose. Monosaccharides with three carbon common nomenclature is different than the larger
atoms, i.e., glyceraldehyde (Fig. 4.1) are named molecules (4–6 carbons). The three carbon
trioses, those with four carbons are tetroses, with aldose is generally referred to as D or
five carbons pentoses, with six carbons hexoses, L-glyceraldehyde (2,3-dihydroxypropanal) and
and so on. Monosaccharides of 5 or 6 carbons, the keto version is dihydroxyacetone (dihydroxy-
however, are the most common. propanone) (Fig. 4.1).
Monosaccharides 167

The most common sugars in food and nutri- After Fischer formulas were introduced for
tional chemistry are pentoses and hexoses. carbohydrates, Haworth representations were
D-glucose is the most important monosaccharide developed to give a more accurate spatial view of
and is derived from the simplest sugar, the molecule (Fig. 4.3). Carbohydrates are either
D-glyceraldehyde, an aldotriose. The designation acyclic or exist as furanosides (f) or pyranosides
of aldose and ketose sugars indicates the chemi- (p), as shown in Fig. 4.3. Because the Haworth
cal character of the reducing form of a sugar and formula does not account for the actual bond
can be indicated by the simple or open-chain for- angles, the modern conformational formulas
mula of Fischer, as shown in Fig. 4.2. (also seen in Fig. 4.3) more accurately represent
the sugar molecule. Pyranose structures exist in
chair conformation, with the bulky –CH2OH
group on carbon 5. A number of chair conforma-
tions of pyranoside sugars are possible
(Shallenberger and Birch 1975), and the two
most important ones for glucose are shown in
Fig. 4.3. These are named the CI D and the IC D
forms (also described as O-outside and O-inside,
respectively). In the CI D form of β-D-gluco-
pyranose, all hydroxyls are in the equatorial posi-
tion, which represents the highest thermodynamic
stability.
Isomers of the same pyranosides are distin-
guishable from the OH in the C-1 position (also
known as anomeric carbon). The two possible
anomeric forms of monosaccharides are desig-
nated by Greek letter prefix α or β. In the
Fig. 4.2  Fischer projection of aldo- and keto structures α-anomer the hydroxyl group points to the right,
for pentoses and hexoses according to the Fischer projection formula; the

Fig. 4.3  Representation fischer, haworth, and conformational representations of α- and β- D-glucose structures
168 4 Carbohydrates

hydroxyl group points to the left in the β-anomer. d­ ifferent compounds have either an all-α or all-β
In Fig. 4.3 the structure marked Cl D represents structure at this position.
the α-anomer, and 1C D represents the β-anomer. Naturally occurring sugars are mostly hex-
The anomeric forms of the sugars are in tauto- oses, but sugars with different numbers of car-
meric equilibrium in solution; and this causes the bons are also present in many food products.
change in optical rotation when a sugar is placed There are also sugars with different functional
in solution. Under normal conditions, it may take groups or substituents, creating diverse com-
several hours or longer before the equilibrium is pounds, including aldoses, ketoses, amino sug-
established and the optical rotation reaches its ars, deoxy sugars, sugar acids, sugar alcohols,
equilibrium value. At room temperature an aque- acetylated or methylated sugars, anhydro-sugars,
ous solution of glucose can exist in four tauto- oligo- and polysaccharides, and glycosides.
meric forms (Angyal 1984): β-furanoside—0.14%, Fructose is the most widely occurring ketose and
acyclic aldehyde—0.0026%, β-pyranoside—62%, is shown in its various representations in Figs. 4.5
and α-pyranoside—38% (Fig. 4.4). Fructose and 4.6. It is the sweetest known natural sugar,
under the same conditions also exists in four tau- and occurs bound to glucose in sucrose or com-
tomeric forms as follows: α-pyranoside—trace, mon table sugar. Of all the other possible hex-
β-pyrano-side—75%, α-furanoside—4%, and oses, only two occur widely—D-mannose and
β-furanoside—21% (Fig. 4.5) (Angyal 1976). D-galactose. Their formulas and relationship to
When the monosaccharides become involved D-glucose are given in Fig. 4.7.
in condensation reactions to produce di-, oligo-, Anomers are also sterioisomers and diastereo-
and polysaccharides, the conformation of the mers that differ in configuration around the
bond on the carbon 1 becomes fixed and the ­anomeric carbon atom as shown in Fig. 4.7. As we

Fig. 4.4  Tautomeric forms of glucose in aqueous solution at room temperature


Monosaccharides 169

Fig. 4.5  Tautomeric forms of fructose in aqueous solution at room temperature

Fig. 4.6 Methods of representation of D-fructose. Carbohydrates and Their Roles, H.W. Shultz, R.F. Cain,
Source: From M.L. Wolfrom, Physical and Chemical and R.W. Wrolstad, eds., 1969, AVI Publishing Co.
Structures of Carbohydrates, in Symposium on Foods:
170 4 Carbohydrates

Fig. 4.7 Relationship of D-aldehyde sugars. Source: and Their Roles, H.W. Schultz, R.F. Cain, and
From M.L. Wolfrom, Physical and Chemical Structures of R.W. Wrolstad, eds., 1969, AVI Publishing Co.
Carbohydrates, in Symposium on Foods: Carbohydrates

will see later, the anomeric forms result in very approximately the same α- and β- anomer distri-
different properties of polymers, such as starch bution as glucose.
and cellulose, which are both glucose polymers. The thermal effects in food are very important
When sugars are in solution and only two tau- in food chemistry, and temperature impacts the
tomers are formed it is considered a simple muta- relative sweetness of sugars in solution. In
rotation, whereas with three or more it is Fig. 4.9, the sweetness of four simple sugars are
considered complex. The furanose and pyranose compared relative to sucrose. It is clear that fruc-
rings are generated from the straight chain inter- tose is much sweeter than sucrose at low temper-
mediate of the sugar. Sugars with the gluco, atures but not as sweet in hot solutions. This is
manno, gulo, and allo configurations exhibit sim- because at low temperatures there of the intensely
ple mutarotation. For example, when glucose is sweet β-D-fructopyranose present. At higher
in solution (Fig. 4.8), only α-D-flucopyranose temperatures there are more open ring structures
and β−D-glucopyranose are present in solution. which are not as sweet. It, therefore, has a greater
The aldehyde content in the solution is estimated effect in a cold beverage than a hot beverage such
to be 0.003%. Maltose and lactose exhibit as coffee or tea.
Related Compounds to Monosaccharides 171

Fig. 4.8 
Conformational isomers
of glucose

Related Compounds
to Monosaccharides

Amino Sugars

Amino sugars usually contain D-glucosamine


(2-deoxy-2-amino glucose). They occur as com-
ponents of high molecular weight compounds
such as the chitin of crustaceans and mollusks,
insects, as well as in certain mushrooms and in
combination with the ovomucin of egg white.
Chitin can be hydrolyzed to produce chitosan
which has potential as a food ingredient, and this
is discussed later in this chapter. After cellulose,
chitin is the second most important natural poly-
mer in the world. Chitin is a largely inert polymer

Fig. 4.9  Effect of temperature on the relative sweetness


of sugars. Modified form Shallenberger and Birch (1975)
172 4 Carbohydrates

Fig. 4.10  Hydrolysis of


chitin removes acetate
groups yielding chitosan

composed of long chains of acetylglucosamine. 7-O-beta-D-glucoside form of genistein and is


Upon hydrolysis the acetates are removed from the predominant form of the isoflavone naturally
the amino group leaving a glucosamine unit. The occurring in plants. The aglycone, genesteine is
long chain polymers are also reduced to various the bioactive form that includes antiatheroscle-
chain lengths of glucosamine polymer. The main rotic, estrogenic and anticancer.
sources exploited are two marine crustaceans,
shrimp and crabs. Chitin can be hydrolyzed with
acid or enzymatically to remove the acetate from Sugar Alcohols
the amino sugar leaving a free amino group in
chitosan as illustrated in Fig. 4.10. Chitosan, These are compounds obtained when aldo- and
which is soluble in acidic aqueous media, is used keto- groups of a sugar are reduced to hydroxyl
in many applications (food, cosmetics, biomedi- groups. Thus, since sugars are polyhydroxy com-
cal, and pharmaceutical applications). pounds, their corresponding sugar alcohols
merely have one more alcohol group. Sugar alco-
hols are also referred to as polyols, polyalcohols,
Glycosides or polyhydric alcohols. (The term “polyol” could
properly cover a much larger group containing
Glycosides are sugars in which the hydrogen of any compound with three or more hydroxy
an anomeric hydroxy group has been replaced by groups, but common usage normally restricts the
an alkyl or aryl group to form a mixed acetal. term to those compounds closely related to sug-
Glycosides are hydrolyzed by acid or enzymes ars and sugar derivatives.) Some sugar alcohols,
but are stable to alkali. Formation of the full ace- particularly pentitols and hexitols, are widely dis-
tal means that glycosides have no reducing tributed in many fruits and vegetables (Washüttl
power. Hydrolysis of glycosides yields sugar and et al. 1973), while others are produced industri-
the aglycone. Genistin is an isoflavone found in a ally from the corresponding sugar by catalytic
number of dietary plants including soy. When hydrogenation as food ingredients (Table 4.2).
genistin is hydrolyzed with either hydrochloric Chemically, physically, and biologically
acid or enzymatically in the gastrointestinal tract, the sugar alcohols resemble sugars to the extent
1 mole each of genistein and glucose are pro- that some are even sweet to the taste and some
duced as shown in Fig. 4.11. Chemically it is the are used as food sweeteners (see Table 4.3),
Related Compounds to Monosaccharides 173

Fig. 4.11  Hydrolysis of


genestin to yield
genestein and glucose

Table 4.2  Occurrence of sugar-alcohols in some fruits and vegetables (expressed as mg/100 g of dry food)
Product Arabinitol Xylitol Mannitol Sorbitol Galactitol
Bananas – 21 – – –
Pears – – – 4600 –
Raspberries – 268 – – –
Strawberries – 362 – – –
Peaches – – – 960 –
Celery – – 4050 – –
Cauliflower – 300 – – –
White mushrooms 340 128 476 – 48
Source: From J. Washüttl, P. Reiderer, and E. Bancher, A Qualitative and Quantitative Study of Sugar-Alcohols in
Several Foods: A Research Role, J. Food Sci., Vol. 38, pp. 1262–1263,1973

Table 4.3  The caloric values, sweetness intensity and application of some key sugar alcohols and common carbohy-
drate sweeteners
Calories Relative sweetness
Type per gram (sucrose = 100%) Typical food applications
Sucrose 4 100 Common beverage and food sweetener
Crystalline fructose 3 180 Occurs naturally in fruits and vegetables.
Common beverage and food sweetener
Fructose (5–15% solution) 115–125 Common beverage and food sweetener
High fructose corn syrup (HFCS) ~2.8 100–130 Common beverage and food sweetener;
market share has decreased since 1999 due to
consumer perception problems
Glucose (8–10% solution) 60–70 Common beverage and food sweetener
Invert syrup 3 105 Common beverage sweetener
Sorbitol 2.6 50–70% Sugar-free candies, chewing gums, frozen
desserts and baked goods
(continued)
174 4 Carbohydrates

Table 4.3 (continued)
Calories Relative sweetness
Type per gram (sucrose = 100%) Typical food applications
Xylitol 2.4 100%a Chewing gum, gum drops and hard candy,
pharmaceuticals and oral health products,
such as throat lozenges, cough syrups,
children’s chewable multivitamins,
toothpastes and mouthwashes; used in foods
for special dietary purposes
Maltitol 2.1 68–75% Hard candies, chewing gum, chocolates,
baked goods and ice cream
Isomalt 2.0 45–65% Candies, toffee, lollipops, fudge, wafers,
cough drops, throat lozenges
Lactitol 2.0 30–40% Chocolate, some baked goods (cookies and
cakes), hard and soft candy and frozen dairy
desserts
Mannitol 1.6 40–70% Dusting powder for chewing gum, ingredient
in chocolate-flavored coating agents for ice
cream and confections
Erythritol 0–0.2b 60–80% Bulk sweetener in low calorie foods
Hydrogenated Starch Hydrolysates 3 25–50% Bulk sweetener in low calorie foods, provide
(HSH) sweetness, texture and bulk to a variety of
sugarless products
Depends on solution concentration
a

FDA accepts 0.2 kcal/g, but some other countries, such as Japan and the European Union, accept 0 kcal/g
b

American Diabetes Association. Nutrition principles and recommendations in diabetes-Position Statement. Diabetes
Care, Jan.2004. Eggleston et al. (2017)

r­epresenting one type of reduced-calorie sweet-


ener. Furthermore, sugar alcohols provide fewer
calories than sugar and have less of an effect on
blood glucose (blood sugar) than other carbohy-
drates. Examples of sugar alcohols are: glycerol,
erythritol, isomalt, lactitol, mannitol, sorbitol,
and xylitol. They can be made by reduction of
free sugars with sodium amalgam, sodium boro-
hydride in water, or by catalytic hydrogenation.
The sugar alcohols are poorly absorbed but are
fermented in the lower gastrointestinal tract
Fig. 4.12  Structures of sorbitol and xylitol
which can result in discomfort. For this reason,
the daily dose needs to be limited. Reduction of
glucose yields glucitol (Fig. 4.12; the trivial name of D-glucose. Mannitol, the reduced form of
is sorbitol). Another commercially produced D-mannose, is found as a component of mush-
sugar alcohol is xylitol, a five-carbon compound, rooms, celery, and olives. Xylitol is obtained from
which is also used in diabetic foods (Fig. 4.12). saccharification of xylan-containing plant materi-
Sorbitol, the most widely distributed natural als; it is a pentitol, being the reduced form of
sugar alcohol, is found in many fruits such as xylose. Sorbitol, mannitol, and xylitol are mono-
plums, berries, cherries, apples, and pears saccharide-derived sugar alcohols with properties
(Table 4.2). It is also a component of fruit juices, that make them valuable for specific applications in
fruit wines, and other fruit products. Sorbitol is foods: they are suitable for diabetics, they are non-
commercially produced by catalytic hydrogenation cariogenic, they possess reduced physiological
Oligosaccharides 175

caloric value, and they are useful as sweeteners Disaccharides


that are non-fermentable by yeasts. Sugar alco-
hols have also gained commercial viability as Two chemically bonded monosaccharides are
sweeteners because they are less expensive than known as disaccharides. The most common disac-
sucrose and corn-based sweeteners, particularly charides in food are sucrose (α-D-glucopyranosyl
high fructose corn syrup (Eggleston et al. 2017). β-D-fructofuranoside; table sugar is sucrose
Furthermore, sugar alcohols are not considered extracted and refined from either sugarcane or
sugars for food labeling purposes, so foods con- sugar beets), lactose (4-O-β-D-galactopyranosyl-
taining them as sweeteners can be named “sugar D-glucopyranose; milk sugar), and maltose
free” or “no sugar added” (Eggleston et al. 2017). (4-O-α-D-glucopyranosyl-D-glucopyranose; from
Most, sugar alcohols, however, are less sweet starch). Disaccharides can be classified into two
than sucrose. Some sugar alcohols give a cooling types: reducing and non-reducing. Disaccharides
sensation on the tongue. Table 4.3 lists the rela- are formed by a glycosidic link between the reduc-
tive sweetness of sugar alcohols and commercial ing groups on one saccharide with the hydroxyl
sugar sweeteners. group of another saccharide. There are multiple
forms possible even with a simple glucose-glucose
disaccharide. Structures of the most abundant
Oligosaccharides disaccharides are illustrated in Fig. 4.13.

When the number of monosaccharides in a gly- Sucrose


cosidic chain is between 2 to 10, the resulting Sucrose, a major sweetener, is a primary oligo-
compound is an oligosaccharide. Oligosaccharides saccharide (or disaccharide) found widely in
can be homologous or heterologous, and occur plants, however, the largest commercial sources
widely in plants, but can also be produced syn- are from sugarcane or sugar beet. In sucrose the
thetically or via microbial fermentative and enzy- reducing groups of the constituent monosaccha-
matic processes. Plant oligosaccharides are often rides (glucose and fructose) are linked by a gly-
grouped into two distinct classes: (1) primary or cosidic bond (Fig. 4.13), thus sucrose is one of
(2) secondary oligosaccharides (Kandler and the few non-reducing disaccharides. As a non-
Hopf 1980). Primary oligosaccharides are those reducing disaccharide, sucrose does not reduce
synthesized in vivo from a mono- or oligosaccha- Fehling solution or form osazones and it does not
ride and a glycosyl donor by the action of a undergo mutarotation in solution. Because of the
glycosyl transferase enzyme (Kandler and
­ unique carbonyl-to-carbonyl linkage, sucrose is
Hopf 1980; Eggleston and Côté 2003). Sucrose is highly labile in acid medium, and acid hydrolysis
the most common primary oligosaccharide in is more rapid than other di- and oligosaccharides.
plants. In comparison, secondary oligosaccha- When sucrose is heated to 210 °C, partial decom-
rides are those formed in vivo or in vitro by position takes place and caramel is formed.
hydrolysis of larger oligosaccharides, polysac- Sucrose is highly soluble over a wide tempera-
charides, glycoproteins, or glycolipids. Common ture range, as is illustrated in Fig. 4.14. This
oligosaccharides occurring in foods are listed in property makes sucrose an excellent ingredient
Table 4.4. for syrups and other sugar-containing foods.

Table 4.4  Common oligosaccharides occurring in foods


Sucrose (α-D-glucopyranosyl β-D-fructofuranoside)
α,α-Trehalose (α-D-glucopyranosyl-α-D-glycopyranoside)
Raffinose [O-α-D-galactopyranosyl-(1 → 6)-O-α-­D-glucopyranosyl-(1  → 2)-β-D-­fructofuranoside]
Stachyose [O-α-D-galactopyranosyl-(1 → 6)-O-α-­D-galactopyranosyl-(1  → 6)-O-α-D-­glucopyranosyl-(1  → 2)-
β-D-­fructofuranoside]
Verbascose [O-α-D-galactopyranosyl-(1 → 6)-O-α-­D-galactopyranosyl-(1  → 6)-O-α-D-­galactopyranosyl-
(1 → 6)-O-α-D-­glucopyranosyl-(1  → 2)-β-D-­fructofuranoside]
Source: From R.S. Shallenberger and G.G. Birch, Sugar Chemistry, 1975, AVI Publishing Co.
176 4 Carbohydrates

Maltose Cellobiose
α-D-glucopyranosyl-(1-4)-α-D-glucopyranose β-D-glucopyranosyl-(1-4)-α-D-glucopyranose

Isomaltose Gentobiose
α-D-glucopyranosyl-(1-6)-β-D-glucopyranose β-D-glucopyranosyl-(1-6)-β-D-glucopyranose

Trehalose Sucrose
α-D-glucopyranosyl-(1-1)-α-D-glucopyranose β-D-Fructofuranosyl α-D-glucopyranoside

Fig. 4.13  Common disaccharides found in foods

Lactose
The sugar in mammalian milk is lactose, which is
normally easily digested and converted to energy.
Some individuals, however, lack the enzyme lac-
tase, which hydrolyzes lactose and are, therefore,
“lactose intolerant.” In such individuals the lac-
tose is fermented in the lower gastrointestinal
tract causing discomfort and diarrhea. Lactose
from cow’s milk is less sweet than sucrose and,
unlike sucrose, is a reducing sugar. With a few
minor exceptions, lactose is the only sugar in the
milk of all mammalian species and does not
occur elsewhere. Lactose is also a major constitu-
ent of the dry matter of cow’s milk, as it repre-
sents close to 50% of the total solids. The lactose
content of cow’s milk ranges from 4.4 to 5.2%,
with an average of 4.8% expressed as anhydrous
Fig. 4.14  Approximate solubility of some sugars at dif-
ferent temperatures. Source: From R.S. Shallenberger and lactose. In comparison, the lactose content of
G.G. Birch, Sugar Chemistry, 1975, AVI Publishing Co. human milk is higher, about 7.0%. Lactose is a
Oligosaccharides 177

disaccharide of D-galactose and D-glucose and is from starch and dextran polysaccharides, and is
designated as 4-O-β-D-galactopyranosyl-D- thus a secondary oligosaccharide; it can also be
glucopyranose (Fig. 4.13). It is hydrolyzed by the found in honey. Gentiobiose occurs as a glyco-
enzyme β-D-galactosidase (lactase) or by dilute side in amygdalin which is found various stone
solutions of strong acids. On the other hand, fruits. Trehalose is found in yeast.
organic acids such as citric acid, which easily Sugars are important in the texture and appear-
hydrolyze sucrose, are unable to hydrolyze lac- ance of foods, particularly confections, cakes and
tose. This difference is the basis of the determina- cookies. The equilibrium between dissolved and
tion of the two sugars in mixtures. crystalline sugars in the cream centers of confec-
tions help control the texture. In cookies, sugar
Maltose can exist in crystalline, glass, and dissolved
Maltose (4-α-D-glucopyranosyl-β-D-glucopyranose) forms. In the crystalline form most sugars are
represents an important disaccharide found single anhydrous anomers. When crystallized at
widely in plants, and is the basic building block temperatures below 50 °C, glucose crystalizes as
of starch and glycogen polysaccharides an α-pyranose monohydrate. D-Lactose crystal-
(Fig.  4.13). When these polysaccharides are lizes as α-D-lactose which is soluble in water (up
hydrolyzed, the primary degradation product is to 20 g/100 mL water), but can be slow to dis-
maltose disaccharide. For example, in brewing, solve. In evaporated milk α-D-lactose crystals
starch is hydrolyzed by amylases to maltose occasionally cause a gritty defect, whereas, the
(maltose has a characteristic flavor of malt) which texture of sweetened condensed milk is depen-
is then available for hydrolysis to glucose by glu- dent on the α-D-lactose crystals.
coamylase, with subsequent conversion to etha- Sugars in general are very soluble in water and
nol by yeast. The α-1 → 4 linkage is broken by are often formulated as sugar syrups. These
amylases and maltases. Maltose is also readily highly concentrated syrups are produced by boil-
broken down in animal gastrointestinal tract and ing or vacuum evaporation. Syrups of reducing
thus provides 4 Kcal energy/g. Maltose is a sugars such as corn syrup (mostly glucose) are
reducing disaccharide, shows mutarotation, is particularly resistant to crystallization. The
fermentable, and is easily soluble in water. worldwide sugar industry takes advantage of the
crystallization of sucrose from concentrated solu-
Cellobiose tions to produce high purity crystals. Impure
Cellobiose (4-β-D-glucopyranosyl-β-D- solutions are also resistant to crystallization.
glucopyranose) is a reducing disaccharide result- Hard candy is produced by boiling an aqueous
ing from the partial hydrolysis of cellulose mixture of sucrose and glucose with flavors and
polysaccharide because it is a building block for colors to produce a glass which is a super-cooled
cellulose (Fig. 4.13). In cellulose and cellobiose liquid. In cakes, the glucose syrup functions to
the β-1 → 4 linkage is not hydrolyzed by animal retain moistness in the crumb and molding char-
and most microbial enzymes, thus making cellu- acteristics of fondants.
lose more stable than starch in the environment.
The cellobiose, its oligomers and cellulose poly-  isaccharide Sugar Alcohols
D
mers can generally be thought of as non-digest- In recent years, disaccharide sugar alcohols have
ible by animals and provide no caloric value. The also become important food ingredients. The
polymeric structure is important for plant cell most common disaccharide alcohols include iso-
wall structure, and fibers such as cotton. malt, maltitol, lactitol, and hydrogenated starch
hydrolyzates (HSH). Maltitol is hydrogenated
 ther Glucose Disaccharides
O maltose, and its structure is shown in Fig. 4.15. It
Other glucose disaccharides are found at relative has the highest sweetness of the disaccharide
low concentrations in nature. Isomaltose is a polyols compared to sugar (Table 4.3) (Heume
breakdown product during production of glucose and Rapaille 1996). It also has a low negative
178 4 Carbohydrates

Legumes contain several primary oligosac-


charides, including raffinose and stachyose,
which are based on sucrose (Fig. 4.16). These
oligosaccharides are poorly absorbed when
ingested, which results in their fermentation in
the large intestine. This leads to gas production
and flatulence, which present a barrier to wider
food use of such legumes. deMan et al. (1975,
Fig. 4.15  Structure of maltitol
1987) analyzed a large number of soybean variet-
ies and found an average content of 1.21% stach-
heat of solution and, therefore, gives no cooling yose, 0.38% raffinose, 3.47% sucrose, and very
effect in contrast to sorbitol and xylitol. Maltitol small amounts of melibose. In soy milk, total
(Fig. 4.15) exhibits a very high viscosity in solu- reducing sugars after inversion amounted to
tion. Sorbitol and maltitol are derived from 11.1% calculated on dry basis.
starch, whereas lactitol is a disaccharide alcohol, Cow’s milk contains traces of oligosaccha-
1 → 4-galactosyl-glucitol, produced by hydroge- rides other than lactose. They are made up of two,
nation of lactose. It has low sweetness and a three, or four units of lactose, glucose, galactose,
lower energy value than other polyols. It has a neuraminic acid, mannose, and acetyl glucos-
calorie value of 2 kcal/g and is non-cariogenic amine. Human milk contains about 1 g/L of these
(Blankers 1995). It can be used in combination oligosaccharides, which are referred to as the
with intense sweeteners like aspartame or acesul- bifidus factor. The oligosaccharides have a bene-
fame-K to produce sweetening power similar to ficial effect on the intestinal flora of infants.
sucrose. These combinations provide a milky, Fructooligosaccharides (FOSs) or “fructans”
sweet taste that allows good perception of other are also primary oligosaccharides based on
flavors. Isomalt, also known as hydrogenated iso- sucrose (Fig. 4.17) where an additional one, two,
maltulose or hydrogenated palatinose, is manu- or three fructose units have been added by a
factured in a two-step process: (1) the enzymatic β-(2 → l)-glucosidic linkage to the fructose unit
transglycosylation of the nonreducing sucrose to of sucrose (Fig. 4.17). The FOSs occur naturally
the reducing sugar isomaltulose; and (2) hydro- as components of edible plants including banana,
genation, which produces isomalt—an equimolar tomato, and onion (Spiegel et al. 1994). FOSs are
mixture of D-glucopyranosyl-α-(l  → l)-D-man- also manufactured commercially by the action of
nitol and D-glucopyranosyl-α-(l  → 6)- a fungal enzyme from Aspergillus niger,
D-sorbitol. Isomalt is extremely stable and has a β-fructofuranosidase, on sucrose. The three pos-
pure, sweet taste. Because it is only half as sweet sible FOSs are lF-(l-β-fructofuranosyl)n-1 sucrose
as sucrose, it can be used as a versatile bulk oligomers with abbreviated and common names
sweetener (Ziesenitz 1996). as follows: GF2 (1-kestose), GF3 (nystose), and
Both monosaccharides and disaccharides con- GF4 (1F-β-fructofuranosylnystose). The com-
tribute flavor and texture in fruits. Vegetables also mercially manufactured product is a mixture of
contribute both sugars and fiber. The sugar in all three FOSs with sucrose, glucose, and fruc-
fruit and fruit juices is the major source of calo- tose. FOSs can also be secondary oligosaccha-
ries, and fruits are also good sources of dietary rides if manufactured from the enzymatic
fiber. Cereal based products contribute primarily breakdown of fructan polysaccharides such as
starch and sugar. Table 4.5 provides examples of inulin extracted from chicory roots. FOSs are
sugar, starch, and fiber from a range of plant- non-digestible by humans. Such prebiotic non-
based foods. It is important to compare the sugars digestible food ingredients beneficially affect the
and starch which provide 4 kcal/g when selecting host animal by selectively stimulating the growth
plant based foods. The dietary fiber which can of bacteria in the colon which is advantageous to
deliver from 0 to 2 kcal/g provides important the host (Eggleston and Côté 2003). They do this
bulk in the lower gastrointestinal tract. by serving as selective substrates for probiotic
Oligosaccharides 179

Table 4.5  Sugars, starches and total fiber in plant based foods (from USDA Nutrient data base)
Glucose Fructose Sucrose Total Total
Food Description (g) (g) (g) Sugars Starch Fiber
Apples, raw, red delicious, with skin 2.71 5.9 1.86 10.48 0.05 2.3
Apricots, raw 2.37 0.94 5.87 9.24 – 2
Bananas, raw 4.98 4.85 2.39 12.23 5.38 2.6
Blueberries, raw 4.88 4.97 0.11 9.96 0.03 2.4
Dates, medjool 33.68 31.95 0.53 66.47 – 6.7
Kiwifruit, green, raw 4.11 4.35 0.15 8.99 0 3
Oranges, raw, navels 1.97 2.25 4.28 8.5 0 2.2
Pineapple, raw, all varieties 1.73 2.12 5.99 9.85 0 1.4
Plums, raw 5.07 3.07 1.57 9.92 0 1.4
Raisins, seedless 27.75 29.68 0.45 59.19 2.70 3.7
Raspberries, raw 1.86 2.35 0.2 4.42 0 6.5
Strawberries, raw 1.99 2.44 0.47 4.42 0 6.5
Watermelon, raw 1.58 3.36 1.21 6.2 0 0.4
Asparagus, cooked, boiled, drained 0.42 0.79 0 1.3 0 2
Beans, snap, green, raw 1.51 1.39 0.36 3.26 0.88 2.7
Broccoli, raw 0.49 0.68 0.1 1.7 0 2.6
Carrots, baby, raw 1.04 1 2.72 4.76 0 2.9
Corn, sweet, white, cooked, boiled, drained, without salt 0.7 1.02 6.02 7.73 4.47 2.7
Onions, sweet, raw 2.26 2.02 0.72 5.02 0 0.9
Peas, green, frozen, unprepared 0.08 0.25 4.6 5 4.17 4.5
Peppers, sweet, green, raw 1.16 1.12 0.11 5.93 – 5.5
Potatoes, flesh and skin, raw 0.33 0.27 0.17 0.78 15.44 2.2
Sweet potato, cooked, baked in skin, flesh, without salt 0.57 0.5 2.28 6.48 7.05 3.3
Tomatoes, red, ripe, raw, year round average 1.25 1.37 0 2.63 0 1.2
Beans, kidney, all types, mature seeds, canned 0 – 1.85 1.85 8.9 4.3
Beans, pinto, canned, drained solids 0 – 0.54 0.54 10.21 5.5
Lentils, raw 0 0.27 1.47 2.03 49.9 10.7
Peanuts, all types, dry-roasted, with salt 0 – 4.9 4.9 4.39 8.4
Macaroni, cooked, enriched 0.04 0.03 0.09 0.56 26.01 1.8
Rice, white, long-grain, precooked or instant, enriched, prepared – – 0 0 26.33 0.6
Spaghetti, cooked, enriched, with added salt 0.04 0.03 0.09 0.56 26.01 1.8
Cereals, oats, unenriched, cooked with water 0 – 0.25 0.27 11.6 1.7
Cereals ready-to-eat, POST, Shredded Wheat, original big biscuit 0 – 0.55 0.94 65.1 12
Cereals ready-to-eat, RALSTON Corn Flakes 1.48 1.34 4.61 7.84 60.42 2.7

bacteria. Many other non-digestible oligosaccha- in specific rotation that eventually reaches the
rides are also used as prebiotics, and include oli- final equilibrium value. The working hypothesis
gosaccharides of D-galactose, D-glucose, for the occurrence of mutarotation has been
D-xylose and combinations. described by Shallenberger and Birch (1975). It
is assumed that five structural isomers are possi-
ble for any given reducing sugar (Fig. 4.18), with
Chemical Reactions of Sugars pyranose and furanose ring structures being gen-
erated from a central straight-chain intermediate.
Mutarotation Mutarotation is the change in the observed opti-
When a crystalline reducing sugar is placed in cal rotation when a reducing sugar is dissolved in
water, an equilibrium is established between iso- water and forms different tautomeric forms. In
mers, as is evidenced by a relatively slow change the crystalline form a sugar will have a specific
180 4 Carbohydrates

Fig. 4.16  Composition of some major oligosaccharides occurring in foods. Source: From R.S. Shallenberger and
G.G. Birch, Sugar Chemistry, 1975, AVI Publishing Co.

Fig. 4.17  Fructooligosaccharides (FOSs) or Fructan oligosaccharides

form, for example, β-D-glucose either as a pyra- There are five possible isomers possible for a
nose or furanose ring. When the sugar dissolves reducing sugar as shown in Fig. 4.18.
in water, the ring structure opens and an equilib- The velocity of the reaction is greatly acceler-
rium is established among the various isomers of ated by acid or base. The rate is at a minimum for
the sugar. Shallenberger and Birch (1975) pyranose-pyranose interconversions in the pH
described the process known as mutarotation. range 2.5–6.5. Table 4.6 contains the distribution
Oligosaccharides 181

Fig. 4.18  Equilibria involved in mutarotation. Source: Wikipedia

Table 4.6  Percentage distribution of isomers of mutarotated sugars at 20 °C.


Sugar α-Pyranose β-Pyranose α-Furanose β-Furanose
D-Glucose 31.1–37.4 64.0–67.9 – –
D-Galactose 29.6–35.0 63.9–70.4 1.0 3.1
D-Mannose 64.0–68.9 31.1–36.0 – –
D-Fructose 4.0 68.4–76.0 – 28.0–31.6
Source: From R.S. Shallenberger and G.G. Birch, Sugar Chemistry, AVI Publishing Co.

of mutarotated sugars at room temperature. Both of this reaction is increased in the presence of
acids and bases accelerate mutarotation rate, with acid or base. The rate is slowest between pH 2.5
bases being more effective. This was expressed and 6.5. At alkaline pH the hydroxyl ion will
by Hudson (1907) in the following equation: increase the rate by as much as 40,000-fold.
Sugars in solution are unstable and undergo a
K 25 ° = 0.0096 + 0.258  H +  + 9.750 OH −  number of reactions, which have been compre-
hensively reviewed by Clarke et al. (1997). In
This indicates that the effect of the hydroxyl addition to mutarotation, which is the first reac-
ion is about 40,000 times greater than that of the tion to occur when a sugar is dissolved, enoliza-
hydrogen ion. The rate of mutarotation is also tion and isomerization, dehydration and
temperature dependent; increases from 1.5 to 3 fragmentation, anhydride formation and polym-
times occur for every 10 °C rise in temperature. erization may all take place. These reactions are
When sugars are in the open ring form they outlined in Fig. 4.19, using glucose as an exam-
tend to be more reactive particularly when amines ple. Compounds (1) and (2) are α and β forms in
are present and the Maillard browning reaction equilibrium during mutarotation with the alde-
takes place. This is discussed in Chap. 3. The rate hydo form (5). Heating results in dehydration of
182 4 Carbohydrates

Fig. 4.19  Reactions of


reducing sugars in
solution. Source: From
R.S. Shallenberger and
G.G. Birch, Sugar
Chemistry, 1975, AVI
Publishing Co.

the IC conformation of β-D glucopyranose (3) of disaccharides. The predominant linkages in


and formation of levoglucosan (4), followed by the newly formed disaccharides are α-D-1 → 6,
the sequence of reactions described under cara- and β-D-1 → 6. A list of reversion disaccharides
melization. Enolization is the formation of an observed by Thompson (1954) in a 0.082 N
enediol (6). These enediols are unstable and can hydrochloric acid solution or in D-glucose is
rearrange in several ways. Since the reactions are shown in Table 4.7.
reversible, the starting material can be regener-
ated. Other possibilities include formation of Caramelization
keto-D-fructose (10) and β-D-fructopyranose Caramelization involves the oxidation of sugars,
(11), and aldehydo-D-mannose (8) and α-D- and is used extensively in cooking to produce
mannopyranose (9). Another possibility is for the nutty flavors and brown color. Moreover, caramel
double bond to move down the carbon chain to food colorants have been a mainstay in the food
form another enediol (7). This compound can and beverage industries for decades (Boyd 2016).
give rise to saccharinic acids (containing one car- During the controlled, heat process volatile
boxyl group) and to 5-(hydroxy)-methylfurfural chemicals are released producing the characteris-
(13). All these reactions are greatly influenced by tic caramel flavor. The reaction involves the
pH. Mutarotation, enolization, and formation of removal of water and the breakdown of the sugar.
succharic acid (containing two carboxyl groups) The caramelization reaction varies depending on
are favored by alkaline pH, formation of anhy- the type of sugar. Sucrose and glucose begin to
drides, and furaldehydes by acid pH. It appears caramelize around 160 °C (320 °F) whereas fruc-
from the aforementioned reactions, that holding a tose caramelizes at a lower temperature of 110 °C
glucose solution at alkaline pH is likely to yield a (230 °F), because it is more labile to acid degra-
mixture of sugars. When an acid solution of sugar dation. This latter effect can be seen in baked
of high concentration is left at ambient tempera- goods made from honey or fructose syrup yield
ture, reversion takes place. This is the formation products with a darker color.
Oligosaccharides 183

Table 4.7  Reversion disaccharides of glucose in 0.082 N HCl


Glucose disaccharides Reversion (%)
β, β-trehalose (β-D-glucopyranosyl β-D-glucopyranoside) 0.1
β-sophorose (2-O-β-D-glucopyranosyl-β-D-­glucopyranose) 0.2
β-maltose (4-O-α-D-glycopyranosyl-β-D-­glycopyranose) 0.4
α-cellobiose (4-O-β-D-glucopyranosyl-α-D-­glucopyranose) 0.1
β-cellobiose (4-O-β-D-glucopyranosyl-β-D-­glucopyranose) 0.3
β-isomaltose (6-O-α-D-glucopyranosyl-β-D-­glucopyranose) 4.2
α-gentiobiose (6-O-β-D-glucopyranosyl-α-D-­glucopyranose) 0.1
β-gentiobiose (6-O-β-D-glucopyranosyl-β-D-­glucopyranose) 3.4
Source: From A. Thompson et al. Acid Reversion Products from D-Glucose, J. Am.
Chem. Soc., Vol. 76, pp. 1309–1311, 1954

Caramelization Products :
The formation of the caramel pigments is con-
2C12 H 22 O11 = 4H 2 O ⋅ C24 H 36 O18Caramelan
sidered a non-enzymatic browning reaction in the
absence of nitrogenous compounds. When sugars 3C12 H 22 O11 = 8H 2 O ⋅ C36 H 50 O 25Caramelen
are subjected to heat in the absence of water or Continued heating yieldss caramelin C125 H188 O80
are heated in concentrated solution, a series of
reactions occurs that finally leads to caramel for- Caramelan is the first product formed through
mation. The initial stage is the formation of loss of water from sucrose molecules as a result
anhydro sugars. Glucose yields glucosan of dehydration and polymerization reactions.
(1,2-anhydro-α-D-glucose) and levoglucosan Caramelan is brown and results in a bitter flavor;
(1,6-anhydro-β-D-glucose), which have widely von Elbe (1936) reported that caramelan was a
differing specific rotations: +69° and −67°, mix of colorless components and a dark brown
respectively. These compounds may dimerize to humin. The 84% alcohol soluble product results
form a number of reversion disaccharides, includ- from an approximate 12% loss in mass as water
ing gentio-biose and sophorose, which are also and the formula C12H18O9. Caramelen is darker
formed when glucose is melted (Shallenberger than caramelan, and is not soluble in 84% etha-
and Birch 1975). nol. Carmelen results from a further 15%, loss in
Caramelization of sucrose starts with the melt- water, and the greater degree of polymerization
ing of the sugar at high temperatures followed by with the formula C36H50O25. Caramelin is gener-
boiling. Sucrose first decomposes into glucose ated at 22% loss of mass as water. Caramelin is a
and fructose, which is followed by a condensa- mix of strongly colored, hot-water-soluble sub-
tion step, in which the individual sugars lose stances more highly polymerized materials with
water and react with each other. Hundreds of new the average formula C96H102O51. These amor-
aromatic compounds are formed having a range phous products are complex mixtures that are
of complex flavors. formed when sucrose is heatied at 180–190 °C
Melting and caramelization of sucrose have (Gelis 1858; Cunningham and Dorée 1917).
been of significant importance to the food and Further work has shown that a reaction of anhy-
confection industries for over 150 years (Gelis drofructose and anhydroglucose (glucosan) pro-
1858; Cunningham and Dorée 1917). Geliś duced isosaccharosan (anhydrous sucrose)(Pictet
(1858) introduced the terms “caramelan”, “cara- and Adrianoff 1924). These studies demonstrated
melen”, and “caramelin”: the complexity of the carmelization reaction
when sugars are heated. Time and temperature
have major effects on the caramel products
formed. Several other sugars exhibit similarities
in thermal behavior to sucrose melting and cara-
184 4 Carbohydrates

melization (Raemy et al. 1983). Other sugars, The pigment caramelan is soluble in water and
such as glucose, fructose, lactose, maltose, and ethanol and has a bitter taste. Its melting point is
xylose form colored products when the sugars are 138 °C. A further 55 min of heating leads to the
heated (von Elbe 1936; Raemy et al. 1983; formation of caramelen. This compound corre-
Feather and Harris 1973; Lappalainen et al. sponds to a weight loss of about 14%, which is
2006). Industrial caramelization processes, as about eight molecules of water from three mole-
related to manufacturing of sugar-based cara- cules of sucrose, as follows:
mels, differ significantly from spontaneous cara- Caramelen is soluble in water only and melts
melization of sugars during heating. Caramels at 154 °C. Additional heating results in the for-
are produced from a mix of caramel ingredients, mation of a very dark, nearly insoluble pigment
as reviewed by Martin (1955) and Sengar and of average molecular composition C125H188O80.
Sharma (2014). This material is called humin or caramelin. The
Caramelization of sucrose requires a tempera- typical caramel flavor is the result of a number of
ture of about 200 °C. At 160 °C, sucrose melts sugar fragmentation and dehydration products,
and forms glucose and fructose anhydrides (levu- including diacetyl, acetic acid, formic acid, and
losans). At 200 °C, the reaction sequence consists two degradation products reported to have typical
of three distinct stages well separated in time. The caramel flavor by Jurch and Tatum (1970),
first step requires 35 min of heating and involves namely, acetylformoin (4-hydroxy-2,3,5-
a weight loss of 4.5%, corresponding to a loss of hexane-trione) and 4-hydroxy-2,5-dimethyl-
one molecule of water per molecule of sucrose. 3(2H)-furanone.
This could involve formation of compounds such
as isosacchrosan. Pictet and Strieker (1924) Crystallization
showed that the composition of this compound is An important characteristic of sugars is their abil-
1,3′; 2,2′-dianhydro-α-D-glucopyranosyl-β-D- ity to form crystals. In the commercial produc-
glucopyranosyl-β-D-fructo-furanose (Fig. 4.20). tion of table sugar (sucrose), crystallization is an
After an additional 55 min of heating, the weight important purification step. Generally, the more
loss amounts to 9%, and the pigment formed is pure a solution of a sugar, the easier and faster it
named caramelan. This corresponds approxi- will crystallize. Non-reducing oligosaccharides
mately to the following equation: also crystallize relatively easily. The fact that cer-
tain reducing sugars crystallize with more diffi-
2C12 H 22 O11 − 4H 2 O ⋅ C24 H 36 O18
culty has been ascribed to the presence of
The reaction is initiated by the formation of anomers and ring isomers in solution, which
anhydro sugars (Shallenberger and Birch 1975). makes these sugars intrinsically “impure”
Glucose is converted to glucosan (1,2,anhydro- (Shallenberger and Birch 1975). Mixtures of sug-
α-D glucose and levoglucosan (1,6-anhydro-β-D ars crystallize less easily than single sugars. In
glucose), which can dimerize to form multiple certain foods, crystallization is undesirable, such
reversion disaccharides such as sophorose and as the crystallization of lactose in sweetened con-
gentiobiose. densed milk or ice cream.

Crystallization of Sucrose in Commercial


Manufacture of Table Sugar
Factors that influence growth of sucrose crystals
have been listed by Smythe (1971). They include
supersaturation of the solution, temperature, rela-
tive velocity of crystal and solution, nature and
concentration of impurities, and nature of the
Fig. 4.20 Structure of isosacchrosan. Source: From
R.S. Shallenberger and G.G. Birch, Sugar Chemistry, crystal surface. Crystal growth of sucrose con-
1975, AVI Publishing Co. sists of two steps: (1) the mass transfer of sucrose
Oligosaccharides 185

molecules to the surface of the crystal, which is a groups of the fructose moiety. As an example of
first-order process; and (2) the incorporation of the type of packing of molecules in a sucrose
the molecules in the crystal surface, a second- crystal, a projection of the crystal structure along
order process. Under typical conditions, overall the a axis is shown in Figs. 4.21 and 4.22. The
growth rate is a function of the rate of both dotted square represents one unit cell. The crystal
processes, with neither being rate-controlling.
­ faces indicated in this figure follow planes
The effect of impurities are various: (1) Viscosity between adjacent sucrose molecules in such a
can increase, thus reducing the rate of mass trans- way that the furanose and pyranose rings are not
fer, (2) impurities can involve adsorption-occlu- intersected.
sion on specific surfaces of the crystal, thereby The processing and refining of sugar (sucrose)
reducing the rate of surface incorporation, (3) represents an excellent example of the seeding
inclusion whereby the impurity is incorporated crystallization process for the large-scale manu-
inside the crystal by the physical capture of syrup facture of sugars. A seed slurry is prepared with
or mother liquor, and (4) co-crystallization can a ball mill whereby powdered sugar (average
occur when there is a different solid impurity 50 μm size) is mixed with isopropyl alcohol and
with a lower free energy (lower solubility) than ground for 24 h. A 1.5–4.0 μm seed slurry is pro-
that of sucrose in the process. duced, however, different sizes are produced for
The crystal structure of sucrose has been different vacuum pan crystallizations depending
established by X-ray diffraction and neutron on the final crystal product. The seed pan is
­diffraction studies. The packing of sucrose mol- added to the vacuum pan containing the correct
ecules in the crystal lattice is determined mainly supersaturated syrup or mother liquor. The
by hydrogen bond formation between hydroxyl growth of sucrose crystals is a two-step process.

Fig. 4.21  The reference molecule (asymmetric unit) in the molecule, projected along a line in the plane of a and
the sucrose crystal structure viewed in projection along b, 30° from a. Numbers attached are bond lengths (A)
the a axis. The insert shows a clearer view of a portion of
186 4 Carbohydrates

Fig. 4.22  Projection of a sucrose crystal along the b axis (Wang et al. 2000)

First the sucrose molecules migrate to the sur- Clarification processes can vary, but the most
face of the growing crystal (a first-order reac- common is hot lime clarification (Eggleston et al.
tion) followed by the incorporation of the sucrose 2003). The clarified juice is then concentrated
molecules into the crystalline matrix (a second through a series of multiple-effect vacuum evapo-
order reaction). rators to syrup of ~65% dissolved solids. Syrup
Commercially available refined sugar (sucrose) from the final evaporator unit is then concentrated,
has a very high purity (>99.9%). To obtain sugar, under vacuum at lower temperature than in evapo-
a pure product from both sugarcane and sugar ration to minimize the chemical and thermal deg-
beet, rather complex isolation and purification radation of sucrose, and crystallized. The vacuum
processes are followed. Essentially there are a pans are seeded with finely ground sucrose to
series of separations of non-sucrose impurities allow larger sucrose crystals to form. A mixture of
from sucrose. Sugarcane processing occurs in two sucrose crystals and mother liquor (massecuites)
stages (Eggleston 2008). Firstly, the juice is is produced that is then separated in centrifuges,
extracted from sugarcane and converted into raw and the mother liquor is re-concentrated and re-
sugar (98–99% purity; golden yellow/brown crys- crystallized to give two more crops of crystals.
tals) at factories (mills) near where sugarcane is The final liquor is the by-product molasses.
grown. Secondly, after raw sugar has been trans- Unlike sugarcane factories that are limited to
ported to the refinery, it is refined using very simi- operation during the sugarcane harvest season,
lar unit processes used in raw sugar manufacture, raw sugar refineries operate year round. Since the
to the white, refined edible sugar. In comparison production of refined sugar from raw sugar at
to sugarcane, sugar beets are grown in temperate the refinery is also a series of separations of non-
areas and are processed directly into white sugar sugars from sucrose (Fig. 4.24), it consists of
at nearby factories. A flow chart of the typical unit many similar processes as in raw sugar manufac-
processes in the manufacture of raw sugar from ture at a factory (Fig. 4.23). There is, however, a
sugarcane at a factory is outlined in Fig. 4.23. greater onus at the refinery to remove color and
Juice is extracted from sugarcane by either tan- ash, so after clarification there are decolorization
dem milling or diffusion. Extracted juice is then or demineralization processes that can differ from
purified in the clarification unit process. refinery to refinery. The first stage of the refining
Oligosaccharides 187

Field Cane

Harvested
Cane

Crushing & Juice


Milling or Diffusion Heating Liming Clarification

Bagasse

syrup Multi-stage
Centrifugation Crystallization
Evaporation

Raw Molasses
Sugar

Fig. 4.23  Basic flow chart of the raw sugar manufacturing process in a sugarcane factory

Raw Sugar

filtration
mixed with washed with water
syrup water

Decolorization
Mingler & or
Melter Clarification
Affination Demineralization
syrup or
Softening

Refined sugar Centrifugation Crystallization

Molasses

Fig. 4.24  Basic scheme of the white, refined sugar manufacturing process in a sugar refinery

process is named affination, where the raw sugar many refiners are trying to eliminate affination
is mixed with syrup (magma) and then centrifuged because it is high energy and cost intensive. The
and washed with water to remove the molasses affined sugar is then melted in water to create a
layer around the raw sugar crystals. Nowadays, melt liquor of ~68% dissolved solids. The melt
188 4 Carbohydrates

liquor is then clarified by either a phosphatation or heated to maintain an 85 °C temperature, before it
carbonation clarification process; both clarifica- is purified with a double-carbonatation clarifica-
tion processes remove turbid particles and some, tion process (Fig. 4.25). In some sugar beet facto-
but not all, colorants. The clarified syrup is then ries, sulfur dioxide is added to filtered, clarified
decolorized with either ion exchange resins or juice to minimize color formation during subse-
granular activated carbon (Eggleston 2008). quent processing. The resulting clarified “thin”
Desalting ion exchange resins can also remove ash juice is then concentrated to 65% dissolved solids
or minerals. Multi-stage crystallization is the final (“thick” juice) across multiple-effect vacuum
purification process at the refinery to produce evaporators, then triple-crystallized and centri-
white, refined sugar sold in solid or liquid form. fuged to produce white refined sugar. In many
Production of refined, white sugar from sugar sugar beet factories additional purification steps
beets has some similarities to refined cane sugar are employed, such as softening, demineralization,
production, as both are a series of process units or color removal with ion-exchange resins or gran-
aimed at separating and removing impurities from ular activated carbon.
sucrose. However, dissimilarities exist as sugar
beet is a tuberous root and sugarcane is a grass  rystallization of Lactose
C
(Eggleston 2008). A basic scheme of white, refined Lactose can occur in two crystalline forms, the
sugar manufacture from sugar beets is illustrated α-hydrate and the β-anhydrous forms and can
in Fig. 4.25. Sugar beets at the factory, are washed, occur in an amorphous or glassy state. The most
and sliced into “V” shaped cossettes. Cossettes are common form is the α-hydrate (C12H22O11·H2O),
added to a counter-current diffuser of a different which can be obtained by crystallization from a
design than sugarcane diffusers. Sucrose and supersaturated solution below 93.5 °C. When
impurities are extracted with hot water at 85 °C in crystallization is carried out above 93.5 °C, the
the diffuser. The generated diffuser juice is then crystals formed are of β-anhydrous type. Some

Field Sugar Beet

Harvested
Sugar Beet

Slicing and juice thin Multi-stage thick


Diffusion Clarification
juice Evaporation juice

Beet Pulp

Decolorization
or
Centrifugation Crystallization Demineralization
or
m Softening
ol
as
se
s SMB Betaine and sugar
White Chromatography
Sugar

Fig. 4.25  Basic scheme of the white, refined sugar manufacturing process in a sugar beet factory
Oligosaccharides 189

Table 4.8  Some physical properties of the two common forms of lactose
Property α-Hydrate β-Anhydride
Melting pointa 202 °C (dec.) 252 °C (dec.)
+89.4° +35°
Specific rotationb [α ]D
20

Solubility (g/100 mL) Water at 20 °C 8 55


Water at 100 °C 70 95
Specific gravity (20 °C) 1.54 1.59
Specific heat 0.299 0.285
Heat of combustion (cal/g) 3761.6 3932.7
Source: From R. Jenness, Principles of Dairy Chemistry, 1959, John Wiley and Sons
a
Values vary with rate of heating, α-hydrate losses H2O (120 °C)
b
Values on anhydrous basis, both forms mutarotate to +55.4°

Fig. 4.26  Solubility of


lactose in water. Source:
From E.O. Whittier,
Lactose and Its
Utilization: A Review,
J. Dairy Sci., Vol. 27,
p. 505, 1944

properties of these forms have been listed by ously change to that form provided sufficient
Jenness (1959) (Table 4.8). water is present. At equilibrium and room tem-
Under normal conditions the α-hydrate form is perature, the β-form is much more soluble and
the stable one, and other solid forms spontane- the amount of α-form is small. However, because
190 4 Carbohydrates

of its lower solubility, the α-hydrate crystallizes β = 109°47′. The crystallographic description of
out and the equilibrium shifts to convert β-into the crystal faces is indicated in Fig. 4.27. These
α-hydrate. The solubility of the two forms faces grow at different rates; the more a face is
and the equilibrium mixture is represented in oriented toward the β direction, the slower it
Fig. 4.26. grows and the (0T0) face does not grow at all.
The solubility of lactose is less than that of Amorphous or glassy lactose is formed when
most other sugars, which may present problems lactose-containing solutions are dried quickly.
in a number of foods containing lactose. When The dry lactose is non-crystalline and contains
milk is concentrated 3:1, the concentration of lac- the same ratio of alpha/beta as the original prod-
tose approaches its final solubility. When this uct. This holds true for spray or roller drying of
product is either cooled or when sucrose is added, milk products and also during drying for moisture
crystals of α-hydrate may develop. Such lactose determination. The glassy lactose is extremely
crystals are very hard and sharp; when left undis- hygroscopic and takes up moisture from the
turbed they may develop to a large size, causing a atmosphere. When the moisture content reaches
sensation of grittiness or sandiness in the mouth. about 8%, the lactose molecules recrystallize and
This same phenomenon limits the amount of milk form α-hydrate crystals. As these crystals grow,
solids that can be incorporated into ice cream. powdered products may cake and become lumpy.
The crystals of α-hydrate lactose usually occur Both lactose and sucrose have been shown to
in a prism or tomahawk shape. The latter is the crystallize in an amorphous form at moisture
basic shape and all other shapes are derived from contents close to the glass transition temperature
it by different relative growth rates of the various (Roos and Karel 1991a,b; Roos and Karel 1992).
faces. The shape of a α-hydrate lactose crystal is When amorphous lactose is held at constant
shown in Fig. 4.27. The crystal has been charac- water content, crystallization releases water to
terized by X-ray diffraction, and the following the remaining amorphous material, which
constants for the dimensions of the unit cell and depresses the glass transition temperature and
one of the axial angles have been established: accelerates crystallization. These authors have
a = 0.798 nm, b = 2.168 nm, c = 0.4836 nm, and done extensive studies on the glass transition of

Fig. 4.27  Crystallographic representation of a tomahawk crystal of α-lactose Monohydrate and photo of R-lactose
crystal (Raghavan et al. 2000)
Polysaccharides 191

amorphous carbohydrate solutions (Roos 1993; chitin the amino group of glucosamine is acety-
Roos and Karel 1991d). lated and exists in polymeric form (Fig. 4.28).
Seeding is a commonly used procedure to pre- Glucosamine is frequently bound to proteins as
vent the slow crystallization of lactose and the part of their glycoprotein structures, i.e., ovomu-
resulting sandiness in some dairy products. Finely cin in egg whites.
ground lactose crystals are introduced into the In glucosamine one of the hydroxyl groups is
concentrated product, and these provide numer- replaced with an amino group. With chitosan the
ous crystal nuclei. Many small crystals are formed amino group is acetylated through addition of an
rapidly; therefore, there is no opportunity for crys- acetate group (Fig. 4.29). Chitosan is also fre-
tals to slowly grow in the supersaturated solution quently partially hydrolyzed to remove the ace-
until they would become noticeable in the mouth. tate group resulting in the formation of chitosan.
Chitin is structural in insects and] crustaceans.
Chitosan has interesting properties existing as a
Compounds Related to Sugars polyamine including absorption and anti-micro-
bial activity.
Sugars can be appended with amino groups in a Galacturonic acid is a sugar where the C6 car-
class of molecules named amino sugars. Amino bon is oxidized to a carboxylic acid (Fig. 4.30).
sugars form the backbone of chitin in insects, D-galactose can also be oxidized at the C1 posi-
crustaceans, mollusks, and some mushrooms. In tion (D-galactonic acid) and at both the C1 and
C6 positions (meso-galactaric acid (mucic acid).
Galacturonic is one of the main monomers in
pectin. In pectin the carboxylic acid is generally
acetylated.

Fig. 4.28  Structure of glucosamine

Fig. 4.29  Repeat unit structures of chitin and chitosan polysaccharides

Fig. 4.30 Different
structures of
galacturonic acids

Galacturonic Galactonic acid meso-


192 4 Carbohydrates

Polysaccharides Starch

Polysaccharides, like oligosaccharides, consist The major carbohydrate polysaccharide in plant


of monosaccharaides bound together by glyco- tubers and seed endosperm is starch (Buléon
sidic bonds. Less than 10 monosaccharaides are et al. 1998; Blazek et al. 2011). Starch is a
considered oligosaccharides and greater than 10 ­hompolymer of D-glucose and is a storage carbo-
are polysaccharides. The number of monosac- hydrate in plants. It occurs as small granules with
charides in the polymer is referred to as the the size range and appearance characteristic to
degree of polymerization (DP). There are only a each botanical plant species. The granules can be
few polysaccharides with DPs less than 100, shown by ordinary and polarized light micros-
most are in the 200–300 DP range. Starches and copy and by X-ray diffraction to have a highly
cellulose can be much larger. Cellulose can have ordered crystalline structure. Morphologies are
DPs from 7000 to 15,000. It has been estimated shown in Fig. 4.31 by confocal laser microscopy.
that over 90% of the carbohydrate mass in the Each granule contains several million amylo-
world is in the form of polysaccharides. pectin molecules packed with a much larger
Homoglycans are polymers like starch and number of smaller amylose molecules. While
cellulose where all of the monosaccharide con- corn is the largest single source of commercial
stituents are the same, in these examples starch, other commonly used sources are wheat,
D-glucopyranosyl units. Homoglycans includes rice, potato, and tapioca (Fig. 4.31). The amylose
polymers that are branched like amylopectin. polymer is composed of glucose units in a linear
Polymers where two types of monosaccharides polymer while amylopectin is a highly branched
are included are called di-heteropolymers. And polymer (Fig. 4.32). Both starch polymers are
those with three different monomers are tri-het- composed of α-D-glucose units in the 4C1 con-
eropolymers, etc. formation. In amylose these are linked -(1 → 4)-,
with the ring oxygen atoms all on the same side,

Fig. 4.31  3-D Images of the starch granules from (a) Velde F, van Riel J, Tromp RH. 2002. Visualisation of
potato, (b) corn, (c) tapioca, (d) wheat, (e) mung bean, (f) starch granule morphologies using confocal scanning
Sweet Potato: Coloring Rhodamine, Objective 63w, laser microscopy (CSLM). J Sci Food Agric. 82, 13,
Electronic Zoom 1, Image size 160 × 160 × 36 μ. van de 1528–1536
Polysaccharides 193

iodine. The length of the chain determines the


color produced (Table 4.9). Many industrial
starch methods are based on this amylose-iodine
reaction, although some starch enzymatic meth-
ods are also available. Such iodometric methods,
measure total starch in food ingredients or food
products, but do not provide information on
how much of the starch is soluble or insoluble
(granular). Just recently, however, a new method
has been reported by Cole et al. (2016), that is
based on microwave-assisted sonication, which
is capable of measuring total, insoluble and solu-
ble starch in food products.
Fig. 4.32  The helical organization of amylase and the Amylose and amylopectin molecules differ
branched structure of amylopectin greatly in structure and function. Fig. 4.33 illus-
trates the linear linkages of amylose that result in
Table 4.9  The color produced by reaction of iodine with a helical structure, as well as the branching of
amyloses of different chain length amylopectin. Amylose has a lower molecular
Chain length No. of helix turns Color produced weight with a relatively extended shape of a heli-
12 2 None cal rod. Amylopectin is an extremely large mol-
12–15 2 Brown ecule but tends to be tightly packed. The presence
20–30 3–5 Red of amylose tends to reduce the crystallinity of the
35–40 6–7 Purple amylopectin and influence the ease of water pen-
<45 9 Blue etration into the granules. Although the α-(1 → 4)
links are capable of relatively free rotation around
whereas in amylopectin about one residue in the (ϕ) phi and (ψ) psi torsions, hydrogen bond-
every 20 is also linked -(1 → 6)- forming branch- ing between the O3′ and O2 oxygen atoms of
points. The degree of branching and amylose to sequential residues tends to encourage a helical
amylopectin ratios are highly variable depending conformation. These helical structures are rela-
on the botanical source of the starch. For exam- tively stiff and result in hydrophobic surfaces.
ple, amylomaizes contain over 50% amylose Amylopectin can be isolated from ‘waxy’
whereas ‘waxy’ maize has almost none (~3%) maize starch whereas amylose (without amylo-
(Li and Yeh 2001; Singh et al. 2003). The number pectin) is best isolated after specifically hydrolyz-
of glucose units may range in various starches ing the amylopectin with pullulanase (Vorwerg
from a few hundred to several thousand units. In et al. 2002). Genetic modification of starch crops
the most common starches, such as corn, rice, has enabled the control of amylose and amylopec-
and potato, the linear fraction is the minor com- tin in crops which improves the ability to control
ponent and represents about 17–30% of the total. functionality of the starches (Gidley et al. 2010).
Some varieties of pea and corn starch may have Starch granules are partially crystalline; native
as much as 75% amylose. The characteristic blue starches contain between 15 and 45% crystalline
color of starch produced with iodine relates material (Oates 1997). The crystallinity can be
exclusively to the linear, amylase fraction. The demonstrated by X-ray diffraction techniques.
polymer chain takes the form of a helix, which Two polymorphic forms, A and B polymorphs,
forms inclusion complexes with iodine mole- have been described. There is also an intermedi-
cules. The inclusions of iodine are due to an ate C form. Crystallinity results from intertwin-
induced dipole effect and consequent resonance ing of amylopectin chains with a linear component
along the helix. Each turn of the helix is made up of over ten glucose units to form a double helix
of six glucose units and encloses one molecule of (Fig. 4.34).
194 4 Carbohydrates

Fig. 4.33  Illustration of the molecular arrangement of amylose and amylopectin starch polysaccharides. Amylose
structure shows the helical nature of the glucose polymers and amylopectin illustrates the branching

Fig. 4.34 Hexagonal
packing of A-type (a)
and B-type (b) starch
crystalline polymorphs.
(a) The dense structure
of the A-type only
allows few structured
water molecules (red
dots). (b) The more open
B-type makes space for
more structured water
molecules (Damager
et al. 2010)

Amylose molecules are composed of mostly Approximately 5% of the residues represent


unbranched chains with 500–20,000 α-(1 → 4)- branch points. There are usually slightly more
D-glucose units. Occasionally amylose will have ‘outer’ unbranched chains (A-chains) than ‘inner’
a few α-1  → 6 branches and phosphate groups branched chains (called B-chains). There is only
bound to –OH groups of some glucose units may one chain (C-chain) containing the single reduc-
be found (Hoover 2001). Amylose can form an ing group (Fig. 4.35). The A-chains contain
extended shape (hydrodynamic radius 7–22 nm; between 13 and 23 residues. There are two main
Parker and Ring 2001). More typically amylose fractions of long and short internal B-chains with
is found as a left-handed single helix or with par- the longer chains (greater than about 23–35 resi-
allel left-handed double helical junction zones dues) connecting between clusters and the shorter
(Imbert et al. 1988). Single helical amylose has chains similar in length to the terminal A-chains.
hydrogen-bonding between the O2 and O6 atoms Each amylopectin molecule contains up to
on outside surface of the helix with only the ring two million glucose residues in a compact struc-
oxygen internal to the helix. ture with a hydrodynamic radius of 21–75 nm
Amylopectin is formed by non-random (Bertoft et al. 2008) (compare with waxy maize
α-1 → 6 branching of the amylose-type α-(1 → 4)- amylopectin >300 nm (Juna et al. 2011). The
D-glucose structure. Typically amylopectin mol- amylopectin molecules are oriented radially in
ecules contain about a million residues. the starch granule and as the radius increases the
Polysaccharides 195

Fig. 4.35 
Representations of key B
elements in starch
structure: Type A
Amylopectin double- Amorphous
helical chains can either regions
form the more open
hydrated. Type B A chains
hexagonal crystallites or
the denser Type A A B chains
crystallites. Type C with Crystalline
staggered monoclinic regions
packing, dependent on
the plant source of the C chains
granules (Parker and
Reducing
Ring 2001). Type A, end
with unbroken chain
lengths of about 23–29
glucose units is found in C Hilum
most cereals 6 glucose
D units

α-1→6
branchpoints

number of branches increases filling the space. in peas and beans (Fig. 4.35). Starch granule
The increased branching results in the formation architecture has been recently comprehensively
of concentric regions of alternating amorphous described (Tang et al. 2006).
and crystalline structure. In Fig. 4.35, A illus- Starch has many important functions in foods.
trates the essential features of amylopectin. B— It is used for water binding and as a thickener, an
the organization of the amorphous and crystalline emulsion stabilizer and gelling agent. An excel-
regions (or domains) of the structure generating lent review of starch functionality in foods can be
the concentric layers that contribute to the found in Copeland et al. (2009). Starch is a major
“growth rings “that are visible by light micros- component in many foods and ingredients such
copy. C—the orientation of the amylopectin mol- as wheat flour, where it is 80% of the flour.
ecules in a cross section of an idealized entire Starch, therefore, inherently delivers function to
granule. D—the likely double helix structure the final product. Refined starches are also added
taken up by neighboring chains and giving rise to to foods to provide functionality such as gelation
the extensive degree of crystallinity in granule. It or thickening. In plant tissue, such as cereals,
is postulated that the crystalline structure consists starch is found with radial, tight packing to form
of parallel left-handed helices with six residues dehydrated granules which contain about one
per turn. An alternative arrangement of intercon- water molecule per glucose. The shape and size
necting clusters has been described for some of the granules in the ingredients are specific to
amylopectins (Bertoft 2004). the botanical source (maize 2–30 μm; wheat
Type B, with slightly longer unbroken chain 1–45 μm; potato 5–100 μm (Jobling 2004). The
lengths of about 30–44 glucose units is found in swelling function is determined by the shape and
banana, some tubers such as potato and high size of the granules. Starch granules are generally
amylose cereal starches. Type C structure, which either larger and lenticular (lens-like, A-starch)
is a combination of types A and B, can be found with large swelling power or smaller and spheri-
196 4 Carbohydrates

Table 4.10  The progressive heating of starch leading to gelatinization


Raw starch that has not had moisture added does not undergo gelatinization. By definition,
gelatinization is a phenomena which takes place in the presence of heat and moisture. The
dry raw starch, if heated, would undergo dextrinization. This certainly would affect the
starch paste viscosity and starch gel strength. The paste viscosity would be decreased and
gel strength decreased
If a “limited amount” of moisture is added to the raw starch you may get partial
gelatinization. This condition exists in baked products
Cornstarch at a 5% level in 95% water would have a slight change occur if heat is initiated.
Water might be slightly ADSORBED onto the surface of the granule. Actually, in the
research from which these images came, I found that I got a difference in paste viscosity
and ultimate op as measured by viscosity if I allowed cornstarch to sit in water at room
temperature. This led me to believe that there is some initiation adsorption upon the
granule at room temperature (27C)
If this 5% dispersion of cornstarch was heated to 40C I would expect more water would be
ADSORBED onto the surface of the granule, the hydrogen bonding between the starch
polymers within the granule might begin to be loosened slightly. In some types of starches
water might even begin to be ABSORBED into the granule

If this 5% dispersion of cornstarch was heated to 50C I would expect more water would be
ADSORBED onto the surface of the granule, the hydrogen bonding between the starch
polymers within the granule would begin to be loosened. This would allow the water to
penetrate into the granule becoming ABSORBED by the granule. Additionally, some of the
amylose may begin to work itself off the granule surface, thus, opening the structure even more
If this 5% dispersion of cornstarch was heated to 60–65C I would expect more water would
be ADSORBED onto the surface of the granule, the hydrogen bonding between the starch
polymers within the granule would loosen. This would allow the water to penetrate into the
granule becoming ABSORBED by the granule. Additionally, some of the amylose would
work itself off the granule surface, thus, opening the structure even more. This in turn
would allow even more of the water to become ABSORBED and more amylose to work
itself out into a colloidal dispersion outside of the granule. The long amylose polymer is a
colloid in characteristics
This is intermediate between 60 and 70C. The precise changes are affected by rate of
heating, condition of the starch and other factors

If this 5% dispersion of cornstarch was heated to 70–90C I would expect more water would be
ADSORBED onto the surface of the granule, the hydrogen bonding between the starch
polymers within the granule would loosen. This would allow the water to penetrate into the
granule becoming ABSORBED by the granule. Additionally, the amylose would work itself
off the granule surface, thus, opening the structure even more. This in turn would allow even
more of the water to become ABSORBED and more amylose to work itself out into a
colloidal dispersion outside of the granule. The long amylose polymer is a colloid in
characteristics
At some point between 60 and 95C we would likely have gelatinization occur. This might
be measured by loss of birefringence, increased viscosity, translucency, increased
susceptibility to enzyme action, X-ray diffraction or some other chemical or physical
means. At this point, the starch granule is swollen as much as possible. It is a starch sol
until you remove it from the heat and begin to allow the amylose and some amylopectin to
recrystallize, i.e. realign
In some instances, when heated to 90C the starch granule could reach optimum
gelatinization and be a nice swollen granule sack. In other cases, this may allow the sack to
“implode” and loose their contents as there is not enough structure and hydrogen bonding
to hold the polymers together. It is interesting that overcooking, as with overstirring, will
decrease the starch paste colloidal sol viscosity
Polysaccharides 197

cal (B-starch) with less swelling power (Ao et al. chains (retrogradation) and its solution can lead
2007). Granules contain regions of amylopectin to an initial loss in viscosity and followed by a
which contain both crystalline (~30%) and amor- more slimy consistency. Mixing with
phous areas. As starch granules become hydrated κ-carrageenan, alginate, xanthan gum and low
they swell, lose crystallinity and leach amylose molecular weight sugars can also reduce retro-
out of the granule. High amylose starches exhibit gradation. At high concentrations, starch gels are
lower swelling and lower gel strength than lower both pseudoplastic and thixotropic with greater
amylose containing starches (Table 4.10). storage stability. Their water binding ability (high
Amylose exhibits the important function of but relatively weak) can provide body and texture
acting as a hydrocolloid. Its extended conforma- to foodstuffs, making it useable as a fat
tion is the cause of the high viscosity of water- replacement.
soluble starch over a broad temperature range. In the native undamaged state starch granules
The extended helical chains possess a relatively are insoluble in water, however, they will revers-
hydrophobic inner surface that does not hold ibly imbibe small amounts of water and swell
water effectively. The hydrophobic core of the slightly. When heated in water the order and crys-
amylose helix serves as a binding site for hydro- tallization within the granule is disrupted in a
phobic molecules such as lipids and aroma com- process referred to as gelatinization. The irrevers-
pounds. Amylose also forms useful gels and ible loss of order in the granule is characterized
films. When heated amylose is cooled the chains by a loss in birefringence, granule swelling and
form an association and crystallization (retrogra- loss of crystallinity. Gelatinization occurs over a
dation) on cooling and storage. When starch gels temperature range (Table 4.10) with the larger
are cooled slowly some starches form a precipi- starch granules gelatinizing first. Throughout the
tate of crystalline like material. This phenome- process there is leaching of amylose.
non is referred to as retrodegradation. It is Measurement of gelatinization and temperature
essentially a precipitation of linear amylose mol- ranges are dependent on the method used for
ecules. Typically starches with smaller amylose measurement and the ratio of water to starch.
polymers (400 glucose units) as in corn starch are Several early stages of gelatinization can be
more prone to retrogradation than starch from observed under a hot stage polarizing light micro-
potato where amylose polymers are longer (2000 scope. The loss of birefringence (disappearance
glucose units). Freezing accelerates the retrogra- of the maltese cross) is a useful means of measur-
dation process. After a freeze thaw cycle starch ing gelatinization. The initial loss of birefrin-
gels frequently become spongy and with slight gence (initiation) is the first stage, the mid-point,
pressure the water will begin to separate from the and the complete loss of birefringence are
gel. Bread staling is a practical example of retro- recorded. The complete loss of birefringence is
degradation. After baking and initial cooling the then defined as complete gelatinization, thus
amylose is already partially retrograded. defining the gelatinization temperature range.
Retrogradation is most readily observed in the When heated in excess water after gelatiniza-
staling of baked products where the texture tion is complete, the starch granules continue to
becomes hard. It can be reversed by reheating the swell. Throughout this process the leaching of
product. The retrogradation decreases storage amylose continues. Eventually the granules will
stability resulting in shrinkage and the release of be completely disrupted. This disruption is
water (syneresis). Increasing amylose concentra- accentuated by applying shear force as one would
tion decreases gel stickiness but increases gel observe during extrusion. This disruption results
firmness. Retrogradation is influenced by lipid in the formation of starch paste (pasting) which is
content, amylose/amylopectin ratio, chain length a solubilization of the amylose and amylopectin
of amylose and amylopectin, and solid concen- in a continuous phase. One can also observe
tration (Chung and Liu 2009). Amylopectin granule fragments or ghosts of starch granules in
interferes with the interaction between amylose the mixture. Complete disruption is rare but there
198 4 Carbohydrates

is more disruption under shear stress as in extru- Potato and tapioca starches form clear, weak gels
sion. When cooled the starch paste forms a firm on cooling. Waxy starches form heavy bodied
rigid gel. stringy clear pastes upon cooling and relatively
Starch gelatinization is an endothermic pro- poor gel formation. High amylose corn starch
cess and the temperatures and enthalpies of gela- requires very high temperatures for gelatinization
tinization are frequently measured by differential resulting in short bodied firm opaque gels on
scanning calorimetry (DSC). In the gelatinization cooling. Starches fill a wide range of roles in food
process water acts as a plasticizer for the starch production; mainly they are used to absorb water
polymers. When starch granules are heated in and produce viscous fluids, pastes and gels for
sufficient water (60%) the plasticized amorphous control of texture in foods. The extent of gelatini-
regions in the starch granule undergo a transition zation of starch in baked goods has a major
from the glassy to the rubbery state. This specific impact on product properties including texture,
transition temperature is the glass transition tem- storage stability and rate of digestion. In high fat
perature (Tg). This is similar to the definition of a low moisture cookies and pie crusts as much as
glass which is a solid capable of supporting its 90% of the starch in the product is not gelati-
own weight against flow, while a rubber is an nized. On the other extreme in white bread and
undercooled liquid that can exhibit viscous flow. angel food cake 96% of the starch is gelatinized
In typical food processing conditions there is and in many cases deformed.
sufficient heat and water for the starch granules
to swell beyond the point where the process can Resistant Starch
be reversed. Water enters the spaces between Resistant starch (RS) is starch that is not digested
chains, disrupts interchain bonding, and estab- in the normal gastric process (Englyst et al. 1987;
lishes hydration layers in the granule separating Sajilata et al. 2010) and as a result falls in the
the macromolecules. This lubrication or plasti- category of dietary fiber. RS is not rapidly
cizing effect causes the starch polymers to be digested like ordinary starch, and imparts the bio-
more separated and enables them to slip or be logical benefits of fermentable dietary fiber. RS is
considered partially dissolved. Ultimately the defined as the fraction of starch, which escapes
starch will swell to several times the original size. digestion in the small intestine, and may be fer-
The properties of cooked starches exhibit a mented by the micribiome in the large intestine
range of characteristics that can be used to further (Englyst et al. 1992). Various factors contribute
classify starches (Table 4.11). Cereal starches to starch’s resistance to digestion, thus there are
(corn, wheat and rice) form viscous short bodied four categories of RS, each with similar resis-
pastes and result in opaque gels on cooling.

Table 4.11  A summary of the size and some physical characteristics of commonly used food starches
Corn Starch Waxy Maize High-Amylose Corn Potato Tapioca Wheat Rice
Granule size (μm) 2–30 2–30 2–24 5–100 4–35 2–55 <1–9
% Amylose 28 <2 50–70 21 17 28 0–21
Gelatinization/Pasting°C 62/80 63/72 66/170a 58-65 52-65 52-85 65–75
Relative viscosity Medium Medium Very Low Very High Low low
High High
Paste Rheology Short Long Short Long Long Long Short
Paste clarity Opaque Cloudy Opaque Clear Clear Opaque Opaque
Retrodegredation High Low High Low Low High High
When choosing a starch for a particular application these characteristics should be considered
Source: From BeMiller JN et al. (2008). Carbohydrates in Fennema’s Food Chemistry, Fourth Edition S. Damodaran,
KL. Parkin, OR. Fennema (eds) p 84–151. Taylor and Francis
a
No increase in viscosity was observed up to 100 °C. Pasting does not occur until temperatures approach 170 °C
Polysaccharides 199

Table 4.12  Types of resistant starch


Type of RS Description Food sources
RS1 Physically protected Whole or partially milled grains and seeds, legumes
RS2 Ungelatinized resistant granules with type B Raw potatoes, green bananas, some legumes, high
crystallinity, slowly hydrolyzed by α-amylase amylose corn starch
RS3 Retrograded starch Cooked and cooled potatoes, bread, corn flakes,
foods with repeated moist heat treatment and cooling
RS4 Chemically modified starches by crosslinking Foods where modified starches are added, i.e. bread,
or acylation cakes

tance properties but very different origins. The however, the bulk consists of polymers, of which
four categories of RS are listed in Table 4.12. retrograded amylose often forms the major frac-
RS1 is the physically protected form of starch tion (Ranhotra et al. 1991a). RS has an impact
found in green bananas, grains, and pulses. It is similar to soluble dietary fiber in the colonic
physically inaccessible because it is not hydrated health by increasing crypt cell production rate, or
and attack by enzymes is prevented by the tissue decreasing the colonic epithelial atrophy in com-
structures. RS2 is raw starch granules, where the parison with no-fiber diets. RS influences tumori-
starch is tightly packed in a radial pattern and is genesis, and reduces serum cholesterol and
relatively dehydrated. This dense structure limits triglycerides. One of the modes of action for RS
the accessibility of digestive enzymes and accounts is that it is fermented to short chain fatty acids by
for the resistant nature. RS2 is typically found in the microbiome. The short chain fatty acids are
raw potato, banana, and high-amylose starch cere- rapidly utilized by the colonic epithelium.
als. RS3 is retrograded starch. The formation of RS3
occurs when the starch granule is completely  ydrolysis of Starch
H
hydrated, the amylose leaches out of the granules Hydrolyzed starches have many applications in
into the solution as a random coil polymer. When foods ranging from limited degrees of hydrolysis
the solution cools the amylose polymer chains for texture modifiers and foam stabilization, to
begin to reassociate as double helices which are sta- greater degrees hydrolysis for use as sweeteners
bilized by hydrogen bonds in a gel (Wu and Sarko and fermentation substrates. Most carbohydrate
1978). A typical example of RS3 formation occurs polymers, including starch, are readily hydro-
when bread or rolls become stale. The stiff texture lyzed by hot acids or enzymes. Greater hydroly-
of stale products is a result of the retrogradation sis of the starch results in the formation of
process. This effect can be partially reversed by dextrins. The dextrins tend to result in less vis-
reheating the product. RS4 includes structures of cous preparations and can be used at higher levels
modified starches obtained by chemical treatments in foods as fillers. They can also be used as coat-
like distarch phosphate ester. Starch with structure ings for panning and carriers of spray dried fla-
intermediate between the more crystalline resistant vors and colors. Some dextrins retain long linear
starch (for example, RS3 in staled bread) and more chains of amylose fragments which result in the
amorphous rapidly digestible starch (for example, formation of strong gels. More extensive hydro-
in boiled potato) is slowly digestible starch lysis of starch with acid or enzymes results in the
(Lehmann and Robin 2007). Slowly digestible formation of maltodextrins. Maltodextrins are
starch results in slower release of glucose causing categorized by their chain length which is
smaller postprandial blood glucose peaks and is expressed as dextrose equivalents. The degree of
therefore useful in the diabetic diet. hydrolysis is expressed as dextrose equivalent
RS is classified as a component of fiber on the (DE), defined as the amount of reducing sugars
basis of the recent definitions of dietary fiber present as dextrose and calculated as a percent-
given by AACC (2000) and NAS (2002). Portions age of the total dry matter. Table 4.13 lists corn
of RS consist of low-molecular-weight dextrins, syrups with various DE. The Dextrose Equivalent
200 4 Carbohydrates

Table 4.13  Composition of representative corn syrups


Saccharides (%)
Type of conversion Dextrose equivalent Mono- Di- Tri- Tetra- Penta Hexa- Hepta- Higher
Acid 30 10.4 9.3 8.6 8.2 7.2 6.0 5.2 45.1
Acid 42 18.5 13.9 11.6 9.9 8.4 6.6 5.7 25.4
Acid-enzyme 43 5.5 46.2 12.3 3.2 1.8 1.5 – 29.5a
Acid 54 29.7 17.8 13.2 9.6 7.3 5.3 4.3 12.8
Acid 60 36.2 19.5 13.2 8.7 6.3 4.4 3.2 8.5
Acid-enzyme 63 38.8 28.1 13.7 4.1 4.5 2.6 – 8.2a
Acid-enzyme 71 43.7 36.7 3.7 3.2 0.8 4.3 – 7.6a
Source: From J.D. Commerford, Corn Sweetener Industry, in Symposium: Sweeteners, I.E. Inglett, ed., 1974, AVI
Publishing Co.
a
Includes heptasaccharides

(DE) is related to the average degree of polymer-


weight can become very hygroscopic.
ization (DP) in the maltodextrin by the equation:
Maltodextrins are excellent bulk fillers for foods
DE = 100/DP. and contribute little or no sweetness. Further
hydrolysis results it mixtures of malto-oligosac-
The DE can also be considered as the percent-
charides, maltose and glucose. Maltodextrins
age of reducing power that would come from
(DE < 20) have compositions that reflect the
pure glucose. The DE, therefore, is inversely
physico-chemical characteristic of the starch
related to average molecular weight of remaining
used, particularly the ­amylose/amylopectin ratio
polymers in the material.
of the starch. A maltodextrin with DE 12 shows
When starches are further hydrolyzed to DE retrogradation in solution, producing cloudiness.
values of 20–60 they are referred to as corn syrup In comparison, a maltodextrin from waxy corn at
solids. These syrups are hygroscopic and rapidly the same DE does not show retrogradation
dissolve in water. Syrups with a DE of 42 are because of the higher level of α-1 → 6 branches.
extensively used in food products (Table 4.13). As the DE decreases, the differences become
The syrups have a high osmolality which inhibits more pronounced. A variety of maltodextrins
growth of most microorganisms yet are highly with different functional properties, such as gel
resistant to crystallization. An example is pan- formation, can be obtained by using different
cake syrup which is colored with caramel and starch raw materials. Maltodextrins of varying
added flavor, but consists primarily of DE 42 molecular weights are plasticized by water and
syrup. The acid conversion process has a practi- decrease the glass transition temperature.
cal limit of 55 DE; above this value, dark color Maltodextrins can retard the crystallization of
and bitter taste become prominent. There is a amorphous sucrose and, at high concentrations,
fairly constant relationship between the composi- totally inhibit sucrose crystallization (Roos and
tion of acid-converted corn syrup and its DE. The Karel 1991c). Maltodextrins with low DE and
composition of syrups made by acid-enzyme or with little or no remaining polysaccharide can be
dual-enzyme processes cannot be as easily pre- produced by using two enzymes. Alpha-amylase
dicted from DE. randomly hydrolyzes 1 → 4 linkages to reduce
Maltodextrins are defined as hydrolyzates the viscosity of the suspension. Pullulanase is
with measurable DE value of less than 20. Low specific for 1 → 6 linkages and acts as a deb-
DE maltodextrins have the highest molecular ranching enzyme. The application of these two
weight and are non-hygroscopic, whereas, high enzymes makes it possible to produce maltodex-
DE maltodextrins have the lowest molecular trins in high yield (Kennedy 1985). The action of
Polysaccharides 201

α-(1,4) links
Pullulanase α-(1,6) links
Exoamylase Target
a-amylase EndoamylaseTarget

a-glucoamylase

Fig. 4.36  Schematic representation of the action of starch-degrading enzymes. β-Amylase can also hydrolyze disac-
charide maltose units from the non-reducing end of amylose and amylopectin

starch degrading enzymes is discussed in more D-glucose syrups and crystalline D-glucose.
detail below. Glucoamylase requires the starch to be gelati-
nized and sequentially hydrolyzes glucose units
of the non-reducing ends amylose and amylopec-
Starch Degrading Enzymes tin. It has the ability to hydrolyze both 1 → 4 and
1  → 6 linkages. Glucoamylase can completely
There are various starch degrading enzymes, hydrolyze starch to glucose, however combing it
with different actions, that are described below with α-amylase makes the process more efficient
and illustrated in Fig. 4.36. by generating more non-reducing ends for
hydrolysis.
α
 -Amylase
The enzymatic hydrolysis of starch by α-amylase β
 -Amylase
is important for both processing and in vivo β-Amylase hydrolyzes disaccharide maltose
digestion. α-Amylase is an endoamylase that units from the non-reducing end of amylose and
cleaves both amylose and amylopectin producing amylopectin. It does not cleave 1 → 6 linkages.
oligosaccharides. The products are the result Therefore, when amylopectin is the substrate, in
α-amylase hydrolyzing the 1 → 4 linkages of the addition to maltose it yields pruned or unhydro-
starch. It does not attack double helix regions or lyzed fragments which are named limit dextrins.
starch helical sections that are complexed with
lipids. The resulting hydrolysis products from  ullulanase and Isoamylase
P
amylopectin remain branched via 1 → 6 linkages. Pullulanase and isoamylase are “debranching”
The bacterial amylases used in processing starch enzymes that specifically hydrolyze 1 → 6 bonds
are similar to the amylases in animal digestive in amylopectin. This produces shorter linear
tracts. This is why starches with large helical 1 → 4 chains.
regions are resistant to digestion.

Glucoamylase
Glucoamylase is an exo-amylase that is used in
combination with α-amylase to produce
202 4 Carbohydrates

High Fructose Corn Syrup (HFCS) sprayed with hydrochloric acid and heat applied.
This can be accomplished by direct spraying with
High fructose corn syrup HFCS) is a major ingre- hydrochloric acid or by moistening the starch and
dient in the food and beverage industry. HFCS applying hydrogen chloride gas. After heat treat-
was first marketed in the 1970s and is less expen- ment the hydrochloric acid is neutralized and the
sive than granulated sugar (sucrose). The major desired starch product is washed with water and
use of HFCS has been in beverages and soft- dried. In lightly treated starches the granules
drinks which, at its peak, accounted for 75–80% remain intact but the starches cook more quickly
use (Eggleston et al. 2017). The use of HFCS, and result in thinner and more clear (less turbid)
however, peaked in 1999 due to consumer and less viscous solutions. These starches are
concern rather than price or technical use. In
­ used for coatings, film formation and pan coating
­particular, consumers are concerned about a pos- of nuts and candies. Corn starch is an excellent
sible link between HFCS and metabolic diseases starting material to produce strong, fast setting
like obesity and diabetes. The flow chart for the gels in products like jelly beans and processed
commercial production of HFCS is illustrated in cheese loafs. A larger variety of products are
Fig. 4.37. The starting material for HFCS produc- obtained from starch hydrolysis by using various
tion is corn starch. This is slurried in water, mixed starches such as corn, wheat, potato, and cassava
with a thermally stable α-amylase and rapidly (tapioca) starch. Glucose syrups, known in the
heated, where the starch is rapidly gelatinized and United States as corn syrups, are hydrolysis prod-
hydrolyzed by the α-amylase. After cooling to ucts of starch with varying amounts of glucose
55–60 °C, the liquefied starch is further hydro- monomer, dimer, oligosaccharides, and polysac-
lyzed with glucoamylase. The hydrolyzed mix- charides. Depending on the method of hydrolysis
ture is then clarified, concentrated, passed through used, different compositions with a broad range
activated carbon and ion exchange beds. Seed of functional properties can be obtained.
crystals are added and D-glucose or D-glucose The initial step in starch hydrolysis by
hydrate crystals are produced. To produce HFCS enzymes is also illustrated in Fig. 4.37, and
the solution of D-glucose is passed through a col- involves the use of a heat-stable endo-α-amylase.
umn containing immobilized glucose isomerase This enzyme randomly attacks α-1  → 4 glyco-
(glucofructoisomerase). Glucose isomerase cata- sidic bonds resulting in rapid decrease in viscos-
lyzes the isomerization of D-glucose to ity. These enzymes can be used at temperatures
D-Fructose through a trans-enediol intermediate, as high as 105 °C. This reaction produces malto-
and it was this reaction that opened the way for dextrins (Fig. 4.37). The next step is saccharifica-
HFCS production (Dziezak 1987). The equilib- tion by using a series of enzymes that hydrolyze
rium of the mixture is approximately 58% glu- either the α-1  → 4 linkages of amylose or the
cose and 42% fructose. Fructose concentrations α-1  → 6 linkages of the branched amylopectin.
are further increased by passing the equilibrium The action of the various starch-degrading
mixture over a cation-exchange column which enzymes is shown in Fig. 4.37 (Olsen 1995). In
retains fructose. The fructose is then eluted and addition to products containing high levels of
can be used to produce the 55% fructose contain- glucose (95–97%), sweeteners with DE of 40–45
ing sweetener that is typically used in beverages. (maltose), 50–55 (high maltose), and 55–70 (high
conversion syrup) can be produced. High dex-
trose syrups can be obtained by saccharification
 tarch Hydrolyzates: Corn
S with amyloglucosidase. At the beginning of the
Sweeteners reaction dextrose formation is rapid but gradually
slows down. This decelleration is caused by for-
As previously stated, starch can be hydrolyzed by mation of branched dextrins by enzyme catalyzed
acid or enzymes or by a combination of acid and transglycosylation reactions, and because at high
enzyme treatments. When acid is used, the corn is
Polysaccharides 203

Fig. 4.37  Major steps in enzymatic starch conversion to Handbook of Starch Hydrolysis Products and Their
high fructose corn syrups. Source: Reprinted from Derivatives, M.W. Kearsley and S.Z. Dziedzic, eds., p. 30,
H.S. Olsen, Enzymic Production0 of Glucose Syrups, in © 1995, Aspen Publishers, Inc.

dextrose levels the repolymerization of dextrose The properties of starches can be modified by
into isomaltose occurs. chemical treatments that result in products suitable
for specific purposes in the food industry (Whistler
and Paschall 1967). Starches are used in food
Modified Starches products to produce viscosity, promote gel forma-
tion, and provide cohesiveness in cooked starches.
Chemical modification and physical modifica- When a slurry of starch granules is heated, the
tions of starch are frequently used to impart granules swell and absorb a large amount of water;
desired functionality in foods. There is a wide this happens at the gelatinization temperature (see
array of modified starches available from differ- Table 4.10), and the viscosity increases to a maxi-
ent starches, as well as various types and extents mum. The swollen granules then start to collapse
of modification. Modification of starches results and break up, and viscosity decreases. Starch can
in pastes that more effectively withstand food also be modified by acid treatment, enzyme treat-
processing conditions, including shear, heat and ment, cross-bonding, substitution, oxidation, or
acidic conditions, compared to native starches. heat (Table 4.14). Acid treatment results in thin
Modifications can be made to reduce the heat boiling starch. The granule structure is weakened
required for gelatinization, increased or decreased or completely destroyed as the acid penetrates into
paste viscosity, gel clarity, gel strength, reduced the intermicellar areas, where a small number of
syneresis, improved film formation and improved bonds are hydrolyzed. When this type of starch is
heat stability. Generally chemical modification of gelatinized, a solution or paste of low viscosity is
starch occurs at low levels (degree of substitu- obtained. A similar result may be obtained by
tion) on the hydroxyl group of the starch. These enzyme treatment. The thin boiling starches yield
degrees of substitution (DS) only range from low-viscosity pastes but retain the ability to form
0.002 to 0.2% but still, however, dramatically gels on cooling. Acid-converted waxy starches
alter the functional attributes of the starch. with low amylose levels, produce stable gels that
remain clear and fluid when cooled. Acid-
Table 4.14  Primary reactions in starch modification
Physical Gelatinization Solubility Functionality
Heat/Moisture Above gelatinization temperature but low Increased paste stability, higher gelatinization temperature Increases resistant starch
(27%) moisture
Annealing 40–60% moisture Higher gelatinization temperature, slower hydration Increases resistant starch
Below gelatinization temperature
Pre-­gelatinization 100 °C excess water Cold water dispersible Instant foods, rapid hydration foods
Drum Drying
Extrusion
Dextrinization Dry roast acidified Starch High solubility, higher reducing sugars Coatings, films, fat replacers in dairy and baked
products
Modification
Partial Acid HCL, H2SO4 Lower molecular weight Confections, batters and coatings
Hydrolysis H3PO4
Enzymatic Hydrolysis Below gelatinization temperature with amylase Lower molecular weight Confections, batters and coatings
Alkaline Treatment NaOH, KOH
Oxidation Peracetic acid Low viscosity, clear gels Batters, coatings binders and film forming
Hydrogen Peroxide
Sodium hypochlorite
Derivatization
Etherification Propylene oxide Improved clarity, less retrogradation. Freeze/thaw stability Gravies, dips sauces, puddings, pie filling
Esterifcation Acetic Anyhdride Lower gelatinization temperature, clear pastes Refrigerated foods, emulsion stabilizers,
Tripolyphosphat Low retrogradation encapsulation
1-Octonyl-succinic anhydride
Acetic + Adipic anhydrides
Sodium ortho-phosphate
Sodium Tripoly phosphate
Phospo-oxy chloride
Crosslinking Sodium ortho-phosphate High stability granules, high gelatinization temperature, Increase viscosity in soups, sauces and dairy
Sodium Tripoly phosphate Resists shear products
Phospho-oxychloride
Polysaccharides 205

converted starches with higher amylose levels are cross-linked starch is below 0.09%. In distarch
more likely to form opaque gels on cooling. The phosphate, the free and combined phosphate,
acid conversion is carried out on aqueous granular expressed as phosphorus, is below 0.04% when
starch slurries with hydrochloric or sulfuric acid at made from cereal starch other than wheat, 0.11%
temperatures of 40–60 °C. The action of acid if made from wheat starch, and 0.14% if made
incorporates a preferential hydrolysis of linkages from potato starch (Wurzburg 1995).
in the non-crystalline areas of the granules. The Substitution of starch is achieved by reacting
granules are weakened and no longer swell; they some of the hydroxyl groups in the starch mole-
take up large amounts of water and produce pastes cules with monofunctional reagents that intro-
of low fluidity. duce different substituents. The action of the
Cross-bonding of starch involves the forma- substituents lowers the ability of the modified
tion of chemical bonds between different areas in starch to associate and form gels. This is achieved
the starch granule. This makes the granules more by preventing the linear portions of the starch
resistant to rupture and degradation on swelling molecules to form crystalline regions. The differ-
and provides a firmer texture. The number of ent types of substituted starch include starch ace-
cross-bonds required to modify the starch granule tates, starch monophosphates, starch sodium
is low; a large change in viscosity can be obtained octenyl succinate, and hydroxypropyl starch
by as few as 1 cross-bond per 100,000 glucose ether. These substitution reactions can be per-
units. Increasing the number of cross-bonds to 1 formed on unmodified starch or in combination
per 10,000 units results in a product that does not with other treatments such as acid hydrolysis or
swell on cooking. There are two methods to cross-linking.
cross-link starch. The first, which gives a product Acetylation of starch is undertaken on suspen-
known as distarch adipate, involves treating an sions of granular starch with acetic anhydride or
aqueous slurry of starch with a mixture of adipic vinyl acetate. No more than 2.5% of acetyl groups
and acetic anhydrides under mildly alkaline con- on a dry starch basis are introduced, which
ditions. After the reaction the starch is neutral- equates to a degree of substitution of about 0.1%.
ized, washed, and dried. The second method, Acetyl substitution reduces the ability of starch
which produces distarch phosphate, involves to produce gels on cooling and also increases the
treating a starch slurry with phosphorous oxy- clarity of the cooled sol.
chloride or sodium trimetaphosphate under alka- Starch phosphates are monophosphate esters,
line conditions. Since the extent of cross-linking i.e., only one hydroxyl group is substituted in
is low, the amount of reaction product in the mod- contrast to the two hydroxyl groups involved in
ified starch is low. Free and combined adipate in production of cross-bonded starch. They are pro-

Fig. 4.38 
Phosphorylation of
starch with sodium
ortho- or triphosphate
206 4 Carbohydrates

Fig. 4.39  Reaction of


starch with octenyl
succinic anhydride

OH

O
Starch + NaoH + Starch - O

Starch plus propylene oxide under alkaline conditions produces hydroxyproplyl starch

Fig. 4.40  Hydroxypropylation of starch

duced by mixing an aqueous solution of ortho-, (Fig. 4.40) is often combined with the introduc-
pyro, or tripolyphosphate with granular starch; tion of distarch cross-links (Wurzburg 1995).
drying the mixture; and subjecting this to dry Oxidized starch is prepared by treating starch
heat at 120–170 °C. The level of phosphorus with the strong oxidant of sodium hypochlorite.
introduced into the starch does not exceed 0.4%. Although this starch is sometimes described as
The introduction of phosphate groups as shown chlorinated starch, no chlorine is introduced into
in Fig. 4.38 gives the product an anionic charge the molecule. The reaction is undertaken by com-
(Wurzburg 1995). Starch monophosphates give bining a starch slurry with a solution of sodium
dispersions with higher viscosity, better clarity, hypochlorite. Under alkaline conditions carboxyl
and better stability than the unmodified starch. groups are formed that modify linear portions of
They also have higher stability at low tempera- the molecule so that association and retrograda-
tures and during freezing. tion are minimized. In addition to the formation
Starch sodium octenyl succinate is a lightly of carboxyl groups, a variety of other oxidative
substituted half ester produced by reacting reactions may occur including the formation of
an aqueous starch slurry with octenyl succinic aldehydic and ketone groups. Oxidation increases
anhydride as shown in Fig. 4.39. The level of the hydrophilic character of starch and lessens
introduction of substituent groups is limited to 1 the tendency toward gel formation.
for about 50 anhydroglucose units. The treatment Dextrinization or pyroconversion of starch
may be combined with other methods of conver- occurs through the action of heat on dry, pow-
sion. The introduction of the hydrophilic car- dered starch. Usually the heat treatment is under-
boxyl group and the lipophilic octenyl group taken with hydrochloric or phosphoric acid at
makes this product amphiphilic and gives it the levels of 0.15% and 0.17%, respectively. After
functionality of an emulsifier (Wurzburg 1995). addition of the acid, the starch is dried and heated
Hydroxypropylated starch is prepared by in a cooker at temperatures ranging from 100 to
reacting an aqueous starch suspension with 200 °C. Two types of reaction occur, hydrolysis
propylenol oxide under alkaline conditions at
­ and transglucosidation. At low degree of conver-
temperatures from 38 to 52 °C. The reaction sion, hydrolysis is the main reaction and the
resulting product is known as white dextrin.
Polysaccharides 207

Table 4.15  Some common starch modification reactions


Modification type
Etherification Hydroxyalkyl
starch (with alkylene oxide)

Esterification Starch acetate


(with vinyl acetate)

Starch acetate
(with acetic anhydride)

Starch phosphate
(with orthophosphates)

Carboxymethyl starch (with


mono chloro acetic acid)

Cross-linking
With PoCI3

With STMP

With EPI

St starch, POCl3 phosphorus oxychloride, STMP sodium tri-meta phosphate

Table 4.16  Properties and applications of modified starches


Process Function/property Application
Acid conversion Viscosity lowering Gum candies, formulated liquid foods
Oxidation Stabilization; adhesion gelling Formulated foods, batters, gum confectionery
clarification
Dextrins Binding; coating; encapsulation; high Confectionery, baking (gloss), flavorings, spices, oils,
solubility fish pastes
Cross-linking Thickening; stabilization; suspension; Pie fillings, breads, frozen bakery products, puddings,
texturizing infant foods, soups, gravies, salad dressings
Esterification Stabilization; thickening; clarification; Candies, emulsions, products gelatinized at lower
when combined with cross-­linking, temperatures
alkali sensitive
Etherification Stabilization; low-temperature storage Soups, puddings, frozen foods
Dual modification Combinations of properties Bakery, soups and sauces, salad dressings, frozen
foods
Source: Reprinted with permission from O.B. Wurzburg, Modified Starches, in Food Polysaccharides and Their
Applications, A.M. Stephen ed., p. 93, 1995. By courtesy of Marcel Dekker, Inc.

Transglucosidation involves initial hydrolysis of more highly converted products known as yellow
α 1 → 4 glucosidic bonds and recombination dextrins. The dextrins have film-forming proper-
with free hydroxyl groups at other locations. In ties and are used for coating and as binders.
this manner new randomly branched structures or The chemical reactions and changes in struc-
dextrins are formed; this reaction happens in the ture of chemically modified starch have been
208 4 Carbohydrates

Fig. 4.41 Schematic
two-dimensional view of
glycogen. A core protein
of glycogenin is
surrounded by branches
of glucose unites. The
entire globular granule
may contain around
30,000 glucose units.
Source: W.D. McArdle,
F. I. Katch; V. L. Katch,
In Exercise Physiology:
Energy, Nutrition, and
Human Performance
(2006). Lippincott
Williams & Wilkins

comprehensively described by Singh et al. linked to the chains with α-1  → 6 glycosidic
(2003). Table 4.15 summarizes the primary starch bonds. The outermost branches of the chains are
modification reactions. These modifications 6–7 glucose units long. Glycogen synthesis is ini-
result in many useful food applications for chem- tiated by autoglucoslylation of a protein, glyco-
ically modified starch. Table 4.16 summarizes genin-1 (Fig. 4.41) (Lomako et al. 2004).
some common applications of chemically modi- In humans, glycogen, hydrated with three or
fied starch. four parts of water, is produced and stored pri-
The properties and applications of modified marily in the cells of the liver and muscles.
starches are summarized in Table 4.10 (Wurzburg Muscle glycogen is converted into glucose by
1995). The application of modified starches as muscle cells, and liver glycogen coverts to glu-
functional food ingredients has been described cose for use throughout the body. Glycogen is
by Luallen (1985). found in the form of granules in the cytosol/cyto-
plasm in many cell types, and plays an important
role in the glucose cycle. Glycogen is similar to
Glycogen starch in is readily rehydrolyzed to produce glu-
cose for the muscle tissue on demand.
Humans, animals, and fungi produce glycogen in In the liver, when a meal containing carbohy-
muscle as an energy reserve. Glycogen is a highly drates or protein is eaten and digested, blood glu-
branched polysaccharide of glucose. Figure 4.41 cose levels rise, and the pancreas secretes insulin.
illustrates the highly branched structure of Insulin acts on blood glucose in the liver to stim-
glycogen. ulate the action of several enzymes including gly-
Glucoses are linked together linearly by cogen synthase. Glucose are added to the chains
α-(1  → 4) glycosidic bonds, and branches are of glycogen as long as both insulin and glucose
Polysaccharides 209

remain plentiful. When it is needed for energy, β-D-glucose joined with β-1 → 4 linkages. In the
glycogen is broken down by glycogen phosphor- native state, cellulose occurs as extended ribbons
ylase and converted again to glucose (Manners of twofold chain geometry stabilized by hydro-
1991). In comparison, muscle cell glycogen gen bonding of each successive residue to its
serves as an immediate reserve source of avail- nearest neighbors. The great strength, fibrous
able glucose for muscle cells only. character, insolubility and inert characteristics of
cellulose, integral to its skeletal function in plant
Glycemic Index cell walls are due to the ordered packing of the
The glycemic index (GI) is a number associated cellulose chains (Rees 1977) The hydrogen bonds
with a particular type of food that indicates the result in a high degree of crystallinity and dense
food’s effect on a person’s blood glucose (also structure contributing to areas that do not absorb
called blood sugar) level). A value of 100 repre- water effectively and exhibit a high degree of
sents the standard, which equivalent to pure glu- enzyme resistance. When foods are dried the
cose. The GI represents the rise in a person’s amorphous regions of cellulose become increas-
blood sugar level 2 h after consumption of the ing crystalline in nature and result in toughness
food, and is useful to understand who the body of dried products.
breaks down available carbohydrates in foods, In food products, the crystalline area of the
i.e., starch. The glycemic effect of foods depends cellulose absorbs water poorly, thus it is highly
on a few factors: type of starch or carbohydrate, resistant to enzymatic attack. The amorphous
physical entrapment of the carbohydrates within regions absorb water and swell although they do
the food, and fat and protein contents of the not dissolve and they remain essentially indigest-
foods (Anon 2017). The GI is typically applied ible. Higher degrees of crystallinity provide
in the context and quantity of the food and the greater tensile strength and increased elastic
amount of carbohydrate in the food that is actu- modulus. An example of this is that when foods
ally consumed. A related measure, the glycemic like carrots are dehydrated, crystallinity increases
load (GL), factors this by multiplying the glyce- and the foods become tougher. Heating cellulose
mic index of the food by the carbohydrate con- causes modest reductions in hydrogen bonding
tent of the serving. Watermelon, for example, which results in some swelling of the cellulose.
had a high GI but a low GL for the quantity usu- Cellulose and modified celluloses are used in
ally consumed. Glycemic index Charts often a wide range of foods to provide important physi-
give only one value per food, but variations cal characteristics including bulk replacements of
occur due to, for example, variety, ripeness, digestible carbohydrates in low calorie foods.
cooking methods, processing, and the length of Unmodified cellulose and its derivatives can be
food storage (Anon 2017). used to provide bulk, imbibe oils or flavors, or
serve as carriers to improve the flow characteris-
tics of intense sweeteners or flavors. The attri-
Cellulose butes of cellulose are based on its physical
characteristics including limited interaction with
Cellulose is one of the most widely distributed water, its insolubility in water, specific rheologi-
compounds in nature generally existing as homo- cal characteristics it brings to the product and the
logus polymers accompanied by other polysac- resulting texture of the product. For example,
charides and lignins. Cellulose can have DP as finely ground or microcrystalline cellulose
high as 10,000 which would translate to a molec- ­provides bulk to low calorie foods. Chemically
ular weight of approximately 1,620,000 Da. It modified celluloses provide emulsification, mod-
provides structural support to most plants and ification of texture, emulsification, foam stabili-
includes the major component in wood. Cotton zation, water binding and ice crystal formation.
cellulose is used for fabric and can be used in With modified cellulose the functions are deter-
some food applications. Cellulose is a polymer of mined by the chain length of the cellulose poly-
210 4 Carbohydrates

Table 4.17  Applications of commercial food cellulose products


Cellulose/modified cellulose Food applications
Hydroxypropyl cellulose Whipped toppings, mousses, extruded foods
Hydroxypropylmethyl cellulose Whipped toppings, mousses, baked goods, bakery fillings, icings, fried foods,
sauces, dressings, frozen desserts, reduced fat foods
Methyl cellulose Sauces, soups, breads, tortillas, fried foods, reduced fat foods, foams
Microcrystalline cellulose Dressings, sauces, baked goods, beverages, whipped toppings, reduced fat foods
Powdered cellulose Breads, cookies, pastries, pasta, imitation cheese, cereals, canned meats
Sodium carboxymethyl cellulose Frozen desserts, baked goods, dressings, sauces, beverages, animal foods,
reduced calorie foods, extruded foods

Fig. 4.42  Carboxymethyl modified cellulose repeating units

mer, the physical characteristics of the particular solutions can remain clear at up to 50% ethanol in
cellulose (amount of amorphous or crystalline the solution. When the degree of substitution is
region), the chemical modification, the degree of lower the gum is less soluble but binds more water.
modification and the interactions with other
ingredients in the food matrix. Microcrystalline Cellulose
The addition of carboxyl groups increases the Microcrystalline cellulose is a type of purified
hydrophilic nature of the cellulose, where the and partially depolymerized cellulose (DP is typi-
addition of hydrocarbons such as methyl or ethyl cally less than 400) that is white, odorless, taste-
groups makes the cellulose more hydrophobic. less, and occurs as a crystalline powder made up
The chain length of the cellulose polymer influ- of porous particles. MCC is synthesized from α−
ences the change in viscosity with longer chains cellulose. It can be synthesized by different pro-
resulting in greater increases in viscosity. When cesses including reactive extrusion, enzyme
producing films or coatings shorter chain length mediated reactions, steam explosion, and acid
is generally preferred. Table 4.17 summarizes the hydrolysis. The role of these reactions is to
applications of some commonly used modified destroy the amorphous regions of cellulose, leav-
celluloses. ing only the crystalline domains. MCC has
Carboxymethyl modification of cellulose is one become a very valuable additive in the food, phar-
of the most common modifications used to pro- maceutical, cosmetic, and other industries. In the
duce cellulose gum (Fig. 4.42). The characteristics food industry, MCC can be used as an important
of the gum are influenced by chain length and base in functional foods to: (1) maintain emulsifi-
degree of substitution. Longer chains result in cation and foam stability, (2) maintain high tem-
higher viscosity. Higher substitution increases perature stability, (3) improve liquid stability, and
water holding capacity. Addition of water soluble (4) act as a nutritional supplement and thickener.
solvents such as glycerine and ethanol increase the
viscosity of the gum. Increasing the degree of sub-
stitution increases the tolerance to dilution of water
by ethanol substantially. Carboxymethylcellulose
Polysaccharides 211

Fig. 4.43  Wheat flour


pentosan repeating unit
of L-arabinofuranosyl
linked by β1 → 4
glycosidic bonds to
D-xylopyranose
backbone

Fig. 4.44  Monomeric components of lignin: (a) trans-coniferyl alcohol, (b) trans-sinapyl alcohol, (c) trans-p-coumaryl alcohol

polysaccharide core with seven to eight xylopy-


Pentosans/Hemicelluloses ranose units and a one D-glucuronic acid attached
by a 1 → 2 linkage at the branch point.
Pentosans and hemicelluloses are groups of non The water-soluble pentosans are highly
–starch and non-cellulosic polysaccharides branched, highly viscous, and gel forming.
which are widely distributed in plants. Because of these properties, it is thought that the
Hemicelluloses are water insoluble polysaccha- pentosans may contribute to the structure of
rides and pentosans are water soluble non-stra- bread dough. Hoseny (1984) has described the
chy polysaccharides (D’Appolonia et al. 1971). functional properties of pentosans in baked
Hemicelluloses are structural plant polysaccha- foods. One of the more significant properties is
rides that are mostly composed of non-glucose due to the water-soluble pentosans, which form
containing sugars. They are classified based on very viscous aqueous solutions. These solutions
the carbohydrate sugars in the polymer. Sugars in are subject to oxidative gelation with certain oxi-
hemicelluloses can be xylose, arabinose, man- dizing agents. The cross-linking of protein and
nose or galactose. The hemicelluloses generally polysaccharide chains creates high molecular
contain polymers of two to four different sugar weight compounds that increase the viscosity and
units. Cereals are common sources of hemi-cel- thereby change the rheological properties of
luloses and pentosans. Arabinose and xylose are dough. Wheat flour contains 2–3% arabinox-
the primary pentoses found in pentosans and ylans. These water soluble pentosans are viscous
hemicellulose. Hexose units include glucose, and gel forming. Wheat flour contains 2–3% of
galactose, rhamnose, glucuronic acid and 4-)- water soluble and insoluble pentosans. The solu-
methyl-D-glucuronic acid. ble portion represents about 0.5–0.8% of the dry
In wheat flour the pentosans are arabinoxylans weight of the flour. It is estimated that the pento-
as shown in Fig. 4.43. The hemicelluloses in sans absorb about one third of the water in a nor-
wheat contain 59% L-arabinose, 38.5% D-xylose, mal dough. Treatment of doughs with pentosanase
and 9% D-glucuronic acid. It is a highly branched hydrolyzes the pentosans to oligomers and mono-
with arabinose on the side chains and an acidic mers of arabinose and xylose thereby drastically
212 4 Carbohydrates

reducing the viscosity of the dough and reducing ents as well as in molecular weight. The polymeric
bake time for cookies and crackers. Pentosanase units have molecular weights between 1000 and
treatment is also important in reducing the vis- 4000 Da. The polymeric units contain numerous
cosity of low/no fat doughs. hydroxylic and ether functions, which provide
opportunities for internal hydrogen bonds. These
properties lend a good deal of rigidity to lignin
Lignin molecules. One of the problems in the study of lig-
nin composition is that separation from the cell
Although lignin is not a polysaccharide, it is wall causes rupturing of lignin-polysaccharide
included in this chapter because it is a component bonds and a reduction in molecular weight so that
of dietary fiber and an important constituent of isolated lignin is never the same as the in situ lig-
plant tissues. Lignin is present in mature plant nin (Sarkanen and Ldwig 1971).
cells and provides mechanical support, conducts
solutes, and provides resistance to microbial deg-
radation (Dreher 1987). Lignin is always associ- Cyclodextrins
ated in the cell wall with cellulose and
hemicelluloses, both in close physical contact but When starch is treated with a glycosyltransferase
also joined by covalent bonds. Lignins are defined enzyme (CGTase), (E.C. 2.4.1.19; CGTase)
as polymeric natural products resulting from cyclic polymers are formed that contain six,
enzyme-initiated dehydrogenative polymerization seven, or eight glucose units. These are known as
of three primary precursors: trans-coniferyl, trans- α-, β-, and γ-cyclodextrins, respectively. The
sinapyl, and trans-p-coumaryl alcohol (Fig. 4.44). structure of β-cyclodextrin is shown in Fig. 4.45.
Lignin occurs in plant cell walls as well as in These ring structures have a hollow cavity that is
wood, with the latter having higher molecular relatively hydrophobic in nature because ­hydrogen
weights. Lignin obtained from different sources atoms and glycosidic oxygen atoms are directed
differs in the relative amounts of the three constitu- to the interior. The outer surfaces of the ring are

Fig. 4.45 Molecular
structure of cyclodextrin
showing linkages
Polysaccharides 213

Table 4.18  Properties of the main cyclodextrins used in food products

Outer Cavity diameter (nm) Cavity volume Solubility, Hydrate H2O


Cyclodextrin Mass diameter (nm) Inner rim Outer rim (mL/g) g/kg H2O Cavity External
α, (glucose)6 972 1.52 0.45 0.53 0.10 129.5 2.0 4.4
β, (glucose)7 1134 1.66 0.60 0.65 0.14 18.4 6.0 3.6
γ, (glucose)8 1296 1.77 0.75 0.85 0.20 249.2 8.8 5.4
Source: From Sabadini 2006; http://www1.lsbu.ac.uk/water/cyclodextrin.html#r915

Fig. 4.46  Structures of α, β and γ cylodextrins

hydrophilic because polar hydroxyl groups are Other possible applications have been described
located on the outer edges. The hydrophobic by Pszczola (1988). A disadvantage of this method
nature of the cavity allows molecules of suitable is that the complexes may become insoluble. This
size to be complexed by hydrophobic interaction. can be overcome by derivatization of the cyclo-
These stable complexes may alter the physical dextrin, for instance, by selective méthylation of
and chemical properties of the guest molecule. the C2 and C3 hydroxyl groups (Szejtli 1984).
For example, vitamin molecules could be com- Cyclodextrins have many applications in the
plexed by cyclodextrin to prevent degradation. food industry (Astray et al. 2009). The three most
214 4 Carbohydrates

widely used cyclodextrins contain six, seven, or with the ether-like anomeric oxygen atoms and
eight D-glucopyranonsyl residues (α-, β-, and the C3-H and C5-H hydrogen atoms. The hydro-
γ-cyclodextrin respectively) linked in a ring by phobic cavity contains about 3 (α-DC), 7 (β-DC)
α-1 → 4 glycosidic bonds. The glucose residues or 9 (γ-DC) poorly held and easily displaceable
have the 4C1(chair) conformation. All three cyclo- water molecules. The hydrophilic cyclodextrin
dextrins have similar structures differing only in molecules may bind non-polar suitably-sized ali-
number of glucose residues (Table 4.18). Their phatic and aromatic compounds such as aroma
shape is like a bottomless bowl. The molecule is compounds and lipophilic drugs in the hydropho-
stiffened by hydrogen bonding between the 3-OH bic cavity. They may bind in 1:1, 2:1 and 1:2
and 2-OH groups around the outer rim. ratios dependent on the molecules involved (for
The different chain lengths of the polymers example, two molecules of γ-cyclodextrin bind
result in different dimensions of the cavities. The well to single C60-fullerene molecules (Buvári-
cavities have different diameters dependent on the Barcza et al. 2001). Such binding also allows
number of glucose units (empty diameters between cyclodextrins to be used to increase the water
anomeric oxygen atoms illustrated in Fig. 4.46. solubility of normally hydrophobic compounds
The side rim depth (shown below in the diagrams) or minimize undesirable properties such as odor
is the same for all three (at about 0.8 nm). or taste in certain food additives. Cyclodextrin
Cyclodextrin rings have a wider rim with the complexes are now widely used in the pharma-
2- and 3-OH groups and the narrower rim with ceutical, food and cosmetic and toiletry fields
6-OH group on its flexible appendage. The (Hedges 1998).
hydrophilic groups are on the outside of cyclo- The cyclodextrins, by themselves, are natural,
dextrin and the inner surface is hydrophobic lined non-toxic additives. The hydroxyl groups may be

Fig. 4.47  Hypothetical structure of polydextrose repeating unit


Polysaccharides 215

derivatized to modify the specificity, physical and Pectins


chemical properties of the cyclodextrins. The
6-OH groups are most easily derivatized. The middle lamellae of plant cell walls contain
pectic materials that serve to cement the cellu-
losic network and help control the movement of
Polydextrose water. These pectic materials can be attached to
cellulose fibers and/or xyloglucans through gly-
Polydextrose is synthetically produced low calo- cosidic bonds. When heated under acidic condi-
rie polymer. It is produced by a random transgly- tions the pectin substances are hydrolyzed to form
cosylation polymerization of glucose with minor pectin and as the reaction continues they become
amounts of sorbitol and citric acid. Polydextrose soluble pectin. As fruit ripens similar hydrolysis
is a randomly bonded condensation polymer of takes place. Pectin is primarily composed of
glucose. It is synthesized in the presence of minor repeating units of α-1 → 4-galacturonic acid. The
amounts of sorbitol and citric acid. The polymer structure is a homologous polymer of 1 → 2-α-D-
contains all possible types of linkages between galactopyranosyluronic acid (Fig. 4.48). Pectins
glucose monomers, resulting in a highly branched can also contain α-D-galactouronan which is a
complex structure (Fig. 4.47). Because of the heteogenious section formed by repeating units of
material’s unusual structure, it is not readily bro- 1 → 2-α-L-rhamnosyl-(1-L)α-D-galactosyluronic
ken down in the human intestinal tract and there- acid. The main blocks in pectin are branched
fore supplies only 1 calorie per g. It is described galacturonan chains that are periodically inter-
as a bulking agent and can be used in low-calorie rupted with rhamnose units which causes bending
diets. It provides no sweetness. When polydex- in the chain. The rhamnose units can carry many
trose use is combined with artificial sweeteners, a sidechain units. The branched units alternate with
reduction in calories of 50% or more can be unbranched blocks. The rhamnose in the branched
achieved (Smiles 1982). blocks form galactan and arabinan blocks or ara-
binogalactan side chains form through a 1 → 4
linkage with the rhamnose. The galacturonic acid
is partially esterified with methyl groups, and the
degree of esterification varies among plant spe-
cies. Pectins are all similar but various species
exhibit differences in chain length, degree of
esterification and in some cases other sugars will
be on the side chains. The number of units
between branch points ranges from 8 to 20
residues.
The most important factors in determining
properties are chain length and degree of meth-
ylation. Pectins range from 9 to 12% methyl
esters, although the theoretical maximum would
be 16%, no pectin has been reported with that
extent of methyoxylation. Treatment of pectin
with alkali results in hydrolysis of the methyl
esters. Complete hydrolysis of the esters results
in forming pectic acid which is completely
Fig. 4.48  A repeating segment of galacturonic acid units insoluble.
in pectin. Source: Reprinted with permission from
In the presence of calcium ions, sugar and acid
D.G. Oakenfull, The Chemistry of High Methoyxl Pectins,
in The Chemistry and Technology of Pectin, R.H. Walter, pectin forms gels. These gels are three dimen-
ed., p. 87, © 1991, Academic Press sional networks of pectins that bind large amounts
216 4 Carbohydrates

of water. The pectins for this purpose should be Table 4.19  Pectin levels in selected fruits and vegetables
by two studies
branched and form interchain associations
through hydrophobic, hydrogen or ionic bonds. Campbell and Palmer 1978 Zilversmit
The ability to form these gels is controlled by the Product Range Average et al. 1979
composition of the side chains, degree of polym- Apples 0.71–0.84 0.78 0.78
erization, composition of the side chains and Apricots 0.71–1.32 1.02 1.00
degree of methylation. Bananas 0.59–1.28 0.94 0.94
Two types of pectins are commonly used in Beans 0.27–1.11 0.69 0.70
foods. They are referred to as High methoxy- Blackberries 0.68–1.19 0.94 0.94
(HM) and Low methoxy- (LM) based on degree Carrots 1.17–2.92 2.04 2.00
of methylation. HM pectin has 50–80% of the Cherries 0.24–0.54 0.39 0.39
Dewberries 0.51–1.00 0.76 nla
acid sites esterified with methyl esters and the
Grapes 0.09–0.28 0.19 0.19
LM has 25–50% esterification. The HM forms
Grapefruit 3.30–4.50 3.90 3.90
acid gels and the LM forms gels with Calcium
Lemons 2.80–2.99 2.90 2.90
ions acting as bridges between exposed carbox-
Loganberries 0.59 0.59 0.59
ylic acid groups. For HM gels to form stable gels
Oranges 2.34–2.38 2.36 2.36
they require the pH to be below 3.6 and the sugar
Raspberries 0.97 0.97 0.97
content to be at least 55% by weight. The HM
Squash 1.00–2.00 1.50 nl
gels are formed as a result of hydrogen and
Sweet 0.78 0.78 0.78
hydrophobic bonding that are stabilized as a Potatoes
result of the high sugar concentration which acts nla not listed
as a dehydrating agent in the system.
Commercially pectins are graded based on the
quantity of sugar required to gel one part pectin of the pectin gel is determined by the ratio of free
to an acceptable firmness. Common conditions carboxylic acids to methyl esters. Furthermore,
for establishing pectin grades are pH 3.2 to 3.5, the occurrence of more free carboxylic acid
sugar 65 to 70% and pectin from 0.2 to 1.5%. groups results in stronger gels.
Commercial pectin grades range from 100 to 500. The pectin content of fruits and vegetables
The degree of methylation in Rapid Set pectin is range widely as shown in Table 4.19. In juice
over 70%. The pectin gels form at a pH 3.0 to 3.4. manufacture pectin can present problems,
The gel strength is determined by chain length: because it holds water and reduces the amount of
the longer the chain the firmer the gel. Degree of juice that can be extracted. This is frequently
methylation does not have a significant effect on resolved by adding pectinases to hydrolyze the
gel strength. Slow set pectin has a degree of pectins in the juice extract.
methylation of 50–70% in a pH range from 2.8 to Pectin is one of the most versatile stabilizers
3.2. These pectins form gels at lower tempera- and gelling agents in food applications.
tures than the rapid set. LM pectins form gels Traditionally, pectin was primarily used in the
with Calcium ions and are not influenced signifi- production of jams and fruit jellies in low as well
cantly by sugar or pH. Gel strength is dependent as high sugar products. It helps develop the
on degree of methylation and molecular weight. desired texture, helps keep fruit distributed in the
The gel formation of pectin (homogalacturo- product and prevents water migration to the sur-
nan) with calcium ions is one of the most com- face. Some typical applications are listed below:
mon uses of pectin in foods. The gel is formed
when calcium ions interact with the free carbox- • Bakery fillings and toppings
ylic acid groups on the galacturonan polymer. –– Fruit preparations for dairy applications
These pectin gels are used in jams, jellies, con- • Fruit applications
fections, desserts, and dairy products such as –– Jams, jellies, and desserts
yogurt. The degree gelatinization and the strength • Dairy application
Polysaccharides 217

Table 4.20  Structure function properties and applications of polysaccharides


Polysaccharide source Molecular structure Function Applications
Agar Red seaweeds Sulfated polymers of (1 → 3)-β-D-­ Gelation Confections, dairy, meat
(Gelidium) Galactose and (1 → 4)-3,6-anhydro-α-­ In vivo –Non-digestible products
L-Galactose alternating; pyruvate and
methyl side chains
Carboxymethyl HO2C-CH2—group ether linked to Water retention, Ice cream, syrups, meat
cellulose Modified O-6 of the linear (1 → 4)-β-D-glucose stabilizer thickener products, cakes
cotton cellulose polymer In vivo—
Carageenans Sulfated polymers of (1 → 3)-β-D-­ Gelation, thickener Ice cream, desserts,
Galactose and (1 → 4)-3,6-anhydro-α-­ Stabilizer instant pudding,
Red Seaweed D-Galactose alternating; pyruvate and In vivo –Non-digestible dressing, meat products
(Gracilaria, methyl side chains
Gigartina,
Eucheuma
Guar Gum/Guar Linear β-D-(1 → 4)-manan changes Water retention, Ice Cream, milk
seeds appended with α-D-galactose side stabilizer, viscosity, products, desserts,
chains retards ice crystal growth bread, cakes, icings,
Locust Bean Gum/ In vivo—bulking agent sauces
Locust bean ~2 kcal/g
Gum Arabic Stems L-Arabino-(1 → 3)and (1 → 6)-β-D-­ Emulsifier, Confections, dairy, meat
of Acacia senegal galactan, branched with L-rhamnose encapsulation, thickener products, beverages
and D-galacturopnic acid Reduces surface tension,
increases fizzing in
carbonated beverages
In vivo—nearly 100%
digestible
Pectins Citrus, apple Partially esterified with methyl groups Gelation, stabilizer, Jams, Jellies, Preserves,
etc. or acetylated Linear and branched thickener dairy, confections,
(1 → 4)-a-D-galacturonic acid; In vivo – beverages
Xanthan gum Cellulosic backbone with D-mannose Thickener, stabilizer Dairy, dressings,
Xanthomonas and D-galacturonic acid side chains; In vivo—Non-digestible beverages
campestris Mannose is pyruvylated and
acetylated

–– Acidified milk and protein drinks gelling, stabilizing, and suspending agents.
–– Yoghurts (thickening) Compounds in this group come from different
• Confectionery sources and may include naturally occurring
–– Fruit jellies compounds as well as their derivatives, such as
–– Neutral jellies exudate gums, seaweed gums, seed gums, micro-
• Beverages bial gums, and starch and cellulose derivatives.
• Nutritional and Health Products Table  4.20 lists the source and molecular
• Pharmaceutical and Medical Applications structure of many of the gums as well as other
polysaccharides (Stephen 1995). These polysac-
charides are extensively used in the food industry
Gums as stabilizers, thickeners, emulsifiers, and gel
formers (Stephen 1995).
This large group of polysaccharides and their Gums have hydrophilic molecules, which can
derivatives is characterized by its ability to give combine with water to form viscous solutions or
highly viscous solutions at low concentrations. gels. The nature of the molecules influences the
Gums are widely used in the food industry as properties of the various gums. Linear polysac-
218 4 Carbohydrates

charide molecules occupy more space and are such as calcium may form salt bridges between
more viscous than highly branched molecules of neighboring molecules, resulting in gel forma-
the same molecular weight. The branched com- tion and—if much calcium is present—precipita-
pounds form gels more easily and are more stable tion. Examples of polysaccharides with strong
because extensive interaction along the chains is acid groups are furcellaran and carrageenan.
not possible. The linear neutral polysaccharides Both are seaweed extractives with sulfuric acid
readily form coherent films when dry, and they ester groups. Because the ionization of sulfuric
are good coating agents. Solutions are not tacky. acid groups is not reduced much at low pH, such
Solutions of branched polysaccharides are tacky gums are stable in solutions of low pH values.
because of extensive entangling of the side chains Gums can be chemically modified by intro-
and because the dried solutions do not form films duction of small amounts of neutral or ionic sub-
readily. The dried material can be more easily stituent groups. Substitution or derivatization to a
redissolved than can the dried linear compounds. degree of substitution (DS) of 0.01 to 0.04 is
Neutral polysaccharides are only slightly often sufficient to completely alter the properties
affected by change in pH, and salts at low con- of a gum. The effect of derivatization is much
centrations also have little effect. High salt con- less dramatic with charged molecules than with
centration may result in removal of the bound neutral ones.
water and precipitation of the polysaccharide. Introduction of neutral substituents along the
Some polysaccharides have long straight chains chains of linear polysaccharides results in
with many short branches. Such compounds, for increased viscosity and solution stability. Some
example, locust bean gum and guar gum, com- of the commonly introduced groups are methyl,
bine many properties of the linear and the ethyl, and hydroxymethyl. Acid groups can be
branched polysaccharides. Some gums have mol- carboxyl, introduced by oxidation, or sulfate and
ecules containing many carboxyl groups along phosphate groups. Introduction of strongly ion-
the chains; examples are pectin and alginate. ized acid groups may make the polysaccharides
These molecules are precipitated below pH 3 mucilaginous.
when free carboxyl groups are formed. At higher
pH values, alkali metal salts of these compounds Gum Arabic
are highly ionized, and the charges keep the mol- Gum arabic is a dried exudate from acacia trees.
ecules in extended form and extensively hydrated. It is a neutral or slightly acidic salt of a complex
This results in stable solutions. Divalent cat-ions polysaccharide containing calcium, magnesium,

Fig. 4.49  Galactomannan structure found in Guar and locust bean


Polysaccharides 219

and potassium anions. The molecule exists in a more soluble than locust bean gum. It is a straight
stiff coil with many side chains and a molecular chain of D-galacto-D-mannogly-can with many
weight of about 300,000 Da. The molecule is single galactose branches. The D-mannopyranose
made up of four sugars, L-arabinose, L-rhamnose, units are joined by β-1 → 4 bonds, and the single
D-galactose, and D-glucuronic acid. It is one of D-galactopyranose units are attached by α-1 → 6
the few gums that require high concentration to linkages. The branches occur at every second
give increased viscosity and is used as crystalli- mannose unit. The compound has a molecular
zation inhibitor and emulsifier. Gum arabic forms weight of 220,000 Da, forms viscous solutions at
coacervates with gelatin and many other low concentration, and hydrates rapidly in either
proteins. hot or cold water. At concentrations of 2–3%,
gels are formed. Guar gum shows no incompati-
 uar and Locust Bean
G bility with proteins or other polysaccharides.
Guar and locust bean gums are composed of Guar gum forms tough, pliable films.
galactomannan polymers, which give solutions Marked synergistic interactions of galacto-
of high viscosity and low polymer concentration, mannans with other specific polysaccharides can
but each has its own characteristics. The galacto- occur (Dea et al. 1977). This interaction is sensi-
mannan polymers are composed of α tive to the mannan/galactose (MG) ratio and thus
(1  → 4)-β-linked D-mannan polymer which is the type of galactomannan. For example, locust
appended with single units of (1 → 6)-α-linked bean gum can form mixed gels with the ordered
galactopyranosyl side chains as shown on helical structures of agars, carageenans, and xan-
Fig. 4.49. Because of the extent of branching the than at polymer concentrations as low as 1 mg/
galactomannans are resistant to crystallization. mL (Dea and Morrison 1975).
Locust bean gum is obtained from the carob
bean (Ceratonia Siliqua), which is cultivated Agar
exclusively around the Mediterranean. The com- Agar is extracted from red algae of the class
mercial gum contains 88% of D-galacto-D- Rhodophyceae. It is soluble in boiling water but
mannoglycan, 4% of pentoglycan, 6% protein, is insoluble in cold water. The gels are heat-resis-
1% cellulose, and 1% ash. The molecular weight tant, and agar is widely used as an emulsifying,
is about 310,000 Da, and the molecule is a linear gelling, and stabilizing agent in foods. The gel
chain of D-mannopyranosyl units linked 1 → 4. formation properties of agar are unique. At ele-
Every fourth or fifth D-mannopyranosyl unit is vated temperatures agarose exists in a “random
substituted on carbon 6 with a D-galactopyranosyl coil” disordered form, but on cooling forms
residue. Locust bean gum is known to form unique turbid, brittle gels at concentrations as
tough, pliable films. low as 0.04% w/w (Dea and Morrison 1975),
Guar gum is obtained from the seed of the which exhibit a marked degree of thermal hyster-
guar plant (Cyamopsis tetragonolubus) and is esis (i.e., gelation takes place at temperatures far

Fig. 4.50  Structure of


agarose
220 4 Carbohydrates

below the gel-melting temperature). Molecular Alginates


weight determinations have given varying results. Alginate gums are salts of alginic acid and they
Osmotic pressure measurements indicate values are obtained from the giant kelp Macrocystis pyr-
from 5000 to 30,000 Da; other methods, as high ifera or large, brown algae. Alginic acid is a
as 110,000. Agar is a mixture of at least two poly- mixed polymer of anhydro 1 → 4-β-D-
saccharides (Glicksman 1969): agarose, a neutral mannuronic acid and L-guluronic acid (Fig. 4.51).
polysaccharide with little or no ester sulfate The most common form is sodium alginate. A
groups, and agaropectin with 5–10% sulfate property of alginates which is of major impor-
groups. The ratio of the two polymers can vary tance both for their biological function as struc-
widely. Agarose consists of a linear chain of aga- tural polysaccharides or food application is the
robiose disaccharide units. The structure, as formation of strong, rigid gels on the addition of
shown in Fig. 4.50, indicates alternating 1 → 4 divalent cations, usually Ca2+ (Smidsrod 1974).
linked, 3,6-anhydro-L-galactose units and This functionality also underpins their thicken-
α1 → 3 linked D-galactose units. Agaropectin is ing, suspending, emulsifying, stabilizing, gel-
a sulfated molecule composed of agarose and forming, and film-forming properties in foods,
ester sulfate, D-glucuronic acid, and small and their solubility in both hot and cold water.
amounts of pyruvic acid. In neutral solutions, When no divalent cations are present, solutions
agar is compatible with proteins and other poly- have long flow properties. Increasing amounts of
saccharides. At pH 3, mixing of warm agar and calcium ions increase viscosity and result in short
gelatin dispersions cause flocculation. Some flow properties.
gums, such as alginate and starch, decrease the
strength of agar gels. Locust bean gum can Carrageenan
improve rupture strain of agar gels several times. Extracted from Irish moss (Chondrus crispus), a
red seaweed, carrageenan consists of salts or sul-
fate esters with a ratio of sulfate to hexose units

Fig. 4.51  Structure of


alginic acid

Fig. 4.52 Idealized
structure of
κ-carrageenan
Polysaccharides 221

Fig. 4.53 Idealized
structure of
λ-carrageenan

Fig. 4.54 Idealized
structure of
ι–carrageenan

of close to unity. Three fractions of carrageenan 3,6-anhydro-D-galactose 2-sulfate (Fig. 4.54). A


have been isolated, named κ-, λ-, and certain amount of 6-sulfate groups present can be
ι-carrageenan. The idealized structure of changed to 3,6-anhydro groups by alkali treat-
κ-carrageenan (Fig. 4.52) is made up of 1 → 3 ment. The comparative properties of the three
linked galactose-4-sulfate units and 1 → 4 linked types of carrageenan have been listed by
3,6-anhydro-D-galactose units. Actually, up to Glicksman (1969). Molecular weights of carra-
20–25% of the 3,6-anhydro-D-galactose units geenan vary from 100,000 to 800,000 Da.
can be sulfated at carbon 2 and some of the Carrageenan can form thermally reversible gels
anhydro-D-galactose may occur as galactose- whose strengths and gelation temperatures are
6-sulfate. The 6-sulfate group can be removed by dependent on the cations potassium and ammo-
heating with lime to yield anhydro residues, and nium. The mechanism has been visualized as a
this treatment results in greatly increased gel zipper arrangement between aligned sections of
strength. The major portion of λ-carrageenan linear polymer sulfates, with the potassium ions
consists of 1 → 3 linked galactose 2-sulfate and locked between alternating sulfate residues.
l → 4 linked galactose 2,6-disulfate (Fig. 4.53); Other monovalent cations, such as sodium, are
about 30% of the 1,3 galactose units are not sul- not effective, probably because of larger ionic
fated. The 6-sulfate group can be removed with diameter. At low concentrations, carrageenan can
lime treatment but does not result in gel forma- alter the degree of agglomeration of caseinate
tion. Iota-carrageenan consists mainly of 1 → 3 particles in milk. It is a highly effective suspend-
linked galactose 4-sulfate and 1 → 4 linked ing agent and is used to suspend cacao particles
222 4 Carbohydrates

Fig. 4.55 Chemical
composition of the
repeating unit of xanthan

in chocolate milk at concentrations as low as than has an ordered conformation (helix) (Morris
0.03%. Carrageenan is often used in combination et al. 1977). The technological use of xanthan as
with starch. The two compounds form complexes a commercial polysaccharide centers on its
that have useful properties in foods. The com- unusual solution properties such as its large vis-
plexes permit a lowering of the starch content by cosity increment in the presence of low molecu-
as much as 50% (Descamps et al. 1986). An lar weight salts (Whitcomb et al. 1977) and
example of mixed gels combining carrageenan having the ability to flow freely with a tenuous,
and whey protein has been described by Mleko gel-like structure at rest, which is capable of
et al. (1997). Optimal gelation occurred at pH 6 holding particles in suspension over long periods
to 7. The shear stress value of whey protein iso- of storage time, for example, in salad dressings.
late at 3% concentration was significantly
enhanced by the presence of 0.5% κ-carrageenan.
Dietary Fiber
Xanthan
Xanthan, is the extracellular polysaccharide from Originally, the fiber content of food was known
the plant pathogen Xanthomas campestris, is one as crude fiber and defined as the residue remain-
of the most commercially important water-solu- ing after acid and alkaline extraction of a defat-
ble polymers. It was discovered by USDA-ARS ted sample. During the 1970s, the physiological
scientist Allene Rosalind Jeanes, and brought effect of dietary fiber began to attract attention
into commercial production by Kelco in the early (Ink and Hurt 1987) and the need for better meth-
1960s (Whistler and BeMiller 1973). It has a ods for the determination of fiber became appar-
pentasaccharide repeating unit (Jansson et al. ent. Dietary fiber can be defined as a complex
1975; Melton et al. 1976) in which charged tri- group of plant substances that are resistant to
saccharide sidechains, which solubilize the poly- mammalian digestive enzymes. Because the defi-
mer, occur on alternate glucose residues of a nition is based on physiological properties rather
cellulose-like backbone, giving rise to a highly than common chemical properties, the analysis
branched molecule (Fig. 4.55). In solution, xan- of dietary fiber is not simple. Included in the defi-
Polysaccharides 223

Fig. 4.56 Association
of Official Analytical
Chemists (AOAC)
method for the
determination of total
dietary fiber. Source:
Reprinted with
permission from AOAC
Collaborative Study,
Total Dietary Fiber
Method, © 1984,
Association of Official
Analytical Chemists

nition of dietary fiber are cellulose, hemicellu- Table 4.21  Differences between crude fiber (CF) and
total dietary fiber (TDF) in some plant materials g/100 g
lose, lignin, cell wall components such as cutin,
minerals, and soluble polysaccharides such as Plant material CF TDF Ratio
pectin. A method for determining total dietary Cellulose 72.5 94.0 1:1.3
fiber (TDF) that is based on enzymatic digestion Pea hulls 36.3 51.8 1:1.4
has been accepted by the Association of Official Corn bran 19.0 88.6 1:4.7
Analytical Chemists (AOAC 1984) and is recog- Distiller’s dried grains 10.9 45.9 1:4.2
nized for labeling food products. To determine White wheat bran 8.7 36.4 1:4.2
the calorie content of a food, the TDF can be sub- Citrus pulp 14.4 24.8 1:1.7
tracted from the total carbohydrate content. The Source: Reprinted with permission from M.L. Dreher,
Handbook of dietary Fiber: An Applied Approach, p. 58
AOAC method for total dietary fiber is outlined 1987. By courtesy of Marcel Dekker, Inc.
in Fig. 4.56.
Earlier literature refers to crude fiber, which
consists of part of the cellulose and lignin only. the difference between the two measurements are
This method is now obsolete. The dietary fiber given in Table 4.20. One of the first alternative
content of foods is usually from 2 to 16 times methods was the acid detergent fiber (ADF)
greater than the crude fiber content. Examples of method developed by van Soest (1963). In
224 4 Carbohydrates

Fig. 4.57  Furda method


for the determination of
insoluble and soluble
dietary fiber. Source:
Reprinted with
permission from
I. Furda, Simultaneous
Analysis of Soluble and
Insoluble Dietary Fiber,
in The Analysis of
Dietary Fiber in Food,
W.P.I. James and
O. Theander, eds., 1981.
By courtesy of Marcel
Dekker, Inc.

this procedure, hemi-cellulose is completely and insoluble non-cellulose polysaccharides


extracted, and the residue contains lignin and cel- (NCP) in terms of hexose, pentose, and uronic
lulose. Thereafter, neutral dietary fiber (NDF) acid units. Examples of the determination of
methods were developed. These methods TDF by the Southgate method are listed in
­measure cellulose, hemicellulose, lignin, cutin, Table 4.21. Finally, enzymatic gravimetric meth-
minerals, and protein, but do not account for sol- ods were developed and adopted to determine
uble polysaccharides such as pectins. One of TDF in food. In these methods the defatted sam-
these methods, enzyme-modified neutral deter- ple is treated with enzymes to degrade proteins
gent fiber (ENDF), has been approved for the and starch. Starch removal is an essential step in
determination of insoluble dietary fiber. Chemical these procedures. The various steps evolved in
methods of determining TDF are known as the AOAC method for the determination of total
Southgate type methods (Southgate 1981). This dietary fiber are shown in Fig. 4.56, which shows
procedure measures cellulose, lignin, and soluble there is no separation of soluble and insoluble
Polysaccharides 225

Table 4.22  Total dietary fiber (TDF) and components as determined by the southgate method (g/100 g dry weight)
Noncellulose polysaccharides
Fiber source Hexose Pentose Uronic acid Cellulose Lignin TDF
Wheat bran 6.9 20.9 1.5 7.6 2.9 39.8
Rye biscuit 7.9 8.0 0.5 2.5 0.9 19.8
Dried apple 1.3 1.8 2.7 3.2 1.0 10.0
Citrus pectin 7.6 7.0 77.3 1.6 – 93.5
Potato powder 11.8 1.3 0.8 3.6 – 17.6
Soya flour 3.3 3.8 1.6 2.1 0.3 11.1
Source: Reprinted with permission from M.L. Dreher, Handbook of Dietary Fiber: An Applied Approach, p. 66, 1987.
By courtesy of Marcel Dekker, Inc.

Table 4.23  Composition of Component %


American Association of
Acid detergent fiber 11.9
Cereal Chemists (AACC)–
certified food-grade wheat Neutral detergent fiber 40.2
bran Total dietary fiber 42.4
Protein 14.3
Lipid 5.2
Ash 5.1
Moisture 10.4
Lignin 3.2
Pectin 3.0
Cutin 0.0
Total starch 17.4
Total sugar 7.0
Pentosan 22.1
Phytic acid 3.4
Source: Reprinted with permission from M.L. Dreher,
Handbook of Dietary Fiber: An Applied Approach, p. 82,
1987. By courtesy of Marcel Dekker, Inc.

Table 4.24  Overview of dietary fiber methods


Method Portion removed during analysis Fiber components determined by method
Crude fiber (CF) 80% lignin, 85% hemicellulose, and Remainder of the lignin, hemicellulose,
20–60% cellulose and cellulose
Acid detergent fiber (ADF) Solubilizes cellular components (starch, Cell wall components, except
sugars, fat, nitrogen compounds, and hemicellulose, as one unit
some minerals) plus hemicellulose
Neutral detergent fiber (NDF) Solubilizes cellular components: Cell wall components as one unit
or enzyme-­modified NDF soluble fiber
NDF-ADF – Hemicellulose
72% sulfuric acid (Klason Cellulose Lignin, insoluble nitrogen compounds,
lignin) cutin, silica
Permanganate oxidation Lignin Lignin (loss in weight)
Southgate-type methods Solubilizes cellular components; Individual chemical components (including
(unavailable carbohydrate) hydrolysis starch soluble and insoluble polysaccharides) = 
total dietary fiber (TDF)
Enzymatic methods Solubilizes cellular components; TDF (indigestible residue); isolation
hydrolysis starch and protein soluble and insoluble fractions
Fractionation methods – Isolation and determination of individual
components
Source: Reprinted with permission from M.L. Dreher, Handbook of Dietary Fiber: An Applied Approach, p. 106, 1987.
By courtesy of Marcel Dekker, Inc.
226 4 Carbohydrates

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Stephen, A. M. (1995). Food polysaccharides and their New York: Academic Press.
applications. New York: Marcel Dekker. Whistler, R. L., & BeMiller, J. (1973). Industrial gums:
Szejtli, J. (1984). Highly soluble β-cyclodextrin deriva- Polysaccharides and their derivatives. New York:
tives. Starch, 36, 429–432. Academic Press.
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Molecular arrangement in blocklets and starch granule Rheology of xanthan gum solutions. Extracellular
architecture. Carbohydrate Polymers, 63, 555–560. Microbial Polysaccharides, ACS Symposium Series,
Thompson, A. (1954). Acid reversion products from 45(12), 160–173.
D-glucose. Journal of the American Chemical Society, Wu, H. C., & Sarko, A. (1978). The double helical molec-
76, 1309–1311. ular structure of crystalline A-amylose. Carbohydrate
van Soest, P. J. (1963). Use of detergents in the analysis of Research, 61, 7.
fibrous seeds. II. A rapid method for the determination Wurzburg, O. B. (1995). Modified starches. In A. M.
of fiber and lignin. Journal of Association of Official Stephen (Ed.), Food polysaccharides and their appli-
Analytical Chemists, 48, 829–835. cations. New York: Marcel Dekker.
van de Velde, F., van Riel, J., & Tromp, R. H. (2002). Ziesenitz, S. C. (1996). Basic structure and metabolism of
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1528–1536. N. Y. (1979). Dietary fiber. In R. Levy, B. Rifkind,
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Journal of the American Chemical Society, 58, coronary heart disease (pp. 149–174). New York, NY:
600–601. Raven Press.
Minerals
5
John W. Finley and John M. deMan

Mineral elements are present in many forms n


Introduction foods including chemical compounds, complexes
and ionic forms (van Dokkum et al. 1989). The
In addition to the major components, all foods diversity of chemical properties among mineral
contain varying amounts of minerals. The min- elements and the vast number of chemical changes
eral material may be present as inorganic or that can occur during processing and storage,
organic salts or may be combined with organic results in a very large number of mineral species in
material, as the phosphorus is combined with foods. Many of the mineral species that occur in
phosphoproteins and metals are combined with foods are unstable making it difficult to isolate and
enzymes. More than 60 elements may be pres- characterize specific mineral species in food.
ent in foods. It is customary to divide the min- All biological systems contain water and most
erals into two groups, the major salt components nutrients are delivered and metabolized in an
and the trace elements. The major salt compo- aqueous environment. Most of the availability
nents include potassium, sodium, calcium, and reactivates of minerals are dependent on sol-
magnesium, chloride, sulfate, phosphate, and ubility in water. Generally, the minerals must be
bicarbonate. Trace elements are all others and in ionic or complexed forms because pure ele-
are usually present in amounts below 50 parts mental minerals are not absorbed by animals.
per million (ppm). Dietary elements in the die The forms of the elements depend on the specific
can be divided into four basic categories as chemical properties of the element. For example,
found in Table 5.1. the highly water soluble elements Na+, K+, Cl−,
A total of 90 naturally occurring elements are and F−, exist in ionic forms in food. Most other
found in the earth’s crust. Approximately 25 ele- minerals are found as complexes, chelates, or
ments are essential for life and are found in living oxygen containing anions.
cells. Because most of our diet is derived from Much of the chemistry of minerals in food and
living material these essential elements are found nutrition can be described in terms of acid/base
in foods in our diet. Foods can also contain non-­ chemistry. The Bronsted theory of acid base
essential nutrients and toxic elements which chemistry states:
often come from contamination during produc-
tion, harvesting, processing and preparation. The A Bronsted acid is any substance capable of
essential elements for humans can be seen on the donating a proton.
naturally occurring portion of the periodic table A Bronsted base is any substance capable of
shown in Fig. 5.1. accepting a proton.

© Springer International Publishing AG 2018 231


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_5
232 5 Minerals

Table 5.1  Elements in human diets


Essential human elements > 50 mg/day Essential trace elements < 50 mg/day
Na, K, Ca, Mg, Cl, P Fe, Cu, I, Co, Mn, Zn, Cr, Ni, Si, F, Mo, Se
Nonnutritive nontoxic elements Toxic elements
Al, B, and Sn Hg, Pb, As, Cd, and Sb

Fig. 5.1  Periodic table of naturally occurring elements. Shaded atoms are of nutritional significance

Foods contain many naturally occurring acids In the Lewis Theory A must possess unpaired
or bases naturally occurring in foods or as and electrons and B must have unpaired electrons.
phosphoric or hydrochloric are common inor- The interaction or bonding occurs when the
ganic acids in foods. Phosphoric acid is a com- orbitals of A and B interact to form new molec-
mon ingredient in foods and represents a tri basic ular orbitals. The stability of the new complex
acid because it contains three available protons depends on the reduction in energy that occurs
for donation. when the new molecule is formed. The product
of Lewis A and B molecules combining is gen-
H 3 PO 4  H 2 PO 4 − + H + pK1 = 2.22 erally referred to as a complex. Understanding
H 2 PO 4 −  HPO 4 2 − + H + pK 2 = 7.10 these interactions is key because metal cations
HPO 4 2 −  PO 4 3− + H + pK1 = 12.40 in foods are classified as Lewis acid which are
bound to Lewis bases. The products from the
The multiple pK values of phosphoric acid reaction to metal cations and various molecules
make it useful as a buffering agent and for pH in food products range from metal hydrates to
control in many foods and beverages. metal containing components such as hemoglo-
The Lewis Theory of acids and bases is based bin and chlorophyll and metalo-enzymes. The
on electrons rather than protons (Shriver et al. number of Lewis base molecules that can bind
1994): to a metal ion is independent of the charge on
the metal. The number of potential binding
A Lewis acid is an electron pair acceptor. molecules is referred to as the coordination
A Lewis base is an electron pair donor. number which can range from 1 to 12, with 6
The conventional expression of a Lewis acid base being the most common number. For example
reaction is: when the Fe+3 ion is in water it binds six water
molecules forming an octahedron structure as
A+ : B → A − B illustrated in Fig. 5.2.
Introduction 233

When ΔG is negative reactions will occur sponta-


neously, and become more favorable as ΔG
becomes more negative. For a spontaneous reac-
tion to occur enthalpy must decrease or entropy
must increase. A decrease in enthalpy represents
a decrease in the electronic energy of the system.
An increase in entropy means that there is an
Fig. 5.2  Ferric ion in acidic conditions, coordinated with increase in the randomness of the system.
six water molecules in an octahedral arrangement A chelate is a complex that forms when a
multi-dentate ligand forms two or more bonds
The electron donating species are referred to with a metal ion. This results in a ring structure
as ligands. A ligand must contain an oxygen which includes the metal ion. For this to occur
donating atom generally they are Oxygen, the chelating ligand must contain at least two
Nitrogen or Sulfur atoms in an organic mole- functional groups capable of donating electrons.
cule. The ligands are classified by the number of These ligands must be spatially close enough so
bonds they can share with a metal ion. The sta- that when the metal is included a ring can form.
bility of the complexes is expressed as stability Chelates are more thermodynamically stable than
constant or formation constant, shown below other simple complexes.
the metal ligand complex can be represented Kratzer and Vohara described critical factors
was M for metal ion, Ln for the number of for the formation of chelates:
ligands and Kn is the stability or formation con-
stant (Shriver et al. 1994). 1. Five and six membered saturated rings tend to
be more stable than smaller rings
ML n
M + L n  ML n K n = 2. The greater the number of rings in a chelate

[ M][ L n ] the greater the stability
3. Stroger Lewis bases tend to form stronger

The stability constants or Log K for some typ- chelates
ical metal complexes are listed in Table 5.2. 4. Charged ligands are more stable than

Many food components can form ligands for uncharged ligands
metal ions, including water, carbohydrates, pro- 5. Relative strengths of ligand bonds:
teins, phospholipids and organic acids. Figure 5.3 Oxygen as donor: H2O > ROH > R2O
illustrates the mon-dentate formation with water Nitrogen as donor: H3N > RN2 > R3N
and homogeneous and heterogonous dentate with Sulfur as donor: R2S > RSH > H2S
oxalate and glycine respectively. 6. Enhanced resonance in the chelate increases
The stabilities pf Lewis acid and base com- stability
plexes are proportional to the driving force of the 7. Large bulky ligands are less stable chelators
reaction. The reaction is described quantitatively
by the Gibbs free energy equation: Chelates are effected by many factors in com-
∆G = ∆H − T∆S. plex systems like foods. The concept of the
Gibb’s free energy can be a useful predictor of
where ΔG is the free energy in the reaction stability. This can be seen in the comparison of
­system, ΔH is the enthalpy change, T is the abso- Cu2+ complexing with ammonia or ethylene
lute temperature and ΔS is the entropy change. diamine (Shriver et al. 1994).
2+
Cu ( H 2 O )6 + NH 3 → Cu ( H 2 O )4 ( NH 3 )2 
2+
+ H2 O

( ∆H = −46 kJmol −1
; ∆S = −8.4 JK −1 mol −1 and log K = 7.7 )
2+
Cu ( H 2 O )6 + NH 2 CH 2 CH 2 NH 2 → Cu ( H 2 O )4 ( NH 2 CH 2 CH 2 NH 2 ) 
2+
+ 2H 2 O

( ∆H = −kJmol −1 −1 −1
;;DS = +23 JK mol ;; log K = 10.1 )
234 5 Minerals

Table 5.2  Major mineral content of some foods


Food Na K Ca Fea P
mg/100 g food
Whole milk 371 1330 912 0.47 776
Butter (unsalted) 11 24 24 0.02 24
Cheddar cheese 653 76 710 0.14 455
Monterey cheese 781 81 705 0.72 444
Yogurt skim milk 77 255 199 0.09 157
Egg whole 142 138 56 1.75 99
Egg white 166 163 7 0.08 15
Beef carcass 59 267 8 1.83 154
Pork carcass 42 253 19 0.69 155
Chicken meat only raw 77 229 12 0.89 173
Turkey whole meat raw 118 235 11 0.86 190
Fish Cod raw 54 413 16 0.38 203
Fish Tuna Yellowfin 45 441 4 0.77 278
Corn flour yellow whole 5 315 7 2.38 272
Wheat whole grain 2 363 34 3.6 357
Asparagras 2 202 24 2.16 70
Broccoli raw flowers 27 325 48 0.88 66
Carrots, baby raw 78 237 32 0.89 28
Kale, raw 38 491 150 1.47 92
Onions, raw 4 146 23 0.21 29
Peas raw 5 244 25 1.47 108
Potatoes fresh raw 6 425 12 0.81 425
Black beans 5 1483 123 5.02 352
Kidney beans, raw 12 1359 83 6.69 406
Apple Fuji raw 1 109 7 0.11 13
Orange, Raw Florida 0 169 43 0.09 12
Peach raw 0 190 9 0.32 31
Digiorno pepperoni pizza 663 268 209 0.9 268
McDonalds Double Cheeseburger 668 216 178 2.24 166
KFC Original Thigh with breading 781 237 64 0.88 230
Iron discussed below as a trace element
a

Both complexes have a single Cu+2 and two


Nitrogens, yet the ethylenediamine complex is
much more stable than the ammonia complex
with the Log formation constant for the ammonia
complex is 7.5 compared to 10.1 for the ethylene-
diamine complex. Both entropy and enthalpy
contribute to the difference, however entropy
change is the major factor. Ammonia forms a
monodentate ligand whereas ethylenediamine
forms a bidentate ligand bond with copper. The
entropy change is a result of the change in the
number of independent molecules in solution.
Fig. 5.3  Examples of metal ions coordinated with differ- The first reaction the number of independent
ent ligands molecules is the same on both sides of the
Introduction 235

Fig. 5.4  Cupric ion


Cu2+ complexed with
ammonia (a) and
chelated with
etylenediamine (b)

Fig. 5.5  EDTA and calcium EDTA chelate

e­quation, thus the entropy change is small. Ethylenediamine tetraacetic acid (EDTA) is
The chelation reaction results in a net increase in an even stronger chelator. EDTA is a hexadentate
the number of independent molecules in solution, chelator displacing six water molecules from a
increasing the entropy (Figs. 5.4 and 5.5). metal.

Ca ( H 2 O )6 + EDTA 4 − → Ca ( EDTA ) + 6H 2 O ∆S = +118 JK −1 mol −1


2+ 2−

The EDTA chelate contains five membered =C=O, =NH, –COOH, and –OH–O–PO(OH)2.
rings, which enhance stability. EDTA is widely Many metal ions, especially the transition metals,
used as a chelater in foods and other biological can serve as acceptors to form chelates with these
systems. ss in food sequester metal ions such as donor groups. Formation of chelates can involve
copper and ion preventing their action as ring systems with four, five, or six members.
proxidants. Some examples of four- five- and six-membered
The chelate’s stability depends on the nature ring structures are given in Fig. 5.6. Other exam-
of the metal ion and is related to the electronega- ples of food components that can be considered
tive chelate normally decreases with decreasing metal chelates are hemoglobin and myoglobin,
pH. In a chelate the donor atoms can be N, O, P, vitamin B12, and calcium caseinate (Pfeilsticker
S, and Cl; some common donor groups are –NH2, 1970). It has also been proposed that the gelation
236 5 Minerals

Fig. 5.6  Examples of metal chelates. With four, five, and six membered rings

of certain polysaccharides, such as alginates and of foods, we must pay great attention to the meth-
pectates, with metal ions occurs through chela- ods of analysis used.
tion involving both hydroxyl and carboxyl groups Some elements appear in plant and animal
(Schweiger 1966). A requirement for the forma- products at relatively constant levels, but in a
tion of chelates by these polysaccharides is that number of cases an abundance of a certain ele-
the OH groups be present in vicinal pairs. ment in the environment may result in a greatly
The minerals in foods are usually determined increased level of that mineral in plant or animal
by ashing or incineration. This destroys the products. Enrichment of elements in a biological
organic compounds and leaves the minerals chain may occur; note, for instance, the high mer-
behind. However, determined in this way, the ash cury levels reported in some large predatory fish
does not include the nitrogen contained in pro- species such as swordfish and tuna.
teins and is in several other respects different
from the real mineral content. Organic anions
disappear during incineration, and metals are I nteractions with Other Food
changed to their oxides. Carbonates in ash may Components
be the result of decomposition of organic mate-
rial. The phosphorus and sulfur of proteins and The behavior of minerals is often influenced by
the phosphorus of lipids are also part of ash. the presence of other food constituents. The
Some of the trace elements and some salts may recent interest in the beneficial effect of dietary
be lost by volatilization during the ashing. fiber has led to studies of the role fiber plays in
Sodium chloride will be lost from the ash if the the absorption of minerals. It has been shown
incineration temperature is over 600 °C. Clearly, (Toma and Curtis 1986) that mineral absorption
when we compare data on mineral composition is decreased by fiber. A study of the behavior of
Major Minerals 237

iron, zinc, and calcium showed that interactions vative in salted meat products. The emergence of
occur with phytate, which is present in fiber. refrigeration and other methods of food preserva-
Phytates can form insoluble complexes with iron tion, has reduced the requirement for salt as a
and zinc and may interfere with the absorption of preservative (He et al. 2007). Salt also provides
calcium by causing formation of fiber-bound cal- flavors that we rapidly become accustomed to
cium in the intestines. and many people develop strong preferences for
Iron bioavailability may be increased in the high salt levels, especially in processed foods.
presence of meat (Politz and Clydesdale 1988). The tastes and flavors associated with historical
This is the so-called meat factor. The exact mech- salt use have come to be expected, and the rela-
anism of this effect is not known, but it has been tively low cost of enhancing the palatability of
suggested that amino acids or polypeptides that processed foods has become a key rationale for
result from digestion are able to chelate nonheme the use of salt in food (Van der Veer 1985).
iron. These complexes would facilitate the Sodium continues to play a significant role in
absorption of iron. In nitrite-cured meats some reducing the growth of pathogens and organisms
factors promote iron bioavailability (the meat that spoil products and reduce their shelf life.
factor), particularly heme iron and ascorbic acid High Sodium levels also result enhanced func-
or erythorbic acid. Negative factors may include tional roles, such as improving texture. In addi-
nitrite and nitrosated heme (Lee and Greger tion to sodium chloride other sodium-containing
1983). compounds are also used for increasing the safety
and shelf life of foods or creating physical
properties.
Major Minerals Salt is effective as a preservative because it
reduces the water activity of foods. Salt’s ability
Foods provide most of the minerals important for to decrease water activity is due to the ability of
good health. Table 5.2 contains some of mineral sodium and chloride ions to associate with water
content of some foods illustrating the range of molecules (Fennema 1996; Potter and Hotchkiss
composition of the macro-minerals in foods. 1995). The addition of salt to foods can also
cause microbial cells to loose water from osmotic
shock, thereby causing cell death or retarded
Sodium growth (Davidson et al. 2001). Because solutes in
aqueous solutions reduce oxygen solubility,
The human body contains approximately 1.4 g/kg interfere with cellular enzymes, or force cells to
Sodium. It is present mostly as an extracellular expend energy to remove sodium ions from the
component maintaining the osmotic pressure of cell, salt can serve to reduce the rate of microbial
extracellular fluid. The U.S. Food and Drug growth (Shelef et al. 2005).
Administration recommends that individuals
consume no more than 2300 mg of sodium per
day, and that certain groups limit intake to Potassium
1500 mg/day. The “Dietary Guidelines for
Americans 2010” addresses sodium intake in The concentration of Potassium in the human
detail. The average daily sodium intake for body is approximately 2 g/kg, making it the most
Americans is 3400 mg/day, an excessive amount predominant cation in intracellular fluid.
that raises blood pressure and poses health risks Potassium is localized mostly within cells where
in some individuals. It is recommended that it regulates intracellular osmotic pressure.
Americans limit daily sodium consumption to Potassium is frequently used as a replacement for
2300 mg. Sodium in salt substitutes. Generally the popula-
Salt provided one of the earliest means of food tion consumes too much Sodium and too little
preservation and remains as a successful preser- potassium.
238 5 Minerals

Recommended dietary allowance guidelines carbohydrates and lipids. The binding of Calcium
for Potassium vary depending on age. Infants is selective based on its binding to neutral oxy-
from 0 to 6 months old should receive 400 mg gens including alcohols and carbonyl groups.
daily, and those from 7 to 12 months old need Because it is a divalent cation Calcium can bind
700 mg. The RDA for children from 1 to 3 years simultaneously to two centers resulting it is cross-
old is 3000 mg each day, those from 4 to 8 years linking function in carbohydrates and proteins.
old warrant 3800 mg and those from 9 to 13 years
old need 4500 mg. Children older than 13 and
adults should get 4700 mg/day, except for lactat- Phosphates
ing women, who require 5100 mg.
Low potassium levels, or hypokalemia, can Phosphates are found in many different forms in
lead to weakness, lack of energy, high blood pres- foods and are both naturally occurring and as
sure, muscle cramping, gastrointestinal distress, food additives. Phosphates play crucial roles in
arrhythmia and abnormal electrocardiograms. living systems. For example adenosine triphos-
phate (ATP) is the primary energy source in liv-
ing cells. Phosphoproteins (ferritins) are critical
Magnesium for iron storage. Calcium as hydroxyapatite
(Ca10(PO4)6(OH)2) comprises the primary struc-
The concentration of Magnesium in the body is ture in bone. Phospholipids are the major compo-
about 250 mg/kg. The daily requirement is 300– nents in membranes and sugar phosphates are
400 mg/day and the daily intake is from 300 to critical in carbohydrate metabolism. Extensive
500 mg/day. It is naturally abundant in many discussions of phosphates can be found in reviews
foods. Magnesium is an activator of many by Ellinger (1972) and Mollins (1990).
enzymes including conversion of energy rich Several phosphate are approved as food addi-
phosphates. Magnesium compounds also are tives and their structures can be found in Fig. 5.7.
involved in intracellular and plasma membranes. Phosphate food additives provide many functions
Membrane involved in many life supporting met- in food including acidification of soft drinks, buff-
abolic functions and thus deficiency can be very
serious. Magnesium is widely distributed in plant
and animal foods.

Calcium

Calcium, the most abundant mineral in the body


is widely distributed in the throughout the body
totally about 1500 g in adults. It is included in the
skeleton bones but also widely distrusted in most
other tissues. Calcium is essential for muscle
function controlling processes like muscle con-
traction. Approximately 99% of the Calcium in
the body is in the skeletal tissue. The Calcium in
soft tissue must be maintained in a narrow physi-
ological range for the body to function. Thus if
the levels of Calcium in the blood and extracel-
lular Calcium are not met by the diet the mineral
will be reabsorbed from the bone.
Calcium has multiple roles in living cells Fig. 5.7  Structures of phosphoric acid and phosphate
related to its complex formation with proteins, ions used as food additives
Major Minerals 239

ering of beverages, anti-caking, stabilizing, leaven- these colloidal particles. In milk the salts of the
ing, emulsification and antioxidants. At pH ranges weak acids (phosphates, citrates, and carbonates)
in most foods, phosphates are negatively charges are distributed among the various possible ionic
and polyphosphates are polyelectrolytes. The nega- forms. As indicated by Jenness and Patton (1959),
tive charges on these molecules result in the phos- the ratios of the ionic species can be calculated by
phates acting like Lewis bases and as a result they using the Henderson–Hasselbach equation,
exhibit strong metal binding characteristics.
pH = pK α + log
[salt ]

[acid ]
Minerals in Milk
The values for the dissociation constants of
The normal levels of the major mineral constitu- the three acids are listed in Table 5.4. When these
ents of cow’s milk are listed in Table 5.3. These are values are substituted in the Henderson–
average values; there is a considerable natural Hasselbach equation for a sample of milk at
variation in the levels of these constituents. pH 6.6, the following ratios will be obtained:
A number of factors influence the variations in
salt composition, such as feed, season, breed and Citrate − Citrate =
= 3000 = 72
individuality of the cow, stage of lactation, and Citric acid Citrate −
udder infections. In all but the last case, the varia- Citrate ≡
tions in individual mineral constituents do not = 16
Citrate =
affect the milk’s osmotic pressure. The ash content
of milk is relatively constant at about 0.7%. An From these ratios we can conclude that in milk
important difference between milk and blood at pH 6.6 no appreciable free citric acid or
plasma is the relative levels of sodium and potas- monocitrate ion is present and that tricitrate and
sium. Blood plasma contains 330 mg/100 mL of dicitrate are the predominant ions, present in a
sodium and only 20 mg/100 mL of potassium. In ratio of about 16 to 1. For phosphates, the follow-
contrast, the potassium level in milk is about three ing ratios are obtained:
times as high as that of sodium. Some of the min-
eral salts of milk are present at levels exceeding H 2 PO 4 − HPO 4 =
= 43, 600 = 0.30
their solubility and therefore occur in the colloidal H 3 PO 4 H 2 PO 4 −
form. Colloidal particles in milk contain calcium, PO 4 ≡
= 0.000002
magnesium, phosphate, and citrate. These colloi- HPO 4 =

dal particles precipitate with the curd when milk is
coagulated with rennin. Dialysis and ultrafiltration This indicates that mono- and diphosphate
are other methods used to obtain a serum free from ions are the predominant species. For carbonates
the ratios are as follows:
Table 5.3  Average values for major mineral content of
cow’s milk (skim milk) HCO3 −
= 1.7
Constituent Normal level (mg/100 mL) H 2 CO3
Sodium 50
CO3 =
Potassium 145 = 0.0002
HCO3 −
Calcium 120
Magnesium 13
Phosphorus (total) 95
Phosphorus (inorganic) 75 Table 5.4  Dissociation constants of weak acids
Chloride 100 Acid pK1 pK2 pK3
Sulfate 10 Citric 3.08 4.74 5.40
Carbonate (as CO2) 20 Phosphoric 1.96 7.12 10.32
Citrate (as citric acid) 175 Carbonic 6.37 10.25 –
240 5 Minerals

The predominant forms are bicarbonates and changes. Gaucheron (2005) has reviewed the
the free acid. changes in detail. Figure 5.8 illustrates some of
Note that milk contains considerably more the important changes. Heat causes a concentra-
cations than anions; Jenness and Patton (1959) tion of Calcium phosphate in the micelle whereas
have suggested that this can be explained by cooling results in migration of calcium phosphate
assuming the formation of complex ions of cal- out of the micelle. Addition of sodium chloride
cium and magnesium with the weak acids. In the results in phosphate coming out of the micelle
case of citrate [Cit] the following equilibria exist: and water migrating into the micelle. Lowering
the pH causes calcium phosphate and water to
H [ Cit ] → [ Cit ]
−3
come out of the micelle along with some of the
[Cit ] + Ca −2 → Ca [Cit ]
−3 −1
caseins prior to isoelectric precipitation.
The equilibria levels of total and soluble cal-
Ca [ Cit ] + H + → CaH [ Cit ]
−1

cium and phosphorus are listed in Table 5.5. The


Ca [ Cit ] + Ca +2 → Ca 3 [ Cit ]2
−1
2C mineral equilibria in milk have been extensively

studied because the ratio of ionic and total calcium
Soluble complex ions such as Ca+2 can account exerts a profound effect on the stability of the
for a considerable portion of the calcium and caseinate particles in milk. Processing conditions
magnesium in milk, and analogous complex ions such as heating and evaporation change the salt
can be formed with phosphate and possibly with equilibria and therefore the protein stability. When
bicarbonate. milk is heated, calcium and phosphate change
Treatments of casein micelles results in a from the soluble to the colloidal phase. Changes in
number of changes in the water, Calcium, phos- pH result in profound changes of all of the salt
phate and other ions which migrate in or out of equilibria in milk. Decreasing the pH results in
the micelle depending on the physiochemical changing calcium and phosphate from the colloi-

Fig. 5.8  Changes in salt equilibrium of casein micelles resulting from physio-chemical treatments (modified from
Gaucheron (2005))
Major Minerals 241

Table 5.5  Total and soluble calcium and phosphorus Table 5.6  Mineral constituents in meat (beef)
content of milk
Constituent mg/100 g
Constituent mg/100 mL Total calcium 8.6
Total calcium 112.5 Soluble calcium 3.8
Soluble calcium 35.2 Total magnesium 24.4
Ionic calcium 27.0 Soluble magnesium 17.7
Total phosphorus 69.6 Total citrate 8.2
Soluble phosphorus 33.3 Soluble citrate 6.6
Total inorganic phosphorus 233.0
Soluble inorganic phosphorus 95.2
Sodium 168
Potassium 244
Chloride 48

tissue contains much more potassium than


sodium. Meat also contains considerably more
magnesium than calcium. Table 5.4 also provides
information about the distribution of these miner-
als between the soluble and non-soluble forms.
Fig. 5.9  Calcium chelate of a polyphosphate
The non-soluble minerals are associated with the
dal to the soluble form. At pH 5.2, all of the cal- proteins. Since the minerals are mainly associated
cium and phosphate of milk becomes soluble. An with the non-fatty portion of meat, the leaner
equilibrium change results from the removal of meats usually have a higher mineral or ash con-
CO2 as milk leaves the cow’s udder. This loss of tent. When liquid is lost from meat (drip loss), the
CO2 by stirring or heating results in an increased major element lost is sodium and, to a lesser
pH. Concentration of milk results in a dual effect. extent, calcium, phosphorus, and potassium.
The reduction in volume leads to a change of cal- Muscle tissue consists of about 40% intracellular
cium and phosphate to the colloidal phase, but this fluid, 20% extracellular fluid, and 40% solids. The
also liberates hydrogen ions, which tend to dis- potassium is found almost entirely in the intracel-
solve some of the colloidal calcium phosphate. lular fluid, as are magnesium, phosphate, and sul-
The net result depends on initial salt balance of the fate. Sodium is mainly present in the extracellular
milk and the nature of the heat treatment. fluid in association with chloride and bicarbonate.
The stability of the caseinate particles in milk During cooking, sodium may be lost, but the other
can be measured by a test such as the heat stabil- minerals are well retained. Processing does not
ity test, rennet coagulation test, or alcohol stabil- usually reduce the mineral content of meat. Many
ity test.Addition of various phosphates—especially processed meats are cured in brine that contains
polyphosphates, which are effective calcium mostly sodium chloride. As a result, the sodium
complexing agents—can increase the caseinate content of cured meats may be increased.
stability of milk. Addition of calcium ions has the Ionic equilibria play an important role in the
opposite effect and decreases the stability of water-binding capacity of meat (Hamm 1971).
milk. Calcium is bound by polyphosphates in the The normal pH of rigor or post-rigor muscle
form of a chelate, as shown in Fig. 5.9. (pH 5.5) is close to the isoelectric point of acto-
myosin. At this point the net charge on the pro-
tein is at a minimum. By addition of an acid or
Minerals in Meat base, a cleavage of salt cross-linkages occurs,
which increases the electrostatic repulsion
The major mineral constituents of meat are listed (Fig. 5.10), loosens the protein network, and thus
in Table 5.6. Sodium, potassium, and phosphorus permits more water to be taken up. Addition of
are present in relatively high amounts. Muscle neutral salts such as sodium chloride to meat
242 5 Minerals

Fig. 5.10 Schematic
representation of the
addition of acid (HA) or
base (B−) to an
isoelectric protein. The
isoelectric protein has
equal numbers of
positive and negative
charges. The acid HA
donates protons, the
base B− accepts protons

increases water-holding capacity and swelling. Table 5.7  Major mineral element components in wheat
grain
The swelling effect has been attributed mainly to
the chloride ion. The existence of intra- and Element mg/100 g
extracellular fluid components has been described Potassium 363
by Merkel (1971) and may explain the effect of Phosphorus 357
salts such as sodium chloride. The proteins inside Calcium 34
the cell membrane are non-diffusible, whereas Magnesium 137
the inorganic ions may move across this semiper- Sodium 2
meable membrane. If a solution of the sodium Source: USDA nutrient database
salt of a protein is on one side of the membrane
and sodium chloride on the other side, diffusion Struvite
will occur until equilibrium has been reached. Occasionally, phosphates can form undesirable
This can be represented as follows: crystals in foods. The most common example is
At equilibrium the product of the concentra- struvite, a magnesium-ammonium phosphate of
tions of diffusable ions on the left side of the the composition Mg(NH4)PO4·6H2O. Struvite
membrane must be equal to the product on the crystals are easily mistaken by consumers for bro-
right side, shown as follows: ken pieces of glass. Most reports of struvite forma-
tion have been related to canned seafood, but
 Na +  Cl −  =  Na +  Cl − 
L L R R occasionally the presence of struvite in other foods
has been reported. It is assumed that in canned sea-
In addition, the sum of the cations on one side food, the struvite is formed from the magnesium
must equal the sum of anions on the other side of sea water and ammonia generated by the effect
and vice versa: of heat on the fish or shellfish muscle protein.
 Na +  =  Pr −  +  Cl −  and  Na +  =  Cl − 
L L L R R

Minerals in Plant Products


This is called the Gibbs–Donnan equilibrium
and provides an insight into the reasons for the Plants generally have a higher content of potas-
higher concentration of sodium ions in the intra- sium than of sodium. The major minerals in wheat
cellular fluid. are listed in Table 5.7 and include potassium,
Major Minerals 243

Table 5.8  Mineral components in endosperm and bran fractions of red winter wheat
P (%) K (%) Na (%) Ca (%) Mg (%) Mn (ppm) Fe (ppm) Cu (ppm)
Total endosperm 0.10 0.13 0.0029 0.017 0.016 2.4 13 8
Total bran wheat kernel 0.38 0.35 0.0067 0.032 0.11 32 31 11
Center section 0.35 0.34 0.0051 0.025 0.086 29 40 7
Germ end 0.55 0.52 0.0036 0.051 0.13 77 81 8
Brush end 0.41 0.41 0.0057 0.036 0.13 44 46 12
Entire kernel 0.44 0.42 0.0064 0.037 0.11 49 54 8
Source: From V.H. Morris et al., Studies on the Composition of the Wheat Kernel. II. Distribution of Certain Inorganic
Elements in Center Sections, Cereal Chem., Vol. 22, pp. 361–372, 1945

phosphorus, calcium, magnesium, and sulfur Table 5.9  Mineral content of soybeans (green raw)
(Schrenk 1964). Sodium in wheat is present at a Mineral Range (%)
level of only about 80 ppm and is considered a Iron 3.5
trace element in this case. The minerals in a wheat Potassium 620
kernel are not uniformly distributed; rather, they Calcium 197
are concentrated in the areas close to the bran coat Magnesium 65
and in the bran itself. The various fractions result- Phosphorus 194
ing from the milling process have quite different Sodium 15
ash contents. The ash content of flour is consid- Source: USDA nutrient database
ered to be related to quality, and the degree of
extraction of wheat in milling can be judged from A major study of the mineral composition of
the ash content of the flour. Wheat flour with high fruits was conducted by Zook and Lehmann
ash content is darker in color; generally, the lower (1968). Some of their findings for the major miner-
the ash content, the whiter the flour. This general als in fruits are listed in Table 5.10. Fruits are gen-
principle applies, but the ash content of wheat erally not as rich in minerals as vegetables are.
may vary within wide limits and is influenced by Apples have the lowest mineral content of the fruits
rainfall, soil conditions, fertilizers, and other fac- analyzed. The mineral levels of all fruits show great
tors. The distribution of mineral components in variation depending on growing region.
the various parts of the wheat kernel is shown in The rate of senescence of fruits and vegetables
Table 5.8. is influenced by the calcium content of the tissue
High-grade patent flour, which is pure endo- (Poovaiah 1986.) When fruits and vegetables are
sperm, has an ash content of 0.30–0.35%, treated with calcium solutions, the quality and
whereas whole wheat meal may have an ash con- storage life of the products can be extended.
tent from 1.35 to 1.80%.
The ash content of soybeans is relatively
high, close to 5%. The ash and major mineral Chloride
levels in soybeans are listed in Table 5.9.
Potassium and phosphorus are the elements Originally, nine of the trace elements were consid-
present in greatest abundance. About 70–80% of ered to be essential to humans: cobalt, copper,
the phosphorus in soybeans is present in the fluorine, iodine, iron, manganese, molybdenum,
form of phytic acid, the phosphoric acid ester of selenium, and zinc. Recently, chromium, silicon,
inositol (Fig. 5.11). Phytin is the calcium-mag- and nickel have been added to this list (Reilly et al.
nesium-potassium salt of inositol hexaphos- 1996). These are mostly metals; some are metal-
phoric acid or phytic acid. The phytates are loids. In addition to essential trace elements, sev-
important because of their effect on protein sol- eral trace elements have no known essentiality and
ubility and because they may interfere with some are toxic (such as lead, mercury, and cad-
absorption of calcium from the diet. Phytic acid mium). These toxic trace elements, which are clas-
is present in many foods of plant origin. sified as contaminants, are dealt with in Chap. 11.
244 5 Minerals

Fig. 5.11  Inositol and


phytic acid

Table 5.10  Mineral content of some fruits


Minerals (mg/100 g)
Fruit Na Ca Mg Fe Zn P K
Orange (Navel) 1 43 11 0.13 0.08 23 166
Apple (raw with skin) 1 6 5 0.12 0.04 11 107
Grape (Thompson) 2 10 7 0.36 0.07 20 191
Cherry (sweet raw) 0 13 11 0.036 0.07 21 222
Pear (fresh raw) 1 9 7 0.18 0.10 12 116
Banana (raw) 1 5 27 0.26 0.15 22 358
Pineapple (Puerto 1 13 12 0.25 0.08 9 125
Rico)
Source: USDA nutrient database

chemicals, and sewage sludge. Sewage sludge is a


Trace Elements good source of nitrogen and phosphate but may
contain high levels of trace minerals, many of
Because trace metals are ubiquitous in our envi- these originating from industrial activities such as
ronment, they are found in all of the foods we eat. electroplating. Trace minerals may also originate
In general, the abundance of trace elements in from food processing and handling equipment,
foods is related to their abundance in the environ- food packaging materials, and food additives.
ment. Table 5.11 presents the recommended daily
allowance (RDA) for some trace elements and
their common food sources. Iron
Trace elements get into foods by different
pathways. The most important source is from the Iron is a component of the heme pigments and of
soil, by absorption of elements in aqueous solu- some enzymes. In spite of the fact that some
tion through the roots. Another, minor, source is foods have high iron levels, much of the popula-
foliar penetration. This is usually associated with tion has frequently been found to be deficient in
industrial air pollution and vehicle emissions. this element. Animal food products may have
Other possible sources are fertilizers, agricultural high levels that are well absorbed; liver may
Trace Elements 245

Table 5.11  Dietary allowances and sources of some important trace elements in the human diet
Element RDA Common sources
Chromium 50–200 mcg Whole grains, brewer’s yeast, nuts and dark chocolate
Copper 1.5–3 mg Seafood, nuts, legumes, green leafy vegetables, dried fruits
(such as prunes and cocoa), yeast, organ meats, nuts, potatoes, grains, beans
Iodine 150 mcg Seafood, Iodized salt
Iron 10–15 mg Heme iron is only present in animal flesh. Beef, liver, clams and oysters.
Non-heme iron is in tofu, legumes, spinach, raisins
Manganese 2.5–5.0 mg Coffee, tea, nuts, whole grains, legumes and some fruits and vegetables
Molybdenum 75–250 mcg Peas, legumes and some breakfast cereals
Selenium 55–70 mcg Seafoods and organ meats, muscle meats, cereals and other grains, and dairy
Fluoride 0.7–3 mg Fluoridated water, tea, sardines, chicken

c­ ontain several thousand ppm of iron. The iron Copper


from other foods such as vegetables and eggs is
more poorly absorbed. In the case of eggs the Copper is present in foods as part of several
uptake is poor because the ferric iron is closely copper-­containing enzymes, including the poly-
bound to the phosphate of the yolk phosphopro- phenolases. Copper is a very powerful prooxi-
teins. Iron is used as a food additive to enrich dant and catalyzes the oxidation of unsaturated
flour and cereal products. The form of iron used fats and oils as well as ascorbic acid. The normal
significantly determines how well it will be taken daily diet contains from 2 to 5 mg of copper,
up by the body. Ferrous sulfate is very well more than ample to cover the daily requirement
absorbed, but will easily discolor or oxidize the of 0.6–2 mg.
food to which it is added. Elemental iron is also
well absorbed and is less likely to change the
food. For these reasons, it is the preferred form of Zinc
iron for the enrichment of flour.
Iron bioavailability may be increased in the Zinc is the second most important of the essential
presence of meat (Politz and Clydesdale 1988). trace elements for humans. It is a constituent of
This is the so-called meat factor. The exact mech- some enzymes, such as carbonic anhydrase. Zinc
anism of this effect is not known, but it has been is sufficiently abundant that deficiencies of zinc
suggested that amino acids or polypeptides that are unknown. The highest levels of zinc are found
result from digestion are able to chelate nonheme in shellfish, which may contain 400 ppm. The
iron. These complexes would facilitate the absorp- level of zinc in cereal grains is 30–40 ppm. When
tion of iron. In nitrite-cured meats some factors acid foods such as fruit juices are stored in galva-
promote iron bioavailability (the meat factor), nized containers, sufficient zinc may be dissolved
particularly heme iron and ascorbic acid or ery- to cause zinc poisoning. The zinc in meat is
thorbic acid. Negative factors may include nitrite tightly bound to the myofibrils and has been
and nitrosated heme (Lee and Greger 1983). speculated to influence meat’s water-binding
capacity (Hamm 1972).

Cobalt
Manganese
Cobalt is an integral part of the only metal con-
taining vitamin B12. The level of cobalt in foods Manganese is present in a wide range of foods
varies widely, from as little as 0.01 ppm in corn but is not easily absorbed. This metal is associ-
and cereals to 1 ppm in some legumes. The ated with the activation of a number of enzymes.
human requirement is very small and deficiencies In wheat, a manganese content of 49 ppm has
do not occur. been reported (Schrenk 1964). This is mostly
246 5 Minerals

concentrated in the germ and bran; the level in Fluorine


the endosperm is only 2.4 ppm. Information on
the manganese content of seafoods has been sup- Fluorine is a constituent of skeletal bone and
plied by Meranger and Somers (1968). Values helps reduce the incidence of dental caries. The
range from a low of 1.1 ppm in salmon to a high fluorine content of drinking water is usually below
of 42 ppm in oyster. 0.2 mg/L but in some locations may be as high as
5 mg/L. The optimal concentration for dental
health is 1 mg/L. The fluoride content of vegeta-
Molybdenum bles is low, with the exception of spinach, which
contains 280 μg/100 g. Milk contains 20 μg/100 g
Molybdenum plays a role in several enzyme and beef about 100 μg/100 g. Fish foods may con-
reactions. Some of the molybdenum-containing tain up to 700 μg/100 g and tea about 100 μg/g.
enzymes are aldehyde oxidase, sulfite oxidase,
xanthine dehydrogenase, and xanthine oxidase.
This metal is found in cereal grains and legumes; Iodine
leafy vegetables, especially those rich in chloro-
phyll; animal organs; and in relatively small Iodine is not present in sufficient amounts in the
amounts, less than 0.1 ppm, in fruits. The molyb- diet in several areas of the world; an iodine defi-
denum content of foods is subject to large ciency results in goiter. The addition of iodine to
variations. table salt has been extremely effective in reduc-
ing the incidence of goiter. The iodine content of
most foods is in the area of a few mg/100 g and is
Selenium subject to great local variations. Fish and shell-
fish have higher levels. Saltwater fish have levels
Selenium has recently been found to protect of about 50–150 mg/100 g and shellfish may
against liver necrosis. It usually occurs bound to have levels as high as 400 mg/100 g.
organic molecules. Different selenium com-
pounds have greater or lesser protective effect.
The most active form of selenium is selenite, Chromium
which is also the least stable chemically. Many
selenium compounds are volatile and can be lost Recent well-controlled studies (Anderson 1988)
by cooking or processing. Kiermeier and have found that dietary intake of chromium is in
Wigand (1969) found about a 5% loss of sele- the order of 50 μg/day. Refining and processing
nium as a result of drying of skim milk. The of foods may lead to loss of chromium. As an
variation in selenium content of milk is wide example, in the milling of flour, recovery of chro-
and undoubtedly associated with the selenium mium in white flour is only 35–44% of that of the
content of the soil. The same authors report fig- parent wheat (Zook et al. 1970). On the other
ures for selenium in milk in various parts of the hand, the widespread use of stainless steel equip-
world ranging from 5 to 1270 μg/kg. The sele- ment in food processing results in leaching of
nium in milk is virtually all bound to the pro- chromium into the food products (Offenbacher
teins. Morris and Levander (1970) determined and Pi-Sunyer 1983). No foods are known to con-
the selenium content of a wide variety of foods. tain higher-than-average levels of chromium. The
Most fruits and vegetables contain less than average daily intake of chromium from various
0.01  μg/g. Grain products range from 0.025 to food groups is shown in Table 5.12. It has been
0.66  μg/g, dried skim milk from 0.095 to suggested that the dietary intake of chromium in
0.24  μg/g, meat from 0.1 to 1.9 μg/g, and sea- most normal individuals is suboptimal and can
food from 0.4 to 0.7 μg/g. lead to nutritional problems (Anderson 1988).
Metal Uptake in Canned Foods 247

Table 5.12  Chromium intake from various food groups


Food group Average daily intake (μg) Comments
Cereal products 3.7 55% from wheat
Meat 5.2 55% from pork
25% from beef
Fish and seafood 0.6
Fruits, vegetables, nuts 6.8 70% from fruits and berries
Dairy products, eggs, margarine 6.2 85% from milk
Beverages, confectionery, sugar, and condiments 6.6 45% from beer, wine, and soft drinks
Total 29.1
Source: Reprinted with permission from R.A. Anderson, Chromium, in Trace Minerals in Foods, K.T. Smith, ed.,
p. 238, 1988, by courtesy of Marcel Dekker, Inc.

Silicon is ubiquitous in the environment and phosphate as a leavening agent and aluminum sul-
present in many foods. Foods of animal origin are fate for pH control. The estimated average daily
relatively low in silicon; foods of plant origin are intake of aluminum is 26.5 mg, with 70% coming
relatively high. Good plant sources are unrefined from grain products (Greger 1985).
grains, cereal products, and root crops. The dietary Fruits contain relatively high levels of organic
intake of silicon is poorly known but appears to be acids, which may combine with metal ions. It is
in the range of 20–50 μg/day. Although silicon is now generally agreed that these compounds
now regarded as an essential mineral for humans, may form chelates of the general formula
a minimum requirement has not been established. MyHpLm(OH)x, where M and L represent the
Silicon Foods, K.T. Smith, ed., p. 385, 1988 metal and the ligand, respectively. According to
Pollard and Timberlake (1971), cupric ions form
strong complexes with acids containing
 dditional Information on Trace
A α-hydroxyl groups. The major fruit acids, citric,
Elements malic, and tartaric, are multidendate ligands
capable of forming polynuclear chelates. Cupric
The variations in trace elements in vegetables may and ferric ions form stronger complexes than fer-
be considerable (Warren 1972) and may depend to rous ions. The strongest complexes are formed by
a large extent on the nature of the soil in which the citrate, followed by malate and then tartrate.
vegetables are grown. Table 5.13 illustrates the
extent of the variability in the content of copper,
zinc, lead, and molybdenum of a number of vege- Metal Uptake in Canned Foods
tables. The range of concentrations of these metals
frequently covers one order of magnitude and Canned foods may take up metals from the con-
occasionally as much as two orders of magnitude. tainer, tin and iron from the tin plate, and tin and
Unusually high concentrations of certain metals lead from the solder. There are several types of
may be associated with the incidence of diseases internal can corrosion. Rapid detinning is one of
such as multiple sclerosis and cancer in humans. the most serious problems of can corrosion. With
Aluminum, which has been assumed to be non- most acid foods, when canned in the absence of
nutritious and nontoxic, has come under increas- oxygen, tin forms the anode of the tin-iron cou-
ing scrutiny. Its presence has been suggested to be ple. The tin under these conditions goes into solu-
involved in several serious conditions, including tion at an extremely slow rate and can provide
Alzheimer’s disease (Greger 1985). Since alumi- product protection for 2 years or longer. There
num is widely used in utensils and packaging are, however, conditions where iron forms the
materials, there is great interest in the aluminum anode, and in the presence of depolarizing or oxi-
content of foods. Several aluminum salts are used dizing agents the dissolution of tin is greatly
as food additives, for example, sodium aluminum accelerated. The food is protected until most of
Table 5.13  Extreme variation in the content of copper, zinc, lead, and molybdenum in some vegetables
“Normal” content in Minimum as fraction Maximum as multiple
ppm wet weight of “Normal” of “Normal” Extreme range
Copper
 Lettuce 0.74 1 8 1–120
15
 Cabbage 0.26 1 2.5 1–15
6
 Potato 0.92 1 4 1–36
9
 Bean (except broad) 0.56 2 2.5 1–22
5
 Carrot 0.52 1 2.5 1–22
9
 Beet 0.78 1 2.5 1–20
9
Zinc
 Lettuce 4.9 1 15 1–90
6
 Cabbage 1.9 ½ 6 1–12
 Potato 2.9 ½ 5 1–10
 Bean (except broad) 3.6 ½ 2 1–4
 Carrot 3.4 ½ 8 1–48
 Beet 4.1 1 12 1–16
4
Lead
 Lettuce 0.25 1 30 1–300
10
 Cabbage 0.10 1 2.5 1–20
8
 Potato 0.40 1 15 1–150
10
 Bean (except broad) 0.24 1 4 1–20
5
 Carrot 0.22 1 9 1–27
3
 Beet 0.20 1 11 1–66
6
Molybdenum
 Lettuce 0.06 1 12 1–96
8
 Cabbage 0.20 1 8 1–240
30
 Potato 0.15 1 7.5 1–120
16
 Bean (except broad) 0.48 1 7 1–210
30
 Carrot 0.22 1 3.5 1–14
4
 Beet 0.04 1 10 1–300
30
Source: From H.V. Warren, Variations in the Trace Element Contents of Some Vegetables, J. Roy. Coll. Gen. Practit.,
Vol. 22, pp. 56–60, 1972
Metal Uptake in Canned Foods 249

the tin is d­ issolved; thereafter, hydrogen is pro- ends. This is usually characterized by detinning
duced and the can swells and becomes a springer. at the headspace interface. Stevenson et al. (1968)
Some foods are more likely to involve rapid found that steam flow closure reduced the detin-
detinning, including spinach, green beans, tomato ning problem, but the best results were obtained
products, potatoes, carrots, vegetable soups, and by complete removal of oxygen through nitrogen
certain fruit juices such as prune and grapefruit closure. Detinning by canned spinach was stud-
juice. ied by Lambeth et al. (1969) and was found to be
Another corrosion problem of cans is sulfide significantly related to the oxalic acid content of
staining. This may happen when the food con- the fresh leaves and the pH of the canned prod-
tains the sulfur-containing amino acids cysteine, uct. High-oxalate spinach caused detinning in
cystine, or methionine. When the food is heated excess of 60% after 9 months’ storage.
or aged, reduction may result in the formation of In some cases the dissolution of tin into a food
sulfide ions, which can then react with tin and may have a beneficial effect on food color, with
iron to form SnS and FeS. The compound SnS is iron having the opposite effect. This is the case
the major component of the sulfide stain. This for canned wax beans (Van Buren and Downing
type of corrosion may occur with foods such as 1969). Stannous ions were effective in preserving
pork, fish, and peas (Seiler 1968). Corrosion of the light color of the beans, whereas small
tin cans depends on the nature of the canned food amounts of iron resulted in considerable darken-
as well as on the type of tin plate used. Formerly, ing. A black discoloration has sometimes been
hot dipped tin plate was used, but this has been observed in canned all-green asparagus after
mostly replaced by electrolytically coated plate. opening of the can. This has been attributed
It has been shown (McKirahan et al. 1959) that (Lueck 1970) to the formation of a black, water-­
the size of the crystals in the tin coating has an insoluble coordination compound of iron and
important effect on corrosion resistance. Tin rutin. The iron is dissolved from the can, and the
plate with small tin crystals easily develops rutin is extracted from the asparagus during the
hydrogen swell, whereas tin plate containing sterilization. Rutin is a flavonol, the 3-rutinoside
large crystals is quite resistant. Seiler (1968) of quercetin. The black discoloration occurs only
found that the orientation of the different crystal after the iron has been oxidized to the ferric state.
planes also significantly affected the ease of Tin forms a yellow, water-soluble complex with
forming sulfide stains. rutin, which does not present a color problem.
The influence of processing techniques for The uptake of iron and tin from canned foods is a
grapefruit juice on the rate of can corrosion was common occurrence, as is demonstrated by Price
studied by Bakal and Mannheim (1966). They and Roos (1969), who studied the presence of
found that the dissolved tin content can serve as a iron and tin in fruit juice (Table 5.14).
corrosion indicator. In Israel the maximum pre-
scribed limit for tin content of canned food is
250 ppm. Deaeration of the juice significantly Table 5.14  Iron and tin content of fruit juices
lowers tin dissolution. In a study of the in-can shelf Product Iron (ppm) Tin (ppm)
life of tomato paste, Van der Merwe and Knock Fresh orange juice 0.5 7.5
(1968) found that, depending on maturity and vari- Bottled orange juice 2.5 25
ety, 1 g of tomato paste stored at 22 °C could cor- Bottled orange juice 2.0 50
rode tin at rates ranging from 9 × 10−6 to Bottled pineapple juice 15.0 50
68 × 10−6 g/month. The useful shelf life could vary Canned orange juice 2.5 60
from 24 months to as few as 3 months. Up to 95% Canned orange juice 0.5 115
of the variation could be related to effects of matu- Canned orange juice 2.5 120
rity and variety and the associated differences in Canned pineapple juice 17.5 135
contents of water-insoluble solids and nitrate. Source: From W.J. Price and J.T.H. Roos, Analysis of
Fruit Juice by Atomic Absorption Spectrophotometry.
Severe detinning has often been observed with I. The Determination of Iron and Tin in Canned Juice,
applesauce packed in plain cans with enameled J. Sci. Food Agric., Vol. 20, pp. 427–439, 1969
250 5 Minerals

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(1968). Interrelations and correlates in children’s learn- tents of some vegetables. The Journal of the Royal
ing and problem solving. Monographs of the Society College of General Practitioners, 22(114), 56–60.
for Research in Child Development, 33(7), 1–68. Zook, E. G., et al. (1970). Nutrient composition of
Toma, R. B., & Curtis, D. J. (1986). Dietary fiber: Effect selected wheat and wheat products. Cereal Chemistry,
on mineral bioavailability. Food Technology, 40(2), 47, 720–727.
111–116. Zook, E. G., & Lehmann, J. (1968). Mineral composition
van Dokkum, W., de Vos, R. H., Muys, T., & Wesstra, of fruits. II. Nitrogen, calcium, magnesium, phospho-
J. A. (1989). Minerals and trace elements in total diets rus, potassium, aluminum, boron, copper, iron, man-
in The Netherlands. The British Journal of Nutrition, ganese, and sodium. Journal of the American Dietetic
61(1), 7–15. Association, 52(3), 225–231.
Color and Food Colorants
6
John W. Finley, John M. deMan,
and Chang Yong Lee

sensor or photocell that can provide consistent


Introduction results. There are several systems of color classi-
fication; the most important is the CIE system
Color is important to many foods, both those that (Commission International de l’Eclairage—
are unprocessed and those that are manufactured. International Commission on Illumination).
Together with flavor and texture, color plays an Other systems used to describe food color are the
important role in food acceptability. In addition, Munsell, Hunter, and Lovibond systems.
color may provide an indication of chemical When the reflectance of different colored objects
changes in a food, such as browning and caramel- is determined by means of spectrophotometry,
ization. For a few clear liquid foods, such as oils curves of the type shown in Fig. 6.1 are obtained.
and beverages, color is mainly a matter of trans- White materials reflect equally over the whole visi-
mission of light. Other foods are opaque—they ble wavelength range, at a high level. Gray and
derive their color mostly from reflection. black materials also reflect equally over this range
Color is the general name for all sensations but to a lower degree. Red materials reflect in the
arising from the activity of the retina of the eye. higher wavelength range and absorb the other wave-
When light reaches the retina, the eye’s neural lengths. Blue materials reflect in the low-wave-
mechanism responds, signaling color among length range and absorb the high-­wavelength light.
other things. Light is the radiant energy in the
wavelength range of about 400–800 nm.
According to this definition, color (like flavor and CIE System
texture) cannot be studied without considering
the human sensory system. The color perceived The spectral energy distribution of CIE light
when the eye views an illuminated object is sources A and C is shown in Fig. 6.2. CIE illumi-
related to the following three factors: the spectral nant A is an incandescent light operated at
composition of the light source, the chemical and 2854 K, and illuminant C is the same light modi-
physical characteristics of the object, and the fied by filters to result in a spectral composition
spectral sensitivity properties of the eye. To eval- that approximates that of normal daylight.
uate the properties of the object, we must stan- Figure 6.2 also shows the luminosity curve of the
dardize the other two factors. Fortunately, the standard observer as specified by CIE. This curve
characteristics of different people’s eyes for indicates how the eyes of normal observers
viewing colors are fairly uniform; it is not too dif- respond to the various spectral light types in the
ficult to replace the eye by some instrumental visible portion of the spectrum. By breaking

© Springer International Publishing AG 2018 253


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_6
254 6  Color and Food Colorants

Fig. 6.1 
Spectrophotometric
reflectance curves of
colored objects
125

White
100

Reflectance (%)
Yellow
75 Blue
Green
Red
50 Gray

25

Black
0

400nm 500nm 600nm 700nm


Wavelength

Fig. 6.2 Spectral
energy distribution of
light sources A and C,
the CIE, and relative
luminosity function γ for 200
the CIE standard
observer

160

120

80

40

400 450 500 550 600 650 700 750


CIE System 255

Fig. 6.3 Spectral
energy distribution of
sunlight, CIE illuminant,
cool white fluorescent
lamp, and sodium light

down the spectrum, complex light types are makes red objects look attractive and blue ones
reduced to their component spectral light types. unattractive. This is called color rendition. The
Each spectral light type is completely determined human eye has the ability to adjust for this effect.
by its wavelength. In some light sources, a great The CIE system is a trichromatic system; its
deal of radiant energy is concentrated in a single basis is the fact that any color can be matched by
spectral light type. An example of this is the a suitable mixture of three primary colors. The
sodium lamp shown in Fig. 6.3, which produces three primary colors, or primaries, are red, green,
monochromatic light. Other light sources, such and blue. Any possible color can be represented
as incandescent lamps, give off a continuous as a point in a triangle. The triangle in Fig. 6.4
spectrum. A fluorescent lamp gives off a continu- shows how colors can be designated as a ratio of
ous spectrum on which is superimposed a line the three primaries. If the red, green, and blue
spectrum of the primary radiation produced by values of a given light type are represented by a,
the gas discharge (Fig. 6.3). b, and c, then the ratios of each to the total light
In the description of light sources, reference is are given by a/(a + b + c), b/(a + b + c), and c/
sometimes made to the black body. This is a radiat- (a + b + c), respectively. Since the sum of these is
ing surface inside a hollow space, and the light one, then only two have to be known to know all
source’s radiation comes out through a small open- three. Color, therefore, is determined by two, not
ing. The radiation is independent of the type of three, of these mutually dependent quantities. In
material the light source is made of. When the tem- Fig.  6.4, a color point is represented by P. By
perature is very high, about 6000 K the maximum determining the distance of P from the right
of the energy distribution will fall about in the angle, the quantities a/(a + b + c) and b/(a + b + c)
middle of the visible spectrum. Such energy distri- are found. The quantity c/(a + b + c) is then
bution corresponds with that of daylight on a found, by first extending the horizontal dotted
cloudy day. At lower temperatures, the maximum line through P until it crosses the hypotenuse at Q
of the energy distribution shifts to longer wave- and by then constructing another right angle tri-
lengths. At 3000 K, the spectral energy distribution angle with Q at the top. All combinations of a, b,
is similar to that of an incandescent lamp; at this and c will be points inside the triangle.
temperature the energy at 380 nm is only one-six- The relative amounts of the three primaries
teenth of that at 780 nm, and most of the energy is required to match a given color are called the tri-
concentrated at higher wavelengths (Fig. 6.3). The stimulus values of the color. The CIE primaries
uneven spectral distribution of incandescent light are imaginary, because there are no real primaries
256 6  Color and Food Colorants

Fig. 6.4 Representation
of a color as a point in a
color triangle

that can be combined to match the highly satu-


rated hues of the spectrum.
In the CIE system the red, green, and blue pri-
maries are indicated by X, Y, and Z. The amount
of each primary at any particular wavelength is
given by the values x , y , and z . These are called
the distribution coefficients or the red, green, and
blue factors. They represent the tristimulus val-
ues for each chosen wavelength. The distribution
coefficients for the visible spectrum are presented
in Fig. 6.5. The values of y correspond with the
luminosity curve of the standard observer
(Fig.  6.2). The distribution coefficients are
dimensionless because they are the numbers by
which radiation energy at each wavelength must
be multiplied to arrive at the X, Y, and Z content.
The amounts of X, Y, and Z primaries required to
produce a given color are calculated as follows:
Fig. 6.5  Distribution coefficients x, y, and z for the visi-
780 ble spectrum. Source: From Hunter Associates Lab., Inc.
X = ∫x IRdh
380
780
where
XY = ∫ y IRdh I = spectral energy distribution of illuminant
380
R = spectral reflectance of sample
780
dh = small wavelength interval
XZ = ∫z IRdh
x , y , z  = red, green, and blue factors
380
CIE System 257

The ratios of the primaries can be expressed as primary at x = 0 and y = 1; and the blue primary at
x = 0 and y = 0. The line connecting the ends of the
X
x= locus represents purples, which are nonspectral
X +Y + Z colors resulting from mixing various amounts of
Y red and blue. All points within the locus represent
y= real colors. All points outside the locus are unreal,
X +Y + Z
including the imaginary primaries X, Y, and Z. At
Z the red end of the locus, there is only one point to
z= represent the wavelength interval of 700–780 nm.
X +Y + Z
This means that all colors in this range can be sim-
The quantities x and y are called the chroma- ply matched by adjustment of luminosity. In the
ticity coordinates and can be calculated for each range of 540–700 nm, the spectrum locus is almost
wavelength from straight; mixtures of two spectral light types along
this line segment will closely match intervening
x = x / (x + y + z )
colors with little loss of purity. In contrast, the
y = y / (x + y + z ) spectrum locus below 540 nm is curved, indicat-
ing that a combination of two spectral lights along
z = 1− (x + y ) this portion of the locus results in colors of

decreased purity.
A plot of x versus y results in the CIE chroma- A pure spectral color is gradually diluted with
ticity diagram (Fig. 6.6). When the chromaticities white when moving from a point on the spectrum
of all of the spectral colors are placed in this graph, locus to the white point P. Such a straight line
they form a line called the locus. Within this locus with purity decreasing from 100 to 0% is known
and the line connecting the ends, represented by as a line of constant dominant wavelength. Each
400 and 700 nm, every point represents a color color, except the purples, has a dominant wave-
that can be made by mixing the three primaries. length. The position of a color on the line con-
The point at which exactly equal amounts of each necting the locus and P is called excitation purity
of the primaries are present is called the equal (pe) and is calculated as follows:
point and is white. This white point represents the x − xw y − yw
chromaticity coordinates of illuminant C. The red Pe = =
x p − xw y p − yw
primary is located at x = 1 and y = 0; the green

Fig. 6.6 CIE
chromaticity diagram
258 6  Color and Food Colorants

where All of the wavelengths between 492.5 and 567 nm


x and y are the chromaticity coordinates of a are complementary to purple. The purples can be
color described in terms of dominant wavelength by
xw and yw are the chromaticity coordinates of using the wavelength complementary to each
the achromatic source purple, and purity can be expressed in a manner
xp and yp are the chromaticity coordinates of similar to that of spectral colors.
the pure spectral color An example of the application of the CIE sys-
tem for color description is shown in Fig. 6.7.
Achromatic colors are white, black, and gray. The curved, dotted line originating from C repre-
Black and gray differ from white only in their sents the locus of the chromaticity coordinates
relative reflection of incident light. The purples of caramel and glycerol solutions. The chroma-
are nonspectral chromatic colors. All other colors ticity coordinates of maple syrup and honey
are chromatic; for example, brown is a yellow of ­follow the same locus. Three triangles on this
low lightness and low saturation. It has a domi- curve represent the chromaticity coordinates of
nant wavelength in the yellow or orange range. U.S. Department of Agriculture (USDA) glass
A color can be specified in terms of the tris- color standards for maple syrup. These are
timulus value Y and the chromaticity coordinates described as light amber, medium amber, and
x and y. The Y value is a measure of luminous dark amber. The six squares are chromaticity
reflectance or transmittance and is expressed in coordinates of honey, designated by USDA as
percent simply as Y/1000. water white, extra white, white, extra light amber,
Another method of expressing color is in terms light amber, and amber. Such specifications are
of luminance, dominant wavelength, and excita- useful in describing color standards for a variety
tion purity. These latter are roughly equivalent to of products. In the case of the light amber stan-
the three recognizable attributes of color: light- dard for maple syrup, the following values apply:
ness, hue, and saturation. Lightness is associated x = 0.486, y = 0.447, and T = 38.9%. In this way,
with the relative luminous flux, reflected or trans- x and y provide a specification for chromaticity
mitted. Hue is associated with the sense of red- and T for luminous transmittance or lightness.
ness, yellowness, blueness, and so forth. Saturation This is easily expressed as the mixture of prima-
is associated with the strength of hue or the rela- ries under illuminant C as follows: 48.6% of red
tive admixture with white. The combination of hue primary, 44.7% of green primary, and 6.7% of
and saturation can be described as chromaticity. blue primary. The light transmittance is 38.9%.
Complementary colors (Table 6.1) are The importance of the light source and other
obtained when a straight line is drawn through conditions that affect viewing of samples cannot
the equal energy point P. When this is done for be overemphasized. Many substances are meta-
the ends of the spectrum locus, the wavelength meric; that is, they may have equal transmittance
complementary to the 700–780 point is at or reflectance at a certain wavelength but possess
492.5 nm, and for the 380–410 point is at 567 nm. noticeably different colors when viewed under
illuminant C.

Table 6.1  Complementary colors


Wavelength (nm) Color Complementary color Munsell System
400 Violet Yellow
450 Blue In the Munsell system of color classification, all
500 Green Orange colors are described by the three attributes of hue,
550 Yellow Red value, and chroma. This can be envisaged as a
Violet three-dimensional system (Fig. 6.8). The hue
600 Orange Blue scale is based on ten hues which are distributed on
650 Red Green the circumference of the hue circle. There are five
700 hues: red, yellow, green, blue, and purple; they are
Munsell System 259

Fig. 6.7 CIE
chromaticity diagram
with color points for
maple syrup and honey
glass color standards

Fig. 6.8  The Munsell


system of color
classification

written as R, Y, G, B, and P. There are also five to 10. The value scale is a lightness scale ranging
intermediate hues, YR, GY, BG, PB, and RP. Each from 0 (black) to 10 (white). This scale is distrib-
of the ten hues is at the midpoint of a scale from 1 uted on a line perpendicular to the plane of the
260 6  Color and Food Colorants

hue circle and intersecting its center. Chroma is a Hunter System


measure of the difference of a color from a gray of
same lightness. It is a measure of purity. The The CIE system of color measurement is based
chroma scale is of irregular length, and begins on the principle of color sensing by the human
with 0 for the central gray. The scale extends out- eye. This accepts that the eyes contain three light-­
ward in steps to the limit of purity obtainable by sensitive receptors—the red, green, and blue
available pigments. The shape of the complete receptors. One problem with this system is that
Munsell color space is indicated in Fig. 6.9. The the X, Y, and Z values have no relationship to
description of a color in the Munsell system is color as perceived, though a color is completely
given as H, V/C. For example, a color indicated as defined. To overcome this problem, other color
5R 2.8/3.7 means a color with a red hue of 5R, a systems have been suggested. One of these,
value of 2.8, and a chroma of 3.7. All colors that widely used for food colorimetry, is the Hunter L,
can be made with available pigments are laid a, b, system. The so-called uniform-color,
down as color chips in the Munsell book of color. opponent-­colors color scales are based on the
opponent-colors theory of color vision. In this
theory, it is assumed that there is an intermediate
signal-switching stage between the light recep-
tors in the retina and the optic nerve, which trans-
mits color signals to the brain. In this switching
mechanism, red responses are compared with
green and result in a red-to-green color dimen-
sion. The green response is compared with blue
to give a yellow-to-blue color dimension. These
two color dimensions are represented by the sym-
bols a and b. The third color dimension is light-
ness L, which is nonlinear and usually indicated
as the square or cube root of Y. This system can
be represented by the color space shown in
Fig. 6.9  The Munsell color space Fig. 6.10. The L, a, b, color solid is similar to the

Fig. 6.10  The Hunter L,


a, b color space. Source:
From Hunter Associates
Lab., Inc.
Lovibond System 261

Munsell color space. The lightness scale is com- Lovibond System


mon to both. The chromatic spacing is different.
In the Munsell system, there are the polar hue and The Lovibond system is widely used for the
chroma coordinates, whereas in the L, a, b, color determination of the color of vegetable oils. The
space, chromaticity is defined by rectangular a method involves the visual comparison of light
and b coordinates. CIE values can be converted to transmitted through a glass cuvette filled with oil
color values by the equations shown in Table 6.2 at one side of an inspection field; at the other
into L, a, b, values and vice versa (MacKinney side, colored glass filters are placed between the
and Little 1962; Clydesdale and Francis 1970). light source and the observer. When the colors on
This is not the case with Munsell values. These each side of the field are matched, the nominal
are obtained from visual comparison with color value of the filters is used to define the color of
chips (called Munsell renotations) or from instru- the oil. Four series of filters are used—red, yel-
mental measurements (called Munsell renota- low, blue, and gray filters. The gray filters are
tions), and conversion is difficult and tedious. used to compensate for intensity when measuring
The Hunter tristimulus data, L (value), a (red- samples with intense chroma (color purity) and
ness or greenness), and b (yellowness or blue- are used in the light path going through the sam-
ness), can be converted to a single color function ple. The red, yellow, and blue filters of increasing
called color difference (∆E) by using the follow- intensity are placed in the light path until a match
ing relationship: with the sample is obtained. Vegetable oil colors
are usually expressed in terms of red and yellow;
∆ E = ( ∆ L ) + ( ∆a ) + ( ∆b )
2 2 2
a typical example of the Lovibond color of an oil

would be R1.7 Y17. The visual determination of
The color difference is a measure of the dis- oil color by the Lovibond method is widely used
tance in color space between two colors. It does in industry and is an official method of the
not indicate the direction in which the colors American Oil Chemists’ Society. Visual methods
differ. of this type are subject to a number of errors, and

Table 6.2  Mathematical relationship between color scales


To convert To L, a, b To X%, Y, Z% Το Υ, x, y
From Χ%, Y Z% Y =Y%
L = 10 Y
17.5 ( X % − Y ) x=
X
a= X +Y + Z
Y
Y
7.0 (Y − Z % ) y=
b= X +Y + Z
Y
From L, a, b Y = 0.01L2 Y = 0.01L2
aL a + 1.75L
X % = 0.01L2 + x=
175 5.645L + a − 3.012b
bL 1.786 L
Z % = 0.01L2 − y=
70 5.645L + a − 3.012b
From Y, x, y L = 10 Y Y
X % = 1.02 ×
1.02 x y
a = 17.5 Y −1
y Y
Z % = .847 1 − ( x + y ) 
2.181 y + x − 1 y
b = 5.929 Y
y

Source: From Hunter Associates Lab., Inc.


262 6  Color and Food Colorants

the results obtained are highly variable. A study may change during processing, storage, and
has been reported (Maes 1997) to calculate CIE preparation that are often perceived as undesir-
and Lovibond color values of oils based on their able, which is an indication of chemical changes.
visible light transmission spectra as measured by Some processed foods are manufactured to be
a spectrophotometer. A computer software has brightly colored to appeal consumers’ eyes,
been developed that can easily convert light especially to children. Therefore, understanding
transmission spectra into CIE and Lovibond how to preserve the natural colors of food during
color indexes. processing is very important for the acceptability
of a food product. In addition, it should also be
noted some substances such as β-carotene or
Gloss riboflavin or anthocyanins are not only colorants
but nutrients and nutraceuticals. Therefore, color
In addition to color, there is another important of foods has multiple effects on foods and to the
aspect of appearance, namely gloss. Gloss can be consumers. Thus controlling, changing, or stabi-
characterized as the reflecting property of a lizing the color of foods is a major objective for
material. Reflection of light can be diffused or food scientists.
undiffused (specular). In specular reflection, the Until the mid-eighteenth century, the only
surface of the object acts as a mirror, and the external sources of coloring used in foods were
light is reflected in a highly directional manner. natural: animal, vegetables, and minerals includ-
Surfaces can range from a perfect mirror with ing saffron, carrots, mulberries, flowers, and
completely specular reflection to a surface copper and iron ores. Since then, food manufac-
reflecting in a completely diffuse manner. In the turers used many different chemicals to color
latter, the light from an incident beam is scat- foods. Pickles were colored green with copper
tered in all directions and the surface is called sulfate. Candy was colored with salts of copper
matte. and lead. Artificial food colors have been
obtained from organic coal tar in the latter part of
the nineteenth century. Addition of colorants to
Food Colorants foods has many commercial reasons: to enhance
colors that occur naturally, to make food more
Color is one of the important quality attributes of attractive and appetizing, to offset color loss dur-
foods. It is because that consumers often use ing processing and storage, to correct natural
color as an index of freshness, wholesomeness, variations in color and to provide color to color-
and overall quality. Specific colors of fruits are less and “fun foods” for kids. Color additives are
often associated with maturity. Consumers use now recognized as an important part of practi-
color as a way to identify a food and a way to cally all processed foods.
judge the quality of a food. No matter how one However, color additives have been controver-
can provide consumers the most nutritious, saf- sial almost since they were first introduced, due
est, and most economical foods, if they do not to the facts that they have the reputation of being
have attractive color, consumers will not accept potentially toxic, they may be used to deceive the
them. Color influence flavor perception: red consumers, and that their primary function is to
color for strawberry, raspberry, or cherry fla- enhance appearance rather than nutritive value,
vored; yellow to be lemon; and green to be lime shelf life, or safety. For example, pickles were
flavored. Color also affects the apparent level of colored with copper sulfate, cheese with lead
sweetness: strongly red-colored strawberry-fla- oxide, candy with compounds containing lead
vored drinks to be sweeter than less strongly col- and mercury. Arsenic salts were also commonly
ored ones. Unfortunately, the color of a food used. The number of synthetic food colorants has
Food Colorants 263

declined over the years as government safety and FD&C number as required by the Food,
regulations and toxicity testing have advanced. Drug and Cosmetic Act: FD&C indicates that
The FDA regulates food color additives under the these colors are approved by FDA for use in
authority of the Color Additives Amendment Act coloring foods, drugs, and cosmetics; and D&C
of 1960 to the Federal Food, Drug, and Cosmetic is considered safe to use with drugs and
Act. FDA’s permitted colors are classified as sub- cosmetics.
ject to certification or exempt from certification, Certified colors may be used as dyes or
both of which are subjected to rigorous safety ­converted to lakes. They can be manufactured
standards prior to their approval and listing for as a powder, granule, or liquid. Lakes are pre-
use in foods: pared by precipitating the soluble dye onto an
approved insoluble base, such as aluminum
+ Certified colors: hydroxide and then dried. Since the lakes are
+ Color that are exempt from certification: dry products providing opacity, they are stable
to heat and light, can be used in dry product and
suitable to use for fats, gums, waxes, oils, and
Certified (Synthetic) Color food packaging materials. They are stable to
heat and light.
Certified colors are synthetically produced dye,
lakes, or pigments and used widely because
they impart an intense, uniform color. They are  olors Exempt from Certification
C
less expensive and blend more easily to create a (Natural)
variety of hues. Those for food use are chemi-
cally classified as azo, xanthene, triphenylmeth- Colors that are exempt from certification are a
ane, and indigoid dyes, which are synthesized group of pigments derived from natural sources
mainly from raw materials obtained from petro- such as animals, plants, or minerals. They always
leum. Currently, there are nine certified color have been part of our diet. They have been isolated
additives that may be added to foods (Table 6.3 and added back to foods for the same reasons as
and Fig. 6.11). Most of the certified colorants the certified colors. However, the exempted natu-
used in the United States have been assigned ral colorants are relatively unstable, easily affected

Table 6.3  Color certified for use in foods


FD&C number Common name Chemical class Characteristics
FD&C Blue No. 1 Brilliant blue FCF Triphenylmethane Stable to heat
Unstable to light Allura
FD&C Blue No. 2 Indigotine (royal blue) Indigoid Stable to light
Unstable to water
D&C Green No. 3 Fast green FCF Triphenylmethane Stable, brilliant color
FD&C Red No. 3 Erythrosine (cherry red) Xanthene Stable to heat
FD&C Red No. 40 Allura red (orange-red) unstable to redox agents
FD&C Yellow No. 5 Tartrazine (lemon yellow) Azo Stable to heat and light
Very soluble in water
FD&C Yellow No. 6 Sunset yellow Azo Fair stability to heat and light
Orange Azo Not soluble in water
Citrus Red No. 2 Azo Use only for orange skins
264 6  Color and Food Colorants

FD&C Blue No. 1 (Brilliant Blue)


Commonly used in ice cream, canned peas, icings, dairy products, sweets and drinks.

FD&C Blue No. 2 (Indigotine, royal blue)


Originally indigo was a natural dye extracted from plants. Today, nearly all indigo dye
is produced by industrial syntheses. It is the blue of blue jeans.

FD&C Green No. 3 (Fast Green)


It can be used for tinned green peas and other vegetables. It is the least used of the
seven main FDA approved dyes. prohibited in European Union and some other
countries.

FD&C Red No. 3 (Erythrosine, Cherry-red)


Known as a organoiodine compound. Primarily used in sweets, such as candies,
cake-decorating gels. Commonly used in many countries of the world, but less
commonly used in the United Stated, the second least used after Fast Green.

FD&C Red No. 40 (Allura Red AC, orange-red)


Has dark red color. Very soluble in watere. The most commonly used red colorant in
the United States, especially in soft drinks.

Fig. 6.11  The chemical structures of FDA certified food colorants. (As noted, most of the FD&C colorants are sodium
salts of sulfonic acids)
Food Colorants 265

FD&C Yellow No. 5 (Tartrazine, lemon yellow)


A commonly used color all over the world for ice cream, confectionery, drink mixes,
corn chips, popcorn, potato chips, mustard, pickles,….

FD&C Yellow No. 6 (Sunset yellow)


It is used in candy, desserts, snacks, sauces and preserved fruits.

Orange B
Restricted use only in frankfurters and sauage casing or surfaces.

Citrus Red No. 2


Restricted use only to color the skins of orange

Fig. 6.11 (continued)

by the food matrix, such as pH, salts, and process- compounds, titanium dioxide. Some of these can
ing conditions, compared to certified synthetic be used only with certain restrictions (Table 6.4).
colorants. However, exempt colors are well Today, some of these are also well perceived by
received by the consumer due to the name of “nat- consumer as the source of antioxidants, derived
ural.” Common natural colorings include annatto, from bioactive green, red, yellow, orange, and
beet root, caramel, carrot oil, grape skins, paprika, blue colors as the sources of lycopene, beta-caro-
saffron, turmeric, and others, such as inorganic tene, lutein, anthocyanin and astaxanthin.
266 6  Color and Food Colorants

Table 6.4  Colorants exempt from certification and cereal products, especially if these products
Exempt color name Uses and restriction have been subjected to heat treatment.
Annatto extract Foods generally
Astaxanthin Salmon fish feed
Beet powder Foods generally Tetrapyrrole Pigments
Canthaxanthin Foods generally (not to exceed
30 mg/lb), and animal feed Myoglobins
Caramel Foods generally
β-apo-carotenal Foods generally The basic unit from which the tetrapyrrole pig-
(not to exceed 15 mg/lb)
ments are derived is pyrrole.
β-carotene Foods generally
Carrot oil Foods generally
Cochineal extract;
carmine
Corn endosperm Chicken feed
oil
Cottonseed flour Foods generally The basic structure of the heme pigments con-
Grape skin extract Carbonated drinks, alcoholic sists of four pyrrole units joined together into a
beverages porphyrin ring as shown in Fig. 6.12. In the heme
Fruit juice Foods generally pigments, the nitrogen atoms are linked to a cen-
Paprika Foods generally tral iron atom. The color of meat is the result of
Riboflavin Foods generally the presence of two pigments, myoglobin and
Saffron Foods generally hemoglobin. Both pigments have globin as the
Titanium dioxide Foods generally (not to exceed 1%)
protein portion, and the heme group is composed
Turmeric Foods generally
of the porphyrin ring system and the central iron
Vegetable juices Foods generally
atom. In myoglobin, the protein portion has a
Source: Griffiths (2005): CFR (2004) molecular weight of about 17,000. In hemoglo-
bin, this is about 67,000—equivalent to four
With few exceptions, the naturally occurring times the size of the myoglobin protein. The cen-
pigments can be divided into the following four tral iron in Fig. 6.12 has six coordination bonds;
groups: each bond represents an electron pair accepted by
the iron from five nitrogen atoms, four from the
1 . Tetrapyrrole compounds: chlorophylls,
hemes, and bilins
2. Isoprenoid derivatives: carotenoids
3. Benzopyran derivatives: anthocyanins and

flavonoids
4. Betalains and other colorants: betacyanins,

betaxanthins, caramels and melanoidins.

The chlorophylls are characteristic of green


vegetables and leaves. The heme pigments are
found in meat and fish. The carotenoids are a
large group of compounds that are widely distrib-
uted in animal and vegetable products; they are
found in fish and crustaceans, vegetables and
fruits, eggs, dairy products, and cereals.
Anthocyanins and flavonoids are found in root
Fig. 6.12  Schematic representation of the heme complex of
vegetables and fruits such as berries and grapes. myoglobin (http://www.wiley.com/college/pratt/0471393878/
Caramels and melanoidins are found in syrups instructor/structure/myoglobin_hemoglobin/index.html)
Tetrapyrrole Pigments 267

porphyrin ring and one from a histidyl residue of


the globin. The sixth bond is available for joining
with any atom that has an electron pair to donate.
The ease with which an electron pair is donated
determines the nature of the bond formed and the
color of the complex. Other factors playing a role
in color formation are the oxidation state of the
iron atom and the physical state of the globin.
In fresh meat and in the presence of oxygen,
there is a dynamic system of three pigments, oxy-
myoglobin, myoglobin, and metmyoglobin. The
reversible reaction with oxygen is
Mb + O 2 → MbO 2

In both pigments, the iron is in the ferrous
form; upon oxidation to the ferric state, the com-
pound becomes metmyoglobin. The bright red
color of fresh meat is due to the presence of oxy-
myoglobin; discoloration to brown occurs in two
stages, as follows:
MbO 2  Mb  MetMb
( red ) ( Purplish ) ( Brownish )

Oxymyoglobin represents a ferrous covalent
complex of myoglobin and oxygen. The absorp-
tion spectra of the three pigments are shown in
Fig. 6.13 (Bodwell and McClain 1971). Myoglobin
forms an ionic complex with water in the absence
of strong electron pair donors that can form cova-
lent complexes. It shows a diffuse absorption band
in the green area of the spectrum at about 555 nm
and has a purple color. In metmyoglobin, the major
absorption peak is shifted toward the blue portion
of the spectrum at about 505 nm with a smaller Fig. 6.13  Absorption spectra of myoglobin, oxymyoglo-
peak at 627 nm. The compound appears brown. bin, and metmyoglobin. Source: From C.E. Bodwell and
As indicated above, oxymyoglobin and myo- P.E. McClain, Proteins, in The Sciences of Meat Products,
2nd ed., J.E. Price and B.S. Schweigert, eds., 1971,
globin exist in a state of equilibrium with oxy- W.H. Freeman & Co.
gen; therefore, the ratio of the pigments is
dependent on oxygen pressure. The oxidized at partial pressures of 1–20 nm of mercury,
form of myoglobin, the metmyoglobin, cannot depending on pigment, pH, and temperature (Fox
bind oxygen. In meat, there is a slow and continu- 1966). When a packaging film with low oxygen
ous oxidation of the heme pigments to the met- permeability is used, the oxygen pressure drops
myoglobin state. Reducing substances in the to the point where oxidation is favored. To pre-
tissue reduce the metmyoglobin to the ferrous vent this, Landrock and Wallace (1955) estab-
form. The oxygen pressure, which is so important lished that oxygen permeability of the packaging
for the state of the equilibrium, is greatly affected film must be at least 5 L of oxygen/m2/day/atm.
by packaging materials used for meats. The max- Fresh meat open to the air displays the bright
imum rate of conversion to metmyoglobin occurs red color of oxymyoglobin on the surface. In the
268 6  Color and Food Colorants

interior, the myoglobin is in the reduced state and that the only correct name is nitric oxide myoglo-
the meat has a dark purple color. As long as reduc- bin. The first reaction of nitrite with myoglobin is
ing substances are present in the meat, the myo- oxidation of the ferrous iron to the ferric form
globin will remain in the reduced form; when they and formation of MetMb. At the same time,
are used up, the brown color of metmyoglobin will nitrate is formed according to the following reac-
predominate. According to Solberg (1970), there tion (Möhler 1974):
is a thin layer a few nanometers below the bright
red surface and just before the myoglobin region, 4 MbO2 + 4 NO2 − + 2H 2 O →
where a definite brown color is visible. This is the
4 MetMbOH + 4 NO3 − + O2
area where the oxygen partial pressure is about
1.4 nm and the brown pigment dominates. The During the formation of the curing pigment,
growth of bacteria at the meat surface may reduce the nitrite content is gradually lowered; there are
the partial oxygen pressure to below the critical no definite theories to account for this loss.
level of 4 nm. Microorganisms entering the loga- The reactions of the heme pigments in meat
rithmic growth phase may change the surface and meat products have been summarized in the
color to that of the purplish-red myoglobin scheme presented in Fig. 6.14 (Fox 1966). Bilin-­
(Solberg 1968). type structures are formed when the porphyrin
In the presence of sulfhydryl as a reducing ring system is broken.
agent, myoglobin may form a green pigment,
called sulfmyoglobin. The pigment is green
because of a strong absorption band in the red Chlorophylls
region of the spectrum at 616 nm. In the presence
of other reducing agents, such as ascorbate, cho- The chlorophylls are green pigments responsible
lemyoglobin is formed. In this pigment, the por- for the color of leafy vegetables and some fruits. In
phyrin ring is oxidized. The conversion into green leaves, the chlorophyll is broken down dur-
sulfmyoglobin is reversible; cholemyoglobin for- ing senescence and the green color tends to disap-
mation is irreversible, and this compound is rap- pear. In many fruits, chlorophyll is present in the
idly oxidized to yield globin, iron, and unripe state and gradually disappears as the yellow
tetrapyrrole. According to Fox (1966), this reac- and red carotenoids take over during ripening. In
tion may happen in the pH range of 5–7. plants, chlorophyll is isolated in the chloroplas-
Heating of meat results in the formation of a tids. These are microscopic particles consisting of
number of pigments. The globin is denatured. In even smaller units, called grana, which are usually
addition, the iron is oxidized to the ferric state. The less than 1 μm in size and at the limit of resolution
pigment of cooked meat is brown and called hemi- of the light microscope. The grana are highly
chrome. In the presence of reducing substances structured and contain laminae between which the
such as those that occur in the interior of cooked chlorophyll molecules are positioned.
meat, the iron may be reduced to the ferrous form; The chlorophylls are tetrapyrrole pigments in
the resulting pigment is pink hemochrome. which the porphyrin ring is in the dihydro form
In the curing of meat, the heme reacts with and the central metal atom is magnesium. There
nitrite of the curing mixture. The nitrite-heme are two chlorophylls, a and b, which occur
complex is called nitrosomyoglobin, which has a together in a ratio of about 1:25. Chlorophyll b
red color but is not particularly stable. On heating differs from chlorophyll a in that the methyl
the more stable nitrosohemochrome, the major group on carbon 3 is replaced with an aldehyde
cured meat pigment is formed, and the globin group. The structural formula of chlorophyll a is
portion of the molecule is denatured. This given in Fig. 6.15. Chlorophyll is a diester of a
requires a temperature of 65 °C. This molecule dicarboxylic acid (chlorophyllin); one group is
has been called nitrosomyoglobin and nitro- esterified with methanol, the other with phytyl
sylmyoglobin, but Möhler (1974) has pointed out alcohol. The magnesium is removed very easily
Tetrapyrrole Pigments 269

Fig. 6.14  Heme pigment reactions in meat and meat globin, the latter two being reaction products of nitrous
products. ChMb cholemyoglobin (oxidized porphyrin acid and the heme portion of the molecule, R reductants,
ring), O2Mb oxymyoglobin (Fe+2), MMb metmyoglobin O strong oxidizing conditions. Source: From J.B. Fox,
(Fe+3), Mb myoglobin (Fe+2), MMb·NO2 metmyoglobin The Chemistry of Meat Pigments, J. Agr. Food Chem.,
nitrate, NOMMb nitrosylmetmyoglobin, NOMb nitro- Vol. 14, no. 3, pp. 207–210, 1966, American Chemical
sylmyoglobin, NMMb nitrimetmyoglobin, NMb nitrimyo- Society

Fig. 6.15  Structure of chlorophyll a. (Chlorophyll b dif- of Enzymology for the Food Sciences, 1972, by courtesy
fers in having a formyl group at carbon 3). Source: of Marcel Dekker, Inc.
Reprinted with permission from J.R. Whitaker, Principles

by acids, giving pheophytins a and b. The action lysis of the phytol group, the water-soluble
of acid is especially important for fruits that are methyl chlorophyllides are formed. This reaction
naturally high in acid. However, it appears that can be catalyzed by the enzyme chlorophyllase.
the chlorophyll in plant tissues is bound to lipo- In the presence of copper or zinc ions, it is pos-
proteins and is protected from the effect of acid. sible to replace the magnesium, and the resulting
Heating coagulates the protein and lowers the zinc or copper complexes are very stable.
protective effect. The color of the pheophytins is Removal of the phytol group and the magnesium
olive-brown. Chlorophyll is stable in alkaline results in pheophorbides. All of these reactions
medium. The phytol chain confers insolubility in are summarized in the scheme presented in
water on the chlorophyll molecule. Upon hydro- Fig. 6.16.
270 6  Color and Food Colorants

Fig. 6.16  Reactions of


chlorophylls

In addition to those reactions described above, (Fig.  6.17), acyclic, monocyclic, and bicyclic.
it appears that chlorophyll can be degraded by yet Examples are lycopene (I)—acyclic; γ-carotene
another pathway. Chichester and McFeeters (II)—monocyclic; and α-carotene and β-carotene
(1971) reported on chlorophyll degradation in (III)—bicyclic.
frozen beans, which they related to fat peroxida- The carotenoids take their name from the
tion. In this reaction, lipoxidase may play a role, major pigments of carrot (Daucus carota). The
and no pheophytins, chlorophyllides, or color is the result of the presence of a system of
pheophorbides are detected. The reaction requires conjugated double bonds. The greater the number
oxygen and is inhibited by antioxidants. of conjugated double bonds present in the mole-
cule, the further the major absorption bands will
be shifted to the region of longer wavelength; as
Isoprenoid Derivative Pigments a result, the hue will become more red. A mini-
mum of seven conjugated double bonds are
Carotenoids required before a perceptible yellow color
appears. Each double bond may occur in either
The naturally occurring carotenoids, with the cis or trans configuration. The carotenoids in
exception of crocetin and bixin, are tetraterpe- foods are usually of the all-trans type and only
noids. They have a basic structure of eight iso- occasionally a mono-cis or di-cis compound
prenoid residues arranged as if two 20-carbon occurs. The prefix neo- is used for stereoisomers
units, formed by head-to-tail condensation of with at least one cis double bond. The prefix pro-
four isoprenoid units, had joined tail to tail. There is for poly-cis carotenoids. The effect of the pres-
are two possible ways of classifying the ence of cis double bonds on the absorption
carotenoids. The first system recognizes two
­ spectrum of β-carotene is shown in Fig. 6.18. The
main classes, the carotenes, which are configuration has an effect on color. The all-trans
­hydrocarbons, and the xanthophylls, which con- compounds have the deepest color; increasing
tain oxygen in the form of hydroxyl, methoxyl, numbers of cis bonds result in gradual lightening
carboxyl, keto, or epoxy groups. The second sys- of the color. Factors that cause change of bonds
tem divides the carotenoids into three types from trans to cis are light, heat, and acid.
Isoprenoid Derivative Pigments 271

Fig. 6.17 The
carotenoids

In the narrower sense, the carotenoids are the


250 four compounds shown in Fig. 6.17—α-, β-, and
T
γ-carotene and lycopene—polyene hydrocarbons
of overall composition C40H56. The relation
between these and carotenoids with fewer than
200
U 40 carbon atoms is shown in Fig. 6.19. The prefix
apo- is used to designate a carotenoid that is
Absorptivity (1/g - cm)

B derived from another one by loss of a structural


150 element through degradation. It has been sug-
gested that some of these smaller carotenoid mol-
a b c d
ecules are formed in nature by oxidative
degradation of C40 carotenoids (Grob 1963).
100

B
Fig. 6.18  Absorption spectra of the three stereoisomers
50 of beta carotene. B = neo-β-carotene; U = neo-β-­
U
carotene-U; T = all-trans-β-carotene. a, b, c, and d indi-
cate the location of the mercury arc lines 334.1 nm,
404.7 nm, 435.8 nm and 491.6 nm, respectively. Source:
T
0 From F. Stitt et al., Spectrophotometric Determination of
320 350 400 450 500 Beta Carotene Stereoisomers in Alfalfa, J. Assoc. Off.
Wavelength (nm) Agric. Chem. Vol. 34, pp. 460–471, 1951
272 6  Color and Food Colorants

Fig. 6.19 Relationship 40C


between the carotene
and carotenoids with CAROTENES
fewer than 40 carbons
20C 20C
VITAMIN A VITAMIN A

8C 24C 8C
METHYL METHYL
BIXIN
HEPTENONE HEPTENONE

10C 20C 10C


PICROCROCIN CROCIN PICROCROCIN

27C 13C
AZAFRIN IONONE

Fig. 6.20  Formation of retinal and vitamin A from β-carotene

Several examples of this possible relation- seed coat of the fruit of a tropical brush, Bixa
ship are found in nature. One of the best known orellana. The pigment bixin is a dicarboxylic
is the formation of retinin and vitamin A from acid esterified with one methanol molecule. A
β-carotene (Fig. 6.20). Another obvious rela- pigment named crocin has been isolated from
tionship is that of lycopene and bixin (Fig. 6.21). saffron. Crocin is a glycoside containing two
Bixin is a food color additive obtained from the molecules of gentiobiose. When these are
Isoprenoid Derivative Pigments 273

Fig. 6.21 Relationship
between lycopene and
bixin. Source: From
E.C. Grob, The
Biogenesis of Carotenes
and Carotenoids, in
Carotenes and
Carotenoids, K. Lang,
ed., 1963, Steinkopff
Verlag

Fig. 6.22 Relationship
between crocin and
picrocrocin and the
carotenoids

removed, the dicarboxylic acid crocetin is because the animal organism has a limited ability
formed (Fig. 6.22). It has the same general to absorb and deposit carotenoids. Some of the
structure as the aliphatic chain of the carotenes. most complex mixtures are found in citrus fruits.
Also obtained from saffron is the bitter com- Beta-carotene as determined in fruits and veg-
pound picrocrocin. It is a glycoside and, after etables is used as a measure of the provitamin A
removal of the glucose, yields saffronal. It is content of foods. The column chromatographic
possible to imagine a combination of two mole- procedure, which determines this content, does
cules of picrocrocin and one of crocin; this not separate α-carotene, β-carotene, and crypto-
would yield protocrocin. Protocrocin, which is xanthin. Provitamin A values of some foods are
directly related to zeaxanthin, has been found in given in Table 6.5. Carotenoids are not synthe-
saffron (Grob 1963). sized by animals, but they may change ingested
The structure of a number of important xan- carotenoids into animal carotenoids—as in, for
thophylls as they relate to the structure of example, salmon, eggs, and crustaceans. Usually
β-carotene is given in Fig. 6.23. Carotenoids may carotenoid content of foods does not exceed 0.1%
occur in foods as relatively simple mixtures of on a dry weight basis.
only a few compounds or as very complex mix- In ripening fruit, carotenoids increase at the
tures of large numbers of carotenoids. The sim- same time chlorophylls decrease. The ratio of
plest mixtures usually exist in animal products carotenes to xanthophylls also increases.
274 6  Color and Food Colorants

Fig. 6.23  Structure of


some of the important
carotenoids. Source:
From B. Borenstein and
R.H. Bunnell,
Carotenoids: Properties,
Occurrence, and
Utilization in Foods, in
Advances in Food
Research, Vol. 15,
C.O. Chichester et al.,
eds., 1967, Academic
Press

Common carotenoids in fruits are α- and there is an increasing proportion of lycopene


γ-carotene and lycopene. Fruit xanthophylls and increasing redness. Many peach varieties
are usually present in esterified form. Oxygen, are devoid of lycopene. Apricots may have
but not light, is required for carotenoid synthe- about 10% and tomatoes up to 90%. The lyco-
sis and the temperature range is critical. The pene content of tomatoes increases during rip-
relative amounts of different carotenoids are ening. As the chlorophyll breaks down during
related to the characteristic color of some fruits. ripening, large amounts of carotenoids are
In the sequence of peach, apricot, and tomato, formed (Table 6.6).
Isoprenoid Derivative Pigments 275

Table 6.5  Provitamin A value of various fruits and Table 6.7  Composition of the carotenes in crude palm oil
vegetables
Carotene % of total carotenes
Product IU/100 g Phytoene 1.27
Carrots, mature 20,000 Cis-β-carotene 0.68
Carrots, young 10,000 Phytofluene 0.06
Spinach 13,000 β-carotene 56.02
Sweet potato 6000 α-carotene 35.06
Broccoli 3500 ζ-carotene 0.69
Apricots 2000 γ-carotene 0.33
Lettuce 2000 δ-carotene 0.83
Tomato 1200 Neurosporene 0.29
Asparagus 1000 β-zeacarotene 0.74
Bean, french 1000 α-zeacarotene 0.23
Cabbage 500 Lycopene 1.30
Peach 800 Source. Reprinted with permission from Choo Yuen May,
Brussels sprouts 700 Carotenoids from Palm Oil, Palm Oil Developments, Vol.
Watermelon 550 22, pp. 1–6, Palm Oil Research Institute of Malaysia
Banana 400
Orange juice 200
Peaches contain violaxanthin, cryptoxanthin,
Source: From B. Borenstein and R.H. Bunnell,
Carotenoids: Properties, Occurrence, and Utilization in
β-carotene, and persicaxanthin as well as 25 other
Foods, in Advances in Food Research, Vol. 15, carotenoids, including neoxanthin. Apricots con-
C.O. Chichester et al., eds., 1967, Academic Press tain mainly β- and γ-carotene, lycopene, and little
if any xanthophyll. Carrots have been found to
have an average of 54 ppm of total carotene
Table 6.6  Development of pigments in the ripening (Borenstein and Bunnell 1967), consisting
tomato
mainly of α-, β, and ζ-carotene and some ­lycopene
Green Half-ripe Ripe and xanthophyll. Canning of carrots resulted in a
Pigment (mg/100 g) (mg/100 g) (mg/100 g) 7–12% loss of provitamin A activity because of
Lycopene 0.11 0.84 7.85 cis-trans isomerization of α- and β-carotene
Carotene 0.16 0.43 0.73
(Weckel 1962). In dehydrated carrots, carotene
Xanthophyll 0.02 0.03 0.06
oxidation and off-flavor development have been
Xanthophyll ester 0 0.02 0.10
correlated (Falconer et al. 1964). Corn contains
about one-third of the total carotenoids as caro-
tenes and two-thirds xanthophylls. Compounds
Color is an important attribute of citrus juice found in corn include zeaxanthin, cryptoxanthin,
and is affected by variety, maturity, and process- β-carotene, and lutein.
ing methods. The carotenoid content of oranges One of the highest known concentrations of
is used as a measure of total color. Curl and carotenoids occurs in crude palm oil. It contains
Bailey (1956) showed that the 5,6-epoxides of about 15–300 times more retinol equivalent than
fresh orange juice isomerize completely to carrots, green leafy vegetables, and tomatoes. All
5,8-epoxides during storage of canned juice. This of the carotenoids in crude palm oil are destroyed
change amounts to the loss of one double bond by the normal processing and refining operations.
from the conjugated double bond system and Recently, improved gentler processes have been
causes a shift in the wavelength of maximum developed that result in a “red palm oil” that retains
absorption as well as a decrease in molar absor- most of the carotenoids. The composition of the
bance. In one year’s storage at 70 °F, an apparent carotenes in crude palm oil with a total carotene
carotenoid loss of 20–30% occurs. concentration of 673 mg/kg is shown in Table 6.7.
276 6  Color and Food Colorants

Milkfat contains carotenoids with seasonal Counsell (1985). Natural carotenoid food colors
variation (related to feed conditions) ranging are annatto, oleoresin of paprika, and unrefined
from 2 to 13 ppm. Egg yolk contains lutein, zea- palm oil.
xanthin, and cryptoxanthin. The total carotenoid
content ranges from 3 to 89 ppm. Crustaceans
contain carotenoids bound to protein resulting in Benzopyran Derivative Pigments
a blue or blue-gray color. When the animal is
immersed in boiling water, the carotenoid-­protein Anthocyanins and Flavonoids
bond is broken and the orange-red color of the
free carotenoid appears. Widely distributed in The anthocyanin pigments are present in the sap
crustaceans is astaxanthin. Red fish contain of plant cells; they take the form of glycosides and
astaxanthin, lutein, and taraxanthin. are responsible for the red, blue, and violet colors
Common unit operations of food processing of many fruits and vegetables. When the sugar
are reported to have only minor effects on the moiety is removed by hydrolysis, the aglucone
carotenoids (Borenstein and Bunnell 1967). The remains and is called anthocyanidin. The sugar
carotenoid-protein complexes are generally more part usually consists of one or two molecules of
stable than the free carotenoids. Because carot- glucose, galactose, and rhamnose. The basic
enoids are highly unsaturated, oxygen and light structure consists of 2-phenyl-­benzopyrylium or
are major factors in their breakdown. Blanching flavylium with a number of hydroxy and methoxy
destroys enzymes that cause carotenoid destruc- substituents. Most of the anthocyanidins are
tion. Carotenoids in frozen or heat-sterilized derived from 3,5,7-trihydroxy-­flavylium chloride
foods are quite stable. The stability of carotenoids (Fig.  6.24) and the sugar moiety is usually
in dehydrated foods is poor, unless the food is attached to the hydroxyl group on carbon 3. The
packaged in inert gas. A notable exception is anthocyanins are highly colored, and their names
dried apricots, which keep their color well. are derived from those of flowers. The structure of
Dehydrated carrots fade rapidly. some of the more important anthocyanidins is
Several of the carotenoids are now commer- shown in Fig. 6.25, and the occurrence of antho-
cially synthesized and used as food colors. A pos- cyanidins in some fruits and vegetables is listed in
sible method of synthesis is described by Table 6.8. Recent studies have indicated that some
Borenstein and Bunnell (1967). Beta-ionone is anthocyanins contain additional components such
obtained from lemon grass oil and converted into as organic acids and metals (Fe, Al, Mg).
a C14 aldehyde. The C14 aldehyde is changed to Substitution of hydroxyl and methoxyl groups
a C16 aldehyde, then to a C19 aldehyde. Two influences the color of the anthocyanins. This
moles of the C19 aldehyde are condensed with effect has been shown by Braverman (1963)
acetylene dimagnesium bromide and, after a (Fig.  6.26). Increase in the number of hydroxyl
series of reactions, yield β-carotene. groups tends to deepen the color to a more bluish
Three synthetically produced carotenoids are used shade. Increase in the number of methoxyl groups
as food colorants, β-carotene, β-apo-8′-carotenal increases redness. The anthocyanins can occur in
(apocarotenal), and canthaxanthin. Because of different forms. In solution, there is an equilib-
their high tinctorial power, they are used at levels rium between the colored cation R+ or oxonium
of 1–25 ppm in foods (Dziezak 1987). They are salt and the colorless pseudobase ROH, which is
unstable in light but otherwise exhibit good stabil- dependent on pH.
ity in food applications. Although they are fat sol-
R + + H 2 O  ROH + H +
uble, water-dispersible forms have been developed
for use in a variety of foods. Beta-carotene imparts As the pH is raised, more pseudobase is
a light yellow to orange color, apocarotenal a light formed and the color becomes weaker. However,
orange to reddish-­ orange, and canthaxanthin, in addition to pH, other factors influence the color
orange-red to red. The application of these com- of anthocyanins, including metal chelation and
pounds in a variety of foods has been described by combination with other flavonoids and tannins.
Benzopyran Derivative Pigments 277

Fig. 6.24  Chemical structure of fruit anthocyanidins

Fig. 6.25  Structure of


some important
anthocyanidins

Anthocyanidins are highly colored in strongly About 16 anthocyanidins have been identified
acid medium. They have two absorption max- in natural products, but only the following six of
ima—one in the visible spectram at 500–550 nm, these occur frequently and in many different prod-
which is responsible for the color, and a second ucts: pelargonidin, cyanidin, delphinidin, peoni-
in the ultraviolet (UV) spectrum at 280 nm. The din, malvidin, and petunidin. The anthocyanin
absorption maxima relate to color. For example, pigments of Red Delicious apples were found to
the relationship in 0.01% HC1 in methanol is as contain mostly cyanidin-3-galactoside, cyanidin-
follows: at 520 nm pelargonidin is scarlet, at 3-arabinoside, and c­yanidin-7-­arabinoside (Sun
535 nm cyanidin is crimson, and at 546 nm del- and Francis 1968). Bing cherries contain primar-
phinidin is blue-mauve (Macheix et al. 1990). ily cyanidin-3-rutinoside, cyanidin-3-glucoside,
278 6  Color and Food Colorants

Table 6.8  Anthocyanidins occurring in some fruits and and small amounts of the pigments cyanidin,
vegetables
peonidin, peonidin-3-­ glucoside, and peonidin-
Fruit or vegetable Anthocyanidin 3-rutinoside (Lynn and Luh 1964). Cranberry
Apple Cyanidin anthocyanins were identified as cyanidin-
Black currant Cyanidin and delphinidin 3-monogalactoside, peonidin-3-­monogalactoside,
Blueberry Cyanidin, delphinidin, malvidin, cyanidin monoarabinoside, and peonidin-3-mono-
petunidin, and peonidin
arabinoside (Zapsalis and Francis 1965). Cabernet
Cabbage (red) Cyanidin
Sauvignon grapes contain four major anthocya-
Cherry Cyanidin and peonidin
nins: delphinidin-­3-­monoglucoside, petunidin-
Grape Malvidin, peonidin, delphinidin,
cyanidin, petunidin, and 3-monoglucoside, malvidin-3-monoglucoside,
pelargonidin and malvidin-­3-­
monoglucoside acetylated with
Orange Cyanidin and delphinidin chlorogenic acid. One of the major pigments is
Peach Cyanidin petunidin (Somaatmadja and Powers 1963).
Plum Cyanidin and peonidin Anthocyanin pigments can easily be destroyed
Radish Pelargonidin when fruits and vegetables are processed. High
Raspberry Cyanidin temperature, increased sugar level, pH, and
Strawberry Pelargonidin and a little cyanidin ascorbic acid can affect the rate of destruction
Source: From P. Markakis, Anthocyanins, in Encyclopedia (Daravingas and Cain 1965). These authors
of Food Technology, A.H. Johnson and M.S. Peterson, ­studied the change in anthocyanin pigments dur-
eds., 1974, AVI Publishing Co.
ing the processing and storage of raspberries.

Fig. 6.26 Effect of substituents on the color of anthocyanidins. Source: Reprinted with permission from
J.B.S. Braverman, Introduction to the Biochemistry of Foods, © 1963, Elsevier Publishing Co.
Benzopyran Derivative Pigments 279

During storage, the absorption maximum of the The colors of the anthocyanins at acid pH val-
pigments shifted, indicating a change in color. ues correspond to those of the oxonium salts. In
The level of pigments was lowered by prolonged slightly alkaline solutions (pH 8–10), highly col-
times and higher temperatures of storage. Higher ored ionized anhydro bases are formed. At pH 12,
concentration of the ingoing sugar syrup and the these hydrolyze rapidly to fully ionized chal-
presence of oxygen resulted in greater pigment cones (Fig. 6.28). Leuco bases are the reduced
destruction. form of the anthocyanins. They are usually with-
The stability of anthocyanins is increased by out much color but are widely distributed in fruits
acylation (Dougall 1997). These acylated antho- and vegetables. Under the influence of oxygen
cyanins may occur naturally as in the case of an and acid hydrolysis, they may develop the char-
anthocyanin from the purple yam (Yoshida et al. acteristic color of the carbonium ion. Canned
1991). This anthocyanin has one sinapic residue pears, for example, may show “pinking”—a
attached through a disaccharide and was found to change from the leuco base to the anthocyanin.
be stable at pH 6.0 compared to other anthocya- The flavonoids or anthoxanthins are glyco-
nins without acylation. Dougall (1997) were able sides with a benzopyrone nucleus. The flavones
to produce stable anthocyanins by acylation of have a double bond between carbons 2 and 3. The
carrot anthocyanins in cell cultures. They found flavonols have an additional hydroyxl group at
that a wide range of aromatic acids could be carbon 3, and the flavanones are saturated at
incorporated into the anthocyanin. ­carbons 2 and 3 (Fig. 6.29). The flavonoids have
Anthocyanins can form purplish or slategray low coloring power but may be involved in dis-
pigments with metals, which are called lakes. This colorations; for example, they can impart
can happen when canned foods take up tin from blue and green colors when combined with iron.
the container. Anthocyanins can be bleached by Some of these compounds are also potential sub-
sulfur dioxide. According to Jurd (1964), this is a strates for enzymic browning and can cause
reversible process that does not involve hydrolysis undesirable discoloration through this mecha-
of the glycosidic linkage, reduction of the pig- nism. The most ubiquitous flavonoid is quercetin,
ment, or addition of bisulfite to a ketonic, chalcone a 3,5,7,3′,4′-pentahydroxy flavone (Fig. 6.30).
derivative. The reactive species was found to be Many flavonoids contain the sugar rutinose, a
the anthocyanin carbonium ion (R+), which reacts disaccharide of glucose and rhamnose. Hesperidin
with a bisulfite ion to form a colorless chromen- is a flavanone occurring in citrus fruits and, at
2(or 4)-sulfonic acid (R–SO3H), similar in struc- pH 12, the inner ring opens to form a chalcone in
ture and properties to an anthocyanin carbinol a similar way as shown for the anthocyanins. The
base (R–OH). This reaction is shown in Fig. 6.27. chalcones are yellow to brown in color.

Fig. 6.27  Reaction of


bisulfite with the
anthocyanin carbonium
ion

Fig. 6.28  Structure of


anhydro base (I) and
chalcone (II)
280 6  Color and Food Colorants

Fig. 6.29  Structure of flavones, flavonals, flavanones, flavanonols, and isoflavones

Gallotannins release gallic acid on hydrolysis,


and ellagitannins produce ellagic acid. Ellagic
acid is the lactone form of hexahydroxydiphenic
acid, which is the compound originally present in
the tannin (Fig. 6.31).
Nonhydrolyzable or condensed tannins are also
Fig. 6.30  Structure of quercetin named proanthocyanidins. These are polymers of
flavan-3-ols, with the flavan bonds most commonly
between C4 and C8 or C6 (Fig. 6.24) (Macheix
Tannins et al. 1990). Many plants contain tannins that are
Tannins are polyphenolic compounds present in polymers of (+)-catechin or (−)-epicatechin. These
many fruits. They are important as color com- are hydrogenated forms of flavonoids or anthocy-
pounds and also for their effect on taste as a fac- anidins. Other monomers occupying places in con-
tor in astringency (see Chap. 7). Tannins can be densed fruit tannins have trihydroxylation in the
divided into two classes—hydrolyzable tannins B-ring: (+)-gallocat-echin and (−)-epigallocate-
and nonhydrolyzable or condensed tannins. The chin. Oligomeric and polymeric procyanidins are
tannins are characterized by the presence of a formed by addition of more flavan-3-ol units and
large number of hydroxyl groups, which provide result in the formation of helical structures. These
the ability to form reversible bonds with other structures can form bonds with proteins.
macromolecules, polysaccharides, and proteins, Tannins are present in the skins of red grapes
as well as other substances such as alkaloids. and play an important part in the flavor profile of
This bond formation may occur during the devel- red wine. Tannins in grapes are usually estimated
opment of the fruit or during the mechanical in terms of the content of gallic acid (Amerine
damage that takes place during processing. and Joslyn 1970).
Hydrolyzable tannins are composed of phenolic Oxidation and polymerization of phenolic
acids and sugars that can be broken down by acid, compounds as a result of enzymic activity of phe-
alkaline, or enzymic hydrolysis. They are polyesters noloxidases or peroxidases may result in the for-
based on gallic acid and/or hexahydroxydiphenic mation of brown pigments. This can take place
acid (Fig. 6.31). The usual sugar is d-glucose and during the growth of fruits (e.g., in dates) or dur-
molecular weights are in the range of 500–2800. ing mechanical damage in processing.
Other Pigments 281

Fig. 6.31  Structure of components of hydrolyzable tannins

Fig. 6.32  Structure of


naturally occurring
betalains in red beets.
Source: From J.H. Von
Elbe and I.-Y. Maing,
Betalains as Possible
Food Colorants of Meat
Substitutes, Cereal Sci.
Today, Vol. 18,
pp. 263–264, 316–317,
1973

Other Pigments red betacyanins and the yellow betaxanthins


(Von Elbe and Maing 1973). The structures of the
Betalains betacyanins are shown in Fig. 6.32. The major
betacyanin is betanin, which accounts for
Table beets are a good source of red pigments; 75–95% of the total pigments of beets. The
these have been increasingly used for food color- remaining pigments contain isobetanin, pre-
ing. The red and yellow pigments obtained from betanin, and isoprebetanin. The latter two are sul-
beets are known as betalains and consist of the fate monoesters of betanin and isobetanin,
282 6  Color and Food Colorants

respectively. The major yellow pigments are vul- air have a degrading effect on betanin, and the
gaxanthin I and vulgaxanthin. II Betanin is the effect is cumulative.
glucoside of betanidin, and isobetanin is the C-15
epimer of betanin.  aramels and Melanoidins
C
Betanidin has three carboxyl groups (pka = 3.4), Caramel color can be produced from a variety of
two phenol groups (pHa = 8.5), and asymmetric carbohydrate sources, but usually corn sugar
carbons at positions 2 and 15. The 15-position is syrup is used. Corn starch is first hydrolyzed with
easily isomerized under acid or basic conditions in acid to a DE of 8–9, followed by hydrolysis with
the absence of oxygen to yield isobetanidin. Under bacterial α-amylase to a DE of 12–14, then with
alkaline conditions and in the presence of gluta- fungal amyloglucosidase up to a DE of 90–95.
mine or glutamic acid, betanin can be converted to Several types of caramel are produced. The larg-
vulgaxanthin (Mabry 1970). est amount is electropositive or positive caramel,
The color of betanin solutions is influenced by which is made with ammonia. Electronegative or
pH. In the range of 3.5–7.0, the spectrum shows a negative caramel is made with ammonium salts.
maximum of 537 nm (Fig. 6.33). Below pH 3, the A slightly electronegative caramel is soluble in
intensity of this maximum decreases and a slight alcohol and is used for coloring beverages
increase in the region of 570–640 nm occurs and (Greenshields 1973). The composition and color-
the color shifts toward violet. At pH values over ing power of caramel depends on the type of raw
7, a shift of the maximum occurs to longer wave- materials and the process used. The melanoidins
length. At pH 9, the maximum is about 544 nm are formed from the reactions between reducing
and the color shifts toward blue. Von Elbe et al. sugars and basic nitrogenous compounds, often
(1974) found that the color of betanin is most called the Maillard reaction. Both Maillard-type
stable between pH 4.0 and 6.0. The thermostabil- reactions and pure caramelizing reactions are
ity is greatest between pH 4.0 and 5.0. Light and thought to be involved commonly in most of heat

Fig. 6.33  Visible spectra of betanin at pH values of 2.0, Food Science, Vol. 39, pp. 334–337, 1974, Institute of
5.0, and 9.0. Source: From J.H. Von Elbe, I.-Y. Maing, and Food Technologists
C.H. Amundson, Color Stability of Betanin, Journal of
References 283

(at high temperatures) processed food products. Fox, J. B. (1966). The chemistry of meat pigments. Journal
of Agricultural and Food Chemistry, 14, 207–210.
The reaction products are extremely complex in
Greenshields, R. N. (1973). Caramel—Part 2.
composition with high and low molecular weight Manufacture, composition and properties. Process
colored compounds, as well as a variety of vola- Biochemistry, 8(4), 17–20.
tile components. Griffiths, J. C. (2005). Natural and synthetic colors play
several roles in food and beverages. Here’s how they
are regulated in the United States. Food Technology,
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Grob, E. C. (1963). The biogenesis of carotenes and carot-
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Flavor
7
Han-Seok Seo, John W. Finley,
and John M. deMan

Our response to the food when we consume is a flavor perception is affected by visual cues (col-
combination of visual, tactile, thermal, taste and ors and images) as well as auditory cues (back-
aroma. Frequently our first interaction with a ground sound and biting/drinking-induced
food is visual or to the aroma of the food. When sound) (DuBose et al. 1980; Spence 2012).
we put it in our mouth we respond to the tempera- The senses of taste and smell give animals or
ture, texture (tactile) and taste response. In this humans the ability to evaluate what they eat and
chapter we will consider the taste and aroma drink. This evaluation helps animals and humans
responses. In recent years, the understanding of to promote ingestion of nutritious substances and
taste and smell has increased exponentially. prevent consumption of potential poisons or tox-
Hall (1968) defined flavor as follows: “Flavor ins. Animals, including humans, develop taste and
is the sensation produced by a material taken in smell preferences, which is the ability to choose
the mouth, perceived principally by the senses of certain types of food in preference to others. Taste
taste and smell, and also by the general pain, tac- and smell preferences can change with differing
tile and temperature receptors in the mouth. body needs and dietary interactions. The senses of
Flavor denotes the sum of the characteristics of taste and smell also motivate us to eat by seeking
the material which produce that sensation.” More the nutrients and energy such as fat and sugar.
recently, the International Organization for However, likings of sugar and fat vary with geno-
Standardization ( 2008) characterized flavor as a type, as well as individual experiences and envi-
“complex combination of the olfactory, gustatory ronmental factors. Animals often develop food
and trigeminal sensations perceived during tast- aversions, particularly if they become ill soon
ing. The flavor may be influenced by tactile, ther- after eating a certain food, even though that food
mal, painful and/or kinaesthetic effects.” was not the cause of the illness. Food preferences
Although the senses of taste and smell are the and aversions involve the senses of taste and
principal systems for distinguishing flavor in smell, and these phenomena are almost certainly
foods, other sensory cues contribute to the overall mediated through the central nervous system. In
sensation of flavor. For example, texture has a addition, by sniffing off-odors or tasting bitterness
very definite effect on our perception of taste. or sourness, the senses of taste and smell help us
Smoothness, roughness, granularity, and viscos- to avoid ingesting harmful foods containing toxic,
ity can all influence flavor, as can hotness of microbes, microbial by products, or chemical
spices, coolness of menthol, brothiness or full- contamination (Reed and Knaapila 2010).
ness of certain amino acids, and the tastes Traditionally we are taught that there are five
described as metallic and alkaline. In addition, basic taste responses on the tongue; salt, acid,

© Springer International Publishing AG 2018 285


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_7
286 7 Flavor

sweet, bitter, and umami. Each taste quality has a causing the pancreas beta cells, located in the
specific role in the detection of nutritious as well islets of Langerhans, to increase the release of
as poisonous substances; sweet taste for carbohy- insulin (Laffitte et al. 2014). Bitter taste receptors
drate sources of calories, umami for protein and (T2Rs) are found in the cilia of human bronchial
amino acid contents, salty for mineral contents, and sinonasal epithelial cells where they can
and sour for fruits ripeness and spoiled foods, serve to cause a response to expel inhaled irri-
and bitter for harmful compounds (Iwata et al. tants (Shah et al. 2009).
2014). Many earlier textbooks and journal arti- Traditionally various areas of the tongue were
cles cite the “tongue map” suggesting that differ- considered to be responsible for the perception of
ent areas of the tongue are sensitive to specific the basic tastes. More recent studies have demon-
tastes as show in Fig. 7.1. However, all taste qual- strated that taste buds contain 50–120 taste cells
ities are sensitive across the area of tounge. with multiple receptors for all five basic tastes.
The interactions of foods with saliva can also Each taste cell has receptors on its apical surface
have a major influence on our perception of tast- that are transmembrane proteins. These proteins
ing substances. Saliva acts as a solvent for taste admit the ions that give rise to the sensations of
substances as well as a diffuser of the solutes to salty and sour tastes as well as bind to the mole-
the taste receptor sites. Salivar also acts as a buf- cules that elicit the sensations of sweet, bitter,
fer for acidic foods and may bind with bitter taste and umami tastes (Engelen 2010).
substances. In addition, some salivary consti- The various types of taste cells are located
tutents alter taste sensitivity by continuously within taste buds. These structures are predomi-
stimulating the taste receptors, and salivar also nantly located on the tongue and soft palate. Most
protect the taste receptors from dryness, bacterial of the taste buds on the tongue are located within
infection, and disuse atrophy (Matsuo 2000). tiny projections on the tongue called papillae.
Taste is the response to dissolved molecules The predominant papillae on the tongue are the
and ions called tastants. Taste is detected when filiform or threadlike structures that do not con-
tastants interact with taste receptor cells. These tain taste buds. The filiform structures are
cells are clustered in taste buds on the tongue and involved in somatosensory and mechanical func-
scattered in other areas of the body, for example tions. The taste buds are found on the fungiform
the nasal epithelium, the trachea, the stomach, papillae on the anterior two-thirds of the tongue.
and the intestines (Finger and Kinnamon 2011). The fungiform papillae are the most noticeable
Sweet taste receptors (T1Rs) are found in cells of and typically contain one or more taste buds. The
the duodenum. When sugars reach the duode- circumvallate papillae are located on the dorsal
num, the cells respond by releasing incretins, side of the tounge and the foliate papillae are
found in small trenches located on the sides of
the rear of the tongue (Fig. 7.2).
Taste buds are onion-shaped structures com-
posed of between 50 and 100 taste cells. Each
taste cell has finger-like projection called a
microvilli. The microvilli protrude through an
opening at the top of the taste bud referred to as
the taste pore. Food chemicals called tastants
which are dissolved in the saliva contact the taste
cells through the taste pore. They then interact
with proteins on the surfaces of the cells known
as taste receptors or with pore like proteins called
ion channels. These interactions cause electrical
Fig. 7.1 Misinterpreted tongue map suggesting four
basic tastes are sensitive on specific regions of the tongue. changes in the taste cells that send chemical sig-
The basic tastes are sensitive on every part of the tongue nals to adjacent neurons ultimately resulting in
7 Flavor 287

Fig. 7.2  Taste buds and the cranial nerves of the tongue. Diagram of a circumvallate papilla showing location of
(a) Distribution of taste papillae on the dorsal surface of individual taste buds. (c) Light micrograph of a taste bud.
the tongue. Different responses to sweet, salty, sour, and (d) Diagram of a taste bud, showing various types of taste
bitter tastants recorded in the three cranial nerves (VII, IX, cells and the associated gustatory nerves. The apical sur-
and X) that innervate the tongue and epiglottis are indi- faces of the receptor cells have microvilli that are oriented
cated at left (a). The size of the circles representing toward the taste pore (reference source of Fig. 7.2 is miss-
sucrose, NaCl, HCl, quinine, and water corresponds to the ing: http://www.uth.tmc.edu/courses/dental/smell-taste/
relative response of the papillae to these stimuli. (b) taste.html?)

impulses to the brain. The electrical changes in The responses to bitter and sweet tastes by the
the taste cells that result in signals to the brain are taste cells are not always closely correlated with
dependent on the concentrations of the ions. Like the chemical structure of the tastant molecule.
neurons, taste cells have a net negative charge Manny carbohydrates, particularly simple sugars
internally and a net positive charge externally. are sweet, but others are not sweet at all. Many
Tastants bind to the taste cells, the concentration non-carbohydrate molecules result in a sweet
of positive ions inside cells increases, eliminating taste response. For example, chloroform, steva-
the net charge differences internally and exter- side, saccharin and aspartame all cause sweet
nally. This depolarization causes the taste cells to responses. None of these molecules has any com-
release chemical signals called neurotransmit- mon structure with sweet sugars. Compounds
ters, which cause neurons connected to the taste that result in salty or sour tastes are less diverse
cells to transmit electrical messages to the brain. because they are generally ions such as hydrogen
288 7 Flavor

ions for sour and sodium for salty taste. The with electrolytes has been proposed by Beidler
chemicals that produce salty and sour tastes act (1957); it is shown in Fig. 7.3. The time required
directly through ion channels, whereas those for taste response to take place is in the order of
responsible for sweet and bitter tastes bind to sur- 25 ms. The taste molecule is weakly adsorbed,
face of the receptor. The receptors then signal the thereby creating a disturbance in the molecular
cells which cause the opening and closing of ion geography of the surface and allowing an inter-
channels. Gustducin is a G-protein that converts change of ions across the surface. This reaction is
the electrical impulse to a signal. Gustducin is followed by an electrical depolarization that initi-
referred to as a G-protein, which is found on the ates a nerve impulse.
underside of many different receptors. The term The taste receptor mechanism has been more
G-protein is used because the activity of such fully described by Kurihara (1987). The process
proteins is regulated by guanosine triphosphate, from chemical stimulation to transmitter release
GTP. When a tastant molecule binds to a taste is schematically presented in Fig. 7.4. The recep-
cell receptor, it prompts the subunits of gustducin tor membranes contain voltage-dependent cal-
to split apart and carry out biochemical reactions cium channels. Taste compounds contact the
that ultimately open and close ion channels and taste cells and depolarize the receptor membrane;
make the cell interior more positively charged. this depolarization spreads to the synaptic area,
Within a taste bud, there is a network of den- activating the voltage-dependent calcium chan-
drites of sensory nerves called “taste nerves.” nels. Influx of calcium triggers the release of the
Taste cells are stimulated by the binding of chem- transmitter norepinephrine.
icals to their receptors causing the taste cell to The relationship between stimulus concentra-
become depoloraized, this depolarization is tion and neural response is not simple. As the
transmitted to the taste nerve fibers resulting in a stimulus concentration increases, the response
potential that is ultimately transmitted to the increases at a decreasing rate until a point is
brain. The nerve transmission rapidly adapts after reached where further increase in stimulus con-
the initial stimulus, and a strong discharge is centration does not produce a further increase in
observed in the taste nerve fibers but within a few response. Beidler (1954) proposed the following
seconds. That response diminishes to a steady-­ equation relating magnitude of response and
state level of much lower amplitude. We know stimulus concentration:
that binding between stimulus and receptor is a
C C 1
weak one because no irreversible effects have = +
been observed. A mechanism of taste stimulation R Rs KRs

Fig. 7.3  Mechanism of


taste stimulation as
proposed by Beidler.
Source: From
L.M. Beidler, Facts and
Theory on the
Mechanism of Taste and
Odor Perception, in
Chemistry of Natural
Food Flavors, 1957,
Quartermaster Food and
Container Institute for
the Armed Forces
7 Flavor 289

Fig. 7.4  Diagram of a


taste cell (reference
source: http://
scienceinit.
in/2016/03/31/
how-do-we-taste)

where There appear to be significant age- or sex-­


C = stimulus concentration related differences in taste sensitivity, and espe-
R = response magnitude cially heavy smoking (more than 20 cigarettes
Rs = maximum response per day) results in a deterioration in taste respon-
K = equilibrium constant for the stimulus- recep- siveness with age.
tor reaction Differences in taste perception between indi-
viduals seem to be common. Peryam (1963) found
K values reported by Beidler for many substances that sweet and salt are usually well recognized.
are in the range of 5–15. However, with sour and bitter taste some difficulty
It appears that the initial step in the stimulus-­ is experienced. Some tasters ascribe a bitter qual-
receptor reaction is the formation of a weak com- ity to citric acid and a sour quality to caffeine.
plex, as evidenced by the small values of K. The Recent studies have demonstrated that taste-­
complex formation results in the initiation of the signalling molecules are distributed not only in the
nerve impulse. Because of the decreasing rate of gustatory epithelium, but also in other tissues,
response, we know that the number of receptor including the gastrointestinal tract, airways, testes
sites is finite. The taste response is a function of and brain. Taste signalling mechanisms in the gas-
the proportion of sites occupied by the stimulus trointestinal tract have been found to participate in
compound. detecting sweet, umami and bitter compounds. It
According to Beidler (1957), the threshold has been proposed that tastant/nutrient detection
value of a substance depends on the equilibrium by other systems contributes to the behavioural
constant and the maximum response. Since K responses to food intake (Iwatsuki and Torii 2012).
and Rs both vary from one substance to another The bitter taste receptor (TAS2R)-family of
and from one species to another, the threshold G-protein-coupled receptors has been identified
also varies between substances and species. The on the tongue as detectors of bitter taste. In the
concentration of the stimulus can be increased in last few years, they have been discovered in
steps just large enough to elicit an increase in extra-oral tissues, including the airways, the gut,
response. This amount is called the just notice- the brain and even the testis. In tissues that
able difference (JND). contact the exterior, protective functions for
­
290 7 Flavor

TAS2Rs have been proposed, in analogy to their Table 7.1  Taste thresholds for basic taste sensations
function on the tongue as toxicity detector. Examples of human taste thresholds
However, TAS2Rs have also been found in inter- Taste Substance Threshold for tasting
nal organs, suggesting other roles for these recep- Salty NaCl 0.01 M
tors, perhaps involving as yet unidentified Sour HCl 0.0009 M
endogenous ligands. The current review gives an Sweet Sucrose 0.01 M
overview of the different proposed functions for Bitter Quinine 0.000008 M
TAS2Rs in tissues other than the oral cavity; Umami Glutamate 0.0007 M
from appetite regulation to the treatment of (Source: [email protected]
asthma, regulation of gastrointestinal motility
and control of airway innate immunity (Avau and The taste cells transduce the stimuli from tas-
Depoortere 2016). tants and provide the identity, concentration, and
Once taste signals are transmitted to the brain, pleasant or unpleasant quality of the tastant. This
several neural pathways are activated that influ- information is translated to the gastrointestinal
ence digestive function. Tasting food is followed system causing salivation and swallowing (or gag-
rapidly by increased salivation and by low level ging and regurgitation if the substance is noxious).
secretory activity in the stomach. The temperature and texture of food is relayed
from the mouth via somatic sensory receptors from
the trigeminal and other sensory cranial nerves to
Taste Sensations the thalamus and somatic sensory cortices. Food is
not simply eaten for nutritional value; taste percep-
The sense of taste is equivalent to excitation of tion also depends on cultural backgrounds and
taste receptors. Taste receptors for a large number psychological factors (Purves et al. 2001).
of specific chemicals have been identified which
contribute to the reception of taste. Five basic
types of tastes are recognized by humans: Chemical Structure and Taste

• Sweet—usually indicates energy rich A first requirement for a substance to produce a


nutrients. taste is that it be water soluble. The relationship
• Bitter—allows sensing of diverse natural between the chemical structure of a compound
toxins. and its taste is more easily established than that
• Salty—allows modulating diet for electrolyte between structure and smell. In general, all acid
balance. substances are sour. Sodium chloride and other
• Sour—typically the taste of acids. salts are salty, but as constituent atoms get bigger,
• Umami—the taste of amino acids (e.g., meat a bitter taste develops. Potassium bromide is both
broth or aged cheese). salty and bitter, and potassium iodide is predomi-
nantly bitter. Sweetness is a property of sugars
None of these tastes are elicited by a single and related compounds but also of lead acetate,
chemical. There are thresholds for detection of beryllium salts, and many other substances such
taste that differ among chemicals that deliver as the artificial sweeteners saccharin and
similar taste. For example, sucrose, 1-propyl-2 cyclamate. Bittemess is exhibited by alkaloids
­
amino-4-nitrobenzene and lactose all taste sweet such as quinine, picric acid, and heavy metal salts.
to humans, but the sweet taste is elicited by these Minor changes in chemical structure may
chemicals at concentrations of roughly 10 mM, 2 change the taste of a compound from sweet to
uM and 30 mM respectively—a range of potency bitter or tasteless. For example, Beidler (1966)
of roughly 15,000-fold. Substances sensed as bit- has examined saccharin and its substitution com-
ter typically have very low thresholds. Table 7.1 pounds. Saccharin is 500 times sweeter than
illustrates the relative threshold concentrations of sugar (Fig. 7.5). Introduction of a methyl group
various types of tastants. or of chloride in the para position reduces the
Chemical Structure and Taste 291

Fig. 7.5  The effect of substitutions in saccharin on sweetness. Source: From L.M. Beidler, Chemical Excitation of
Taste and Odor Receptors, in Flavor Chemistry, I. Hornstein, ed., 1966, American Chemistry Society

these are probably derived from sulfur-­containing


decomposition products. Seven amino acids have
a bitter taste in the L form or a sweet taste in the
D form, except for L-alanine, which has a sweet
taste (Table 7.2). Solms et al. (1965) reported on
the taste intensity, especially of aromatic amino
acids. L-tryptophan is about half as bitter as caf-
Fig. 7.6  Taste of nitrotoluidine isomers feine; D-tryptophan is 35 times sweeter than
sucrose and 1.7 times sweeter than calcium
cyclamate. L-phenylalanine is about one-fourth
sweetness by half. Placing a nitro group in the as bitter as caffeine; the D form is about seven
meta position makes the compound very bitter. times sweeter than sucrose. L-tyrosine is about
Introduction of an amino group in the para posi- one-twentieth as bitter as caffeine, but D-tyrosine
tion retains the sweetness. Substitutions at the is still 5.5 times sweeter than sucrose.
imino group by methyl, ethyl, or bromoethyl Some researchers claim that differences exist
groups all result in tasteless compounds. between the L and D forms of some sugars. They
However, introduction of sodium at this location propose that L-glucose is slightly salty and not
yields sodium saccharin, which is very sweet. sweet, whereas D-glucose is sweet. There is even
The compound 5-nitro-o-toluidine is sweet. a difference in taste between the two anomers of
The positional isomers 3-nitro-o-toluidine and D-mannose. The α form is sweet as sugar, and the
3-nitro-p-toluidine are both tasteless (Fig. 7.6). β form is bitter as quinine.
Teranishi et al. (1971) provided another example Optical isomers of carvone have totally dif-
of change in taste resulting from the position of ferent flavors. The D+ form is characteristic of
substituent group: 2-amino-4-nitro-propoxyben- caraway; the L− form is characteristic of
zene is 4000 times sweeter than sugar, 2-nitro-­4-­ spearmint.
amino-propoxybenzene is tasteless, and The ability to taste certain substances is genet-
2,4-dinitro-propoxybenzene is bitter (Fig. 7.7). ically determined and has been studied with
Dulcin (p-ethoxyphenylurea) is extremely sweet, phenylthiourea. At low concentrations, about
the thiourea analog is bitter, and the o-­ 25% of subjects tested do not taste this com-
ethoxyphenylurea is tasteless (Fig. 7.8). pound; for the other 75%, the taste is bitter. The
Just as positional isomers affect taste, so do inability to taste phenylthiourea is probably due
different stereoisomers. There are eight amino to a recessive gene. The compounds by which
acids that are practically tasteless. A group of tasters and nontasters can be differentiated all
three has varying tastes; except for glutamic acid, contain the following isothiocyanate group:
292 7 Flavor

Fig. 7.7 Taste
of substituted
propoxybenzenes

Fig. 7.8  Taste of


substituted
ethoxybenenes

Table 7.2  Difference in taste between the L-and D-forms


of amino acids
Taste of D
Amino acid Taste of L isomer isomer
Asparagine Insipid Sweet
Glutamic acid Unique Almost tasteless
Phenylalanine Faintly bitter Sweet, bitter
aftertaste
Leucine Flat, faintly bitter Strikingly sweet
Valine Slightly sweet, Strikingly sweet S
bitter Fig. 7.9  Compounds containing the || group by
Serine Faintly sweet, stale Strikingly sweet -C - N-
after taste which tasters and nontasters can be differentiated
Histidine Tasteless to bitter Sweet
Isoleucine Bitter Sweet
Methionine Flat Sweet
Tryptophan Bitter Very sweet

phenylurea, urea, and uracil, do not show this


phenomenon. Another compound containing the
isothiocyanate group has been found in many spe-
cies of the Cruciferae family; this family includes
These compounds—phenylthiourea, thiourea, cabbage, turnips, and rapeseed and is well known
and thiouracil—are illustrated in Fig. 7.9. The for its goitrogenic effect. The compound is goi-
corresponding compounds that contain the group, trin, 5-vinyloxazolidine-2-thione (Fig. 7.10).
Sweet Taste 293

Sweet Taste the sweetness of benzyl alcohol and the sweet-


ness of the anti isomer of anisaldehyde oxime, as
Many investigators have attempted to relate the well as the lack of sweetness of the syn isomer.
chemical structure of sweet tasting compounds to The structure of these compounds is given in
the taste effect, and a series of theories have been Fig.  7.12. The AH,B system present in sweet
proposed (Shallenberger 1971). Shallenberger compounds is, according to Shallenberger, able
and Acree (1967, 1969) proposed a theory that to react with a similar AH,B unit that exists at the
can be considered a refinement of some of the taste bud receptor site through the formation of
ideas incorporated in previous theories. simultaneous hydrogen bonds. The relatively
According to this theory, called the AH,B theory, strong nature of such bonds could explain why
all compounds that bring about a sweet taste the sense of sweetness is a lingering sensation.
response possess an electronegative atom A, such According to the AH,B theory, there should not
as oxygen or nitrogen. This atom also possesses a be a difference in sweetness between the L and D
proton attached to it by a single covalent bond; isomers of sugars. Experiments by Shallenberger
therefore, AH can represent a hydroxyl group, an (1971) indicated that a panel could not distin-
imine or amine group, or a methine group. Within guish among the sweet taste of the enantiomor-
a distance of about 0.3 nm from the AH proton, phic forms of glucose, galactose, mannose,
there must be a second electronegative atom B, arabinose, xylose, rhamnose, and glucoheptu-
which again can be oxygen or nitrogen (Fig. 7.11). lose. This suggests that the notion that L sugars
Investigators have recognized that sugars that are tasteless is a myth.
occur in a favored chair conformation yield a gly- Spillane (1996) has pointed out that the AH,B
col unit conformation with the proton of one theory appears to work quite well, although spatial,
hydroxyl group at a distance of about 0.3 nm hydrophobic/hydrophilic, and electronic effects
from the oxygen of the next hydroxyl group; this are also important. Shallenberger (1998) describes
unit can be considered as an AH,B system. It was the initiation of sweetness as being due to a con-
also found that the π bonding cloud of the ben- certed intermolecular, antiparallel hydrogen-­
zene ring could serve as a B moiety. This explains bonding interaction between the glycophore
(Greek glyks, sweet; phoros, to carry) and receptor
dipoles. The difficulty in explaining the sweetness
of compounds with different chemical structures is
also covered by Shallenberger (1998) and how this
has resulted in alternative taste theories. The appli-
cation of sweetness theory is shown to have impor-
tant applications in the food industry.
Extensive experiments with a large number of
sugars by Birch and Lee (1971) support
Shallenberger’s theory of sweetness and indicate
that the fourth hydroxyl group of glucopyrano-
Fig. 7.10  5-vinyloxazoli ine-2-thione sides is of unique importance in determining

Fig. 7.11  The AH,B


theory of sweet taste
perception
294 7 Flavor

Table 7.3 Relative sweetness of sugars and other


sweeteners
Compound Relative sweetness
Sucrose 1
Lactose 0.27
Maltose 0.5
Sorbitol 0.5
Galactose 0.6
Glucose 0.5–0.7
Mannitol 0.7
Glycerol 0.8
Fig. 7.12  Anti-anisaldehyde oxime, sweet; and syn-­ Fructose 1.1–1.5
anisaldehyde oxime, tasteless Cyclamate 30–80
Glycyrrhizin 50
Aspartyl-phenylalanine methylester 100–200
sweetness, possibly by donating the proton as the Stevioside 300
AH group. Apparently the primary alcohol group Naringin dihydrochalcone 300
is of little importance for sweetness. Substitution Saccharin 500–700
of acetyl or azide groups confers intense bitter- Neohesperidin dihydrochalcone 1000–1500
ness to sugars, whereas substitution of benzoyl Source: From J. Solms, Nonvolatile Compounds and the
groups causes tastelessness. Flavor of Foods, in Gustation and Olfaction, G Ohloff and
As the molecular weight of saccharides A.F. Thomas, eds., 1971, Academic Press
increases, their sweetness decreases. This is best
explained by the decrease in solubility and increase
in size of the molecule. Apparently, only one sugar
residue in each oligosaccharide is involved in the Sour Taste
interaction at the taste bud receptor site.
The relative sweetness of a number of sugars Although it is generally recognized that sour
and other sweeteners has been reported by Solms taste is a property of the hydrogen ion, there is no
(1971) and is given in Table 7.3. These figures simple relationship between sourness and acid
apply to compounds tasted singly and do not nec- concentration. Acids have different tastes; the
essarily apply to sugars in foods, except in a gen- sourness as experienced in the mouth may depend
eral sense. The relative sweetness of mixtures of on the nature of the acid group, pH, titratable
sugars changes with the concentration of the acidity, buffering effects and the presence of
components. Synergistic effects may increase the other compounds, especially sugars. Organic
sweetness by as much as 20–30% in such mix- acids have a greater taste effect than inorganic
tures (Stone and Oliver 1969). acids (such as hydrochloric acid) at the same
Steroidal alkaloids (SAs) and their glycosyl- pH. Information on a number of the most
ated forms (SGAs) found in the nightshade fam- ­common acids found in foods and phosphoric
ily are toxic to humans and animals. These acid (which is also used in soft drinks) has been
compounds are produced by members of the collected by Solms (1971) and compared with
Solanaceae and Liliaceae plant families. In the hydrochloric acid. This information is presented
plants these metabolites serves as a chemical bar- in Table 7.4.
riers against a broad range of pests and patho- According to Beatty and Cragg (1935), rela-
gens. In humans and animals, SAs are considered tive sourness in unbuffered solutions of acids is
anti-nutritional factors because they affect the not a function of molarity but is proportional to
digestion and absorption of nutrients from food the amount of phosphate buffer required to bring
and in some cases they can cause poisoning the pH to 4.4. Ough (1963) determined relative
(Cardenas et al. 2015). sourness of four organic acids added to wine and
Salty Taste 295

Table 7.4  Properties of some acids, arranged in order of decreasing acid taste and with tartaric acid as reference
Properties of 0.05 N solutions Ionization Taste
Acid Taste Total acid (g/L) pH constant sensation Found In
Hydrochloric +1.43 1.85 1.70 – – –
Tartaric 0 3.75 2.45 1.04 × 10–3 Hard Grape
Malic −0.43 3.35 2.65 3.9 × 10–4 Green Apple, pear, prune, grape, cherry,
apricot
Phosphoric −1.14 1.65 2.25 7.52 × 10–3 Intense Orange, grapefruit
Acetic −1.14 3.00 2.95 1.75 × 10–5 Vinegar –
Lactic −1.14 4.50 2.60 1.26 × 10–4 Sour, tart –
Citric −1.28 3.50 2.60 8.4 × 10–4 Fresh Berries, citrus, pineapple
Propionic −1.85 3.70 2.90 1.34 × 10–5 Sour, cheesy –
Source: From J. Solms, Nonvolatile Compounds and the Flavor of Foods, in Gustation and Olfaction, G. Ohloff and
A.F. Thomas, eds., 1971, Academic Press

also preference for these acids. Citric acid was


judged the most sour, fumaric and tartaric about Salty Taste
equal, and adipic least sour. The tastes of citric
and tartaric acids were preferred over those of The salty taste is best exhibited by sodium chlo-
fumaric and adipic acids. ride. It is sometimes claimed that the taste of salt
Pangborn (1963) determined the relative by itself is unpleasant and that the main purpose
sourness of lactic, tartaric, acetic, and citric acid of salt as a food component is to act as a flavor
and found no relation between pH, total acidity, enhancer or flavor potentiator. The taste of salts
and relative sourness. It was also found that depends on the nature of both cation and anion.
there may be considerable differences in taste As the molecular weight of either cation or
effects between sugars and acids when they are anion—or both—increases, salts are likely to
tested in aqueous solutions and in actual food taste bitter. The lead and beryllium salts of acetic
products. acid have a sweet taste. The taste of a number of
Buffering action appears to help determine the salts is presented in Table 7.5.
sourness of various acids; this may explain why The current trend of reducing sodium intake in
weak organic acids taste more sour than mineral the diet has resulted in the formulation of low-­
acids of the same pH. It is suggested that the buff- sodium or reduced-sodium foods. It has been
ering capacity of saliva may play a role, and shown (Gillette 1985) that sodium chloride
foods contain many substances that could have a enhances mouthfeel, sweetness, balance, and
buffering capacity. saltiness, and also masks or decreases off-notes.
Wucherpfennig (1969) examined the sour taste Salt substitutes based on potassium chloride do
in wine and found that alcohol may decrease the not enhance mouthfeel or balance and increase
sourness of organic acids. He examined the rela- bitter or metallic off-notes.
tive sourness of 17 organic acids and found that Some individuals are sensitive are sensi-
the acids tasted at the same level of undissociated tive and need to reduce the sodium content of
acid have greatly different intensities of sourness. their diet. Salt sensitivity individuals experience
Partially neutralized acids taste more sour than increases in blood pressure in response to salt
pure acids containing the same amount of undis- intake, Salt sensitive individuals are more likely
sociated acids. The change of malic into lactic to have high blood pressure than those who are
acid during the malolactic fermentation of wines resistant to salt. Salt-sensitive individuals are at
leads to a decrease in sourness, thus making the higher risk for high blood pressure, cardiovascu-
flavor of the wine milder. lar disease and lower survival rate later in life if
296 7 Flavor

Table 7.5  Taste sensations of salts gene, and the AGT gene, Table 7.7 lists the fre-
Taste Salts quency of risk variants associated with increased
Salty LiCl, LiBr, Lil, NaNO3, NaCl, NaBr, risk for salt sensitivity and hypertension.
Nal, KNO3, KCl To help consumers reduce or control sodium
Salty and KBr, NH4l, KCl intake, many salt substitutes with low sodium
bitter
content have been designed to reduce the risk of
Bitter CsCl, CsBr, Kl, MgSO4
high blood pressure and cardiovascular disease
Sweet Lead acetate,a beryllium acetatea
associated with a high intake of sodium chloride,
Taste Salts
while delivering similar taste [Scientific Advisory
a
Extremely toxic
Committee on Nutrition Salt and Health (2003)].
The increase in sodium consumption is consid-
Table 7.6  Percentage of salt-sensitive people in different ered a potential health threat for some individu-
populations (data from Sullivan 1991) als. The Institute of Medicine of the National
Population Academy of Sciences has established adequate
Blood pressure White (%) Black (%) daily intakes (AIs) for sodium and potassium and
Normal 15 27 a tolerable upper intake level (UL) for sodium,
Hypertension 29 50 based on its effects on blood pressure (Table 7.8;
IOM 2004). Persons with a greater risk for hyper-
tension (adults who are Black, over 40 years old,
they continuously live an unhealthy lifestyle or or already have hypertension or prehypertension)
have a high-sodium diet (Weinberger et al. 2001). have been urged to consume no more than the AI
A study by Sullivan 1991), salt-sensitive individ- level of sodium each day (CDCP 2009; Doyle
uals are more likely to have hypertension, as are and Glass 2010). These products are predomi-
blacks more than whites (Table 7.6). Another nantly potassium chloride (KCl). Potassium
study reports that approximately 60% of Chinese Chloride’s toxicity is similar to Sodium Chloride
who have high blood pressure are salt-sensitive in healthy individuals; the LD50 is about 2.5 g/
(Li 2012). kg. Potassium lactate is frequently used to reduce
Sodium homeostasis in the human body is sodium levels in meat and poultry products. The
regulated mainly by the renin-angiotensin-­ recommended daily allowance of potassium is
aldosterone system. This system operates mainly higher than that for sodium (Caggiula et al. 1985).
in the kidney and in vascular smooth muscle Sodium is an essential micronutrient and, via
cells. Variations in this system, due to genetic salt taste, appetitive. High consumption of
background, age, race, gender and medical his- sodium is, however, related to negative health
tory, cause the kidney of salt-sensitive individu- effects such as hypertension, cardiovascular dis-
als to handle excess sodium less efficiently. Asian eases and stroke. In industrialized countries,
or African ancestry, older age, female gender, about 75% of sodium in the diet comes from pro-
high blood pressure, and kidney disease are all cessed foods and foods eaten away from home.
associated with salt-sensitivity. Reducing sodium in processed foods will be,
Salt sensitive individuals exhibit variations in however, challenging due to sodium’s specific
genes involved in the renin-angiotensin-­functionality in terms of flavor and associated
aldosterone system which predispose them to salt palatability of foods (i.e., increase of saltiness,
sensitivity (Sanada et al. 2011). About 38% of the reduced bitterness, enhancement of sweetness
general population carries an ACE gene variant and other congruent flavors). Salt has many ben-
that causes increased activity of the system lead- eficial properties for both preservation and mul-
ing to blood pressure increase in response to tiple culinary benefits. Salt improves the sensory
higher sodium levels in the blood. This part of the properties of nearly all foods. The principle rea-
population becomes salt-sensitive. Two other son for adding salt to food is that enhances the
genes associated with salt sensitivity are the NOS3 positive sensory attributes of foods. Salt makes
Salty Taste 297

Table 7.7  Percentage gene variants associated with salt sensitivity in different populations
Gene symbol All (%) African (%) American (%) Asian (%) European (%)
ACE 38 17 40 31 56
ADD1 27 17 19 50 20
ADRB1 30 40 21 21 34
AGT 66 88 64 83 41
AGTR1 16 3 23 7 27
CYP11B2 36 17 43 31 49
GNB3 48 79 42 47 31
NOS3 26 50 50 20 50
(Source: http://www.gbhealthwatch.com/Trait-Salt-Sensitivity.php?)
ALL general population, AFR Africans, AMR Americans, ASN Asians, EUR Europeans. Data are from 1000 genome project

Table 7.8  Daily sodium and potassium intakes and recommended intakes in the U.S. (IOM 2004)
Sodium Sodium chloride (g) Potassium
AI (adequate intake): 19–50 years 1.5 g/d (65 mmol)  3.8 4.7 g/d (120 mmol)
AI: 51–70 years 1.3 g/d (55 mmol)  3.3 4.7 g/d (120 mmol)
AI: >71 years 1.2 g/d (50 mmol)    3 4.7 g/d (120 mmol)
UL: tolerable upper intake level 2.3 g/d (95 mmol)  5.8 Not established
Median intake (males)   4.2 g (183 mmol) 10.6 2.9–3.2 g/d (74–82 mmol)
Median intake (females)   3.3 g (142 mmol)  8.3 2.1–2.3 g/d (54–59 mmol)
AI (adequate intake): 19–50 years 1.5 g/d (65 mmol)  3.8 4.7 g/d (120 mmol)
(Source: IOM 2004)

foods “taste” better. Consumers who are accus- sodium in the saliva. The results correlated l with
tomed to higher levels of salt in their foods find the sensory perceived saltiness, where the small-
foods without salt unpalatable. Reductions in lev- est crystal size fraction resulted in the fastest
els of salt in their food therefore must be gradual. salty perception, highest maximum saltiness
In order to lower salt consumption in the popula- intensity and maximum total saltiness. The dif-
tion as a whole, it will be necessary to reduce salt ferent delivery rates can be explained by differen-
levels in the human food supply with careful tial dissolution kinetics and enhanced mass
attention to their flavor-enhancing properties transfer of sodium into the saliva. The sodium
(Liem et al. 2011). concentration in the saliva from the various crys-
Rama et al. (2013) demonstrated that salt tal sizes salt are shown in Fig. 7.13.
crystal size impacted upon the rate of initial The results demonstrate that when salt is
response and perceived saltiness. They studied placed on the surface of foods the total salt added
three different sizes of salt crystals on potato can be reduced by using smaller crystal size salt.
crisps to measure the rate of solubilisation of the Salt substitutes offer alternatives to enhance fla-
salt crystals. A single sample of salt was ground vor while reducing sodium content of the food.
in a mortar and pestle and mechanically sieved to Frequently low-sodium products have been
produce three sizes of salt particles: S1 ­formulated with a blend of sodium and potassium
(<106 μm), S2 (106–425 μm), S3 (425–710 μm). chlorides, but potassium causes bitter and metal-
The smallest crystal size salt dissolved and dif- lic tastes. Many methods have been developed to
fused throughout the mouth to the tongue saliva improve foods made with low or reduced sodium.
faster than the medium and the larger crystals Some food manufacturers have reduced
ones; the smallest crystal size delivered the high- sodium in foods like salty snacks. AkzoNobel
est maximum concentration and greatest total (www.akzonobel.com/saltspecialties) developed
298 7 Flavor

Fig. 7.13  Salivary sodium concentration after chewing and false chew (n = 8). Error bars indicate standard devia-
crisps with differing salt crystal fractions, S1 (<106 μm), tion. (Adapted from: Rama et al. 2013)
S2 (106–425 μm), and S3 (425–710 μm), Blank (no salt)

its OneGrain technology to combine regular salt, ride and potassium chloride with 25% less
a salt replacer, and taste-enhancing flavors in sin- sodium than regular salt (Nachay 2013).
gle salt grains to achieve up to 50% sodium reduc- Tate & Lyle (www.tateandlyle.com) offers
tion. Suprasel Loso OneGrain, produced using SODA-LO™, which is manufactured using pro-
the technology, can provide a one-for-one replace- prietary technology that turns salt crystals into
ment for regular salt, and the company says that free-flowing crystalline microspheres. The bene-
the ingredient is a genuine replacement for salt in fit of this ingredient, according to the company, is
terms of taste and functionality (Nachay 2013). that the smaller crystals optimize saltiness per-
Reducing sodium in baked goods is challeng- ception in foods by maximizing the surface area
ing because of the important roles that sodium relative to volume, allowing for an up to 50%
plays. Innophos (www.innophos.com) offers cal- reduction in sodium in some applications. The
cium phosphates and sodium aluminum phos- company also emphasizes that since the ingredi-
phates that can be used to reduce sodium in ent is made from salt, it does not impart any off-
chemically leavened bakery products. These tastes. It functions well in breads, breadings, and
ingredients like Cal-Rise® calcium acid pyro- coatings, and salty snacks (Nachay 2013).
phosphate, Regent 12XX® monocalcium phos-
phate, monohydrate, Levair® sodium aluminum
phosphate, and more replace some or all of the Bitter Taste
traditional leavening agents in a variety of baked
goods applications. Each ingredient has its own Bitter taste is characteristic of many foods and
benefits, some of which are improved texture, can be attributed to a great variety of inorganic
resilient crumb structure, and better stability and organic compounds. Many substances of
(Nachay 2013). plant origin are bitter. Although bitter taste by
Morton Salt (www.mortonsalt.com) offers itself is usually considered to be unpleasant, it is a
Morton® LiteSalt™ Mixture, a blend of sodium component of the taste of many foods, usually
chloride and potassium chloride that contains those foods that are sweet or sour. Inorganic salts
50% less sodium than regular salt, and Morton can have a bitter taste (Table 7.5). Some amino
Salt Balance® Salt Blend, a blend of sodium chlo- acids may be bitter (Table 7.2). Bitter peptides
Bitter Taste 299

Table 7.9  Taste of some selected peptides


Taste Composition of peptides
Flat L-Lys-L-Glu, L-PhE-L-Phe, Gly-Gly-Gly-Gly
Sour L-Ala-L-Asp, γ-L-Glu-L-Glu,
Gly- L-Asp-L-Ser-Gly
Bitter L-Leu-L-Leu,
L-Arg-L-Pro, L-Val- L-Val-L-Val
Sweet L-Asp-L-Phe-OMe, L-Asp-L- Met-OMe Fig. 7.14  Structure of quinine. This has an intensely bit-
Biting γ-L-Glutamyl-S-(prop-1-enyl)-L-cysteln ter taste
Source: From J. Solms, Nonvolatile Compounds and the
Flavor of Foods, in Gustation and Olfaction, G. Ohloff
and A. F. Thomas, eds., 1971, Academic Press

may be formed during the partial enzymic hydro-


lysis of proteins—for example, during the ripen-
ing of cheese. Solms (1969) has given a list of
peptides with different taste sensations (Table 7.9).
Bitter taste is an important evolutionary sys- Fig. 7.15  Caffeine and theobromine
tem that helps prevents mammal from ingesting
food containing bitter-tasting toxins, which response to bitter-tasting compounds (Bartoshuk
include a wide range of structurally diverse mol- et al. 1994). There are several members of the
ecules. Bitter taste mediated by a family of hepta- TAS2R receptor gene family that encode taste
helical G protein-coupled receptors, called taste 2 receptors on the tongue, and genetic polymor-
receptors or TAS2Rs or T2Rs. The ability of phisms of TAS2R38 have been associated with
TAS2Rs to recognize a broad range of bitter com- differences in the perception of PTC and PROP
pounds provides us with the ability to detect the (El-Sohemy et al. 2007; Hayes and Keast 2011).
wide range of bitter substances in foods and bev- Alkaloids and glycosides are the most common
erages. Individual TAS2Rs possess only one bitter compounds in foods. Alkaloids are basic
binding site, in which they accommodate their nitrogen-containing organic compounds that are
ligands by contacting different but overlapping derived from pyridine, pyrrolidine, quinoline, iso-
sets of amino acids on the protein in the trans- quinoline, or purine. Quinine is often used as a
membrane portion of the cell. There is a large standard for testing bitterness (Fig. 7.14). The bit-
genetic variability in TAS2Rss in humans includ- terness of quinine hydrochloride is detectable in a
ing single nucleotide polymorphisms, variations solution as dilute as 0.00004 M, or 0.0016%. If
in copy numbers and receptor functionally which 5-mL of this solution is tasted, the amount of sub-
cause variability in the sensitivity to the bitterness stance a person detects would be 0.08 mg
of specific compounds (Meyerhof et al. 2011). (Moncrieff 1951). Our sensitivity to bitterness is
Food preferences are influenced by many more extreme than our sensitivity to other tastes;
factors including personal experiences, cultural
­ the order of sensitivity is from bitter to sour to
adaptations and perceived health benefits. Taste is salty and our least sensitivity is to sweet taste.
the most important determinant effecting whether Threshold values reported by Moncrieff are as fol-
a food is liked or disliked. Based on the response lows: sour—0.007% HC1; salt—0.25% NaCl;
to bitter-tasting compounds, such as phenylthio- and sweet—0.5% sucrose. If the artificial sweet-
carbamide (PTC) or 6-n-­propylthiouracil (PROP), eners such as saccharine are considered, the sweet
individuals can be classified as supertasters, tast- sensitivity is second to bitter. Quinine is used as a
ers, or nontasters. Genetic differences in bitter component of some soft drinks to produce bitter-
taste perception may account for many individual ness. Other alkaloids occurring as natural bitter
differences in food preferences. Other factors such constituents of foods are caffeine and theobro-
as age, sex and ethnicity may also modify the mine (Fig. 7.15), which are derivatives of purine.
300 7 Flavor

Fig. 7.16  Naringin, hesperidin, rutinose, 6-O-α-L-rharnnopyranosyl-D-glucopyranose

Another naturally occurring bitter substance is the and glucose is 1 → 6, the compound is tasteless as
glycoside naringin, which occurs in grapefruit and in hesperidin, where the sugar part, rutinose, is
some other citrus fruits. Naringin in pure form is 6-O-α-L-rhamnopyranosyl-D-glucose.
more bitter than quinine and can be detected in Bitterness occurs as a defect in dairy products
concentrations of less than 0.002%. Naringin as a result of casein proteolysis by enzymes that
(Fig.  7.16) contains the sugar moiety rutinose produce bitter peptides. Bitter peptides are pro-
(L-rhamnose-­D-glucose), which can be removed duced in cheese because of an undesirable pattern
by hydrolysis with boiling mineral acid. The aglu- of hydrolysis of milk casein (Habibi-Najafi and
cose is called naringenin, and it lacks the bitter- Lee 1996). According to Ney (1979), bitterness in
ness of naringin. Since naringin is only slightly amino acids and peptides is related to hydropho-
soluble in water (0.05% at 20 °C), it may crystal- bicity. Each amino acid has a hydrophobicity
lize out when grapefruit is subjected to below-­ value (Δf), which is defined as the free energy of
freezing temperatures. Hesperidin (Fig. 7.16) transfer of the side chains and is based on solubil-
occurs widely in citrus fruits and is also a rutinose ity properties (Table 7.10). The average hydro-
glycoside. It occurs in oranges and lemons. Dried phobicity of a peptide, Q, is obtained as the sum
orange peel may contain as much as 8% hesperi- of the Δf of component amino acids divided by
din. The aglycone of hesperidin is called hespere- the number of amino acid residues. Ney (1979)
tin. The sugar moiety is attached to carbon 7. reported that bittemess is found only in peptides
Horowitz and Gentili (1969) have studied the rela- with molecular weights below 6000 Da when
tionship between bitterness and the structure of their Q value is greater than 1400. These findings
7-­rhamnoglycosides of citrus fruits; they found indicate the importance of molecular weight and
that the structure of the disaccharide moiety plays hydrophobicity. In a more detailed study of the
an important role in bitterness. The point of attach- composition of bitter peptides, Kanehisa (1984)
ment of rhamnose to glucose determines whether reported that at least six amino acids are required
the substance will be bitter or tasteless. Thus, neo- for strong bitterness. A bitter peptide requires the
hesperidin contains the disaccharide neohespori- presence of a basic amino acid at the N-terminal
dose, which contains rhamnose linked 1 → 2 to position and a hydrophobic one at the C-terminal
glucose; therefore, the sugar moiety is 2-O-α-L- position. It appears that at least two hydrophobic
rhamnopyranosyl-­D-glucose. Glycosides contain- amino acids are required in the C-terminal area of
ing this sugar, including neohesperidin, have a the peptide to produce intense bitterness. The
bitter taste. When the linkage between rhamnose high hydrophobicity of leucine and the number of
Other Aspects of Taste 301

leucine and possibly proline residues in the pep- flavor modifiers and/or stabilizers. The polyphe-
tide probably play a role in the bitterness. nols can also bind with protein contributing to
Flavan-3-ols and their condensation products haze in beer (Aron and Shellhammer 2010).
are the most common flavonoids consumed in the The antioxidant capacity of polyphenols
American diet. The flavan-3-ols and their poly- from green tea, grape juice, and chocolate, as
meric condensation products, the proanthocyani- well as a wide range of fresh fruits and vegeta-
dins, are regarded as functional ingredients in bles is well established. Fresh produce is a good
various beverages, whole and processed foods, source of polyphenols which influence the
herbal remedies and supplements. They are pres- sensory and nutritional qualities of produce.
­
ence in food influence several taste parameters The astringency and bitterness of foods and
including astringency, bitterness, sourness, ­beverages are largely due to their polyphenolic
sweetness, salivary viscosity, aroma, and color content. Considerable variation is found in mea-
formation. Some foods contain only monomeric suring the polyphenolic content of produce. The
flavan-3-ols [(−)-epicatechin predominates] and polyphenolic content of produce seems to be
dimeric proanthocyanidins, most foods contain primarily influenced by genetics, but numerous
oligomers of d.p. values ranging from 1 to 10 or other factors including degree of ripeness, cli-
greater than 10. Flavan-3-ols have been reported mate, storage and processing can also influence
to exhibit several health beneficial effects by act- phenolic content (Hughes 2010).
ing as antioxidant, anticarcinogen, cardiopreven-
tive, antimicrobial, anti-viral, and neuro-protective
agents Aron and Kennedy (2008). Other Aspects of Taste
Beer flavonoids such as the flavan-3-ols and
their condensed products, the proanthocyanidins The basic sensations—sweet, sour, salty, and bit-
are easily oxidized. As a result they are capable of ter—account for the major part of the taste
hindering or preventing the oxididation of other response. However, it is generally agreed that
components present in beer. Flavan-3-ols and pro- these basic tastes alone cannot completely
anthocyanidin improve oxidative stability in food describe taste. In addition to the four individual
systems, and thus theyalso can function as beer tastes, there are important interrelationships
among them. One of the most important in foods
is the interrelationship between sweet and sour.
Table 7.10 Hydrophobicity values (Δf) of the side
chains of amino acids
The sugar-acid ratio plays an important part in
many foods, especially fruits. Kushman and
Amino acid Abbreviation Δf (cal/mol)
Ballinger (1968) have demonstrated the change
Glycine Gly 0
in sugar-acid ratio in ripening blueberries
Serine Ser 40
(Table 7.11). Sugar-acid ratios play an important
Threonine Thr 440
role in the flavor quality of fruit juices and wines
Histidine His 500
Aspartic acid Asp 540
Glutamic acid Glu 550
Table 7.11  Change in sugar-acid ratio during ripening of
Arginine Arg 730
blueberriesa
Alanine Ala 730
Methionine Met 1300 Unripe Ripe Overripe
Lysine Lys 1500 Total sugar (%) 5.8 7.9 12.4
Valine Val 1690 pH 2.83 3.91 3.76
Leucine Leu 2420 Titratable acidity 23.9 12.9 7.5
Proline Pro 2620 Sugar-acid ratio 3.8 9.5 25.8
Phenylalanine Phe 2650 Source: From L.J. Kushman and W.E. Ballinger, Acid and
Tyrosine Tyr 2870 Sugar Changes During Ripening in Wolcott Blueberries,
Proc. Amer. Soc. Hort. Soc., Vol. 92, pp. 290–295, 1968
Isoleucine lle 2970 a
The sugars are mainly glucose and fructose, and the acid-
Tryptophan Trp 3000 ity is expressed as citric acid
302 7 Flavor

Fig. 7.17  Isomeric forms of menthol

(Ough 1963). Alkaline taste has been attributed


to the hydroxyl ion. Caustic compounds can be
detected in solutions containing only 0.01% of
the alkali. Probably the major effect of alkali is
irritation of the general nerve endings in the
mouth. Another effect that is difficult to describe
is astringency. Borax is known for its ability to
produce this effect, as are the tannins present in
Fig. 7.18  Pipeline, responsible for the hotness of pepper
foods, especially those that occur in tea. Even if
astringency is not considered a part of the taste
sense, it must still be considered a feature of food
flavor.
Another important taste sensation is coolness,
which is a characteristic of menthol. The cooling
effect of menthol is part of the mint flavor com-
plex and is exhibited by only some of the possible
Fig. 7.19  Capsaicin, the pungent principle of red pepper
isomeric forms. Only (−) and (+) menthol show
the cooling effect, the former to a higher degree
than the latter, but the isomers isomenthol, neom- can be measured by an organoleptic threshold
enthol, and neoisomenthol do not give a cooling method (Rogers 1966) and expressed in heat
effect (Fig. 7.17) (Kulka 1967). Hotness is a units. The pungent principle of capsicum is cap-
property associated with spices and is also saicin. The structure of capsaicin is given in
referred to as pungency. The compound primarily Fig. 7.19. Capsaicin shows similarity to the com-
responsible for the hotness of black pepper is pip- pound zingerone, the pungent principle of ginger
erine (Fig. 7.18). In red pepper or capsicum, non- (Fig. 7.20).
volatile amides are responsible for the heat effect. Govindarajan (1979) has described the rela-
The heat effect of spices and their constituents tionship between pungency and chemical struc-
Taste Inhibition and Modification 303

ture of pungent compounds. There are three in orange juice. Copper is well known for its abil-
groups of natural pungent compounds—the capsa- ity to catalyze oxidation reactions. Stark and
icinoids, piperine, and the gingerols. These have Forss (1962) have isolated and identified oct-1-­
some common structural aspects, including an en-3-one as the compound responsible for the
aromatic ring and an alkyl side chain with a car- metallic flavor in dairy products.
bonyl function (Figs. 7.19 and 7.20). Structural
variations in these compounds affect the intensity
of the pungent response. These structural ­variations Taste Inhibition and Modification
include the length of the alkyl side chain, the posi-
tion of the amide group near the polar aromatic Some substances have the ability to modify our
end, the nature of the groupings at the alkyl end, perception of taste qualities. Two such com-
and the unsaturation of the alkyl chain. pounds are gymnemagenin, which is able to sup-
The metallic taste has been described by press the ability to taste sweetness, and the
Moncrieff (1964). There are no receptor sites for protein from miracle fruit, which changes the
this taste or for the alkaline and meaty tastes. perception of sour to sweet. Both compounds are
However, according to Moncrieff, there is no obtained from tropical plants.
doubt that the metallic taste is a real one. It is The leaves of the tropical plant Gymnema syl-
observable over a wide area of the surface of the vestre, when chewed, suppress the ability to taste
tongue and mouth and, like irritation and pain, sweetness. The effect lasts for hours, and sugar
appears to be a modality of the common chemical seems like sand in the mouth. The ability to taste
sense. The metallic taste can be generated by salts other sweeteners such as saccharin is equally
of metals such as mercury and silver (which are suppressed. There is also a decrease in the ability
most potent) but normally by salts of iron, cop- to taste bitterness. The active principle of leaves
per, and tin. The threshold concentration is in the has been named gymnemic acid and has been
order of 20–30 ppm of the metal ion. In canned found (Stöcklin et al. 1967) to consist of four
foods, considerable metal uptake may occur and components, designated as gymnemic acids, A1,
the threshold could be exceeded in such cases. A2, A3, and A4. These are D-glucuronides of acet-
Moncrieff (1964) also mentions the possibility of ylated gymnemagenins. The unacetylated
metallic ion exchange between the food and the gymnemagenin is a hexahydroxy pentacyclic tri-
container. The threshold concentration of copper terpene; its structure is given in Fig. 7.21.
is increased by salt, sugar, citric acid, and alco- The berries of a West African shrub (Syn-­
hol. Tannin, on the other hand, lowers the thresh- sepalum dulcificum) contain a substance that has
old value and makes the copper taste more the ability to make sour substances taste sweet. The
noticeable. The metallic taste is frequently berry, also known as miracle fruit, has been shown
observed as an aftertaste. The lead salt of saccha-
rin gives an impression of intense sweetness, fol-
lowed by a metallic aftertaste. Interestingly, the
metallic taste is frequently associated with oxi-
dized products. Tressler and Joslyn (1954) indi-
cate that 20 ppm of copper is detectable by taste

Fig. 7.20  Zingerone, the pungent principle of ginger Fig. 7.21  Structure of gymnemagenin
304 7 Flavor

to contain a taste-modifying protein (Kurihara and Monosodium glutamate has long been recog-
Beidler 1968, 1969). The protein is a basic glyco- nized as a flavor enhancer and is now being con-
protein with a molecular weight of 44,000. It is sidered a primary taste, umami. The flavor
suggested that the protein binds to the receptor potentiation capacity of monosodium glutamate
membrane near the sweet receptor site. The low in foods is not the result of an intensifying effect
pH changes the conformation of the membrane so of the four primary tastes. Glutamate may exist in
that the sugar part of the protein fits into the sweet the L and D forms and as a racemic mixture. The
receptor site. The taste-modifying protein was L form is the naturally occurring isomer that has
found to contain 6.7% of arabinose and xylose. a flavor-enhancing property. The D form is inert.
These taste-modifying substances provide an Although glutamic acid was first isolated in 1866,
insight into the mechanism of the production of the flavor-enhancing properties of the sodium salt
taste sensations and, therefore, are a valuable tool were not discovered until 1909 by the Japanese
in the study of the interrelationship between taste chemist Kikunae Ikeda. Almost immediately,
and chemical structure. commercial production of the compound started
and total production for the year 1954 was esti-
mated at 13000,000 pounds. The product as first
Flavor Enhancement—Umami described by Ikeda was made by neutralizing a
hydrolysate of the seaweed Laminaria japonica
A number of compounds have the ability to with soda. Monosodium glutamate is now pro-
enhance or improve the flavor of foods. It has duced from wheat gluten, beet sugar waste, and
often been suggested that these compounds do soy protein and is used in the form of the pure
not have a particular taste of their own. Evidence crystallized compound. It can also be used in the
now suggests that there is a basic taste response form of protein hydrolysates derived from pro-
to amino acids, especially glutamic acid. This teins that contain 16% or more of glutamic acid.
taste is sometimes described by the word umami, Wheat gluten, casein, and soy flour are good
derived from the Japanese for deliciousness sources of glutamic acid and are used to produce
(Kawamura and Kare 1987). It is suggested that a protein hydrolysates. The glutamic acid content
primary taste has the following characteristics: of some proteins is listed in Table 7.12 (Hall
1948). The protein is hydrolyzed with hydrochlo-
• The receptor site for a primary taste chemical ric acid, and the neutralized hydrolysate is used
is different from those of other primary tastes. in liquid form or as a dry powder. Soy sauce,
• The taste quality is different from others.
• The taste cannot be reproduced by a mixture
Table 7.12  Glutamic acid content of some proteins
of chemicals of different primary tastes.
Protein source Glutamic acid (%)
From these criteria, we can deduce that the Wheat gluten 36.0
glutamic acid taste is a primary taste for the fol- Corn gluten 24.5
Zein 36.0
lowing reasons:
Peanut flour 19.5
Cottonseed flour 17.6
• The receptor for glutamic acid is different
Soybean flour 21.0
from the receptors for sweet, sour, salty, and
Casein 22.0
bitter. Rice 24.1
• Glutamic acid does not affect the taste of the Egg albumin 16.0
four primary tastes. Yeast 18.5
• The taste quality of glutamic acid is different Source: From L.A. Hall, Protein Hydrolysates as a Source
from that of the four primary tastes. of Glutamate Flavors, in Monosodium Glutamate—A
• Umami cannot be reproduced by mixing any Symposium, 1948, Quartermaster Food and Container
of the four primary tastes. Institute for the Armed Forces
Flavor Enhancement—Umami 305

which is similar to these hydrolysates, is pro- Table 7.13  Threshold levels of flavor enhancers alone
and in mixtures in aqueous solution
duced wholly or partially by enzymic hydrolysis.
This results in the formation of ammonia from Threshold level (%)
acid amides; soy sauce contains ammonium com- Disodium Disodium Monosodium
Solvent 5′-inosinate 5′-guanylate L-glutamate
plexes of amino acids, including ammonium
Water 0.012 0.0035 0.03
glutamate.
0.1% 0.0001 0.00003 –
The flavor of glutamate is difficult to describe. glutamate
It has sometimes been suggested that glutamate 0.01% – – 0.002
has a meaty or chickeny taste, but it is now gener- inosinate
ally agreed that glutamate flavor is unique and Source: From A. Kuninaka, Recent Studies of
has no similarity to meat. Pure sodium glutamate 5′-Nucleotides as New Flavor Enhancers, in Flavor
is detectable in concentrations as low as 0.03%; Chemistry, I. Hornstein, ed., 1966, American Chemical
Society
at 0.05% the taste is very strong and does not
increase at higher concentrations. The taste has
been described as a mixture of the four tastes kind of fish). However, they were not produced
(Crocker 1948). At about 2 threshold values of commercially and used as flavor enhancers until
glutamate concentration, it could be well matched recently, when technical problems in their pro-
by a solution containing 0.6 threshold of sweet, duction were solved. The general structure of the
0.7 of salty, 0.3 of sour, and 0.9 of bitter. In addi- nucleotides with flavor activity is presented in
tion, glutamate is said to cause a tingling feeling Fig. 7.22. There are three types of inosinic acid,
and a marked persistency of taste sensation. This 2′-, 3′-, and 5′-isomers; only the 5′-isomer has
feeling is present in the whole of the mouth and flavor activity. Both riboside and 5′-phospho-
provides a feeling of satisfaction or fullness. monoester linkages are required for flavor activ-
Apparently glutamate stimulates our tactile sense ity, which is also the case for the OH group at the
as well as our taste receptors. The presence of salt 6-position of the ring. Replacing the OH group
is required to produce the glutamate effect. with other groups, such as an amino group,
Glutamate taste is most effective in the pH range sharply reduces flavor activity but this is not true
of 6–8 and decreases at lower pH values. Sugar for the group at the 2-position. Hydrogen at the
content also affects glutamate taste. The taste in a 2-position corresponds with inosinate and an
complex food, therefore, depends on a complex amino group with guanylate; both have compa-
interaction of sweet, sour, and salty, as well as the rable flavor activity, and the effect of the two
added glutamate. compounds is additive.
Monosodium glutamate improves the flavor of The synergistic effect of umami substances is
many food products and is therefore widely used exceptional. The subjective taste intensity of a
in processed foods. Products benefiting from the blend of monosodium glutamate and disodium
addition of glutamate include meat and poultry, 5′-inosinate was found to be 16 times stronger
soups, vegetables, and seafood. than that of the glutamate by itself at the same
For many years glutamate was the only known total concentration (Yamaguchi 1979).
flavor enhancer, but recently a number of com- 5′-nucleotides can be produced by degradation
pounds that act similarly have been discovered. of ribonucleic acid. The problem is that most
The 5′-nucleotides, especially 5′-inosinate and enzymes split the molecule at the 3′-phosphodies-
5′-guanylate, have enhancement properties and ter linkages, resulting in nucleotides without flavor
also show a synergistic effect in the presence of activity. Suitable enzymes were found in strains of
glutamate. This synergistic effect has been dem- Penicillium and Streptomyces. With the aid of
onstrated by determining the threshold levels of these enzymes, the 5′-nucleotides can be manufac-
the compounds alone and in mixtures. The data tured industrially from yeast ribonucleic acid.
in Table 7.13 are quoted from Kuninaka (1966). Another process produces the nucleoside inosine
The 5′-nucleotides were discovered many years by fermentation, followed by chemical phosphor-
ago in Japan as components of dried bonito (a ylation to 5′-inosinic acid (Kuninaka 1966).
306 7 Flavor

Fig. 7.22  Structure of nucleotides with flavor activity

Maltol has antioxidant properties. It has been


found to prolong storage life of coffee and roasted
cereal products. Maltol is used as a flavor
enhancer in chocolate and candies, ice cream,
baked products, instant coffee and tea, liqueurs,
and flavorings. It is used in concentrations of
50–250 ppm and is commercially produced by a
Fig. 7.23  Tricholomic and ibotenic acid
fermentation process.

The search for other flavor enhancers has


brought to light two new amino acids, tricholo- Odor
mic acid and ibotenic acid, obtained from fungi
(Fig. 7.23). These amino acids have flavor activi- The olfactory mechanism is both more complex
ties similar to that of monosodium glutamate. and more sensitive than the process of gustation.
Apparently, the flavor enhancers can be divided Olfaction, the sense of smell is a primal sense for
into two groups; the first consists of 5′-inosinate all humans and animals. Olfaction helps both
and 5′-guanylate with the same kind of activity vertebrates and other organisms with olfactory
and an additive relationship. The other group receptors to identify food, mates, and predators.
consists of glutamate, tricholomic, and ibotenic It also can provide sensual pleasure (the odor of
acid, which are additive in action. Between the flowers and perfume) or threats from spoiled
members of the two groups, the activity is food or chemical dangers. Thus, olfaction is one
synergistic. of our principal means to communicate with our
A different type of flavor enhancer is maltol, environment. There are thousands of odors, and
which has the ability to enhance sweetness pro- the sensitivity of the smell organ is about 10,000
duced by sugars. Maltol is formed during roasting times greater than that of the taste organ.
of malt, coffee, cacao, and grains. During the bak- Odorants are volatile chemical compounds
ing process, maltol is formed in the crust of bread. that are carried by inhaled air to the Regio olfac-
It is also found in many dairy products that have toria (olfactory region) which are located in the
been heated, as a product of decomposition of the roof of the two nasal cavities of the human nose
casein-lactose system. Maltol (Fig. 7.24) is (Fig.  7.25). The molecular properties of an
formed from di-, tri-, and tetrasaccharides includ- odorant determine the sensory properties of
­
ing isomaltose, maltotretraose, and panose but not the compound. The odorant must be at least par-
from maltotriose. Formation of maltol is brought tially water solubility, have a sufficiently high
about by high temperatures and is catalyzed by vapor pressure, low polarity, some ability to dis-
metals such as iron, nickel, and calcium. solve in fat (lipophilicity), and surface activity
Odor 307

Fig. 7.24 Some
furanones (1,2,3),
isomaltol (4), and maltol
(5)

Fig. 7.25  Structure of Olfactory Nerve Olfactory Bulb


olifactory cells and Mitral Cell
olifactory bulb Olfactory Tract
(Leffingwell 2001). The
diagrams of the
“Olfactory Region” and
the “Olfactory Bulb” are
adapted from a drawing Glomeruli
that appears at http:// Olfactory Nerve filaments
www.sciencenet.org.uk/ Cribriform (bone)
database/Social/Senses/
s00122b.html Axons

Olfactory Receptor Neurons Olfactory


Epithelium
Cilia in Mucosa Mucosa

Air and
Odorant Molecules

(Leffingwell 2001; Greenman and Benkara soluble in the mucous can interact with the olfac-
Mostefa Saad 2009). tory receptors and produce the signals that our
The olfactory sense is able to distinguish brain interprets as odor. Each olfactory receptor
among numerous chemical compounds at very neuron has 8–20 cilia that are whip-like exten-
low concentrations (Ohloff 1994). The olfactory sions 30–200 μ in length. The olfactory cilia are
region in the two nasal passages of humans is a the sites where molecular reception with the
small area (about 2.5 cm2) containing approxi- odorant occurs and sensory transduction (i.e.,
mately 50 million primary sensory receptor cells. transmission) starts (Leffingwell 2001).
The olfactory region consists of cilia projecting The base olfactory epithelium consists
out of the olfactory epithelium into a layer of partially of basal cells located in the lowest cel-
mucous which is about 60 μ thick. This mucous lular layer of the olfactory epithelium just above
layer is a lipid-rich secretion that bathes the sur- the mucous layer. These basal cells undergo
face of the receptors at the epithelium surface. mitotic cell division to form olfactory receptor
The mucous layer is produced by the Bowman’s neurons. Olfactory receptor neurons turnover
glands which reside in the olfactory epithelium. approximately every 40 days. The olfactory
­
The mucous lipids assist in transporting the odor- receptor neurons extend through the epithelium
ant molecules as only volatile materials that are to contact odorants in the atmosphere. Within the
308 7 Flavor

epithelium, the neuronal cells form axons that are Most odorous compounds are soluble in a
bundled in groups of 10–100 and these bundles variety of solvents, but it appears that solubility is
penetrate the bone, ending in the olfactory bulb less important than type of molecular arrange-
of the brain. In the olfactory bulb of the brain, the ment, which confers both solubility and chemical
neurons converge and terminate with post-­ reactivity (Moncrieff 1951). Some degree of
synaptic cells to form synaptic structures called water solubility does appear to contribute.
glomeruli. The glomeruli are connected in groups There are two common ways to establish rela-
that converge into mitral cells. In rabbits, there tionships between a sample odor and a chemical.
are 26,000 receptor neurons that converge onto One is to find statistical associations between the
200 glomeruli, which then converge at 25:1 onto data obtained from sensory analysis and GC-MS
each mitral cell. The total convergence is esti- analysis for the specific sample or volatile com-
mated to be about 1000:1. From the mitral cells, pounds. The other is to use trained panelists sniff-
the message is sent directly to the higher levels of ing at GC-MS ports to detect and identify volatile
the central nervous system in the corticomedial compounds that are also identified by sensors and
amygdala portion of the brain (via the olfactory computer programs (GC-MS Olfactormetry).
nerve tract) where the signaling process is Moreover, direct relationships to identify an odor
decoded and olfactory interpretation and response compound can be done by comparing the sample
occurs (Leffingwell 2001). odor to a number of volatile compounds that may
The olfactory epithelium contains another have a similar odor description, which may
sensory system in the form of “trigeminal nerve” require knowledges of odorous volatile com-
receptors. Some chemicals are trigeminal stimu- pounds. Instrumental measurements are useful
lants that produce sensations described as hot, when detecting and identifying specific com-
cold, tingling or irritating. For example, leavo-­ pounds. GC-MS may be used in combination
menthol or (−)-menthol produces the trigeminal with the sensory aromatic profile analysis to
feeling of cold at moderate concentrations and identify the volatile compounds present in the
“hot” at high concentrations in the nasal cavity. sample. More specifically, GC-MS sniff ports
Similarly, camphor which possesses markedly may be used to identify volatile compounds that
more aroma than menthol, also produces the have an odor detectable by the human nose. In
“cold” sensation via interaction with trigeminal this way, compounds identified by GC-MS can
receptors. Other commonly encountered trigemi- be related to the sensory aromatic results.
nal stimulants include the chemicals allyl Drawing correlations is frequently complicated
­isothiocyanate (mustard, mustard oil), capsiacin since perceived intensity and aroma concentra-
(hot chile powder, mace spray) and Diallyl sul- tion are rarely linear. Frequently the detection or
fide (onion) (Leffingwell 2001). See Fig. 7.26. recognition threasholds are rapidly reached and

Fig. 7.26 Common
food compounds that are
trigeminal stimulants
Odor 309

Fig. 7.27 Schematic
diagram of potential
relationships between
chemical concentration
and sensory intensity Below Threshold
(from Chambers and

Sensory intensity
Koppel 2013)
Detection Threshold
Terminal Threshold
Recognition Threshold

Assumed Linear Correlation

Chemical concentration

at higher levels the perceived intensity levels off Table 7.14  Odor threshold concentrations of odorous
reaching a terminal threshold as shown in substances perceived during normal inspiration
Fig. 7.27 (Chambers and Koppel 2013). Threshold concentration
The number of volatile compounds occurring Compound (molecules/cc)
in foods is very high. Maarse (1991) has given Allyl mercaptan 6 × 107
the following numbers for some foods: beef Sec. butyl mercaptan 1 × 108
(boiled, cooked)—486; beer—562; butter—257; Isopropyl mercaptan 1 × 108
coffee—790; grape—466; orange—203; tea— Isobutyl mercaptan 4 × 108
Tert. butyl mercaptan 6 × 108
541; tomato—387; and wine (white)—644. Not
Thiophenol 8 × 108
all of these substances may be essential in deter-
Ethyl mercaptan 1 × 109
mining the odor of a product. Usually, the relative
1,3-Xylen-4-ol 2 × 1012
amounts of a limited number of these volatile
μ-Xylene 2 × 1012
compounds are important in establishing the
Acetone 6 × 1013
characteristic odor and flavor of a food product.
Source: From K.B. Döving, Problems in the Physiology
The sensitivity of the human olfactory organ is of Olfaction, in Symposium on Foods: The Chemistry and
inferior to that of many animals. Dogs and rats Physiology of Flavors, H.W. Schultz et al., eds., 1967,
can detect odorous compounds at threshold con- AVI Publishing
centrations 100 times lower than man. When air
is breathed in, only a small part of it is likely to
flow over the olfactory epithelium in the upper so that more odors can reach the olfactory tissue.
nasal cavity. When a smell is perceived, sniffing What is remarkable about the olfactory
may increase the amount reaching the olfactory ­mechanism is not only that thousands of odors
tissue. When foods are eaten, the passage of can be recognized, but that it is possible to store
breath during exhalation reaches the nasal cavity the information in the brain for retrieval after
from the back. Döving (1967) has quoted the long periods of time. The ability to smell is
threshold concentrations of odorous substances affected by several conditions, such as colds,
listed in Table 7.14. Apparently, it is possible to menstrual cycle, and drugs such as penicillin.
change odor thresholds by a factor of 100 or more Odors are usually the result of the presence of
by stimulating the sympathetic nervous system mixtures of several, sometimes many, different
310 7 Flavor

odorous compounds. The combined effect cre- The musks are a common illustration of com-
ates an impression that may be very different pounds with different structures that all give simi-
from that of the individual components. Many lar odors. These may include tricyclic compounds,
food flavors, natural as well as artificial, are of macrocyclic ketones and lactones, steroids, nitro-
this compound nature. cyclohexanes, indanes, tetrahydronaphthalenes,
and acetophenoses. Small changes in the struc-
ture of these molecules may significantly change
Odor and Molecular Structure in potency but will not affect quality, since all are
musky. There are also some compounds that have
M. Stoll wrote in 1957: “The whole subject of the similar structures and very different odors, such
relation between molecular structure and odor is as nootkatone and related compounds (Teranishi
very perplexing, as there is no doubt that there 1971). Nootkatone is a flavor compound from
exist as many relationships of structure and odor grapefruit oil. This compound and
as there are structures of odorous substances.” In 1,10-­dihydronootkatone have a grapefruity flavor
1971 (referring to Stoll 1957), Teranishi wrote: (Fig. 7.28). Several other related compounds have
“The relation between molecular structure and a woody flavor. The odor character of stereoiso-
odor was perplexing then. It is now.” We can mers may be quite different. The case of menthol
observe a number of similarities between the has already been described. Only menthol iso-
chemical structure of compounds and their odors. mers have peppermint aroma. The iso-, neo-, and
However, the field of food flavors, as is the field neoisomenthols have an unpleasant musty flavor.
of perfumery, is still very much an art, albeit one Naves (1957) describes the difference between
greatly supported by scientists’ advancing ability the cis- and trans- forms of 3-hexenol (CH2OH–
to classify structures and identify the effect of CH2–CH = CH–CH2CH3). The cis-isomer has a
certain molecular configurations. The odor fresh green odor, whereas the trans-isomer has a
potency of various compounds ranges widely. scent reminiscent of chrysanthemum. The
Table 7.15 indicates a range of about eight orders 2-trans-6-­cis nonadienal smells of cucumber and
of magnitude (Teranishi 1971). This indicates is quite different from the smell of the 2-trans-6-­
that volatile flavor compounds may be present in trans isomer (nonadienal, CHO–CH = CH–
greatly differing quantities, from traces to rela- (CH2)2–CH = CH–CH2-CH3). Lengthening of the
tively large amounts. carbon chain may affect odorous properties. The
odor of saturated acids changes remarkably as
Table 7.15  Odor thresholds of compounds covering a chain length increases. The lower fatty acids,
wide range of intensity especially butyric, have very intense and unpleas-
Threshold ant flavors, because an increased chain length
Odorant (μg/L of water) changes flavor character (Table 7.16) and lessens
Ethanol 100,000 intensity. The fatty acids with 16 or 18 carbon
Butyric acid 240 atoms have only a faint flavor.
Nootkatone 170 Another example is given by Kulka (1967).
Humulene 160
Gamma-nonalactone has a strong coconut-like
Myrcene 15
flavor; γ-undecalactone has a peach aroma. As
n-Amyl acetate 5
the chain length is increased by one more carbon
n-Decanal 0.0
atom, the flavor character becomes peach-musk.
α- and β-Sinensal 0.05
The lactones are compounds of widely differing
Methyl mercaptan 0.02
β-Ionone 0.007
structure and odor quality and are found as com-
2-methoxy-3- isobutylpyra- zine 0.002 ponents of many food flavors. Gamma- and
Source: From R. Teranishi, Odor and Molecular Structure,
δ-lactones with 10–16 carbon atoms have been
in Gustation and Olfaction, G. Ohloff and A.F. Thomas, reported (Jurriens and Oelej 1965) as flavor com-
eds., 1971, Academic Press ponents of butter, contributing to the butter flavor
Odor and Molecular Structure 311

Fig. 7.28  Odor character of nootkatone and related compounds

Table 7.16  Flavor character of some N-carboxylic acids of phthalides (phthalides are lactones of phthalic
Acid Flavor character acid, lactones are internal esters of hydroxy
Formic Acid, pungent acids). These include the following:
Acetic Acid, vinegary, pungent
Propionic Acid, pungent, rancid, cheesy • 3-isobutyliden-3a,4-dihydrophthalide (Fig. 7.30).
Butyric Acid, rancid • 3-isovalidene-3a,4-dihydrophthalide.
Hexanoic Sweaty, goaty • 3-isobutylidene phthalide.
Octanoic Rancid • 3-isovalidene phthalide.
Decanoic Waxy
Lauric Tallowy These compounds exhibit celery-like odors at
Myristic Soapy, cardboard
levels of 0.1 ppm in water. Pyrazines have been
Palmitic Soapy
identified as the compounds giving the character-
istic intense odor of green peppers (Seifert et al.
in concentrations of only parts per million. The 1970). A number of pyrazine derivatives were
flavor character and chemical structure of some tested and, within this single class of compounds,
γ-lactones as reported by Teranishi (1971) are odor potencies showed a range of eight orders of
shown in Fig. 7.29. One of these, the γ-lactone magnitude equal to that of the widely varying
with a total chain length of 10 carbons, has peach compounds listed in Table 7.15. The compounds
flavor. The α-hydroxy-β-methyl-γ-carboxy-Δα–β-­ examined by Seifert et al. (1970) are listed in
γ-­hex-eno-lactone occurs in protein hydrolysate Table  7.17. 2-methoxy-3-isobutylpyrazine
and has very strong odor and flavor of beef bouil- appears to be the compound responsible for the
lon. Gold and Wilson (1963) found that the vola- green pepper odor. Removal of the methoxy- or
tile flavor compounds of celery contain a number alkyl-groups reduces the odor potency by 105–
312 7 Flavor

Fig. 7.29 Flavor
character of some
lactones. Source: From
R. Teranishi, Odor and
Molecular Structure, in
Gustation and Olfaction,
G. Ohloff and
A.F. Thomas, eds., 1971,
Academic Press

Fig. 7.30  Phthalides of


celery volatiles

106 times, as is the case with 2-methoxypyrazine, mass spectrometry. The components are methyl
2-iosbutylpyrazine, and 2,5-dimethylpyrazine. pyrazine; 2,3-dimethylpyrazine; 2-ethyl-­
5-­
Thus, small changes in molecular structure may methyl-pyrazine; trimethylpyrazine;
greatly affect flavor potency. The odors of isobu- 2,5-dimethyl-3-ethylpyrazine; 2,6-dimethyl-­3-­
tyl, propyl, and hexyl methoxypyrazines are sim- ethylpyrazine; and tetramethylpyrazine. Other
ilar to that of green peppers. The isopropyl researchers (Flament et al. 1967; Marion et al.
compound is moderately similar to peppers and 1967) have isolated these and other pyrazines
its odor is somewhat similar to raw potato. The from the aroma components of cocoa. Pyrazines
ethyl compound is even more similar to raw are also aroma constituents of coffee. Goldman
potato and less to pepper. In fact, this compound et al. (1967) isolated and identified 24 pyrazines
can be isolated from potatoes. The methyl com- and pyridines and revealed the presence of pos-
pound has an odor like roasted peanuts. The sibly 10 more. Bondarovich et al. (1967) isolated
structure of some of the pyrazines is shown in and identified a large number of pyrazines from
Fig. 7.31. Pyrazines have been identified as flavor coffee aroma and drew attention to the impor-
components in a number of foods that are nor- tance of pyrazines and dihydropyrazines to the
mally heated during processing. Rizzi (1967) flavor of roasted or otherwise cooked foods.
demonstrated the presence of seven alkyl-­ These authors also drew attention to the instabil-
substituted pyrazines in chocolate aroma. These ity of the dihydropyrazines. This instability not
were isolated by steam distillation, separated by only makes their detection and isolation difficult,
gas-liquid chromatography, and identified by but may help explain why flavors such as that of
Odor and Molecular Structure 313

Fig. 7.31 (a) Pyrazine,


(b) 2-methoxypyrazine,
and (c) 2-methoxy-
3-hexylpyrazine

Table 7.17  Odor threshold of pyrazine and derivatives Table 7.18  Primary odors for humans and compounds
eliciting these odors
Odor threshold (parts
Compound per 1012 parts of water) Primary odor Odor compounds
2-methoxy-3-hexylpyrazine 1 Camphoraceous Borneol, tert-butyl alcohol
2-methoxy-3-­isobutylpyrazine 2 d-camphor, cineol, pentamethyl ethyl
2-methoxy-3-­propylpyrazine 6 alcohol
2-methoxy-3-­ 2 Pungent Allyl alcohol, cyanogen,
isopropylpyrazine formaldehyde, formic acid,
methylisothiocyanate
2-methoxy-3-ethylpyrazine 400
Ethereal Acetylene, carbon tetrachloride,
2-methoxy-3-­methylpyrazine 4000
chloroform, ethylene dichloride,
2-methoxypyrazine 700,000 propyl alcohol
2-isobutylpyrazine 400,000 Floral Benzyl acetate, geraniol, α-ionone,
2–5-dimethylpyrazine 1,800,000 phenylethyl alcohol, terpineol
Pyrazine 175,000,000 Pepperminty tert-butylcarbinol, cyclohexanone,
Source: From R.M. Seifert et al., Synthesis of Some menthone, piperitol,
2-Methoxy-3-Alkylpyrazines with Strong Bell Pepper– 1,1,3-trimethyl-cyclo-5-hexanone
Like Odors, J. Agric. Food Chem., Vol. 18, pp. 246–249, Musky Androstan-3α-ol (strong),
1970, American Chemical Society cyclohexadecanone, ethylene
cebacate, 17- methylandrostan-­3α-­ol,
pentadecanolactone
Putrid Amylmercaptan, cadaverine,
roasted coffee rapidly change with time. Another hydrogen sulfide, indole (when
roasted product from which pyrazines have been concentrated, floral when dilute),
skatole
isolated is peanuts. Mason et al. (1966) found
Source: From J.E. Amoore et al., The Stereochemical
methylpyrazine; 2,5-dimethylpyrazine; trimeth-
Theory of Odor, Sci. Am., Vol. 210, No. 2, pp. 42–49, 1964
ylpyrazine; methylethylpyrazine; and dimethyle-
thylpyrazine in the flavor of roasted peanuts. The
pyrazines appear to be present in unprocessed as Amoore (Amoore et al. 1964; Amoore 1967)
well as in heated foods. The thresholds of some compared the various odor qualities that have
pyrazines in water are found in Table 7.17. been used to characterize odors and concluded
Another group of compounds that have been that seven primary odors would suffice to cover
related to the aroma of heated foods is the fura- them all: camphoraceous, pungent, ethereal, flo-
nones. Teranishi (1971) summarized the findings ral, pepperminty, musky, and putrid. Table 7.18
on several of the furanones (see Fig. 7.24). The lists some of the chemical compounds that can be
4-hydroxy-2, 5-dimethyl-3-dihydrofuranone (1) used to demonstrate these primary odors. The
has a caramel or burnt pineapple odor. The theory is based on the assumption that all odor-
4-hydroxy-5-methyl-3-dihydrofuranone (2) has a ous compounds have a distinctive molecular
roasted chicory root odor. Both compounds may shape and size that fit into a socket on the recep-
contribute to beef broth flavor. The 2,5-dimethyl-­ tor site. This would be similar to the “lock-and-­
3-dihydrofuranone (3) has the odor of freshly key” concept of enzyme action.
baked bread. Isomaltol (4) and maltol (5) are The suggestion that odorous character is
products of the caramelization and pyrolysis of related to vibrational specificity of odor mole-
carbohydrates. cules has led to the vibrational theory of olfaction
314 7 Flavor

(Wright 1957). Vibrational energy levels can to use pattern recognition techniques to further
be derived from the infrared or Raman spectra. describe the flavor. The pattern recognition method
The spectral area of greatest interest is that below involves the application of computer analysis of
700 cm−1, which is related to vibrations of chains complex mixtures of compounds. Computer mul-
and flexing or twisting of bonds between groups tivariate analysis has been used for the detection of
of atoms in the molecule. Wright and others have adulteration of orange juice and Spanish sherries
demonstrated that correlations exist between (Maarse et al. 1987).
spectral properties and odor quality in a number Flavors are often described by using the
of cases, but inconsistencies in other cases have human senses on the basis of widely recognized
yet to be explained. taste and smell sensations. A proposed wine
Obviously, none of the many theories of olfac- aroma description system is shown in Fig. 7.32
tion proposed so far have been entirely satisfac- (Noble et al. 1987). Such systems attempt to pro-
tory. It might be better to speak of hypotheses vide an orderly and reliable basis for comparison
rather than of theories. Most of these theories of flavor descriptions by different tasters.
deal with the explanation of odor quality and do The aroma is divided into first-, second-, and
not account for the quantitative aspects of the third-tier terms, with the first-tier terms in the
mechanism of olfaction. The classification of center. Examination of the descriptors in the
odor and the correlation of chemical structure aroma wheel shows that they can be divided into
and odor remain difficult to resolve. two types, flavors and off-flavors. Thus, it would
be useful to divide the flavor wheel into two
tables—one for flavors and one for off-flavors, as
 escription of Food and Beverage
D shown in Tables 7.19 and 7.20.
Flavors The difficulty in relating chemical composi-
tion and structure to the aroma of a food that con-
The flavor impression of a food is influenced by tains a multitude of flavor compounds is evident
compounds that affect both taste and odor. The from the work of Meyboom and Jongenotter
analysis and identification of many volatile flavor (1981). They studied the flavor of straight-chain,
compounds in a large variety of food products unsaturated aldehydes as a function of double-­
have been assisted by the development of powerful bond position and geometry. Some of their results
analytical techniques. Gas-liquid chromatography are presented in Table 7.21. Flavors of unsatu-
was widely used in the early 1950s when commer- rated aldehydes of different chain length and
cial instruments became available. Introduction of geometry may vary from bitter almond to lemon
the flame ionization detector increased sensitivity and cucumber when tasted separately.
by a factor of 100 and, together with mass spec- A method of flavor description, developed by
trometers, gave a method for rapid identification of researchers at A.D. Little Inc. (Sjöström 1972),
many components in complex mixtures. These has been named the flavor profile method. The
methods have been described by Teranishi et al. flavor profile method uses the recognition,
(1971). As a result, a great deal of information on description, and comparison of aroma and flavor
volatile flavor components has been obtained in by a trained panel of four to six people. Through
recent years for a variety of food products. The training, the panel members are made familiar
combination of gas chromatography and mass with the terminology used in describing flavor
spectrometry can provide identification and quan- qualities. In addition to describing flavor quality,
titation of flavor compounds. However, when the intensity values are assigned to each of the qual-
flavor consists of many compounds, sometimes ity aspects. The intensity scale is threshold,
several hundred, it is impossible to evaluate a fla- slight, moderate, and strong, and these are repre-
vor from this information alone. It is then possible sented by the symbols)(, 1, 2, and 3, respectively.
Description of Food and Beverage Flavors 315

Fig. 7.32  Modified wine aroma wheel for the description Terminology, Am. J. Enol. Vitic., Vol. 38, pp. 143–146,
of wine aroma. Source: From A.C. Noble et al., 1987, American Society of Enology and Viticulture
Modification of a Standardized System of Wine Aroma

With the exception of threshold value, the units a­ nalysis, called “flavor by mouth,” a specialists’
are ranges and can be more precisely defined by description of what a consumer would experi-
the use of reference standards. In the panel work, ence eating the food. Flavor analysis includes
the evaluation of aroma is conducted first such factors as taste, aroma, feeling, and after-
because odor notes can be overpowered when taste. A sample flavor profile of margarine is
the food is eaten. This is followed by flavor given in Table 7.22.
316 7 Flavor

Table 7.19  Aroma description of wine as listed in the aroma wheel, listing only the flavor contribution
First tier Second tier Third tier First tier Second tier Third tier
Floral Floral Geranium Canned/ Green beans
Violet cooked Asparagus
Rose Green olive
Orange blossom Black olive
Linalool Artichoke
Spicy Spicy Licorice anise Dried Hay/straw
Black pepper Tea
Cloves Tobacco
Fruity Citrus Grapefruit Nutty Nutty Walnut
Lemon Hazelnut
Berry Blackberry Almond
Raspberry Caramelized Caramelized Honey
Strawberry Butterscotch
Black currant Diacetyl (butter)
Tree fruit Cherry Soy sauce
Apricot Chocolate
Peach Molasses
Apple Woody Phenolic Phenolic
Tropical fruit Pineapple Vanilla
Melon Resinous Cedar
Banana Oak
Dried fruit Strawberry jam Burned Smoky
Raisin Burnt toast/charred
Prune Coffee
Fig
Other Artificial fruit Methyl anthranilate
Vegetative Fresh Stemmy
Grass, cut green
Bell pepper
Eucalyptus
Mint

Astringency in the mouth, followed by a physiological


response. The astringent reaction has been found
The sensation of astringency is considered to be to occur between salivary proteins that are rich in
related more to touch than to taste. Astringency proline (Luck et al. 1994). These proline-rich
causes a drying and puckering over the whole proteins (PRPs) have a high affinity for polyphe-
surface of the mouth and tongue. This sensation nols. The effect of the structure of PRP is two-
is caused by interaction of astringent compounds fold: (1) proline causes the protein to have an
with proteins and glycoproteins in the mouth. open and flexible structure, and (2) the proline
Astringent compounds are present in fruits and residue itself plays an important role in recogniz-
beverages derived from fruit (such as juice, wine, ing the polyphenols involved in the complex for-
and cider), in tea and cocoa, and in beverages mation. The complex formation between PRP
matured in oak casks. Astringency is caused by and polyphenol has been represented by Luck
tannins, either those present in the food or et al. (1994) in pictorial form (Fig. 7.33). The
extracted from the wood of oak barrels. The reaction is mediated by hydrophobic effects and
astringent reaction involves a bonding to proteins hydrogen bonding on protein sites close to prolyl
Flavor and Off-Flavor 317

Table 7.20  Aroma description of wine as listed in the Table 7.21  Flavor description of unsaturated aldehydes
aroma wheel, listing only the off-flavors dissolved in paraffin oil
First tier Second tier Third tier Aldehyde Flavor description
Earthy Moldy Moldy cork trans-3-hexenal Green, odor of pine tree needles
Musty (mildew) cis-3-hexenal Green beans, tomato green
Earthy Mushroom trans-2-heptenal Bitter almonds
Dusty
cis-6-heptenal Green, melon
Chemical Petroleum Diesel
trans-2-octenal Nutty
Kerosene
Plastic trans-5-octenal Cucumber
Tar cis-5-octenal Cucumber
Sulfur Wet wool, wet dog trans-2-nonenal Starch, glue
Sulfur dioxide trans-7-nonenal Melon
Burnt match
Source: From P.W. Meyboom and G.A. Jongenotter, Flavor
Cabbage
Perceptibility of Straight Chain, Unsaturated Aldehydes
Skunk
as a Function of Double Bond Position and Geometry,
Garlic
J. Am. Oil Chem. Soc., Vol. 58, pp. 680–682, 1981
Mercaptan
Hydrogen sulfide
Rubbery Table 7.22  Flavor profile of margarine
Papery Wet cardboard Aroma Flavor by mouth
Filterpad Amplitude 2 Amplitude 2½
Pungent Sulfur dioxide Sweet cream ½ Sweet cream 1½
Ethanol Oil )( Oil ½
Acetic acid
Sour ½ Salt 1½
Ethyl acetate
Vanillin sweet )( Butter mouthfeel 2
Other Fusel alcohol
Sorbate soapy Sour 1
Fishy Note:)( = threshold; 1 = slight; 2 = moderate; 3 = strong.
Pungent Cool Menthol Source: Reprinted with permission from L.B. Sjöström,
Hot Alcohol The Flavor Profile, © 1972, A.D. Little, Inc.
Oxidized Oxidized Acetaldehyde
Microbiological Yeasty Leesy
Flor yeast Flavor and Off-Flavor
Lactic Lactic acid
Sweaty It is impossible to deal with the subject of flavor
Butyric acid without considering off-flavors. In many cases,
Sauerkraut the same chemical compounds are involved in
Other Mousey
both flavors and off-flavors. The only distinction
Horsey
appears to be whether a flavor is judged to be
pleasant or unpleasant. This amounts to a per-
sonal judgment, although many unpleasant fla-
residues in the PRP. The resulting cross-linking, vors (or off-flavors) are universally found to be
aggregation, and precipitation of the PRP causes unpleasant. A distinction is sometimes made
the sensation of astringency. between off-flavors—defined as unpleasant odors
Some anthocyanins are both bitter and astrin- or flavors imparted to food through internal dete-
gent. Bitter compounds such as quinine and caf- riorative change—and taints—defined as
feine compete with the tannins in complexing unpleasant odors or flavors imparted to food
with buccal proteins and thereby lower the astrin- through external sources (Saxby 1996). Off-­
gent response. Astringency is caused by higher flavors in animal products, meat and milk, may
molecular weight tannins, whereas the lower be caused by transfer of substances from feed.
molecular weight tannins up to tetrameres are Off-flavors in otherwise sound foods can be
associated with bitterness (Macheix et al. 1990). caused by heat, oxidation, light, or enzymic
318 7 Flavor

Fig. 7.33  Complex formation between proline-rich pro- InTech, DOI: https://doi.org/10.5772/67452. Available
teins and polyphenols L. Federico Casassa (2017). from: https://www.intechopen.com/books/phenolic-com-
Flavonoid Phenolics in Red Winemaking, Phenolic pounds-natural-sources-importance-and-applications/
Compounds - Natural Sources, Importance and flavonoid-phenolic
Applications, Prof. Marcos Soto-Hernández (Ed.),

action. The perception of taste and flavor can be


defined for a given group of people by the
International Standards Organization (ISO) 5492
standard (ISO 1992) as follows: The odor or taste
threshold is the lowest concentration of a com-
pound detectable by a certain proportion (usually
50%) of a given group of people. A graphic rep-
resentation of this relationship has been given by
Saxby (1996). The graph in Fig. 7.34 relates the
percentage of people within a given group to the
ability to detect a substance at varying concentra-
tions. Of the population, 50% can detect the com-
pound at the concentration of one unit. At a
concentration of the compound 10 times greater
than the mean threshold, about 10% of the popu-
lation is still not able to detect it. At the other end
of the spectrum, 5% of the population can still
detect the compound at a concentration 10 times
less than the mean threshold. These findings have
important consequences for the presence of com-
Fig. 7.34  Variation of taste threshold within a given pop-
pounds causing off-flavors. Even very low levels ulation. Adapted from M.J. Saxby, Food Taints and Off-­
of a chemical that produces off-flavors may cause Flavors, p. 43, © 1996, Aspen Publishers, Inc.
a significant number of people to complain.
Flavor of Some Foods 319

Certain flavor compounds may appear quite Flavor of Some Foods


pleasant in one case and extremely unpleasant in
another. Many examples of this can be cited. One As indicated previously, the two main factors
of the well-known cases is that of short-chain free affecting flavor are taste and odor. In a general
fatty acids in certain dairy products. Many cheese way, food flavors can be divided into two groups.
flavors contain volatile fatty acids as flavor con- The first consists of foods whose flavor cannot be
tributors (Day 1967). Yet, the same fatty acids in attributed to one or a few outstanding flavor
very low concentrations in milk and other dairy notes; their flavor is the result of the complex
products cause a very unpleasant, rancid off-­ interaction of a variety of taste and odor compo-
flavor (Patton 1964). Forss (1969) has drawn nents. Examples include bread, meat, and cheese.
attention to the compound non-2-enal. During The second group consists of foods in which the
studies of dairy product off-flavor, this compound flavor can be related to one or a few easily recog-
was ­isolated as a component of the oxidation off- nized components (contributory flavor com-
flavor and was found to have an odor reminiscent pounds). Examples include certain fruits,
of cucumbers. The same compound was isolated vegetables, and spices. Another way of differenti-
from cucumbers, and the cucumber-like flavor ating food flavors is by considering one group in
was assigned to the molecular structure of a which the flavor compounds are naturally present
2-trans-enal with 9 or 10 carbon atoms. Further and another group in which the flavor compounds
unsaturation and conjugation to give a 2,4-dienal are produced by processing methods.
produces flavors reminiscent of cardboard or
linoleum. Lactones were isolated by Keeney
and Patton (1956) and Tharp and Patton (1960) Bread
and were considered to be the cause of stale
­off-­flavors in certain dairy products. The same The flavor of white bread is formed mainly from
lactones, including δ-decalactone and the fermentation and baking processes. Freshly
δ-dodecalactone, were subsequently recognized baked bread has a delightful aroma that is rapidly
as contributors to the pleasant aroma of butter lost on cooling and storage. It has been suggested
(Day 1966). Dimethylsulfide is a component of that this loss of flavor is the result of disappear-
the agreeable aroma of meat and fish but has also ance of volatile flavor components. However, it is
been found to cause an off-flavor in canned well known that the aroma may be at least par-
salmon (Tarr 1966). Acetaldehyde occurs naturally tially regenerated by simply heating the bread.
in many foods, especially fruits, and is reported Schoch (1965) suggested that volatile flavor
to be essential for imparting the taste of freshness compounds may become locked in by the linear
(Byrne and Sherman 1984). The same compound fraction of wheat starch. The change in texture
is responsible for a very unpleasant oxidized fla- upon aging may be a contributory factor in the
vor in wine. Sinki (1988) has discussed the prob- loss of flavor. During fermentation, a number of
lems involved in creating a universally acceptable alcohols are formed, including ethanol, n-­
taste, and has stated that most individual flavor propanol, isoamyl and amyl alcohol, isobutyl
chemicals are either repugnant or painful outside alcohol, and β-phenol alcohol. The importance of
their proper formulations. This complex interaction the alcohols to bread flavor is a matter of contro-
between flavor chemicals, and between flavors versy. Much of the alcohols are lost to the oven
and the individual, makes the creation of a flavor- air during baking. A large number of organic
ful product both a science and an art, according to acids are also formed (Johnson et al. 1966).
Sinki. The subject of pleasantness and unpleas- These include many of the odd and even carbon
antness of flavors is the basis of a chapter in number saturated aliphatic acids, from formic to
Odour Description and Odour Classification by capric, as well as lactic, succinic, pyruvic, hydro-
Harper et al. (1968) and is the main subject of cinnamic, benzilic, itaconic, and levulinic acid. A
Moncrieff’s Odour Preferences (1966). large number of carbonyl compounds have been
320 7 Flavor

identified in bread, and these are believed to be Meat


important flavor components. Johnson et al.
(1966) list the carbonyl compounds isolated by Meat is another food in which the flavor is devel-
various workers from bread; this list includes 14 oped by heating from precursors present in the
aldehydes and 6 ketones. In white bread made meat; this occurs in a Maillard-type browning
with glucose, the prevalent carbonyl compound is reaction. The overall flavor impression is the
hydroxymethylfurfural (Linko et al. 1962). The result of the presence of a large number of non-
formation of the crust and browning during bak- volatile compounds and the volatiles produced
ing appear to be primary contributors to bread during heating. The contribution of nonvolatile
flavor. The browning is mainly the result of a compounds in meat flavor has been summarized
Maillard-type browning reaction rather than car- by Solms (1971). Meat extracts contain a large
amelization. This accounts for the presence of the number of amino acids, peptides, nucleotides,
carbonyl compounds, especially furfural, acids, and sugars. The presence of relatively large
hydroxymethylfurfural, and other aldehydes. In amounts of inosine-5'-monophosphate has been
the Maillard reaction, the amino acids are trans- the reason for considering this compound as a
formed into aldehydes with one less carbon atom. basic flavor component. In combination with
Specific aldehydes can thus be formed in bread other compounds, this nucleotide would be
crust if the necessary amino acids are present. responsible for the meaty taste. Living muscle
The formation of aldehydes in bread crust is contains adenosine-5'-triphosphate; this is con-
accompanied by a lowering of the amino acid verted after slaughter into adenosine-5'-mono-
content compared to that in the crumb. Johnson phosphate, which is deaminated to form
et al. (1966) have listed the aldehydes that can be inosine-5'-monophosphate (Jones 1969). The
formed from amino acids in bread crust as a volatile compounds produced on heating can be
result of the Strecker degradation (Table 7.23). accounted for by reactions involving amino acids
Grosch and Schieberle (1991) reported the and sugars present in meat extract. Lean beef,
aroma of wheat bread to include ethanol, pork, and lamb are surprisingly similar in flavor;
2-­methylpropanal, 3-methylbutanal, this reflects the similarity in composition of
2,3-­butanedione, and 3-methylbutanol. These extracts in terms of amino acid and sugar compo-
compounds contribute significantly to bread nents. The fats of these different species may
aroma, whereas other compounds are of minor account for some of the normal differences in fla-
importance. vor. In the volatile fractions of meat aroma,
hydrogen sulfide and methyl mercaptan have
been found; these may be important contributors
Table 7.23  Aldehydes that can be formed from amino
acids in bread crust as a result of the strecker degradation to meat flavor. Other volatiles that have been iso-
lated include a variety of carbonyls such as acet-
Amino acid Aldehyde
aldehyde, propionaldehyde, 2-methylpropanal,
Alanine Acetaldehyde
3-methylbutanal, acetone, 2-butanone, n-hexanal,
Glycine Formaldehyde
and 3-methyl-2-butanone (Moody 1983).
Isoleucine 2-Methylbutanal
Leucine Isovaleraldehyde
Methionine Methional
Phenylalanine Phenylacetaldehyde
Fish
Threonine 2-Hydroxypropanal
Serine Glyoxal
Fish contains sugars and amino acids that may be
involved in Maillard-type reactions during heat
Source: From J.A. Johnson et al., Chemistry of Bread
Flavor, in Flavor Chemistry, 1. Hornstein, ed., 1966, processing (canning). Proline is a prominent amino
American Chemical Society acid in fish and may contribute to sweetness.
Flavor of Some Foods 321

The sugars ribose, glucose, and glucose-­ 6-­ canned salmon that is related to dimethylsulfide.
phosphate are flavor contributors, as is 5′-inosinic The salmon was found to feed on zooplankton
acid, which contributes a meaty flavor note. containing large amounts of dimethyl-2-­
Volatile sulfur compounds contribute to the flavor carboxyethyl sulfonium chloride. This compound
of fish; hydrogen sulfide, methylmercaptan, and became part of the liver and flesh of the salmon
dimethylsulfide may contribute to the aroma of and in canning degraded to dimethylsulfide
fish. Tarr (1966) described an off-flavor problem in according to the following equation:

( CH3 )2 − SH − CH 2 − CH 2 − COOH → ( CH3 )2 S + CH3 − CH 2 − COOH

The flavor of cooked, fresh fish is caused by the Some of these sulfur compounds are also produced
presence of sugars, including glucose and fruc- from methionine when milk is exposed to light.
tose, giving a sweet impression as well as a umami Heterocyclic compounds are produced by nonen-
component arising from the synergism between zymatic browning reactions. Bitter peptides can be
inosine monophosphate and free amino acids. The formed by milk or bacterial proteinases.
fresh flavor of fish is rapidly lost by bacterial spoil- The basic taste of milk is very bland, slightly
age. In fresh fish, a small amount of free ammonia, sweet, and salty. Processing conditions influence
which has a pH level of below seven, exists in pro- flavor profiles. The extent of heat treatment deter-
tonated form. As spoilage increases, the pH rises mines the type of flavor produced. Low heat treat-
and ammonia is released. The main source of ment produces traces of hydrogen sulfide.
ammonia is trimethylamine, produced as a degra- Ultra-high temperature treatment results in a slight
dation product of trimethyl-amineoxide. fruity, ketone-like flavor. Sterilization results in
The taste-producing properties of hypoxan- strong ketone-like and caramelization/sterilization
thine and histidine in fish have been described by flavors. Sterilization flavors of milk are caused by
Konosu (1979). 5′-inosinate accumulates in fish the presence of 2-alkanones and heterocyclic
muscle as a postmortem degradation product of compounds resulting from the Maillard reaction.
ATP. The inosinate slowly degrades into hypo- Because of the bland flavor of milk, it is relatively
xanthine, which has a strong bitter taste. Some easy for off-flavors to take over.
kinds of fish, such as tuna and mackerel, contain
very high levels of free histidine, which has been
postulated to contribute to the flavor of these fish. Cheese

The flavor of cheese largely results from the fer-


Milk mentation process that is common to most variet-
ies of cheese. The microorganisms used as
The flavor of normal fresh milk is probably pro- cultures in the manufacture of cheese act on
duced by the cow’s metabolism and is comprised many of the milk components and produce a
of free fatty acids, carbonyl compounds, alkanols, large variety of metabolites. Depending on the
and sulfur compounds. Free fatty acids may result type of culture used and the duration of the ripen-
from the action of milk lipase or bacterial lipase. ing process, the cheese may vary in flavor from
Other decomposition products of lipids may be mild to extremely powerful. Casein, the main
produced by the action of heat. In addition to lip- protein in cheese, is hydrolyzed in a pattem and
ids, proteins and lactose may be precursors of fla- at a rate that is characteristic for each type of
vor compounds in milk (Badings 1991). Sulfur cheese. Proteolytic enzymes produce a range
compounds that can be formed by heat from of peptides of specific composition that are
β-lactoglobulin include dimethyl sulfide, hydro- related to the specificity of the enzymes present.
gen sulfide, dimethyl disulfide, and methanethiol. Under certain conditions bitter peptides may be
322 7 Flavor

formed, which produce an off-flavor. Continued hydrocarbons include mainly D-limonene,


hydrolysis yields amino acids. The range of pep- β-myrcene, and a compound of composition
tides and amino acids provides a “brothy” taste C15H24. The esters include isovalerate, methyl
background to the aroma of cheese. Some of alphaethyl-n-caproate, citronellyl acetate, and
these compounds may function as flavor enhanc- terpinyl acetate. In the group of carbonyls, the
ers. Breakdown of the lipids is essential for the following compounds were identified: n-hexanal,
production of cheese aroma since cheese made n-octanal, n-decanal, and citronella; and in the
from skim milk never develops the full aroma of group of alcohols, linalool, α-terpineol, n-­
normal cheese. The lipases elaborated by the cul- hexane-­1-ol, n-octan-1-ol, n-decan-1-ol, and
ture organisms hydrolyze the triglycerides to 3-hexen-1-ol were identified. The flavor deterio-
form fatty acids and partial glycerides. The par- ration of canned orange juice during storage
ticular flavor of some Italian cheeses can be results in stale off-flavors. This is due to reactions
enhanced by adding enzymes during the cheese-­ of the nonvolatile water-soluble constituents. As
making process that cause preferential hydrolysis in the case of citrus fruits, no single compound is
of short-­chain fatty acids. Apparently, a variety of completely responsible for any single fruit aroma.
minor components are important in producing However, some organoleptically important com-
the characteristic flavor of cheese. Carbonyls, pounds characteristic for particular fruits have
esters, and sulfur compounds are included in this been found. These include amyl esters in banana
group. The relative importance of many of these aroma, citral in lemon, and lactones in peaches.
constituents is still uncertain. Sulfur compounds The major flavor component of Bartlett pears was
found in cheese include hydrogen sulfide, identified by Jennings and Sevenants (1964) as
dimethylsulfide, methional, and methyl mercap- ethyl trans-2-cis-4-decadienoate.
tan. All of these compounds are derived from
sulfur-­containing amino acids. The flavor of blue
cheese is mainly the result of the presence of a Vegetables
number of methyl ketones with odd carbon num-
bers ranging in chain length from 3 to 15 carbons Vegetables contain an extensive array of volatile
(Day 1967). The most important of these are flavor compounds, either in original form or pro-
2-heptanone and 2-nonanone. The methyl ketones duced by enzyme action from precursors. Maarse
are formed by β-oxidation of fatty acids by the (1991) has reviewed these in detail. Onion and gar-
spores of P. roqueforti. lic have distinctive and pungent aromas that result
mostly from the presence of sulfur-­ containing
compounds. A large number of flavor compounds
Fruits in vegetables are formed after cooking or frying. In
raw onions, an important compound is thio-­
The flavor of many fruits appears to be a combi- propanal s-oxide—the lachrymatory factor. The
nation of a delicate balance of sweet and sour distinctive odor of freshly cut onions involves two
taste and the odor of a number of volatile com- main compounds, propyl methane-­ thiosulfonate
pounds. The characteristic flavor of citrus prod- and propyl propanethiosulfonate. Raw garlic con-
ucts is largely due to essential oils contained in tains virtually exclusively sulfur compounds: four
the peel. The essential oil of citrus fruits contains thiols, three sulfides, seven disulfides, three trisul-
a group of terpenes and sesquiterpenes and a fides, and six dialkylthio-sulfinates.
group of oxygenated compounds. Only the latter
are important as contributors to the citrus flavor.
The volatile oil of orange juice was found to be Tea
91.6 mg per kg, of which 88.4 was hydrocarbons
(Kefford 1959). The volatile water-soluble con- The flavor of black tea is the result of a number of
stituents of orange juice consist mainly of acetal- compounds formed during the processing of
dehyde, ethanol, methanol, and acetic acid. The green tea leaves. The processing involves wither-
Flavor of Some Foods 323

ing, fermentation, and firing. Bokuchava and to its content of lactones, aldehydes, alcohols,
Skobeleva (1969) indicate that the formation of acids, and pyridines.
the aroma occurs mainly during firing. Aromatic
compounds isolated and identified from black tea
include acrolein, n-butyric aldehyde, ethanol, Coffee
n-butanol, isobutanol, hexanal, pentanal,
2-­hexanol, 3-hexen-1-ol, benzaldehyde, linalool, The flavor of coffee is developed during the
terpeneol, methylsalicylate, benzyl alcohol, roasting of the green coffee bean. Gas-liquid
β-phenylethanol, isobutyric aldehyde, geraniol, chromatography can be used to demonstrate
and acetophenone. The flavor substances of tea (Fig. 7.35) the development of volatile constitu-
can be divided into the following four fractions: a ents in increasing amounts as intensity of roast-
carbonyl-free neutral fraction including a number ing increases (Gianturco 1967). The total number
of alcohols, a carbonyl fraction, a carboxylic acid of volatile compounds that have been isolated is
fraction, and a phenolic fraction. A compilation in the hundreds, and many of these have been
(Maarse 1991) identifies a total of 467 flavor con- identified. To determine the flavor contribution of
stituents in tea. The distinctive flavor of tea is due each of these is a Herculean task. Many com-
pounds result from the pyrolytic decomposition

Fig. 7.35 Development
of volatile constituents
during roasting of
coffee. From top to
bottom: green coffee
after 2, 6, 8, 11, and
15 min of roasting. The
gas chromatograms
show increasing
concentrations of
volatile compounds.
Source: From
M.A. Gianturco, Coffee
Flavor, in Symposium on
Foods: The Chemistry
and Physiology of
Flavors, H.W. Schultz
et al., eds., 1967, AVI
Publishing Co.
324 7 Flavor

of carbohydrates into units of 2, 3, 4, or 5 car- cal treatments. Biggers et al. (1969) differenti-
bons. Other compounds of carbohydrate origin ated the beverage quality of two varieties of
are 16 furanic compounds, cyclic diketones, and coffee (arabica and robusta) on the basis of con-
maltol. Roasting of the proteins of the coffee tributions of flavor compounds.
bean can yield low molecular weight products Recent studies have identified 655 compounds
such as amino acids, ammonia, amines, hydrogen in the flavor of coffee, the principal ones being
sulfide, methyl mercaptan, dimethylsulfide, and furans, pyrazines, pyrroles, and ketones (Maarse
dimethyl disulfide. A series of furanic and pyrro- 1991). The distinctiveness of coffee flavor is
lic compounds identified include the following: related to the fact that it contains a large percent-
furan, furfural, acetylfuran, 5-methylfuran, age of thiophenes, furans, pyrroles, as well as
5-methylfurfural, 5-methyl-2-acetylfuran and oxazoles, thiazoles, and phenols.
pyrrole, 2-pyrrolaldehyde, 2-acetylpyrrole,
N-methylpyrrole, N-methyl-2-pyrrolaldehyde,
and N-methyl-2-acetylpyrrole. Differences in the Alcoholic Beverages
aroma of different coffees can be related to quan-
titative differences in some of the compounds In distilled beverages, one of the major flavor
isolated by gas chromatography, and ratios and compounds is acetaldehyde. Acetaldehyde repre-
amounts of these compounds may be different. sents about 90% of the total aldehydes present in
Pyrazines, furanes, pyrroles, and thiophen beverages like whiskey, cognac, and rum.
derivatives are particularly abundant in coffee
­ Together with other short-chain aliphatic alde-
aroma. Furfuryl-methyl-sulfide and its homologs hydes, it produces a pungent odor and sharp fla-
are important contributors to the aroma of coffee. vor, which is masked by other flavor components
The structures of some of the important aroma in cognac, fruit brandies, rum, and whiskey. In
contributors are presented in Fig. 7.36. The com- vodka the presence of acetaldehyde may result in
pounds identified in coffee aroma are listed and an off-flavor. Propanol and 2-methylpropanol, as
differentiated on the basis of functional groups in well as unsaturated aldehydes, are also present in
Table  7.24. It is, of course, impossible to com- distilled beverages. The aldehydes are very reac-
pare the aroma of different coffees on the basis of tive and can form acetals by reacting with etha-
one or a few of the flavor constituents. Computer-­ nol. This reaction results in a smoother flavor
generated histograms can be used for compari- profile. Another important flavor compound in
sons after selection of important regions of distilled beverages is the diketone,
gas-liquid chromatograms by using mathemati- 2,3-­butanedione (diacetyl), which is a product of
fermentation. Depending on fermentation and
distillation conditions, the level of diacetyl varies
widely in different beverages.
Fusel alcohols, which are present in most dis-
tilled beverages, influence flavor. They are
formed during fermentation from amino acids
through decarboxylation and deamination, and
include 1-propanol, 2-methylpropanol,
2-­methylbutanol, 3-methylbutanol, and
2-phenylethanol.
Distilled beverages also contain fatty acids—
from acetic acid (which is one of the major fatty
acids) to long-chain unsaturated fatty acids.
Maturation in oak barrels has a major effect on
Fig. 7.36  Structure of some important constituents of the flavor of distilled beverages. Maturing fresh distil-
aroma of coffee lates in oak barrels can transform a raw-tasting
References 325

Table 7.24  Volatile compounds in roasted coffee aroma


Functional Group
—O —O
−C—— −C——
Compound Type None —OH —O— OH C=O —CO—CO— OR —S— Other
Aliphatic 17 19 — 13 30 10 16 9 25
Isocyclic 3 1 — — 6 6 — — —
Benzenic 20 — 6 4 5 1 5 2 16
Furanic 15 1 4 3 13 4 11 7 3
Thiophenic 6 1 — 2 6 2 3 20 —
Pyrrolic 10 — — 8 5 1 — — —
Pyrazinic 27 — — — — — — — —
Other 5 2 8 — 7 — 1 5 13
Total number 103 24 18 30 72 24 36 43 57

product into a mellow, well-rounded beverage. Badings, H. T. (1991). Milk. In Volatile compounds in
The reactions that take place during maturation foods and beverages. New York: Marcel Dekker.
Bartoshuk, L. M., Duffy, V. B., & Miller, I. J. (1994).
involve reactions between components of the dis- PTC/PROP tasting: Anatomy, psychophysics, and sex
tillate and reactions between distillate components effects. Physiology & Behavior, 56, 1165–1171.
and compounds present in the oak wood. The Beatty, R. M., & Cragg, L. H. (1935). The sourness of
alcoholic solution in the barrel extracts lignin acids. Journal of the American Chemical Society, 57,
2347–2351.
from the oak to form an alcohol-soluble ethanol-­ Beidler, L. M. (1954). A theory of taste stimulation. The
lignin. Alcoholysis converts this to coniferic alco- Journal of General Physiology, 38, 133–139.
hol and then by oxidation to coniferaldehyde. Beidler, L. M. (1957). Facts and theory on the mechanism
Similarly, sinapic alcohol is converted to sinapal- of taste and odor perception. In Chemistry of natu-
ral food flavors. Chicago: Quartermaster Food and
dehyde. These aldehydes then produce syringal- Container Institute for the Armed Forces.
dehyde and vanillin. The latter compound is Beidler, L. M. (1966). Chemical excitation of taste and
important in the flavor of cognac and whiskey. A odor receptors. In I. Horn-stein (Ed.), Flavor chemis-
similar process occurs in the aging of wines in oak try. Washington: American Chemical Society.
Biggers, R. E., Hilton, J. J., & Gianturco, M. A. (1969).
barrels to produce the distinctive smoothness of Differentiation between Coffea arabica and Coffea
oak-aged wines. robusta by computer evaluation of gas chromato-
graphic profiles: Comparison of numerically derived
quality predictions with organoleptic evaluations.
Journal of Chromatographic Science, 7, 453–472.
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Texture
8
Harry Levine and John W. Finley

and Schleining 2011), and fat replacement


Introduction alters biscuit quality (Sudha et al. 2007). The
texture characteristics of foods can be evaluated
Texture is one of the key attributes of foods, by descriptive sensory (subjective) or instrumen-
which is used to define product quality and tal (objective) analyses. Sensory analysis can be
acceptability. Food texture can be defined by the time-­ consuming and expensive; consequently,
way in which the various constituents and struc- many empirical mechanical tests have been
tural elements are arranged and combined into a developed, the results of which correlate with the
micro- and macrostructure, and by the external sensory analysis of food texture.
manifestations of this structure, in terms of flow Food texture can be defined as “all the rheo-
and deformation. Texture measurements are used logical and structural (geometric and surface)
throughout the food value chain to monitor and attributes of the product perceptible by means
control quality, from harvest to assessing the of mechanical, tactile, and where appropriate,
impacts of postharvest handling and processing visual and auditory receptors” (Lawless and
on shelf life and consumer acceptance. Heymann 1998). Most of our foods are com-
Postharvest handling and processing conditions plex physico-­ chemical structures, and, as a
such as storage temperature can have a signifi- result, the physical properties cover a wide
cant influence on, e.g., the textural properties of range—from fluid, Newtonian materials to the
meat (Farag et al. 2009), apple quality and most complex disperse systems with semi-
acceptability (Konopacka and Plocharski 2004), solid character. There is a direct relationship
and storage and ripening of cut tomatoes (Lana between the chemical composition of a food,
et al. 2005). Addition of new ingredients, replace- its physical structure, and the resulting textural
ment of existing ones, or changes in processing properties. Chemical composition determines
conditions can result in both desirable and/or the basic physical structure, and this in turn
undesirable changes in texture. For example, influences the texture of the food, whether
added whey proteins and carbohydrate polymers measured mechanically or by sensory evalua-
alter food microstructure (Foegeding et al. 2010), tion. These interrelationships, with examples
added inulin and fructo-oligosaccharides alter of testing methods, are illustrated in Fig. 8.1.
bread texture (Morris and Morris 2012), pre-­ Food texture can be evaluated by a range of
drying alters texture and oil uptake in potato mechanical tests (instrumental methods) or by
chips (Pedreschia and Moyano 2005), frying sensory analysis. In the latter case, we use the
conditions alter crispness of crackers (Saeleaw human sense organs as analytical tools. A proper

© Springer International Publishing AG 2018 329


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_8
330 8 Texture

Fig. 8.1 The
interrelationships of
chemical composition,
physical structure, and
the texture
characteristics of a food.
Each area corresponds
to a range of applicable
measurement methods.
Such test results are then
related to those from the
sensory evaluation of the
food

understanding of textural properties often Most foods are complex systems, in which
requires study of the physical structure. This is many textural characteristics are experienced
most often accomplished by light and/or elec- simultaneously when we eat. Examples include
tron microscopy, as well as by several other sandwich cookies (with a viscoelastic filling and
physical methods. X-ray diffraction analysis a brittle base cake), vegetables with cream sauce,
provides information about crystalline struc- and macaroni and cheese. Such multi-texture sys-
ture, differential scanning calorimetry provides tems present great difficulty in developing suit-
information about melting, solidification, and able mechanical testing methods. Generally, the
other phase or state transitions, and particle different phases must be considered separately.
size analysis and sedimentation methods pro- As a result, it is difficult in many cases to relate
vide information about particle size distribu- results obtained by instrumental measurement
tion and particle shape. techniques to the types of responses obtained by
A discussion of food texture can be divided sensory panel tests.
into two parts: fluids and solids. Chemical
composition and chemical interactions ulti-
mately drive the characteristics of both solids Fluids
and fluids. Different testing techniques are
employed to obtain objective data to help Fluid dynamics are best described as the flow and
explain the sensory responses to foods. deformation of a fluid under stress. Viscosity (η)
Figure  8.1 illustrates the different approaches is a measure of the resistance to flow of a fluid
used to define physical parameters. under stress. The higher the viscosity of a fluid,
In the study of food texture, attention is given the generally greater is the resistance to flow.
to two interdependent areas: the flow and defor- Fluids can be grouped into Newtonian and non-­
mation properties and the macro- and microstruc- Newtonian fluids. In 1687, Sir Isaac Newton
ture. The study of food texture is carried out for developed his three laws of motion:
three important reasons:
1. A body continues in its state of rest or uniform
1. to evaluate the resistance of products against motion in a straight line, unless it is acted
mechanical action, such as in mechanical har- upon by another force.
vesting of fruits and vegetables; 2. The rate of change in momentum is propor-
2. to determine the flow properties of products tional to the applied force and in the direction
during processing, handling, and storage; of the applied force.
3. to understand the mechanical behavior of a 3. For every action, there is an equal and oppo-
food when consumed. site reaction.
Fluids 331

Fig. 8.2  The effect of Plate moving at velocity D


shear force exerted by a Shear
moving plate on the flow
of a fluid at increasing Force
distances from the
surface where the force
is applied

Infinitesimal
layers of liquid

Stationary Plate

The application with relevance to fluids is


illustrated in Fig. 8.2. This model illustrates the
shear force exerted by a moving plate, against the
infinitesimal layers of liquid between the moving
plate at velocity ν and a stationary plate. It can be
seen that, the further from the moving plate, the
less the liquid moves with the force.
Consider a liquid contained between two par-
allel plates, each of area A cm2 (Fig. 8.2). The
plates are h cm apart, and a force of P dynes is
applied horizontally to the upper plate. This
shearing stress causes it to move, with respect to
the lower plate, with a velocity of v cm s−1. The
shearing stress τ acts throughout the liquid con-
tained between the plates, and can be defined as
the shearing force P divided by the area A, or P/A
dynes/cm2. The deformation can be expressed as Fig. 8.3  Shear stress-shear rate diagrams: (A) Newtonian
liquid, viscous flow; (B) dilatant flow; (C) pseudoplastic
the mean rate of shear γ or the velocity gradient, flow; (D) plastic flow
and is equal to the velocity difference divided by
the distance between the plates, γ = v / h ,
expressed in units of s−1. The relationship between shear stress and
In Newtonian flow, it is assumed that, at a shear rate can be used to define the flow proper-
given temperature, and for all such materials, vis- ties of materials. In the simplest case, the shear
cosity is independent of shear rate. In such sys- stress is directly proportional to the mean shear
tems, twice the force would cause the fluid to rate, τ = ηγ (Fig. 8.3). The proportionality con-
move twice as fast. Shear is the movement of the stant η is called the viscosity coefficient, or
fluid relative to the parallel surface, shear stress is dynamic viscosity, or simply the viscosity of the liq-
the force per unit area to cause flow, and shear uid. The metric unit of viscosity is the dyne.s cm−2,
rate is the difference in flow between two layers or Poise (P). The commonly used unit is 100
of liquid, as illustrated by the arrows in Fig. 8.2. times smaller than P, and called centiPoise (cP).
332 8 Texture

Table 8.1  Viscosity coefficients of some foods Generally, there are three classes of non-­
Temperature Viscosity Newtonian systems that must be considered: (1)
Product (°C) (cP) plastic flow, (2) pseudoplastic flow, and (3) dilat-
Water 0 1.79 ant flow.
Water 20 1.00 Plastic flow occurs when a preparation under-
Beer 4 1.1 goes flocculation or aggregation of particles in a
Milk, whole 18 2.0 suspension. These are referred to as Bingham
Milk, whole 49 10 bodies. These bodies do not begin to flow, until a
Cream (30% fat) 16 14 shear force or stress corresponding to a yield
Cream (50% fat) 32 55 value is exceeded. The yield value is the force
Soybean oil 24 60 that must be applied to convert the system to a
Soybean oil 80 12
Newtonian fluid, as seen for line D in Fig. 8.3.
Sucrose solution 21 60.2
Examples of such materials are tooth paste in a
(60%)
tube, chili paste, or tomato paste. The ratio of
Olive oil 30 84.0
Cottonseed oil 16 91.0
shear stress to shear rate in such materials is not a
Mayonnaise 21 20,000
constant value, so the value is designated as
Molasses 21 1400–13,000 apparent viscosity. To be useful, a reported value
for the apparent viscosity of a non-Newtonian
material should be given together with the value
In the SI system, η is expressed in N.s/m2, or Pa.s. of shear rate or shear stress used in the determi-
Therefore, 1 Pa.s = 10 P = 1000 cP. Some instru- nation. The relationship between shear stress and
ments measure kinematic viscosity, which is shear rate for non-Newtonian materials, such as
equal to dynamic viscosity × density and is the dilatant and pseudoplastic bodies in Fig. 8.3,
expressed in units of Stokes. The viscosity of can be represented by a power law as follows:
water at room temperature is about
τ = Aγ n
1.0 cP. Table 8.1 lists the absolute viscosities for
a range of fluid food materials, expressed in where A and n are constants. A is the consis-
centipoise. tency index or apparent viscosity, and n is the
Materials that exhibit a direct proportionality flow behavior index. The exponent is n = 1 for
between shear stress and shear rate are called Newtonian liquids; for dilatant materials, it is
Newtonian fluids. These include water and aque- greater than 1; and for pseudoplastic materials, it
ous solutions, simple organic liquids, and dilute is less than 1. In its logarithmic form,
suspensions and emulsions. In contrast, most
log τ = log A + n logγ
foods are non-Newtonian in character, and their
shear stress–shear rate curves are either not straight A plot of log τ versus log γ will yield a
lines, or do not go through the origin, or both. This straight line with a slope of n.
introduces a considerable difficulty, because their For non-Newtonian materials that have a yield
flow behavior cannot be expressed by a single stress, the Casson or Hershel-Bulkley models can
value, as is the case for Newtonian liquids. be used. The Casson model is represented by the
In non-Newtonian fluids, the relationship equation,
between force and shear is not constant. When
the shear rate is varied, the shear stress does not τ = τ 0 + A γ

respond in a linear fashion. The viscosity of such
liquids will change as the shear rate is varied. where τ0 = yield stress.
Such phenomena can be seen in heterogeneous This model has been found useful for several
liquid and solid dispersions, such as colloidal food products, especially chocolate (Kleinert
suspensions, emulsions, and liquid suspensions. 1976).
Texture of Solids 333

Fig. 8.4  The interaction of shear stress, flow, and viscosity for the four different types of fluids

The Hershel-Bulkley model describes a mate- • Hardness has been defined as resistance to
rial with a yield stress and a linear relationship deformation.
between log shear stress and log shear rate: • Firmness is essentially identical to hardness,
but is occasionally used to describe the prop-
τ = τ 0 + Aγ n
erty of a substance able to resist deformation
The value of n indicates how close the plot of under its own weight.
shear stress vs. shear rate is to being a straight • Brittleness is the property of fracturing before
line. Fig. 8.4 represents the four types of flow, significant flow has occurred.
illustrating the responses of different types of flu- • Stickiness is a surface property related to the
ids to shear stress and how the flow and viscosity adhesion between material and adjoining sur-
are affected. face. When the two surfaces are of identical
material, we use the term cohesion.

Texture of Solids A variety of other words and expressions are


used to describe textural characteristics, such as
The terms for the textural properties of foods body, crisp, greasy, brittle, tender, juicy, mealy,
have a long history. Many of the terms are flaky, crunchy, and so forth. Many of these terms
accepted, but are often poorly defined descrip- have been discussed by Szczesniak (1963) and
tive terms. The following are some examples of Sherman (1969); most have no objective physical
such terms: meaning and cannot be expressed in units of
measurement that are universally applicable.
• Consistency denotes those aspects of texture Kokini (1985) has attempted to relate some of
that relate to flow and deformation. It can be these ill-defined terms to the physical properties
said to encompass all of the rheological prop- involved in their evaluation. Through the years,
erties of a product. many types of instruments have been developed
334 8 Texture

for measuring certain aspects of food texture. 1. Brittleness—the force with which the material
More recently, instruments have been developed fractures. This is related to hardness and cohe-
that are more widely applicable and are based on siveness. In brittle materials, cohesiveness is
sound physical and engineering principles. low, and hardness can be either low or high.
Brittle materials often create sound effects
when masticated (e.g., toast, carrots, celery);
Texture Profile 2. Chewiness—the energy required to masticate
a solid food product to a state ready for swal-
Texture is an important aspect of food quality, lowing. It is related to hardness, cohesiveness,
sometimes even more important than flavor and and elasticity;
color. Szczesniak and Kleyn (1963) conducted a 3. Gumminess—the energy required to disinte-
consumer-awareness study of texture and found grate a semi-solid food to a state ready for
that texture significantly influences people’s swallowing. It is related to hardness and
image of food. Texture was most important in cohesiveness.
bland foods and foods that are crunchy or crisp.
The characteristics most often referred to were Table 8.2  Classification of textural characteristics of
hardness, cohesiveness, and moisture content. foods
Several attempts have been made to develop a Mechanical characteristics
classification system for textural characteristics. Secondary
Szczesniak (1963) divided textural characteris- Primary parameters parameters Popular terms
tics into three main classes, as follows: Hardness Soft → firm →
hard
1. mechanical characteristics; Cohesiveness Brittleness Crumbly →
crunchy →
2. geometrical characteristics; brittle
3. other characteristics, related mainly to mois- Chewiness Tender →
ture and fat content. chewy → tough
Gumminess Short → mealy
Mechanical characteristics include five basic → pasty →
gummy
parameters.
Viscosity Thin → viscous
Elasticity Plastic →
1. Hardness—the force necessary to attain a elastic
given deformation. Adhesiveness Sticky → tacky
2. Cohesiveness—the strength of the internal → gooey
bonds making up the body of the product. Geometrical characteristics
3. Viscosity—the rate of flow per unit force. Class Examples
4. Elasticity—the rate at which a deformed Particle size and Gritty, grainy, coarse, etc.
material reverts to its undeformed condition, shape
after the deforming force is removed. Particle shape and Fibrous, cellular, crystalline, etc.
orientation
5. Adhesiveness—the work necessary to over-
Other characteristics
come the attractive forces between the surface
Primary Secondary Popular terms
of a food and the surfaces of other materials parameters parameters
with which the food comes in contact (e.g., Moisture content Dry → moist →
tongue, teeth, and palate). wet → watery
Fat content Oiliness Oily
In addition, there are in this class the three fol- Greasiness Greasy
lowing secondary parameters: Source: from Szczesniak (1963)
Measurement of Texture 335

Geometrical characteristics include two gen- analytical characteristics from which all other
eral groups: those related to size and shape of the attributes are derived. The basic rheological
particles, and those related to shape and orienta- parameters—elasticity, viscosity, and adhesion—
tion. Terms for geometrical characteristics form the secondary category. The remaining
include smooth, cellular, fibrous, and so on. The attributes form the tertiary category, since they
group of other characteristics in this system is are a complex mixture of these secondary param-
related to moisture and fat content, and includes eters. This system is interesting, because it
qualities such as moist, oily, and greasy. A sum- attempts to relate sensory responses to mechani-
mary of this system is given in Table 8.2. cal strain-time tests. Sensory panel responses,
Based on the Szczesniak system of textural associated with masticatory tertiary characteris-
characteristics, Brandt et al. (1963) developed a tics of the Sherman texture profile for solid, semi-­
method for profiling texture, so that a sensory solid, and liquid foods, are given in Fig. 8.6.
evaluation could be given that would assess the
entire texture of a food. This texture profile
method was based on the earlier development of Measurement of  Texture
the flavor profile (Cairncross and Sjöström 1950).
The Szczesniak system was critically exam- The objective measurement of texture belongs in
ined by Sherman (1969), who proposed some the area of rheology, which is the science of flow
modifications. In this improved system, no dis- and deformation of matter. However, determining
tinction is drawn among analytical, geometrical, the rheological properties of a food does not nec-
and mechanical attributes. Instead, the only crite- essarily mean that the complete texture of the
rion is whether a characteristic is a fundamental product is determined. Nevertheless, knowledge
property or is derived by a combination of two or of some of the rheological properties of a food
more attributes in unknown proportions. The may give important clues as to its acceptability and
Sherman system contains three groups of charac- may be important in determining the nature and
teristics (Fig. 8.5). The primary category includes design of processing methods and equipment.

Fig. 8.5  The modified texture profile (Sherman 1969)


336 8 Texture

Fig. 8.6  Panel responses associated with masticatory tertiary characteristics of the modified texture profile

Food rheology is mainly concerned with or inertia of a body may constitute a force leading
forces and deformations. In addition, time is an to deformation. Generally, however, the exter-
important factor; many rheological phenomena nally applied forces are of much greater magni-
are time-dependent. Temperature is another tude, and the effect of weight is usually neglected.
important variable. Many products show impor- The forces acting on a body can be expressed in
tant changes in rheological behavior as a result of grams or in pounds. Stress is the intensity factor
changes in temperature. In addition to flow and of force and is expressed as force per unit area; it
deformation of cohesive bodies, food rheology is similar to pressure. There are several types of
includes such phenomena as the breakup or rup- stress: compressive stress (with the stress compo-
ture of solid materials and surface phenomena nents directed at right angles toward the plane on
such as stickiness (adhesion). which they act); tensile stress (in which the stress
Deformation may be of one or both of two components are directed away from the plane on
types, irreversible deformation, called flow, which they act); and shearing stress (in which the
and reversible deformation, called elasticity. stress components act tangentially to the plane on
The energy used in irreversible deformation is which they act). A uniaxial stress is usually desig-
dissipated as heat, and the body is permanently nated by the symbol σ, a shearing stress by τ.
deformed. The energy used in reversible defor- Shear stress is expressed in dynes/cm2, when
mation is recovered upon release of the using the metric system of measurement; in the SI
deforming stress, when the body regains its system, it is expressed in N/m2 or Pascals (Pa).
original shape.

Deformation and Strain
Force and Stress
When the dimensions of a body change, we speak
When a force acts externally on a body, several of deformation. Deformation can be linear, as in
different cases may be distinguished: tension, a tensile test, when a body of original length L is
compression, and shear. Bending involves tension subjected to a tensile stress. The linear deforma-
and compression, torque involves shear, and tion ΔL can then be expressed as strain ε = ΔL/L.
hydrostatic compression involves all three. All Strain can be expressed as a ratio or percent;
other cases may involve one of these three factors inches per inch or centimeters per centimeter. In
or a combination of them. In addition, the weight addition to linear deformations, there are other
Different Types of Bodies 337

Fig. 8.7  Relaxation curve (relationship of stress to time


under constant strain) Fig. 8.8  Creep curve (relationship of strain to time under
constant stress)

types of deformation, such as in a hydrostatic


test, where there will be a volumetric strain ΔV/V. ing creep curve has the shape indicated in Fig. 8.8.
For certain materials, the deformation result- At time zero, the applied load results in a strain εo,
ing from an applied force can be very large; this which increases with time. When the load is
indicates the material is a liquid. In such cases, removed at time T, the strain immediately
we deal with rate of deformation, or shear rate: decreases, as indicated by the vertical straight por-
dγ/dt or γ . This is the velocity difference per tion of the curve at T; the strain continues to
unit thickness of the liquid; γ is expressed in decrease thereafter with time. In many materials,
units of s−1. the value of ε never reaches zero, and we know,
therefore, that a permanent deformation εp has
resulted. The ratio of strain to applied stress in a
Principles of Measurement creep experiment is a function of time and is called
the creep compliance (J). Creep experiments are
For Newtonian fluids, it is sufficient to measure sometimes plotted as graphs relating J to time.
the ratio of shear stress to shear rate, from which
the viscosity can be calculated. This can be done in
a viscometer, which can be one of various types, Different Types of Bodies
including capillary, rotational, falling-ball, and so
on. For non-Newtonian materials, such as the The Elastic Body
dilatant, pseudoplastic, and plastic bodies shown
in Fig. 8.4, the problem is more difficult. With For certain solid bodies, the relationship between
non-Newtonian materials, several methods of stress and strain is represented by a straight line
measurement involve the ratio of shear stress to through the origin (Fig. 8.9), up to the so-called
shear rate, the relationship of stress to time under limit of elasticity, according to the law of Hooke,
constant strain (relaxation), and the relationship σ = Εε. The proportionality factor E for uniaxial
of strain to time under constant stress (creep). In stress is called modulus of elasticity, or Young’s
relaxation measurements, a material is subjected modulus. For a shear stress, the modulus is G, or
to a sudden deformation εo, which is held con- Coulomb modulus. Note that a modulus is the
stant. In many materials, the stress will decay ratio of stress to strain, E = σ/ε. The behavior of a
with time, according to the curve of Fig. 8.7. The Hookean body is further exemplified by the
point at which the stress has decayed to σ/e, or stress-time and strain-time curves of Fig. 8.10.
36.7% of the original value of σo, is called the When a Hookean body is subjected to a constant
relaxation time T. When the strain is removed at strain εo, the stress σ will remain constant with
time T, the stress returns to zero. In a creep experi- time, and will return to zero, when the strain is
ment, a material is subjected to the instantaneous removed at time T. The strain ε will follow the
application of a constant load or stress, and the same pattern, when a constant stress is applied,
strain is measured as a function of time. The result- and then released at time T.
338 8 Texture

Fig. 8.12 (a) Stress-time and (b) strain-time curves for a


retarded elastic body

Fig. 8.9  Stress-strain curve for a perfectly elastic body


slowly return to zero. There is no permanent
deformation. The corresponding relaxation
(stress-time) and creep (strain-time) curves for
this type of body are given in Fig. 8.12.

The Viscous Body

A viscous or Newtonian liquid is one showing a


direct proportionality between shear stress and
shear rate, as indicated by curve A in Fig. 8.3.
Fig. 8.10 (a) Stress-time and (b) strain-time curves for a
Hookean body
The Viscoelastic Body

Certain bodies combine the properties of both


viscous and elastic materials. The elastic compo-
nent can be partially retarded elasticity.
Viscoelastic bodies may flow slowly and non-­
reversibly under the influence of a small stress.
Under larger stresses, the elastic component
becomes apparent. The relaxation curve for vis-
coelastic materials has the shape indicated in
Fig.  8.13a. The curve has the tendency to
approach the time axis. The creep curve indicates
Fig. 8.11  Stress-strain curve for a retarded elastic body that the strain increases for as long as the stress is
applied (Fig. 8.13b). The magnitude of the per-
manent deformation of the body increases with
The Retarded Elastic Body the applied stress and with the length of time of
application.
In bodies showing retarded elasticity, the defor- Mechanical models can be used to visualize
mation is a function of time as well as stress. the behavior of different bodies. Thus, a spring
Such a stress-strain curve is shown in Fig. 8.11. denotes a Hookean body, and a dash-pot denotes
The upward part of the curve represents increas- a purely viscous body or Newtonian fluid. These
ing values of stress; when the stress is reduced, elements can be combined in a variety of ways to
the corresponding strains are greater on the represent the rheological behavior of complex
downward part of the curve. When the stress substances. Two basic viscoelastic models are the
reaches 0, the strain has a finite value, which will Voigt-Kelvin and the Maxwell bodies. The Voigt-­
Different Types of Bodies 339

rather, instantly. The Voigt-Kelvin body, there-


fore, shows no stress relaxation, but the Maxwell
body does. A variety of models can be con-
structed to represent the rheological behavior of
viscoelastic materials. By placing a number of
Kelvin models in series, a so-called generalized
Kelvin model is obtained. Similarly, a general-
ized Maxwell model is obtained by placing a
Fig. 8.13 (a) Stress-time and (b) strain-time curves for a number of Maxwell models in parallel. The com-
viscoelastic body bination of a Kelvin and a Maxwell model in
series (Fig. 8.14c) is called a Burgers model.
For ideal viscoelastic materials, the initial
elastic deformation at the time the load is applied
should equal the instantaneous elastic deforma-
tion when the load is removed (Fig. 8.15). For
most food products, this is not the case. As is
shown by the example of butter in Fig. 8.15, the
initial deformation is greater than the elastic
recovery at time t. This may result from the fact
that such foods are plastic as well as viscoelastic,
which means they have a yield value. Therefore,
the initial deformation consists of both an instan-
taneous elastic deformation and a permanent
Fig. 8.14 (a) Voigt-Kelvin, (b) Maxwell, and (c) Burgers deformation (viscous flow component). It has
models also been found (deMan et al. 1985) that the mag-
nitude of the instantaneous elastic recovery in fat
Kelvin model employs a spring and dashpot in products is time-dependent and decreases as the
parallel; the Maxwell model employs a spring time of application of the load increases. It
and dashpot in series (Fig. 8.14). In the Voigt-­ appears that the fat crystal network gradually col-
Kelvin body, the stress is the sum of two compo- lapses, as the load remains on the sample.
nents, where one is proportional to the strain and
the other to the rate of shear. Because the ele-
ments are in parallel, they must move together. In The Plastic Body
the Maxwell model, the deformation is composed
of two parts—one purely viscous, the other A plastic material is defined as one that does not
purely elastic. Although both the Voigt-Kelvin undergo a permanent deformation, until a certain
and Maxwell bodies represent viscoelasticity, yield stress has been exceeded. A perfectly plastic
they react differently in relaxation and creep body showing no elasticity would have the stress-
experiments. When a constant load is applied in a strain behavior depicted in Fig. 8.16. Under the
creep test to a Voigt-Kelvin model, a final steady-­ influence of a small stress, no deformation occurs;
state deformation is obtained, because the com- when the stress is increased, the material will sud-
pressed spring element resists further movement. denly start to flow at applied stress σo (the yield
The Maxwell model will give continuing flow stress). The material will then continue to flow at
under these conditions, because the viscous ele- the same stress, until this is removed; the material
ment is not limited by the spring element. When retains its total deformation. In reality, few bodies
the load is removed, the Voigt-Kelvin model are perfectly plastic; rather, they are plasto-elastic
recovers completely, but not instantaneously. The or plasto-­ viscoelastic. The mechanical model
Maxwell body does not recover completely, but, used to represent a plastic body, also called a
340 8 Texture

Fig. 8.15  Creep curve for an ideal viscoelastic body, and creep curve for butter

Fig. 8.16  Stress-strain curve for an ideal plastic body

St. Venant body, is a friction element. The model


is analogous to a block of solid material that rests
Fig. 8.17  Mechanical models for a plastic body: (a) St.
on a flat horizontal surface. The block will not Venant body, (b) plasto-elastic body, and (c) plasto-­
move, when a force is applied to it, until the force viscoelastic or Bingham body
exceeds the friction existing between block and
surface. The models for ideal plastic and plasto-­
elastic bodies are shown in Fig. 8.17a, b. creep experiment with stress not exceeding the
A more common body is the plasto-­viscoelastic, yield stress, the creep curve would be similar to the
or Bingham body. Its mechanical model is shown one for a Hookean body (Fig. 8.10b). When the
in Fig. 8.17c. When a stress below the yield stress shear stress is greater than the yield stress, the
is applied, the Bingham body reacts as an elastic strain increases with time, similar to the behavior
body. At stress values beyond the yield stress, of a Maxwell body (Fig. 8.18). Upon removal of
there are two components, one of which is con- the stress at time T, the strain decreases instanta-
stant and is represented by the friction element, neously and remains constant thereafter. The
and the other, which is proportional to the shear decrease represents the elastic component; the
rate and represents the viscous flow element. In a plastic deformation is permanent. The relationship
Different Types of Bodies 341

Fig. 8.18  Creep curve for a Bingham body subjected to a Fig. 8.20  Shear stress–shear rate diagram for a thixo-
stress greater than the yield stress tropic body. Source: from deMan and Wood (1959)

The Thixotropic Body

Thixotropy can be defined as an isothermal,


reversible, sol-gel transformation, and is a behav-
ior common to many foods. Thixotropy is an
effect brought about by mechanical action, and it
results in a lowered apparent viscosity. When the
body is allowed sufficient time, the apparent vis-
cosity will return to its original value. Such
behavior would result in a shear stress-shear rate
Fig. 8.19  Shear rate-shear stress diagrams for Bingham
bodies: (a) ideal case, and (b) practical case. The yield diagram, as shown in Fig. 8.20. Increasing shear
values are as follows: lower yield value (1), upper yield rate results in increased shear stress, up to a max-
value (2), and Bingham yield value (3) imum; after the maximum is reached, decreasing
shear rates will result in substantially lower shear
stress.
of shear rate to shear stress for a Bingham body
would have the form shown in Fig. 8.19a. When
flow occurs, the relationship between shear stress Dynamic Behavior
and shear rate is given by
Viscoelastic materials are often characterized by

σ − σ o = UD
their dynamic behavior. Because viscoelastic
where materials are prone to structural breakdown when
σo = yield stress subjected to large strains, it is useful to analyze
U = proportionality constant them by small amplitude sinusoidal strain. The
D = mean rate of shear relationship between stress and strain under these
conditions can be evaluated from Fig. 8.21 (Bell
The constant U can be termed plastic viscos- 1989). The applied stress is alternating at a
ity, and its reciprocal 1/U is referred to as mobil- selected frequency and is expressed in cycles s−1,
ity. In reality, plastic materials are more likely to or ω in radians s−1. The response of a purely elas-
have a curve similar to the one in Fig. 8.19b. The tic material will show a stress and strain response
yield stress or yield value can be taken at three that is in phase, with the phase angle δ = 0°. A
different points: the lower yield value, at the purely viscous material will show the stress being
point where the curve starts on the stress axis; the out of phase by 90°, and a viscoelastic material
upper yield value, where the curve becomes will show intermediate behavior, with δ between
straight; and the Bingham yield value, which is 0° and 90°. The viscoelastic dynamic response is
found by extrapolating the straight portion of the composed of an in-phase component (sin ωt) and
curve to the stress axis. an out-of-phase component (cos ωt). The energy
342 8 Texture

Rheology Applications in Foods

Rheology has multiple applications in consider-


ation of food acceptability, food processing, and
food handling (Barbosa-Canovas et al. 1996).
Foods are complex materials often composed of
mixtures of solids and fluids (Finney 1972).
Rheology measures the flow and deformation of
substances in the transient region between solids
and fluids. Rheology defines the relationship
between the stress applied to a given material and
the resulting deformation and/or flow. Rheological
instruments measure the force and deformation
of materials as a function of time.
Fundamental rheological methods account for
the magnitude and direction of forces and defor-
mations, placing restrictions on acceptance of
sample shapes and compositions. Fundamental
tests have the advantage of being based on known
concepts and equations of physics. Empirical
methods are applied when sample composition or
geometry is too complex to account for forces
and deformations. Empirical methods correlate
with a particular property of interest, whereas
fundamental tests determine fundamental physi-
cal properties. Rheology assesses the responses
of materials to applied forces and deformations.
Basic concepts of stress (force per area) and
strain (deformation per length) are key to all rhe-
ological evaluations. Stress (τ) is always a mea-
surement of force per unit of surface area and is
expressed in units of Pascals (Pa). The direction
of the force with respect to the impacted surface
area determines the type of stress. Normal stress
Fig. 8.21  Dynamic (oscillation) measurement of visco- occurs when the force is directly perpendicular to
elastic materials. As an oscillating strain is applied, the a surface, and can be achieved during tension or
resulting stress values are recorded; δ is the phase angle,
and its value indicates whether the material is viscous, compression. Shear stress occurs when the forces
elastic, or viscoelastic. Source: reprinted from Bell (1989) act parallel to a surface (Tabilo-Munizaga and
Barbosa-Canovas 2005). Many of the rheological
properties of complex biological materials are
time-dependent, and Mohsenin (1970) has sug-
used for the viscous component is lost as heat; gested that many foods can be regarded as visco-
that used for the elastic component is retained as elastic materials. Many foods are disperse
stored energy. This results in two moduli, the systems of interacting non-spherical particles and
storage modulus (G') and the loss modulus (G''). show thixotropic behavior. Such particles may
The ratio of the two moduli is known as tan δ and interact to form a three-dimensional network that
is given by tan δ = G''/G'. imparts rigidity to a system. The interaction may
Rheology Applications in Foods 343

Fig. 8.22 Thixotropic
hardness change in
butter: (a) freshly
worked butter left
undisturbed for 4 weeks
at 5 °C; (b) the same
butter stored at −20 °C
for 3 weeks, then left at
5 °C; (c) the same butter
left at 5 °C for 3 weeks,
then frozen for 3 weeks,
and again placed at 5 °C.
Source: from deMan and
Wood (1959)

be the result of ionic forces in aqueous systems, is more difficult, because single-point determina-
or of hydrophobic or van der Waals interactions tions (i.e., at one single shear stress) will yield no
in systems that contain fat crystals in liquid oil useful information. We can visualize the rate of
(e.g., butter, margarine, and shortening). shear dependence for Newtonian fluids by con-
Mechanical action, such as agitation, kneading, sidering a diagram of two fluids, as shown in
or working, results in disruption of the network Fig. 8.23 (Sherman 1973). The behavior of these
structure and a corresponding loss in hardness. fluids is represented by two straight lines parallel
When such a system is then left undisturbed, the to the shear-rate axis. With non-Newtonian flu-
bonds between particles will reform, and hard- ids, a situation as shown in Fig. 8.24 may arise.
ness will increase with time, until maximum The fluids 3 and 4 have curves that intersect.
hardness is reached. The nature of thixotropy was Below this point of intersection, fluid 4 will appear
demonstrated for butter by deMan and Wood more viscous; beyond the intersection, fluid 3 will
(1959). Hardness of freshly worked butter was appear more viscous. Fluids 5 and 6 do not inter-
determined over a period of 3 weeks (Fig. 8.22). sect, and the problem does not arise. In spite of the
The same butter was frozen and then removed possibility of such problems, many practical appli-
from frozen storage after 3 weeks. No thixotropic cations of rheological measurements for non-
change had occurred with the frozen sample. The Newtonian fluids are carried out at only one shear
freezing had completely immobilized the crystal rate. Note that results obtained in this way should
particles. Thixotropy is important in many food be interpreted with caution. Shoemaker et al.
products; great care must be exercised, so that (1987) have given an overview of the application
measurements are not influenced by thixotropic of rheological techniques for foods.
changes. Probably the most widely used type of vis-
The viscosity of Newtonian liquids can be cometer in the food industry is the Brookfield
measured simply, by one-point determinations rotational viscometer. An example of this instru-
with viscometers, such as rotational, capillary, or ment’s application to a non-Newtonian food
falling-ball viscometers. For non-Newtonian product was given in the work of Saravacos and
materials, measurement of rheological properties Moyer (1967) on fruit purees. Viscometer scale
344 8 Texture

Fig. 8.23  Rate of shear


dependence of the
viscosity of two
Newtonian fluids.
Adopted from Sherman
(1973)

Fig. 8.24  Rate of shear


dependence of the
apparent viscosity of
several non-Newtonian
fluids. Adopted from
Sherman (1973)
Rheology Applications in Foods 345

Fig. 8.25 Apparent
viscosities of fruit
purees determined at
86 °C. Modified from
Saravacos and Moyer
(1967)

readings were plotted against rotational speed Apparent viscosities for fruit purees, deter-
on a logarithmic scale, and the slope of the mined in this manner, are shown in Fig. 8.25.
straight line obtained was taken as the exponent Factors have been reported in the literature
n in the following equation for pseudoplastic (Johnston and Brower 1966) for the conversion
materials: of Brookfield viscometer scale readings to yield
values or viscosities. Saravacos (1968) has also
τ = K γ n
used capillary viscometers for rheological mea-
where surements of fruit purees.
τ = shear stress (dyne/cm2) For products not sufficiently fluid to be stud-
K = constant ied with viscometers, a variety of texture-­
γ = shear rate (s−1) measuring devices is available. These range from
simple penetrometers, such as the Magness-­
The instrument readings were converted into Taylor fruit pressure tester, to complex universal
shear-stress values by using an oil of known vis- testing machines, such as the Instron. All these
cosity. The shear rate at a given rotational speed instruments either apply a known and constant
N was calculated from stress and measure deformation, or cause a con-
stant deformation and measure stress. Some of

γ = 4π N n
the more sophisticated instruments can do both.
When shear stress τ was plotted against shear In the Instron Universal Testing Machine, the
rate γ on a double-logarithmic scale, the intercept crosshead moves at a speed that can be selected
of the straight line on the τ axis at γ = 1 s−1 was by changing gears. The drive is by rotating
taken as the value of the constant K. The apparent screws, and the force measurement is done with
viscosity μapp at a given shear rate was then calcu- load cells. Mohsenin (1970) and coworkers have
lated from the equation developed a type of universal testing machine, in
which the movement is achieved by air pressure.
µapp = K γ n −1
The Kramer shear press uses a hydraulic system
for movement of the crosshead.
346 8 Texture

Texture-measuring instruments can be classi- The advantage of this conversion is that


fied according to their use of penetration, com- changes in hardness are more uniform than
pression, shear, or flow. changes in penetration depth. With the latter, a
Penetrometers come in a variety of types. One difference of an equal number of units, at the tip
of the most widely utilized is the Precision pene- of the cone and higher up on the cone, is not at all
trometer, which is used for measuring the consis- comparable.
tency of fats. The procedure and cone dimensions Many penetrometers use punches of various
are standardized and described in the Official shapes and sizes as penetrating bodies. Little
Methods and Recommended Practices of the was known about the relationship between
American Oil Chemists’ Society. According to shape and size and penetrating force until
this method, the results are expressed in mm/10 Bourne’s (1966) work. He postulated that when
of penetration depth. Haighton (1959) proposed a punch penetrates a food, both compression
the following formula for the conversion of pen- and shear occur. Shear, in this case, is defined
etration depth into yield value: as the movement of interfaces in opposite direc-
tions. Bourne suggested that compression is
C = K W P1.6 proportional to the area under the punch and to
the compressive strength of the food, and also
where that the shear force is proportional to the perim-
C = yield value eter of the punch and to the shear strength of
K = constant dependent on the angle of the cone the food (Fig. 8.26). The following equation
p = penetration depth was suggested:
W = weight of cone
F = Kc A + Ks P +C

Vasic and deMan (1968) suggested a conver-
sion of the penetration-depth readings into hard- where
ness, using the formula F = measured force
Kc = compression coefficient of the tested food
H=G A Ks = shear coefficient of the tested food

where A = area of the punch
H = ha.rdness P = perimeter of the punch
G = total weight of the cone assembly C = constant
A = area of impression

Fig. 8.26  Compression and shear components in penetration tests. Adopted: from Bourne (1966)
Rheology Applications in Foods 347

The relationship between penetration force and shear press was designed to be a versatile and
the cross-sectional area of a cylindrical punch has widely applicable instrument for texture measure-
been established by Kamel and deMan (1975). ment on a variety of products. The shear press is
Bourne (1966) showed that, for a variety of essentially a hydraulically driven piston, to which
foods, the relationships between punch area and a standard ten-blade shear cell or a variety of other
force and between punch perimeter and force specialized devices can be attached. Force mea-
were represented by straight lines. deMan (1969) surement is achieved either by a direct-­reading
later showed that, for certain products such as proving ring or by an electronic recording device.
butter and margarine, the penetrating force was The results obtained with the shear press are influ-
dependent only on area and was not influenced enced by the weight of the sample and the speed
by perimeter. deMan suggested that, in such of the crosshead. These factors have been exhaus-
products, flow is the only factor affecting force tively studied by Szczesniak et al. (1970). The
readings. It appears that useful conclusions can relationship between maximum force value and
be drawn, regarding the textural characteristics of sample weight was found to be different for dif-
a food, by using penetration tests. ferent foods. Products fell into three categories:
A variation on the penetration method is the those having a constant force-­ to-­
weight ratio
back-extrusion technique, where the sample is (e.g., white bread, sponge cake); those having a
contained in a cylinder, and the penetrating body continuously decreasing force-to-­ weight ratio
leaves only a small annular gap for the product to (e.g., raw apples, cooked white beans); and those
flow. The application of the back-extrusion giving a constant force value, independent of
method to non-Newtonian fluids has been sample weight beyond a certain fill level (e.g.,
described by Steffe and Osorio (1987). canned beets, canned and frozen peas). This is
Many instruments combine shear and com- demonstrated by the curves in Fig. 8.27. Some of
pression testing. One of the most widely used is the available attachments to the shear press are the
the Kramer shear press. Based on the principle of succulometer cell, the single-­ blade meat shear
the shear cell used in the pea tenderometer, the cell, and the compression cell.

Fig. 8.27  Effect of


sample weight on
maximum force
registered with the shear
press and using the
ten-blade standard cell:
(1) white bread and
sponge cake, (2) raw
apples and cooked white
beans, (3) canned beets
and peas and frozen
peas. Adopted from
Szczesniak (1973)
348 8 Texture

Fig. 8.28 Typical
texturometer curve

Based on the Szczesniak classification of tex- Textural Properties of some Foods


tural characteristics, a new instrument was devel-
oped in the General Foods research laboratories; Meat Texture
it was called the General Foods Texturometer.
This device was an improved version of the MIT Meat texture is usually described in terms of ten-
denture tenderometer (Proctor et al. 1956). From derness or the lack of it—toughness. This obvi-
the reciprocating motion of a deforming body on ously is related to the ease with which a piece of
a sample, which was contained in a tray equipped meat can be cut with a knife or with the teeth. The
with strain gages, a force record called a texture oldest and most widely used device for measuring
profile curve (Fig. 8.28) was obtained. From this meat tenderness is the Warner-Bratzler shear
texturometer curve, a variety of rheological device (Bratzler 1932). In this device, a cylindri-
parameters can be obtained. Hardness is mea- cal core of cooked meat is subjected to the shear-
sured from the height of the first peak. ing action of a steel blade, and the maximum
Cohesiveness is expressed as the ratio of the force is indicated by a spring-loaded mechanism.
areas under the second and first peaks. Elasticity A considerable improvement was provided by the
is measured as the difference between distance B, shear apparatus described by Voisey and Hansen
measured from initial sample contact to sample (1967). In this apparatus, the shearing force is
contact on the second “chew,” and the same dis- sensed by a strain-gage transducer, and a com-
tance (distance B) measured for a completely plete shear force-time curve is recorded on a strip
inelastic material such as clay. Adhesiveness is chart. The Warner-Bratzler shear method has sev-
measured as the area of the negative peak, A3, eral disadvantages. It is very difficult to obtain
beneath the baseline. In addition, other parame- uniform meat cores. Cores from different posi-
ters can be derived from the curve, such as brittle- tions in one cut of meat may vary in tenderness,
ness, chewiness, and gumminess. and cooking method may affect tenderness.
Textural Properties of some Foods 349

Meat tenderness has been measured with the


Kramer shear press. This can be done with the
ten-blade universal cell or with the single-blade
meat-shear attachment. There is no standard pro-
cedure for measuring meat tenderness with the
shear press; sample size, sample preparation, and
shear rate are factors that may affect the results.
A pressure method for measuring meat ten-
derness has been described by Sperring et al.
(1959). A sample of raw meat is contained in a
cylinder that has a small hole in its bottom. A
hydraulic press forces a plunger into the cylinder,
and the pressure required to squeeze the meat
through the hole is taken as a measure of
tenderness.
A portable rotating-knife tenderometer has
been described by Bjorksten et al. (1967). A rotat-
ing blunt knife is forced into the meat sample, and
a tracing of the area traversed by a recording pen is
used as a measure of tenderness.
A meat-grinder technique for measuring meat
Fig. 8.29  Mechanical model for postmortem striated
tenderness was reported by Miyada and Tappel muscle. Source: from C.W. Brabender Instruments, Inc.,
(1956); in this method, power consumption of the South Hackensack, New Jersey
meat-grinder motor was used as a measure of
meat tenderness. The electronic-recording food
grinder described by Voisey and Voisey and Wheat Flour Dough
deMan (1970) measured the torque exerted on a
strain-gage transducer. This apparatus was used The rheological properties of wheat flour dough
successfully for measuring meat tenderness. are important in determining the baking quality of
Other methods used for meat tenderness eval- wheat flour. For many years, the Farinograph was
uation have included measurement of sarcomere used to measure the physical properties of wheat
length (Howard and Judge 1968) and determina- flour dough. The Farinograph is a dough mixer
tion of the amount of connective tissue present. hooked up to a dynamometer for recording torque.
Stoner et al. (1974) proposed a mechanical The instrument can measure the water absorption
model for postmortem striated muscle; it is of a wheat flour. A typical Farinograph curve is
shown in Fig. 8.29. The model is a combination presented in Fig. 8.30. The amount of water,
of the Voigt model with a four-element viscoelas- added from a buret, required to bring the middle
tic model. The former includes a contractile ele- of the curve to 500 units is a measure of the water
ment (CE), which is the force generator. The absorption of a wheat flour dough. The measure-
element SE is a spring that is passively elongated ment from zero to the point where the top of the
by the shortening of the CE, and thus develops an curve first intersects the 500-unit line on the chart
internal force. The parallel elastic component is called the arrival time; the measurement from
(PE) contributes to the resting tension of the mus- zero to maximum consistency is called the peak
cle. The combination of elements PE, CE, and SE time; the point where the top of the curve leaves
represents the purely elastic properties of the the 500-unit line is called the departure time; and
muscle as the fourth component of a four-element the difference between the departure and arrival
model (of which E2, η3, and η2 are the other three times is taken as the stability. The elasticity of a
components). wheat flour dough is measured with the
350 8 Texture

Fig. 8.30  Typical Farinograph curve: (A) arrival time, (B) peak time, (C) stability, (D) departure, (E) mixing tolerance
index, (F) 20-min drop. Source: from Wehrli and Pomeranz (1969)

Extensigraph, which records the force required to flour is strongly influenced by its gluten protein
stretch a piece of dough of standard dimensions. quality and content and the disulfide/sulfhydryl
The unique viscoelastic properties of wheat ratio. A schematic diagram of the bonds within
flour dough are the result of the presence of a and between polypeptide chains in a wheat flour
three-dimensional network of gluten proteins. dough is given in Fig. 8.31.
The network is formed by thiol-disulfide
exchange reactions among gluten proteins.
Peptide disulfides can interfere in a thiol-­disulfide Fats
exchange system, by reacting with a protein
(PR)-thiol to liberate a peptide (R)-thiol and form The consistency of fats is commonly determined
a mixed disulfide, as follows: with a cone penetrometer, as specified in the
Official Methods and Recommended Practices of
PR – SH + R – SS – R → R – H + PR – SS – R the American Oil Chemists’ Society (Method Cc
Disulfide bonds between proteins have an 16–60). Other methods that have frequently been
energy of 49 kcal/mol and are not broken at room employed involve extrusion; they include the
temperature, except as the result of a chemical extrusion attachment to the Kramer shear press
reaction. The effects of oxidizing agents on the (Vasic and deMan 1967), an extrusion rheometer
rheological properties of a wheat flour dough used with the Instron Universal Testing Machine
may be quantitatively explained in terms of the (Scherr and Wittnauer 1967), and the FIRA-­
breaking of disulfide cross-links; their reforma- NIRD extruder (Prentice 1954).
tion may be explained as due to exchange reac- Other devices used for fat-consistency mea-
tions with sulfhydryl groups (Wehrli and surements include wire-cutting instruments (sec-
Pomeranz 1969). The baking quality of a wheat tilometers), penetration of a probe when a sample
Textural Properties of some Foods 351

Fig. 8.31 Schematic diagram of bonds within and sulfhydryl group, (3) intermolecular disulfide bond, (4)
between polypeptide chains in a wheat flour dough. Solid ionic bond, (5) van der Waals bond, (6) interpeptide
lines represent covalent bonds; dotted lines represent hydrogen bond, (7) side-chain hydrogen bond. Source:
other bonds: (1) intramolecular disulfide bond, (2) free from Bloksma (1972)

Fig. 8.32  Examples of


compression curves for
shortenings: (1) and (2)
soy-palm; (3) soy-­
canola-­palm; (4) soy
only; (5) tallow-lard; (6)
lard; (7) palm-vegetable;
(8) palm-palm kernel;
a = elastic non-­
recoverable deformation;
b = viscous flow;
B = breaking force;
P = plateau force;
distance between B and
P is an indication of
brittleness

is contained in a small cup, and compression of a brittleness. These characteristics are important in
cylindrical sample between two parallel plates. shortenings destined for use in cakes and puff
The compression method reveals detailed infor- pastries. Compression curves for a variety of
mation about plastic fats (deMan et al. 1991), shortenings are displayed in Fig. 8.32.
including elasticity, viscous flow, and degree of Temperature treatment of a fat has a profound
352 8 Texture

effect on its texture. deMan and deMan (1996)


studied the effects of crystallization temperature
and tempering temperature on the textures of
palm oil and hydrogenated fats, using the com-
pression method, and found that lowering the
crystallization temperature from 10 to 0 °C
resulted in softer textures, especially for palm oil.
Increasing the tempering temperature from 25° to
30 °C also resulted in softer textures, especially
for hydrogenated fats.
The hardness or consistency of fats is a result
of the presence of a three-dimensional network Fig. 8.33  Mechanical model for foods as viscoplastic
of fat crystals. All fat products, such as marga- materials. Source: from Tanaka et al. (1971)
rine, shortening, and butter, are mixtures of
solid fat, in crystallized form, and liquid oil. viscoplastic materials. The model consists of a
Because the individual glycerides in fats have a dashpot representing the viscous element, in par-
wide range of melting points, the ratio of solid- allel with a friction element representing the
to-liquid fat is highly temperature-dependent. yield value.
The crystal particles are linked by weak van der The theory of bond formation between the
Waals forces. These bonds are easily broken by crystal particles in plastic fats needs revision. It
mechanical action during processing, and the has been proposed that a process of “sinter-
consistency may be greatly influenced by such ing”—the formation of solid bridges between fat
mechanical forces. After a rest period, some of crystals—occurs during post-crystallization
these bonds are reformed, and the reversible hardening (Heertje et al. 1987; Johansson and
sol-gel transformation taking place is called Bergenstahl 1995). The use of the word sintering
thixotropy. There appear to be two types of is unfortunate, since it ordinarily describes the
bonds in fats—those that are reformed after fusion of small particles into a solid block; this is
mechanical action, and those that do not reform. not the case, however, because the fat crystals in
The latter result in a portion of the hardness fats can be suspended in a solvent such as isobu-
loss that is irreversible. The nature of these tanol (Chawla and deMan 1990), without show-
bonds has not been established with certainty, ing any sign of fusion into larger aggregates. At
but it is assumed to mainly involve van der the solid-fat level found in plastic fats (15–35%),
Waals forces. The hardness loss of fats, as a the crystals are tightly packed together and are
result of working, is called work softening and suggested to exist in a state of entanglement
can be expressed as follows: (deMan 1997). Entanglement of crystals is a
more realistic description of the structure forma-
H o − Hw tion process in fats. The sintering process
WS = ×100%
Ho described above can be considered to be a form

of entanglement.
where Ho and Hw are the hardness before and Many interrelated factors influence the texture
after working. of plastic fats. Fatty acid and glyceride composi-
The work softening is influenced not only by tion are basic factors in establishing the proper-
the nature of the mechanical treatment, but also ties of a fat. These factors, in turn, are related to
by the temperature conditions and the size and solid-fat content, crystal size and shape, and
quantity of fat crystals. polymorphic behavior. Once a crystal network is
Tanaka et al. (1971) used a two-element formed, mechanical treatment and temperature
mechanical model (Fig. 8.33) to represent fats as history may influence the texture.
Textural Properties of some Foods 353

The network systems in plastic fats differ from


those in protein or carbohydrate systems. Fat
crystals are embedded in liquid oil, and the crys-
tals have no ionized groups. Therefore, the inter-
active forces in fat crystal networks are weak.
The minimum concentration of solid particles in
a fat, required to provide a yield value, is in the
range of 10–15%.

Fruits and  Vegetables

Much of the texture work with fruits and vegeta-


bles has been done with the Kramer shear press.
The shear press was developed because of the
tenderometer’s limitations, and has been widely
used for measuring the tenderness of peas for
processing (Kramer 1961). The shear press has
also been used, for example, in the quality method
suggested for raw and canned sweet com (Kramer
and Cooler 1962). This procedure determines
shear force with the standard shear cell, and the
amount of juice pressed out with the juice extrac-
tion cell. It is possible to relate the quality of corn Fig. 8.34  Mechanical model proposed by Morrow and
to these parameters. In addition to the shear press, Mohsenin: (a) viscoelastic foods, (b) the strain-time, and
the Instron Universal Testing Machine and others (c) stress-time, characteristics of the system
based on the same principle are popular for use
on fruit and vegetable products. A special testing Morrow and Mohsenin (1966) have studied
machine has been developed by Mohsenin the physical properties of a variety of vegetables;
(1970). This machine uses an air motor for move- they assumed these products to behave as vis-
ment of the crosshead, but is otherwise similar to coelastic materials and thus according to the
other universal testing machines. A mechanically three-­element model represented in Fig. 8.34a.
driven test system has been developed by Voisey Such viscoelastic materials are characterized by
(1971). Voisey (1970) has also described a num- the strain-time and stress-time relationships
ber of test cells that are simpler in design than the shown in Fig. 8.34b, c, respectively.
standard shear cell of the Kramer shear press.
The texture of fruit and vegetable products is
related to the cellular structure of these materials. Starch
Reeve and Brown (1968a, b) studied the develop-
ment of cellular structure in the green bean pod, as The texture of starch suspensions in water is
it relates to texture and eating quality. Sterling determined by the source of the starch, the chem-
(1968) studied the effect of solutes and pH on the ical and/or physical modification of the starch
structure and firmness of cooked carrot. Sterling granules, and the cooking conditions of the starch
also related histological changes, such as cellular (Kruger and Murray 1976). The texture of starch
separation and collapse, to product texture. In fruits suspensions can be measured by means of a vis-
and vegetables, the relationship between physical coamylograph. The viscosity is recorded, while
structure and physical properties is probably more the temperature of the suspension is raised from
evident than in many other types of products. 30° to 95 °C, held at 95 °C for 30 min, lowered to
354 8 Texture

30° 95° 95° 25° 25°C


VISCOSITY POISES

10
A

C
0

a b c d

Fig. 8.35  Viscosity and granule appearance in a vis-


coamylograph test of a 5% suspension of waxy corn
starch in water: A = viscosity curve, B = starch granule
Fig. 8.36  Viscosity and granule appearance in a vis-
shape and size, C = magnified portion of curve to indicate
coamylograph test of a 5% aqueous suspension of waxy
cohesiveness; a = unswollen granule, b = swollen granule,
corn starch with 1 cross-bond per 100,000 glucose units:
c = collapsed granule, d = entwined collapsed granules.
A = viscosity curve, B = granule appearance. Source:
Source: reprinted from Kruger and Murray (1976)
reprinted from Kruger and Murray (1976)

25 °C, and held at that temperature for 30 min. is decreased. Most food starches, used at pH val-
The viscosity of a 5% suspension of waxy corn in ues from 4 to 8, have 2 to 3 cross-bonds per
water is shown in Fig. 8.35. Initially, the viscosity 10,000 glucose units.
is low, but it increases rapidly at the gelatiniza- Waxy corn starch contains only amylopectin;
tion temperature of about 73 °C. As the granules normal corn starch contains both amylopectin
swell, they become more fragile and start to dis- and amylose. This results in different viscosity
integrate, causing the viscosity to drop. When the profiles (Fig. 8.38). Normal corn starch shows a
temperature is lowered to 25 °C, there is another lower peak viscosity and less breakdown during
increase in viscosity, caused by the interaction of heating. After cooling, the viscosity continues to
the broken and deformed granules. This phenom- increase, possibly because the amylose interlinks
enon is demonstrated by the width and irregular- with the amylopectin. On further storage at
ity of the recorded line, which is indicative of the 25 °C, the slurry sets to a firm gel. Tapioca starch
cohesiveness of the starch particles. shows behavior intermediate between that of nor-
Modification of a starch has a profound effect mal corn starch and waxy corn starch (Fig. 8.38).
on the texture of its suspensions. Introduction of This could be explained by the fact that tapioca
as little as 1 cross-bond per 100,000 glucose units amylose molecules are larger than those in nor-
slows the breakdown of the swollen granules dur- mal corn starch.
ing and after cooking (Fig. 8.36). This results in a Starches can be modified with non-ionic or
higher final viscosity. Increasing the cross-­ ionic groups. The latter can be made anionic by
bonding to 1, 3, or 6 cross-bonds per 10,000 glu- the introduction of phosphate or succinate groups.
cose units results in no breakdown during the Such modified starches have lower gelatinization
heating cycle (Fig. 8.37). As the cross-bonding temperature, higher peak viscosity, and higher
increases, the granule is strengthened and does final cold viscosity than non-ionically modified
not swell much during heating, but the viscosity starches (Fig. 8.39).
Microstructure 355

Fig. 8.37  Viscosity and granule appearance in a vis-


coamylograph test of a 5% aqueous suspension of cross-­
bonded waxy corn starch: A = 1 cross-bond per 10,000 Fig. 8.38  Viscosity and granule appearance in viscoam-
glucose units, B = 3 cross-bonds, C = 6 cross-bonds, and ylograph tests of aqueous suspensions of corn and tapioca
D = starch granule appearance. Source: reprinted from starches: A = 6% corn starch, B = 5% tapioca starch,
Kruger and Murray (1976) C = corn starch granule appearance, and D = tapioca
starch granule appearance. Source: reprinted from Kruger
and Murray (1976)

Microstructure associate to form network structures that may


entrap large volumes of the continuous phase.
With only a few exceptions, food products are The isothermal, reversible sol-gel transformation
non-Newtonian and possess a variety of internal exhibited by many foods is called thixotropy.
structures. Cellular and fibrous structures are Disperse systems can be classified on the basis of
found in fruits and vegetables; fibrous structures particle size. Coarse dispersions have particle
are found in meats; and many manufactured sizes greater than 0.5 μm. Their particles can be
foods contain protein, carbohydrate, or fat-­crystal seen in a light microscope, can be filtered over a
networks. paper filter, and will sediment rapidly. Colloidal
Many such food systems are dispersions that dispersions have particle sizes in the range of
fall within the realm of colloids. Colloids are 0.5 μm to 1 nm. Such particles remain in suspen-
defined as heterogeneous or dispersed systems sion by Brownian movement, and can run through
that contain at least two phases—a dispersed a paper filter, but cannot run through a membrane
phase and a continuous phase. Colloids are char- filter. Particles smaller than these are molecular
acterized by their ability to exist in either a sol or dispersions or solutions. Depending on the nature
a gel form. In the former, the dispersed particles of the two phases, disperse systems can be classi-
exist as independent entities; in the latter, they fied into a number of types. A solid dispersed in a
356 8 Texture

An important aspect of the subdivision of a


disperse phase is the enormous increase in spe-
cific surface area. If a sphere with a radius
R = 1 cm is dispersed into particles with radius
r = 10−6 cm, the area of the interface will increase
by a factor of 106. The mechanical work, dA,
needed to increase the interfacial area is propor-
tional to the area increase, as follows:

dA = σ dO
where O = total interfacial area. The proportional-
ity factor σ is the. surface tension. In the production
of emulsions, the surface tension is reduced
by using surface-active agents (see Chap. 2).
As particle size is reduced to colloidal dimen-
sions, the particles become subject to Brownian
movement. Brownian movement is a result of the
random thermal movement of molecules, which
impact on colloidal particles to give them a ran-
dom movement as well (Fig. 8.40). The size of
dispersed particles has a profound effect on the
properties of dispersions (Schubert 1987).
Fig. 8.39  Viscoamylograph viscosity curves for modified Figure  8.41 shows the qualitative relationships
waxy corn starches: A = cross-bonded waxy corn starch, between particle size and system properties. As
B = non-ionically modified cross-bonded waxy corn particle size decreases, fracture resistance
starch, and C = anionically modified cross-bonded waxy increases. The particles become increasingly uni-
corn starch. Source: reprinted from Kruger and Murray
(1976) form, which results in a grinding limit, below
which particles cannot be further reduced in size.
The terminal settling rate, illustrated by a flour
liquid is called a sol; for example, margarine, particle falling through air, increases rapidly as a
which has solid-fat crystals dispersed in liquid function of increasing particle size. According to
oil, is a sol. Dispersions of liquid in liquid are Schubert (1987), a flour particle of 1 μm in size
called emulsions; many examples of these are takes more than 6 h to fall a distance of 1 m in
found among foods, such as milk and mayon- still air. Wetting becomes more difficult as parti-
naise. Dispersions of gas in liquid are called cle size decreases. The specific surface area (the
foams (e.g., whipped cream). In many cases, surface per unit volume) increases rapidly with
such dispersions are more complex, and have decreasing particle size.
more than one disperse phase. In fact, many Colloidal systems, because of their large num-
foods have several dispersed phases. For instance, ber of dispersed particles, show non-Newtonian
in chocolate, both solid cocoa particles and fat flow behavior. For a highly dilute dispersion of
crystals are dispersed phases. spherical particles, the following equation was
The production of disperse systems is often proposed by Einstein:
achieved by dispersion methods, in which the
η = ηo (1 + 2.5φ )
disperse phase is subdivided into small particles
by mechanical means. Liquids are emulsified by
stirring and homogenization; solids are subdi- where
vided by grinding, as, for instance, roller mills ηo = viscosity of the continuous phase
are used in chocolate making and colloid mills phi = ratio of the volumes of the disperse and
are used in other food preparations. continuous phases
Microstructure 357

Fig. 8.40 Flat-plane
projection of the
location of a colloidal
particle subject to
Brownian movement.
Source: from Schubert
(1987)

Fig. 8.41  Relationships between particle size and system Sv = surface area per unit volume, W = particle weight,
properties: D = particle deposition in fibrous fillers, Vg = terminal setting rate, σs  = particle fracture resis-
F = adhesion force, H = homogeneity of a particle, tance. Source: from Schubert (1987)

In this equation, viscosity is independent of


the degree of dispersion. As soon as the ratio of
disperse and continuous phases increases to the
point where particles start to interact, the flow
behavior becomes more complex. The effect of
increasing the concentration of the disperse phase
on the flow behavior of a disperse system is
shown in Fig. 8.42. The disperse phase, as well as
the low-solids dispersion (curves 1 and 2), shows
Newtonian flow behavior. As the solids content
increases, the flow behavior becomes non-­
Newtonian (curves 3 and 4). Especially with
anisotropic particles, interactions between them
Fig. 8.42  Effect of increasing the concentration of a dis-
will result in the formation of three-dimensional perse phase on the flow behavior of a disperse system:
network structures. These network structures 1—continuous phase, 2—low solids content, 3—medium
usually show non-Newtonian flow behavior and solids content, 4—high solids content
viscoelastic properties, and often have a yield
value. Network structure formation may occur in result in the formation of networks, may be van
emulsions (Fig. 8.43), as well as in particulate der Waals forces, hydrophobic interactions, or
systems. The forces between particles, which covalent bonds. Network formation may result
358 8 Texture

Fig. 8.43 Structure
formation in particulate
systems: (a) flocculation
of an emulsion, (b)
network formation in
crystallized fat

Table 8.3  Relationship between critical particle fraction


(αc) and number of bond sites (f)
αc f
0.05 21
0.10 11
0.15 8
0.20 6
0.30 4
0.50 3

from heating or from chemical reactions that


occur spontaneously, involving either compo-
nents already present in a food or added enzymes
or coagulants. The formation of networks requires Fig. 8.44  Microstructure of soybean curd (tofu), as seen
a minimal fraction of particles to be present, the in a scanning electron microscope
critical fraction αc, and the larger the number of
sites f, used for bond formation, the sooner a net- ture formation in acidic milk gels and found
work will form. These two quantities are related that the final texture of such products was influ-
as follows: enced by many factors, including heat, salt bal-
ance, pH, culture, and thickening agents.
α c = 1 / ( f − 1)
Structure formation in soy milk, induced by
coagulants in the form of calcium or magne-
At particle concentrations below 10%, sium salts, results in a semi-solid food called
numerous contact points are required to form a tofu, which has a fine internal protein network
network structure (Table 8.3). This means that structure (Fig. 8.44). Hermansson and Larsson
only certain types of molecules or particles can (1986) reported on the structure of gluten gels
form networks at such concentrations. As a net- and concluded that such gels consist of a con-
work is formed, the viscosity increases, until, at tinuous phase of densely packed protein units.
a certain point, the system acquires plastic and/ DeMan and Beers (1987) reviewed the factors
or viscoelastic properties. Network formation that influence the formation of three-dimensional
thus depends on particle concentration, reactive fat-crystal networks. The fat-crystal networks in
sites on the particles, and particle size and plastic fats are highly thixotropic, and mechani-
shape. Heertje et al. (1985) investigated struc- cal action on such systems will result in a drastic
Water Activity and Texture 359

Fig. 8.45  Effect of cooling rate on crystal size for milk cal-laser-microscopy-principles-and-applications-in-
fat, after 90 min at 35 °C and 24 h cooling at 10 °C/min: medicine-biology-and-the-food-sciences/
(a) slow cooling (0.1 °C/min), and (b) fast cooling applications-of-confocal-laser-scanning-microscopy-
(5.5 °C/min). http://www.intechopen.com/books/confo- clsm-in-foods

reduction of hardness. Figure 8.45 illustrates the Water Activity and Texture


influence of cooling rate on the degree of crystal-
lization in butterfat, which impacts the plasticity Water activity (aw) and water content have a pro-
of the fat. found influence on the textural properties of
A variety of rheological tests can be used to foods. The three regions of a typical sorption iso-
evaluate the nature and properties of different therm can be used to classify foods on the basis
network structures in foods. The strength of of their textural properties (Fig. 8.46). Region 3
bonds in a fat-crystal network can be evaluated is the high-moisture area, which includes many
by stress relaxation, and by the decrease in elastic soft foods. Foods in the intermediate-moisture
recovery in creep tests as a function of loading area (region 2) appear dry and firm. At the lowest
time (deMan et al. 1985). Van Kleef et al. (1978) values of aw (region 1), most products are hard
reported on a determination of the number of and crisp (Bourne 1987).
cross-links in a protein gel, from its mechanical Katz and Labuza (1981) examined the rela-
and swelling properties. Oakenfull (1984) used tionship between aw and crispness, in a study of
shear-modulus measurements to estimate the size the crispness of popcorn (Fig. 8.47). They found
and thermodynamic stability of junction zones in a direct relationship between crispness and aw.
non-covalently cross-linked gels. Many foods contain biopolymers and low-­
Dynamic measurements of gels can provide molecular-­weight carbohydrates. These materi-
information on the extent of cross-linking (Bell als can be present in a metastable amorphous
1989). Systems with a relatively high storage state that is sensitive to temperature and the state
modulus G’ show a low value for G”/G’, which of the water present. The amorphous state can
indicates a highly cross-linked system, such as exist in the form of a rubbery liquid structure or a
an agar gel. very viscous, glassy solid, as shown in Fig. 8.48
360 8 Texture

Fig. 8.46  The three


regions of a schematic
sorption isotherm, as
related to the textural
properties of food
systems. Source:
reprinted with
permission from Bourne
(1987)

Fig. 8.48 Rubbery liquid and glassy solid states of


moisture-­containing foods, as affected by temperature:
Fig. 8.47  Relationship between the water activity and the Tm = melting point; Tg = glass transition temperature
crispness of popcorn. Source: reprinted with permission
from Katz and Labuza (1981) melting temperature (Tm), the material enters a
state of rubbery liquid flow. As the temperature
(Slade and Levine 1991; Levine and Slade 1992; is lowered further, a leathery state is observed. In
Roos and Karel 1991). A more detailed analysis the leathery region, the modulus increases
of the effect of temperature on textural properties, sharply, until the glass transition temperature
expressed as modulus, is presented in Fig. 8.49 (Tg) is reached, and the material transforms to a
(Slade and Levine 1991). Below the crystalline glassy solid.
References 361

Fig. 8.49  Effect of


temperature on texture,
expressed in terms
of modulus: Tm =
crystalline melting
temperature, Tg = glass
transition temperature.
Source: from Slade and
Levine (1991)

Bratzler, L. J. (1932). Measuring the tenderness of meat


Kapsalis et al. (1970) reported on a study of by means of a mechanical shear. Master’s thesis,
the textural properties of freeze-dried beef at dif- Kansas State College.
ferent points on the moisture-sorption isotherm, Cairncross, S. E., & Sjöström, L. B. (1950). Flavor
over the entire range of water activity. Important profiles—A new approach to flavor problems. Food
Technology, 4, 308–311.
changes in textural properties were observed at Chawla, P., & deMan, J. M. (1990). Measurement of
aw values of 0.85 and at 0.15–0.30. the size distribution of fat crystals using a laser par-
ticle counter. Journal of the American Oil Chemists'
Society, 67, 329–332.
deMan, J. M. (1969). Food texture measurements with
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based on oral processing considerations. International Chapman and Hall.
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Haighton, A. J. (1959). The measurement of hardness of In H. G. Schwartzberg & R. W. Hartel (Eds.), Physical
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sarcomere length to other predictors of beef tender- and their role in weight management: A review. Food
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Journal of the American Oil Chemists' Society, 72, rials. Journal of Food Science, 31, 686–698.
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Johnston, C. W., & Brower, C. H. (1966). Rheological of shear modulus to estimate the size and thermody-
and gelation measurements of plastisols. Society of namic stability of junction zones in non-covalently
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Wissenschaft und Technologie, 8, 123–127. on texture and oil uptake of potato chips. LWT - Food
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of low and intermediate moisture foods. Journal of the spreadability of butter. Laboratory Practice, 3,
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ity on the sensory crispness and mechanical deforma- recording strain-gage denture tenderometer for foods.
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age conditions on relation between apple firmness ture. 2 structure and composition of edible maturity.
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of a capillary extrusion rheometer to the determina- shear compression cell of the shear press and the effect
tion of the flow characteristics of lard. Journal of the of sample weight on peak area and maximum force.
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Schubert, H. (1987). Food particle technology. Part 1: Szczesniak, A. S., & Kleyn, D. H. (1963). Consumer
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Instrumentation for rheological measurements of M. (1978). Determination of the number of cross-links
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Vitamins
9
John W. Finley and John M. deMan

c­ ommonly consumed foods are reported. This data


Introduction is from the USDA Nutrient data base and more
data can be found on raw, and processed foods
Vitamins are minor components of foods that including various commercial preparations.
play an essential role in human nutrition. Many Healthy levels of vitamins in the diet refers to
vitamins are unstable under certain conditions of the absence of disease based on clinical signs and
processing and storage (Table 9.1), and their lev- symptoms of deficiency or excess, and normal
els in processed foods, therefore, may be consid- function of the individual. The concept of protec-
erably reduced. Synthetic vitamins are used tive nutrient intake for some vitamins refers to an
extensively to compensate for these losses and to amount which may be protective against a speci-
restore vitamin levels in foods. The vitamins are fied health or nutritional risk, for example vitamin
usually divided into two main groups, the water-­ C intake of 25 mg with each meal to enhances iron
soluble and the fat-soluble vitamins. The occur- absorption and prevent anemia) (Cook and Reddy
rence of the vitamins in the various food groups 2001). Protective intake levels for a specific vita-
is related to their water-or fat-solubility. The con- min can be expressed either as a daily value or as
tribution of foods to vitamins in the diet is varies an amount to be consumed with a meal.
significantly between food groups and specific In Fig. 9.1 the acceptable range of intake in the
products within the food groups. Some vitamins shaded ranges correspond to approaches defining
function as part of a coenzyme, without which requirements to prevent deficit and excess. The
the enzyme would be ineffective as a biocatalyst. estimated average requirement is the average
Frequently, such coenzymes are phosphorylated daily intake required to prevent deficit in half of
forms of vitamins and play a role in the metabo- the population. The recommended nutrient intake
lism of fats, proteins, and carbohydrates. Some is the amount necessary to meet the needs of most
vitamins occur in foods as provitamins—com- (97.5%) of the population, which is defined as the
pounds that are not vitamins but can be changed set as the estimated average requirement plus two
by the body into vitamins. Vitamers are members standard deviations. The tolerable upper intake
of the same vitamin family. level is the level at which no evidence of toxicity
The contribution of vitamins by foods varies is observed. In Fig. 9.1 the risk function of defi-
greatly among food groups. Water soluble vita- ciency and excess for individuals in a population
mins are found in fruits and vegetables. Vitamin D related to food intake, assuming a Gaussian distri-
is found almost exclusively in animal sources. bution of requirements to prevent deficit and
In Table 9.2 the vitamins found in a range of avoid excess. The upper tolerable nutrient intake

© Springer International Publishing AG 2018 365


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_9
366 9 Vitamins

Table 9.1  Stability of vitamins under different conditions


Unstable to:
Vitamin Vitamer UV Light Heata O2 Acid Base Metalsb Most stable
Vitamin A Retinol + + + + Dark, seal
Retinal + + + Seal
Retinoic acid Good stability
Dehydroret. + Seal
Ret. esters Good stability
β-carotene + Seal
Vitamin D D2 + + + + + Dark, cool, seal
D3 + + + + + Dark, cool, seal
Vitamin E Tocopherols + + + + + Cool, neutral pH
Tocopherol esters + + Good stability
Vitamin K K + + + + Avoid reductants
MK + + + + Avoid reductants
Menadione + + + Avoid reductants
Vitamin C Ascorbic acid +b + + Seal, neutral pH
Thiamine Disulfide form + + + + + Neutral pH
Hydrochlorided + + + + + Seal, neutral pH
Riboflavin Riboflavin +e + + + Dark, pH 1.5–4
Niacin Nicotinic acid Good stability
Nicotinamide Good stability
Vitamin B6 Pyridoxal + + Cool
Pyridoxol (HCl) Good stability
Biotin Biotin + + Seal, neutral pH
Pantothenic Free acidf + + + Cool, neutral pH
Acid Ca saltyd + Seal, pH 6–7
Folate fh4 + + + +g + Good stability
Vitamin B12 Cn-b12 + +h +i Good stability
a
i.e., 100°C
b
In solution with Fe+++ and Cu++
c
Unstable to reducing agents
d
Slightly hygroscopic
e
Especially in alkaline solution
f
Very hygroscopic
g
pH < 5
h
pH < 3
i
pH > 9
Source: Reprinted with permission from G.F. Combs, The Vitamins: Fundamental Aspects in Nutrition and Health,
p. 449, © 1992, Academic Press

have been set for some vitamins and other micro- their absorption and/or excretion are regulated.
nutrients and are defined as the maximum intake The special situation of consumption of nutri-
from food, water and supplements that is unlikely tional supplements which, when added to the
to pose risk of adverse health effects from excess nutrient intake from food, may result in a total
in almost all (97.5%) of healthy individuals in intake in excess of the upper limit of the nutrient.
specific age groups. ULs should be based on long- Lack of vitamins has long been recognized to
term exposure to all foods, including fortified result in serious deficiency diseases. It is now
food products. Most vitamins cause no adverse also recognized that overdoses of certain vita-
effects when they are consumed as foods because mins, especially some of the fat-soluble vitamins
Fat-Soluble Vitamins 367

which can result in serious toxic effects. For this molecule of β-carotene could yield two mole-
reason, the addition of vitamins to foods should cules of vitamin A. The enzyme 15-15'-dioxy-
be carefully controlled. genase is able to cleave a β-carotene molecule
Our requirements for vitamins changes as a symmetrically to produce two molecules of vita-
function of age. Table 9.3 illustrates the recom- min A (Fig. 9.4). This enzyme occurs in intestinal
mended daily allowance for the water and fat mucosa, but the actual conversion is much less
soluble vitamins. Vitamin requirements are listed efficient. As shown in Fig. 9.4, there are other
either in mg/day or in International Units per day reactions that may cause the yield of retinol to be
(IU). The international units are used in cases less than two. After cleavage of the β-carotene,
where the actual amounts per day are small and the first reaction product is retinal, which is
more accurately expressed in activity units. reduced to retinol (Rouseff and Nagy 1994). A
general requirement for the conversion of a carot-
enoid to vitamin A is an unsubstituted β-ionone
Fat-Soluble Vitamins ring. Citrus fruits are a good source of provitamin
A, which results mostly from the presence of
Vitamin A (Retinol) β-cryptoxanthin, β-carotene, and α-carotene.
Gross (1987) reported a total of 16 carotenoids
The structural formula of vitamin A is shown in with provitamin A activity in citrus fruits.
Fig. 9.2. It is an alcohol that occurs in nature pre- Vitamin A levels are frequently expressed in
dominantly in the form of fatty acid esters. International Units (IU). One IU equals 0.344 μg
Highest levels of vitamin A are found in certain of crystalline vitamin A acetate, or 0.300 μg vita-
fish liver oils, such as cod and tuna. Other impor- min A alcohol, or 0.600 μg β-carotene. The rec-
tant sources are mammalian liver, egg yolk, and ommended daily allowance (RDA) of vitamin A
milk and milk products. The levels of vitamin A of the National Research Council Food and
and its provitamin carotene in some foods are Nutrition Board as seen in Table 9.2. Other
listed in Table 9.4. The structural formula of sources quote the human requirement at about
Fig. 9.2 shows the unsaturated character of vita- 1 μg/day. Conditions of rapid growth, pregnancy,
min A. The all-trans form is the most active bio- or lactation increase the need for vitamin A.
logically. The 13-cis isomer is known as Vitamin A, or retinol, is also known as vitamin
neovitamin A; its biological activity is only about A1. Another form, vitamin A2, is found in fish
75% of that of the all-trans form. The amount of liver oils and is 3-dehydroretinol.
neo-vitamin A in natural vitamin A preparations The Food and Agriculture Organization and
is about one-third of the total. The amount is usu- the World Health Organization of the United
ally much less in synthetic vitamin A. The syn- Nations (FAO/WHO) and the National Academy
thetic vitamin A is made as acetate or palmitate of Sciences of the United States ( 1974a) have
and marketed commercially in the form of oil recommended that vitamin A activity be reported
solutions, stabilized powders, or aqueous emul- as the equivalent weight of retinol. To calculate
sions. The compounds are insoluble in water but total retinol equivalents, it is proposed that food
soluble in fats, oils, and fat solvents. analyses list retinol, carotene, and other provita-
There are several provitamins A; these belong min A carotenoids separately. It is also desirable
to the carotenoid pigments. The most important to distinguish between the cis- and trans- forms
one is β-carotene, and some of the pigments that of the provitamins in cooked vegetables. By defi-
can be derived from it are of practical importance. nition, 1 retinol equivalent is equal to 1 μg of
These are β-apo-8'-carotenal and β-apo-8'- retinol, or 6 μg of β-carotene, or 12 μg of other
carotenoic acid ethyl ester (Fig. 9.3). Other provi- provitamin A carotenoids. The National
tamins are α- and γ-carotene and cryptoxanthin. Academy of Sciences (1974a) states that 1 reti-
Beta-carotene occurs widely in plant products nol equivalent is equal to 3.3 IU of retinol or
and has a high vitamin A activity. In theory, one 10 IU of β-carotene.
368

Table 9.2  Vitamin content of some commonly consumed foods Data from USDA Nutrient Data base
Water soluble vitamins Fat soluble vitamins
100 g food 100 g food
Ascorbic Thiamin Riboflavin Niacin B-6 Folate B-12
Food acid (mg) (mg) (mg) (mg) (mg) (μg) (μg) A (IU) E (mg) D (IU) K (μg)
Vegetables
Asparagus 5.6 1.143 0.141 0.978 0.091 52 0.00 756 1.13 0 41.6
Pinto beans raw, sprouted 21.7 0.230 0.175 2.280 0.171 118 0.00 2 – 0 –
Green beans, snap raw 12.2 0.082 0.104 0.734 0.141 33 0.00 690 0.41 0 43
Beets, raw 4.9 0.031 0.040 0.334 0.067 109 0.00 33 0.04 0 0.20
Broccoli, raw 89.2 0.071 0.117 0.639 0.175 63 0.00 623 0.78 0 101.6
Carrots, raw 5.9 0.066 0.058 0.983 0.138 19 0.00 16,706 0.66 0 13.2
Corn, sweet yellow 6.8 0.155 0.055 1.770 0.093 42 0.00 187 0.07 0 0.30
Lettuce, iceberg 2.8 0.041 0.025 0.123 0.042 29 0.00 502 0.18 0 24.1
Onions, raw 7.4 0.046 0.027 0.116 0.120 19 0.00 2 0.02 0 0.4
Green peas, raw 40 0.266 0.132 2.090 0.169 65 0.00 765 0.13 0 24.8
Green peppers, raw 80.4 0.057 0.028 0.480 0.224 10 0.00 370 0.37 0 7.4
Potatoes, fresh with skin, raw 19.7 0.081 0.032 1.061 0.298 15 0 2 0.01 0 2.0
Spinach, raw 28.1 0.078 0.189 0.724 0.195 194 0 9377 2.03 0 482.9
Summer squash, all varieties, raw 17.0 0.48 0.142 0.487 0.218 29 0 200 0.12 0 3.0
Fruits
Apples, raw with skin 4.6 0.017 0.026 0.091 0.041 3 0 54 0.18 0 2.2
Avocados, raw 10 0.067 0.130 1.738 0.277 81 0 146 2.07 0 21.0
Blueberries, raw 9.7 0.037 0.041 0.418 0.052 6 0 3 0.57 0 19.30
9 Vitamins
Cranberries, raw 14 0.012 0.020 0.101 0.057 1 0 63 1.32 0 5.0
Oranges, Florida, raw 45 0.100 0.040 0.400 0.051 17 0 225 0.18 0 0.0
Pomegranates, raw 10.2 0.067 0.053 0.293 0.075 38 0 0 0.60 0 16.4
Strawberries, raw 58.8 0.024 0.022 0.386 0.047 24 0 12 0.29 0 2.2
Cereals and grains
Corn flour, whole grain-yellow 0 0.246 0.080 1.90 0.370 25 0 214 0.42 0 0.3
Fat-Soluble Vitamins

Pasta, dry enriched 0 0.90 0.060 1.70 18 18 0 0 0.11 0 0.1


Rice, brown long grain 0 0.541 0.095 6.494 0.477 23 0 0 0.60 0 0.6
Rice, long grain, unenriched 0 0.224 0.050 5.048 0.452 8 0 0 0.03 0 0.1
Wheat, bread flour unenriched 0 0.080 0.060 1.000 0.037 33 0 2 0.40 0 0.3
Meat, eggs and dairy
Ground beef, 20% fat uncooked 0 0.43 0.151 4.227 0.323 7 2.14 14 0.17 3 1.8
Beef tenderloin steak 0 0.071 0.441 6.235 0.765 7 4.27 5 0.28 3 1.4
Lamb, whole leg 0 0.141 0.287 4.964 0.359 2.70 2.70 – – – –
Pork, fresh, composite of trimmed retail cuts 0.5 0.841 0.254 4.504 0.445 5 – 7 – – –
(leg, loin, shoulder, and spareribs), raw
Chicken, broilers or fryers, back, meat and skin, raw 1.6 0.051 0.116 4.935 0.190 6 0.25 251 0.37 – 2.4
Cheese, cheddar 0 0.29 4.28 0.059 0.066 27 1.10 1242 0.71 24 4.2
Milk, whole, 3.25% milkfat 0 0.046 0.169 0.089 0.036 5 0.45 46 162 2 0.3
Not fortified
Egg, raw. fresh 0 0.004 0.4390 0.105 0.005 4 0.09 0 0 0 0
369
370 9 Vitamins

Fig. 9.1  Risk function of deficiency or excess for individuals in a population related to vitamin intake assuming a
Gaussian distribution of requirements to prevent deficit and avoid excess

Vitamin A occurs only in animals and not in various food processing operations. Losses may
plants. The A1 form occurs in all animals and fish, occur at high temperatures in the presence of
the A2 form in freshwater fish and not in land ani- oxygen. These compounds are also susceptible to
mals. The biological value of the A2 form is only oxidation by lipid peroxides, and conditions
about 40% of that of A1. Good sources of provita- favoring lipid oxidation also result in vitamin A
min A in vegetable products are carrots, sweet breakdown. The prooxidant copper is especially
potatoes, tomatoes, and broccoli. In milk and harmful, as is iron to a lesser extent. Pasteurization
milk products, vitamin A and carotene levels are of milk does not result in vitamin A loss, but
subject to seasonal variations. Hartman and exposure to light does. It is essential, therefore,
Dryden (1978) report the levels of vitamin A in that sterilized milk be packaged in light-­
fluid whole milk in winter at 1083 IU/L and in impervious containers. Possible losses during
summer at 1786 IU/L. Butter contains an average storage of foods are more affected by duration of
of 2.7 μg of carotene and 5.0 μg of vitamin A per storage than by storage temperature. Blanching
g during winter and 6.1 μg of carotene and 7.6 μg of fruits and vegetables helps prevent losses dur-
of vitamin A per g during summer. ing frozen storage.
Vitamin A is used to fortify margarine and Vitamin A added to milk is more easily
skim milk. It is added to margarine at a level destroyed by light than the native vitamin A. This
of 3525 IU per 100 g. Some of the carotenoids is not because natural and synthetic vitamin A are
(provitamin A) are used as food colors. different, but because these two types of vitamin
Vitamin A is relatively stable to heat in the A are dispersed differently in the milk (deMan
absence of oxygen (Table 9.5). Because of the 1981). The form in which vitamin A is added to
highly unsaturated character of the molecule, it is food products may influence its stability. Vitamin
quite susceptible to oxidation—especially under A in beadlet form is more stable than that added
the influence of light, whether sunlight or artifi- as a solution in oil. The beadlets are stabilized by
cial light. Vitamin A is unstable in the presence a protective coating. If this coating is damaged by
of mineral acids but stable in alkali. Vitamin A water, the stability of the vitamin is greatly
and the carotenoids have good stability during reduced (deMan 1986).
Fat-Soluble Vitamins

Table 9.3  Relative daily requirements of water and fat soluble vitamins as a function of age (https://ods.od.nih.gov/Health_Information/Dietary_Reference_Intakes.aspx)
Water soluble vitamins Fat soluble vitamins
Vitamin C Folate Niacin Pantothenic Riboflavin B-12
Age (mg) (mcg) (mg) acid (mg) (mg) B-6 (mg) (mcg) A (IU) D (IU) E (IU) K (mcg)
1–3 15 150 6 2 0.5 0.5 0.9 1000 600 13 (S) 30
9 (N)
4–8 25 200 8 3 0.6 0.6 1.2 1300 600 16 (S) 55
10(N)
9–13 45 300 12 4 0.9 1.0 1.8 2000 600 24 (S) 60
16(N)
14–18 75(m) 400 16 (m) 5 1.3 (m) 1.3 (m) 2.4 1000 600 33 (S) 75
65 (f) 14 (f) 1 (f) 1.2 (f) 22(N)
Adult 90 (m) 400 16 (m) 5 1.3 (m) 1.3 (19–50) 2.4 3000 (m) 600 33 (S) 120 (m)
75 (f) 14 (f) 1.1 (f) 1.5 (50+) 2300 (f) 800 (51+) 22(N) 90 (f)
m male, f female, S synthetic, N natural
371
372 9 Vitamins

deficiency. Enrichment of some foods with vita-


min D has significantly helped to eradicate rick-
ets, which is a vitamin D deficiency disease.
Margarine and milk are the foods commonly
used as carrier for added vitamin D.
Fig. 9.2  Structural formula of vitamin A. Acetate: R=CO The unit of activity of vitamin D is the IU,
CH3. Palmitate: R=CO (CH2)14 CH3 which is equivalent to the activity of 1 mg of a
standard preparation issued by the WHO. One IU
is also equivalent to the activity of 0.025 μg of
Table 9.4  Vitamin A and carotene content of some foods
pure crystalline vitamin D2 or D3. The human
Vitamin A Carotene requirement amounts to 400–500 IU but increases
Product (IU/100 g) (mg/100 g)
to 1000 IU during pregnancy and lactation.
Beef (grilled sirloin) 37 0.04
Adults who are regularly exposed to sunlight are
Butter (May–November) 2363–3452 0.43–0.77
likely to have a sufficient supply of vitamin
Cheddar cheese 553–1078 0.07–0.71
D. Excessive intakes are toxic.
Eggs (boiled) 165–488 0.01–0.15
Vitamin D is extremely stable, and little or no
Herring (canned) 178 0.07
loss is experienced in processing and storage.
Milk 110–307 0.01–0.06
Vitamin D in milk is not affected by pasteuriza-
Tomato (canned) 0 0.5
tion, boiling, or sterilization (Hartman and
Peach 0 0.34
Dryden 1978). Frozen storage of milk or butter
Cabbage 0 0.3
Broccoli (boiled) 0 2.5
also has little or no effect on vitamin D levels,
Spinach (boiled) 0 6.0
and the same result is obtained during storage of
dry milk.
The vitamin D potency of milk can be
Vitamin D increased in several ways: by feeding cows sub-
stances that are high in vitamin D activity, such
This vitamin occurs in several forms; the two as irradiated yeast; by irradiating milk; and by
most important are vitamin D2, or ergocalciferol, adding vitamin D concentrates. The latter method
and vitamin D3, or cholecalciferol. The structural is now the only commonly used procedure. The
formulas of these compounds are presented in practice of irradiating milk to increase the vita-
Fig. 9.5. Vitamin D does not occur in plant prod- min D potency has been discontinued, undoubt-
ucts. Vitamin D2 occurs in small amounts in fish edly because of the deteriorative action of the
liver oils; vitamin D3 is widely distributed in ani- radiation on other milk components. Vitamin D is
mal products, but large amounts occur only in added to milk to provide a concentration of
fish liver oils. Smaller quantities of vitamin D3 400 IU per quart. Addition of vitamin D to mar-
occur in eggs, milk, butter, and cheese (Table 9.6). garine is at a level of 550 IU per 100 g.
The precursors of vitamins D2 and D3 are
ergosterol and 7-dehydrocholesterol, respec-
tively. These precursors or provitamins can be Tocopherols (Vitamin E)
converted into the respective D vitamins by irra-
diation with ultraviolet light. In addition to the The tocopherols are derivatives of tocol, and the
two major provitamins, there are several other occurrence of a number of related substances in
sterols that can acquire vitamin D activity when animal and vegetable products has been demon-
irradiated. The provitamins can be converted to strated. Cottonseed oil was found to contain α-,
vitamin D in the human skin by exposure to sun- β-, and γ-tocopherol, and a fourth, δ-tocopherol,
light. Because very few foods are good sources of was isolated from soybean oil. Several other
vitamin D, humans have a greater likelihood of tocopherols have been found in other products,
vitamin D deficiency than of any other vitamin and Morton (1967) suggests that there are
Fat-Soluble Vitamins 373

Fig. 9.3 Structural
formulas of some
provitamins A. (a)
β-carotene, and (b)
apocarotenal (R=CHO)
and apocarotenoic acid
ester (R=COOC2H5)

Fig. 9.4 Conversion
of beta-carotene
to vitamin A

four tocopherols and four tocotrienols. The toco- The carbons at locations 4′ and 8′ in the side
trienols have three unsaturated isoprenoid groups chains of the tocopherols are asymmetric, as is
in the side chain. The structure of tocol is given the number 2 carbon in the chroman ring. The
in Fig. 9.6 and the structures of the tocopherols resulting ­possible isomers are described as hav-
and tocotrienols in Fig. 9.7. The four tocopherols ing R or S rotation. The natural tocopherols and
are characterized by a saturated side chain con- tocotrienols are predominantly RRR isomers.
sisting of three isoprenoid units. The tocotrienols Stocker and Kearney (2004) have summarized
have three double bonds at the 3′, 7′, and 11′ the chemistry of the oxidation of tocopherols as
­carbons of the isoprenoid side chain (Fig. 9.7). shown in Fig. 9.8.
374 9 Vitamins

Table 9.5  Vitamin A and carotene stability in foods


Product Nutrient content Storage conditions Retention (%)
Vitamin A
 Butter 17,000–30,000 IU/Ib 12 months @ 5 °C 66–98
5 months @ 28 °C 64–68
 Margarine 15,000 IU/Ib 6 months @ 5 °C 89–100
6 months @ 23 °C 83–100
 Nonfat dry milk 10,000 IU/Ib 3 months @ 37 °C 94–100
12 months @ 23 °C 69–89
 Fortified ready-to-eat cereal 4000 IU/oz 6 months @ 23 °C 83
 Fortified potato chips 700 IU/100 g 2 months @ 23 °C 100
Carotene
 Margarine 3 mg/Ib 6 months @ 5 °C 98
6 months @ 23 °C 89
 Lard 3.3 mg/lb 6 months @ 5 °C 100
6 months @ 23 °C 100
 Dried egg yolk 35.2 mg/100 g 3 months @ 37 °C 94
12 months @ 23 °C 80
 Carbonated beverage 7.6 mg/29 oz 2 months @ 30 °C 94
2 months @ 23 °C 94
 Canned juice drinks 0.6–1.3 mg/8 fl oz 12 months @ 23 °C 85–100
Source: From E. deRitter, Stability Characteristics of Vitamins in Processed Foods, Food Technol., Vol. 30, pp. 48–51,
54, 1976

Table 9.6  Vitamin D content of some foods


Product Vitamin D (μg/1000 g edible portion)
Liver (beef, pork) 2–5
Eggs 44
Milk 0.9
Butter 2–40
Cheese 12–47
Herring oil 2500

quinone yields a quinol. Tocopherolquinones


occur naturally. Oxidation with nitric acid yields
the o-quinone or tocopherol red, which is not
found in nature. Alpha-tocopheronic acid and
α-tocopheronolactone are some of the products
of metabolism of tocopherol. Much of the bio-
logical activity of the tocopherols is related to
their antioxidant activity. Because α-tocopherol
is the most abundant of the different tocopherols,
Fig. 9.5  Structural formulas of (a) vitamin D2 and (b) and because it appears to have the greatest bio-
vitamin D3 logical activity, the α-tocopherol content of foods
is usually considered to be most important.
On oxidation, α-tocopherol can form a The biological activity of the tocopherols and
­ eta-­stable epoxide that can be irreversibly con-
m tocotrienols varies with the number and position
verted to α-tocopherolquinone. Reduction of the of the methyl groups on the chroman ring and by
Fat-Soluble Vitamins 375

Fig. 9.6 Structural
formula of tocol and
α-tocopherol

Fig. 9.7  Chemical structure of the tocopherols and tocotrienols

the configuration of the asymmetric carbons in proportion of α-tocopherol, which amounts to


the side chain. The R configuration at each chiral about 60% of the total tocopherols. Refining of
center has the highest biological activity. Because vegetable oils, carried out under normal precau-
the different isomers have different activities, it is tions (such as excluding air), appears to result in
necessary to measure each homolog and convert little destruction of tocopherol. The tocopherol
these to RRR-α-tocopherol equivalents (α-TE). and tocotrienol content of selected fats and oils
One α-TE is the activity of 1 mg of RRR-α-­ and their primary homologs are listed in
tocopherol (Eitenmiller 1997). The vitamin E Table 9.8. The seed oils contain only tocopherol.
activity of α-tocopherol isomers and synthetic Tree oils, palm, palm kernel, coconut oil, and rice
tocopherols is listed in Table 9.7. bran oil also contain major amounts of tocotri-
Tocopherols are important as antioxidants in enols. The processing of vegetable oils by
foods, especially in vegetable oils. With few deodorization or physical refining removes a con-
exceptions, animal and vegetable products con- siderable portion of the tocopherols, and these
tain from about 0.5 to 1.5 mg/100 g; vegetable steam-volatile compounds accumulate in the
oils from 10 to 60 mg/100 g; and cereal germ fatty acid distillate (Ong 1993). This product is
oils, which are a very good source, from 150 to an important source of natural vitamin E prepara-
500 mg/100 g. Vegetable oils have the highest tions. Baltes (1967) carried out tests in which two
376 9 Vitamins

Fig. 9.8  The major pathways of α-TOH oxidation. Two-­ further scavenge radicals, to produce 8a-substituted
electron oxidants such as HOCl and ONOO− oxidize tocopherone adducts, or scavenge LOO, to produce
α-TOH to the intermediate tocopheroxylium cation 8a-hydroperoxy-epoxytocopherones. These hydrolyze to
(α-TO+), which hydrolyzes to α-tocopherylquinone α-TQ and α-tocopheryl quinone epoxides (TQEs), respec-
(α-TQ). Radical oxidants (R) generate the α-TO that can tively (Stocker and Kearney 2004)

Table 9.7  Vitamin E activity of α-tocopherol isomers easily oxidizable fats, lard and partially hydroge-
and synthetic tocopherols nated whale oil, were stabilized with α-tocopherol
Name IU/mg and ascorbyl palmitate and citric acid as syner-
d-α-tocopherol (2R4′R8′R) 1.49 gists. Without antioxidants, these fats cannot be
RRR-α-tocopherol used in the commercial food chain. Amounts of
1-α-tocopherol (2S4′R8′R) 0.46 α-tocopherol ranging from 0.5 to 10 mg/100 g
dl-α-tocopherol all-rac-α-tocopherol 1.10 were effective in prolonging the storage life of
2R4′R8′S-α-tocopherol 1.34 some samples up to 2 years.
2S4′R8′S-α-tocopherol 0.55 The tocopherol content of some animal and
2S4′S8′S-α-tocopherol 1.09 vegetable products as reported by Thaler (1967)
2S4′S8′R-α-tocopherol 0.31 is listed in Table 9.9. Cereals and cereal products
2R4′S8′R-α-tocopherol 0.85 are good sources of tocopherol (Table 9.10). The
2S4′S8′S-α-tocopherol 1.10 distribution of tocopherol throughout the kernels
d-α-tocopheryl acetate RRR-α-tocopheryl 1.36 is not uniform, and flour of different degrees of
acetate
extraction can have different tocopherol levels.
dl-α-tocopherol all-rac-α-tocopherol acetate 1.00
This was shown by Menger (1957) in a study of
Source: Reprinted with permission from R.R. Eiten-miller,
Vitamin E Content of Fats and Oils: Nutritional
wheat flour (Table 9.11).
Implications, Food Technol., Vol. 51, no. 5, p. 79, © 1997, Processing and storage of foods can result in
Institute of Food Technologists substantial tocopherol losses. An example is
Fat-Soluble Vitamins 377

Table 9.8  Tocopherol (T) and tocotrienol (T3) content of vegetable oils and their primary homologs
Fats and oils Total T + T3 (mg/100 g) α-TE/100 g %T %T3 Primary homologs
Sunflower 46–67 35–63 100 0 α-T, γ-Τ
Cottonseed 78 43 100 0 α-T, γ-Τ
Safflower 49–80 41–46 100 0 α-T, δ-Τ, γ-Τ, β-Τ
Safflower—high 41 41 100 0 α-T, β-Τ
linolenic
Safflower—high oleic 32 31 100 0 α-T, β-Τ, γ-Τ
Palm 89–117 21–34 17–55 45–83 α-Τ, α-Τ3, δ-Τ3, α-Τ, δ-Τ3
Canola 65 25 100 0 γ-Τ, α-Τ, δ-Τ, α-T3(Tr), β-T(Tr)
Corn 78–109 20–34 95 5 γ-Τ, α-Τ, δ-Τ, γ-Τ3, δ-Τ3
Soybean 96–115 17–20 100 0 γ-Τ, δ-Τ, α-Τ
Rice bran 9–160 0.9–41 19–49 51–81 γ-Τ3, αΤ, α-Τ3, β-Τ, β-Τ3
Peanut 37 16 100 0 γ-Τ, α-Τ, δ-Τ
Olive 5.1 5.1 100 0 α-Τ
Cocoa butter 20 3.0 99 1 γ-Τ, δ-Τ, α-Τ, α-Τ3
Palm kernel 3.4 1.9 38 62 α-Τ3, α-Τ
Butter 1.1–2.3 1.1–2.3 100 0 α-Τ
Lard 0.6 0.6 100 0 α-Τ
Coconut 1.0–3.6 0.3–0.7 31 69 γ-Τ3, α-Τ3, δ-Τ, α-Τ, β-Τ3
Source: Reprinted with permission from R.R. Eitenmiller, Vitamin E Content of Fats and Oils: Nutritional Implications,
Food Technol., Vol. 51, no. 5, p. 80, © 1997, Institute of Food Technologists

Table 9.9  Tocopherol content of some animal and vege- Table 9.10  Tocopherol content of cereals and cereal
table food products products
Total tocopherol as Total tocopherol
Product α-tocopherol (mg/100 g) Product as α-tocopherol (mg/100 g)
Beef liver 0.9–1.6 Wheat 7–10
Veal, lean 0.9 Rye 2.2–5.7
Herring 1.8 Oats 1.8–4.9
Mackerel 1.6 Rice (with hulls) 2.9
Crab, frozen 5.9 Rice (polished) 0.4
Milk 0.02–0.15 Corn 9.5
Cheese 0.4 Whole wheat meal 3.7
Egg 0.5–1.5 Wheat flour 2.3–5.4
Egg yolk 3.0 Whole rye meal 2.0–4.5
Cabbage 2–3 Oat flakes 3.85
Spinach 0.2–6.0 Corn grits 1.17
Beans 1–4 Corn flakes 0.43
Lettuce 0.2–0.8 (0.06) White bread 2.15
Peas 4–6 Whole rye bread 1.3
Tomato 0.9 (0.4) Crisp bread 4.0
Carrots 0.2 (0.11) Source: From H. Thaler, Concentration and Stability of
Onion 0.3 (0.22) Tocopherols in Foods, in Tocopherols, K. Lang, ed., 1967,
Potato (0.12) Steinkopff Verlag, Darmstadt, Germany
Mushrooms 0.08
Source: From H. Thaler, Concentration and Stability of
given in Table 9.12, where the loss of tocopherol
Tocopherols in Foods, in Tocopherols, K. Lang, ed., 1967, during frying of potato chips is reported. After
Steinkopff Verlag, Darmstadt, Germany only 2 weeks’ storage of the chips at room
378 9 Vitamins

Table 9.11  Tocopherol content of wheat and its milling double bond in the side chain. The vitamins K2
products
have a side chain consisting of a number of regu-
Tocopherol mg/100 g lar units of the type.
Product Ash (%) (dry basis)
Whole wheat 2.05 5.04
Flour 1 (fine) 1.68 5.90
Flour 2 1.14 4.27
Flour 3 0.84 3.48
Flour 4 0.59 2.55
where n can equal 4, 5, 6, 7, and so forth.
Flour 5 0.47 2.35
Vitamin K1 is slowly decomposed by atmo-
Flour 6 (coarse) 0.48 2.13
spheric oxygen but is readily destroyed by light.
Germ 4.10 25.0
It is stable against heat, but unstable against
Source: From A. Menger, Investigation of the Stability of
Vitamin E in Cereal Milling Products and Baked Goods,
alkali.
Brot. Gebäck, Vol. 11, pp. 167–173, 1957 (German) The human adult requirement is estimated at
about 4 mg per day. Menadione (2-methyl
1,4-naphtoquinone) is a synthetic product and
Table 9.12  Tocopherol losses during processing and
storage of potato chips has about twice the activity of naturally occurring
vitamin K.
Tocopherol
(mg/100 g) Loss (%) Vitamin K occurs widely in foods and is also
Oil before use 82 – synthesized by the intestinal flora. Good sources
Oil after use 73 11 of vitamin K are dark green vegetables such as
Oil from fresh chips 75 – spinach and cabbage leaves, and also cauliflower,
After 2 weeks at room 39 48 peas, and cereals. Animal products contain little
temperature vitamin K1, except for pork liver, which is a good
After 1 month at room 22 71 source.
temperature The Vitamin K levels in some foods, expressed
After 2 months at room 17 77 in menadione units, are given in Table 9.13.
temperature
After 1 month at −12 °C 28 63
After 2 months at −12 °C 24 68
Water-Soluble Vitamins

t­emperature, nearly half of the tocopherol was Vitamin C (L-Ascorbic Acid)


lost. The losses were only slightly smaller during
storage at freezer temperature. Boiling of vegeta- This vitamin occurs in all living tissues, where it
bles in water for up to 30 min results in only influences oxidation-reduction reactions. The
minor losses of tocopherol. Baking of white major source of L-ascorbic acid in foods is veg-
bread results in a loss of about 5% of the tocoph- etables and fruits (Table 9.14).
erol in the crumb. L-ascorbic acid (Fig. 9.10) is a lactone (inter-
The human daily requirement of vitamin E is nal ester of a hydroxycarboxylic acid) and is
estimated at 30 IU. Increased intake of polyun- characterized by the enediol group, which makes
saturated fatty acids increases the need for this it a strongly reducing compound. The D form has
vitamin. no biological activity. One of the isomers,
D-isoascorbic acid, or erythorbic acid, is
­produced commercially for use as a food addi-
Vitamin K tive. L-ascorbic acid is readily and reversibly oxi-
dized to dehydro-L-ascorbic acid (Fig. 9.11),
This vitamin occurs in a series of different forms, which retains vitamin C activity. This compound
and these can be divided into two groups. The can be further oxidized to diketo-L-gulonic acid,
first is vitamin K1 (Fig. 9.9), characterized by one in a nonreversible reaction. Diketo-L-gulonic
Water-Soluble Vitamins 379

Fig. 9.9  Structural formula of vitamin K1

Table 9.13  Vitamin K in some foods (expressed as men- acid has no biological activity, is unstable, and is
adione units per 100 g of edible portion)
further oxidized to several possible compounds,
Product Units/100 g including 1-threonic acid. Dehydration and
Cabbage, white 70 decarboxylation can lead to the formation of fur-
Cabbage, red 18 fural, which can polymerize to form brown pig-
Cauliflower 23 ments or combine with amino acids in the
Carrots 5 Strecker degradation.
Honey 25 Humans and guinea pigs are the only primates
Liver (chicken) 13 unable to synthesize vitamin C. The human
Liver (pork) 111 requirement of vitamin C is not well defined.
Milk 8 Figures ranging from 45 to 75 mg/day have been
Peas 50 listed as daily needs. Continued stress and drug
Potatoes 10
therapy may increase the need for this vitamin.
Spinach 161
Vitamin C is widely distributed in nature,
Tomatoes (green) 24
mostly in plant products such as fruits (especially
Tomatoes (ripe) 12
citrus fruits), green vegetables, tomatoes, pota-
Wheat 17
toes, and berries. The only animal sources of this
Wheat bran 36
vitamin are milk and liver. Although widely dis-
Wheat germ 18
tributed, very high levels of the vitamin occur
only in a few products, such as rose hips and
West Indian cherries. The concentration varies
Table 9.14  Vitamin C content of some foods widely in different tissues of fruits; for example,
Product Ascorbic acid (mg/100 g) in apples, the concentration of vitamin C is two to
Black currants 200 three times as great in the peel as in the pulp.
Brussels sprouts 100 Vitamin C is the least stable of all vitamins
Cauliflower 70 and is easily destroyed during processing and
Cabbage 60 storage. The rate of destruction is increased by
Spinach 60 the action of metals, especially copper and iron,
Orange 50 and by the action of enzymes. Exposure to oxy-
Orange juice 40–50 gen, prolonged heating in the presence of oxy-
Lemon 50 gen, and exposure to light are all harmful to the
Peas 25 vitamin C content of foods. Enzymes containing
Tomato 20 copper or iron in their prosthetic groups are effi-
Apple 5 cient catalysts of ascorbic acid decomposition.
Lettuce 15 The most important enzymes of this group are
Carrots 6 ascorbic acid oxidase, phenolase, cytochrome
Milk 2.1–2.7 oxidase, and peroxidase. Only ascorbic acid oxi-
Potatoes 30 dase involves a direct reaction among enzyme,
380 9 Vitamins

Fig. 9.10  Structural formulas of L-ascorbic acid and its stereoisomers

Fig. 9.11  Oxidation of L-ascorbic acid

substrate, and molecular oxygen. The other Table 9.15  Effect of blanching method on ascorbic acid
levels of broccoli
enzymes oxidize the vitamin indirectly. Phenolase
catalyzes the oxidation of mono and dihydroxy Ascorbic acid (mg/100 g)
phenols to quinones. The quinones react directly Factor effect Reduced Dehydro Total
with the ascorbic acid. Cytochrome oxidase Raw 94.0 4.0 98.2
­oxidizes cytochrome to the oxidized form and Water blanch 45.3 5.7 51.0
this reacts with L-ascorbic acid. Steam blanch 48.8 7.4 56.2
Peroxidase, in combination with phenolic Source: From D. Ödland and M.S. Eheart, Ascorbic Acid,
Mineral and Quality Retention in Frozen Broccoli
compounds, utilizes hydrogen peroxide to bring Blanched in Water, Steam, and Ammonia-Steam, J. Food
about oxidation. The enzymes do not act in intact Sci., Vol. 40, pp. 1004–1007, 1975
fruits because of the physical separation of
enzyme and substrate. Mechanical damage, rot,
or senescence lead to cellular disorganization and smaller losses of ascorbic acid (Table 9.15). The
initiate decomposition. Inhibition of the enzymes retention of ascorbic acid in frozen spinach
in vegetables is achieved by blanching with steam depends on storage temperature. At a very low
or by electronic heating. Blanching is necessary temperature (−29 °C), only 10% of the initially
before vegetables are dried or frozen. In fruit present ascorbic acid was lost after 1 year. At
juices, the enzymes can be inhibited by pasteuri- −12°, the loss after 1 year was much higher,
zation, deaeration, or holding at low temperature 55%. The presence of metal chelating compounds
for a short period. The effect of blanching meth- stabilizes vitamin C. These compounds include
ods on the ascorbic acid content of broccoli was anthocyanins and flavonols, polybasic or polyhy-
reported by Odland and Eheart (1975). Steam droxy acids such as malic and citric acids, and
blanching was found to result in significantly polyphosphates.
Water-Soluble Vitamins 381

Ascorbic acid is oxidized in the presence of judged by the percentage of ascorbic acid that has
air under neutral and alkaline conditions. At acid been lost. The extent of loss depends on the amount
pH (for example, in citrus juice), the vitamin is of water used. During blanching, vegetables that
more stable. Because oxygen is required for the are covered with water may lose 80%; half cov-
breakdown, removal of oxygen should have a sta- ered, 40%; and quarter covered, 40% of the ascor-
bilizing effect. For the production of fruit drinks, bic acid. Particle size affects the size of the loss;
the water should be deaerated to minimize vita- for example, in blanching small pieces of carrots,
min C loss. The type of container may also affect losses may range from 32 to 50%, and in blanch-
the extent of ascorbic acid destruction. Use of tin ing large pieces, only 22 to 33%. Blanching of
cans for fruit juices results in rapid depletion of cabbage may result in a 20% loss of ascorbic acid,
oxygen by the electrochemical process of corro- and subsequent dehydration may increase this to a
sion. In bottles, all of the residual oxygen is avail- total of 50%. In the processing of milk, losses may
able for ascorbic acid oxidation. To account for occur at various stages. From an initial level of
processing and storage losses, it is common to about 22 mg/L in raw milk, the content in the
allow for a loss of 7–14 mg of ascorbic acid per product reaching the consumer may be well below
100 mL of fruit juice. Light results in rapid 10 mg/L. Further losses may occur in the house-
destruction of ascorbic acid in milk. It has been hold during storage of the opened container.
shown (Sattar and deMan 1973) that transparent The processing of milk into various dairy
packaging materials permit rapid destruction of products may result in vitamin C losses. Ice cream
vitamin C (Fig. 9.12). The extent of ascorbic acid contains no vitamin C, nor does cheese. The pro-
destruction is closely parallel to the development duction of powdered milk involves a 20–30%
of off-flavors. The destruction of ascorbic acid in loss, evaporated milk a 50–90% loss. Bullock
milk by light occurs under the influence of ribo- (1968) studied the stability of added vitamin C in
flavin as a sensitizer. The reaction occurs in the evaporated milk and found that adding 266 mg of
presence of light and oxygen, and the riboflavin sodium ascorbate per kg was sufficient to ensure
is converted to lumichrome. the presence of at least 140 mg/L of ascorbic acid
Factors that affect vitamin C destruction during during 12 months of storage at 21 °C. Data on the
processing include heat treatment and leaching. stability of vitamin C in fortified foods have been
The severity of processing conditions can often be assembled by deRitter (1976) (Table 9.16).

Fig. 9.12  Effect of


exposure time at light
intensity of 200 Ft-C on
the loss of ascorbic acid
in milk. Packaging
materials: (1) clear
plastic pouch, (2)
laminated
nontransparent pouch,
(3) carton, (4) plastic
3-quart jug. Source:
From A. Sattar and J.M.
deMan, Effect of
Packaging Material on
Light Induced Quality
Deterioration of Milk,
Can. Inst. Food Sci.
Technol. J., Vol. 6,
pp. 170–174, 1973
382 9 Vitamins

Table 9.16  Vitamin C stability in fortified foods and as cocarboxylase. Thiamin is available in the form
beverages after storage at 23 °C for 12 months, except as
of its chloride or nitrate, and its structural formula
noted
is shown in Fig. 9.13. The molecule contains two
Retention basic nitrogen atoms; one is in the primary amino
No. of Mean Range
group, the other in the quaternary ammonium
Product samples (%) (%)
group. It forms salts with inorganic and organic
Ready-to-eat cereal 4 71 60–87
acids. The vitamin contains a primary alcohol
Dry fruit drink mix 3 94 91–97
Cocoa powder 3 97 80–100
group, which is usually present in the naturally
Dry whole milk, air pack 2 75 65–84
occurring vitamin in esterified form with ortho-,
Dry whole milk, gas pack 1 93 – di-, or triphosphoric acid. In aqueous solution, the
Dry soy powder 1 81 – compound may occur in different forms, depend-
Potato flakesa 3 85 73–92 ing on pH. In acid solution, the equilibrium favors
Frozen peaches 1 80 – the formation of positive ions (Fig. 9.14). The
Frozen apricotsb 1 80 – thiol- form is favored in alkaline medium. This
Apple juice 5 68 58–76 form can react with compounds containing sulfhy-
Cranberry juice 2 81 78–83 dryl groups to form disulfide bridges. It has been
Grapefruit juice 5 81 73–86 suggested that thiamin occurs in some foods linked
Pineapple juice 2 78 74–82 to protein by disulfide bridges.
Tomato juice 4 80 64–93 Small quantities of thiamin are present in
Vegetable juice 2 68 66–69 almost all foods of plant and animal origin. Good
Grape drink 3 76 65–94 sources are whole cereal grains; organ meats
Orange drink 5 80 75–83 such as liver, heart, and kidney; lean pork; eggs;
Carbonated beverage 3 60 54–64 nuts; and potatoes (Table 9.17). Although thia-
Evaporated milk 4 75 70–82 min content is usually measured in mg per 100 g
a
Stored for 6 months at 23 °C of a food, another unit has been used occasion-
b
Thawed after storage in freezer for 5 months ally, the IU corresponding to 3 μg of thiamin-­
Source: From E. deRitter, Stability Characteristics of
hydrochloride. The human daily requirement is
Vitamins in Processed Foods, Food Technol., Vol. 30,
pp. 48–51, 54, 1976 related to the carbohydrate level of the diet. A
minimum intake of 1 mg per 2000 kcal is consid-
ered essential. Increased metabolic activity, such
There are many technical uses of ascorbic acid as that which results from heavy work, preg-
in food processing. It is used to prevent browning nancy, or disease, requires higher intake.
and discoloration in vegetables and fruit p­ roducts; Thiamin is one of the more unstable vitamins.
as an antioxidant in fats, fish products, and dairy Various food processing operations may consider-
products; as a stabilizer of color in meat; as an ably reduce thiamin levels. Heat, oxygen, sulfur
improver of flour; as an oxygen acceptor in beer dioxide, leaching, and neutral or alkaline pH may
processing; as a reducing agent in wine, partially all result in destruction of thiamin. Light has no
replacing sulfur dioxide; and as an added nutri- effect. The enzyme is stable under acid conditions;
ent. The vitamin is protected by sulfur dioxide, at pH values of 3.5 or below, foods can be auto-
presumably by inhibiting polyphenolase. claved at 120 °C with little or no loss of thiamin.
At neutral or alkaline pH, the vitamin is destroyed
by boiling or even by storage at room temperature.
Vitamin B1 (Thiamin) Even the slight alkalinity of water used for pro-
cessing may have an important effect. Bender
This vitamin acts as a coenzyme in the metabolism (1971) reports that cooking rice in distilled water
of carbohydrates and is present in all living tissues. reduced thiamin content negligibly, whereas cook-
It acts in the form of thiamin diphosphate in the ing in tap water caused an 8–10% loss, and cook-
decarboxylation of α-keto acids and is referred to ing in well water caused a loss of up to 36%.
Water-Soluble Vitamins 383

Fig. 9.13 Structural
formula of thiamin.
Hydrochloride: X=Cl−,
HCl;mononitrate:X=NO3−

Fig. 9.14  Behavior of thiamin in aqueous solutions

Table 9.17  Thiamin content of some foods related to size of cut, fat content, and so on. Boiling
Thiamin (mg/100 g) loss is 15–40%; frying, 40–50%; roasting, 30–60%;
Product edible portion and canning, 50–75%. Similar losses apply to fish.
Almonds 0.24 Because thiamin and other vitamins are located
Corn 0.37 near the bran of cereal grains, there is a great loss
Egg 0.11 during milling. White flour, therefore, has a greatly
Filberts 0.46
reduced content of B vitamins and vitamin E
Beef heart 0.53
(Fig. 9.15). Not only is thiamin content lowered by
Beef liver 0.25
milling, but also storage of whole grain may result
Macaroni (enriched) 0.88
in losses. This depends on moisture content. At
Macaroni (not enriched) 0.09
normal moisture level of 12%, 5 months’ storage
Milk 0.03
results in a 12% loss; at 17% moisture, a 30% loss;
Peas 0.28
and at 6% moisture, no loss at all. Because of the
Pork, lean 0.87
losses that are likely to occur in cereal grain pro-
Potatoes 0.10
cessing and in the processing of other foods, a pro-
Wheat (hard red spring) 0.57
Wheat flour (enriched) 0.44
gram of fortification of flour is an important factor
Wheat flour (not enriched) 0.08
in preventing vitamin deficiencies. Table 9.18 lists
the nutrients and recommended levels for grain
products fortification (National Academy of
Some fish species contain an enzyme that can Sciences 1974b). A summary of data relating pro-
destroy thiamin. Sulfur dioxide rapidly destroys cessing treatment to thiamin stability has been
thiamin. For this reason, sulfur dioxide is not per- given by deRitter (1976) (Table 9.19).
mitted as an additive in foods that contain appre-
ciable amounts of thiamin.
Baking of white bread may result in thiamin Vitamin B2 (Riboflavin)
loss of 20%. Thiamin loss in milk processing is as
follows: pasteurization, 3–20%; sterilization, The molecule consists of a d-ribitol unit attached
30–50%; spray drying, 10%; and roller drying, to an isoalloxazine ring (Fig. 9.16). Anything
20–30%. Cooking of meat causes losses that are more than a minor change in the molecule results
384 9 Vitamins

Fig. 9.15  Typical losses of vitamins during flour milling

Table 9.18  Nutrients and levels recommended for inclu- Table 9.19  Thiamin stability in foods
sion in fortification of cereal-grain productsa
Product Treatment Retention (%)
Level Nine canned Processing 31–89
Nutrient (mg/Ib) (mg/100 g) vegetables
Vitamin Ab 2.2 0.48 Four canned Storage, 2–3 months 73–94
Thiamin 2.9 0.64 vegetables @ room temperature
Riboflavin 1.8 0.40 Cereals Extrusion cooking 48–90
Niacin 24.0 5.29 Fortified ready- Storage, 12 months 100
to-eat cereal @ 23 °C
Vitamin B6 2.0 0.44
Bread (white, Commercial baking 74–79
Folic acid 0.3 0.07
whole wheat)
Iron 40 8.81
Devil’s food Baking 0–7
Calcium 900 198.2 cake (pH 9)
Magnesium 200 44.1
Source: From E. deRitter, Stability Characteristics of
Zinc 10 2.2 Vitamins in Processed Foods, Food Technol., Vol. 30,
a
Wheat flour, corn grits, cornmeal, rice. Other cereal-grain pp. 48–51, 54, 1976
products in proportion to their cereal-grain content
b
Retinol equivalent
Source: Reprinted with permission from National coenzymes, flavin mononucleotide (FMN) and
Academy of Sciences, Recommended Dietary Allowances, flavin adenine dinucleotide (FAD). FMN is
8th rev. ed., © 1974, National Academy of Sciences
riboflavin-5′-phosphate and forms part of several
enzymes, including cytochrome c reductase. The
in a loss of vitamin activity. Aqueous solutions of flavoproteins serve as electron carriers and are
riboflavin are yellow with a yellowish-green fluo- involved in the oxidation of glucose, fatty acids,
rescence. The vitamin is a constituent of two amino acids, and purines.
Water-Soluble Vitamins 385

bound to protein. Only in milk does riboflavin


occur mostly in the free form.
Under the influence of light and alkaline pH,
riboflavin is transformed into lumiflavin, an inac-
tive compound with a yellowish green fluores-
cence. Under acid conditions, riboflavin is
transformed into another inactive derivative, lumi-
chrome, and ribitol. This compound has a blue
fluorescence. The transformation into lumiflavin
in milk results in the destruction of ascorbic acid.
The light sensitivity of riboflavin results in
Fig. 9.16  Structural formula of riboflavin. Riboflavin:
losses of up to 50% when milk is exposed to sun-
R=OH; Riboflavin phosphate: R=PO3NaOH
light for 2 h. The nature of the packaging material
significantly affects the extent of riboflavin
Table 9.20  Riboflavin content of some foods destruction. It appears that the wavelengths of
Riboflavin (mg/100 g) light responsible for the riboflavin destruction are
Product edible portion in the visible spectrum below 500–520 nm.
Beef 0.16 Ultraviolet light has been reported to have no
Cabbage 0.05 destructive effect on riboflavin (Hartman and
Eggs 0.30 Dryden 1978). Riboflavin is stable in dry milk for
Chicken 0.19 storage periods of up to 16 months. Pasteurization
Beef liver 3.26 of milk causes only minor losses of riboflavin.
Chicken liver 2.49
Beef kidney 2.55
Peas 0.29 Vitamin B6 (Pyridoxine)
Spinach 0.20
Tomato 0.04 There are three compounds with vitamin B6
Yeast (dry) 5.41 activity. The structural formula of pyridoxine is
Milk 0.17 presented in Fig. 9.17. The other two forms of
Nonfat dry milk 1.78 this vitamin are different from pyridoxine—they
have another substituent on carbon 4 of the ben-
zene ring. Pyridoxal has a –CHO group in this
Very good sources of riboflavin are milk and position and pyridoxamine has a –CH2NH2
milk products; other sources are beef muscle, group. All three compounds can occur as salts.
liver, kidney, poultry, tomatoes, eggs, green veg- Vitamin B6 plays an important role in the metab-
etables, and yeast (Table 9.20). olism of amino acids, where it is active in the
Riboflavin is stable to oxygen and acid pH but
is unstable in alkaline medium and is very
­sensitive to light. When exposed to light, the rate
of destruction increases as pH and temperature
increase. Heating under neutral or acidic condi-
tions does not destroy the vitamin.
The human requirement for riboflavin varies
with metabolic activity and body weight and
ranges from 1 to 3 mg per day. Normal adult
requirement is 1.1–1.6 mg per day. In most cases,
the riboflavin of foods is present in the form of
the dinucleotide, the phosphoric acid ester, or is Fig. 9.17  Structural formula of pyridoxine
386 9 Vitamins

coenzyme form pyridoxal-5-phosphate. The milk as 0.54 mg per L. Other sources are meats,
three forms of vitamin B6 are equally active in liver, vegetables, whole grain cereals, and egg
rats; although it can be expected that the same yolk.
applies for humans, this has not been definitely The effects of processing on pyridoxine levels
established. in milk and milk products have been reviewed by
Vitamin B6 is widely distributed in many Hartman and Dryden (1978). No significant
foods (Table 9.21), and deficiencies of this vita- losses have been reported to result from pasteuri-
min are uncommon. The recommended allow- zation, homogenization, or production of dried
ance for adults has been established at 2 mg per milk. Heat sterilization of milk, however, has
day. The requirement appears to increase with the been reported to result in losses ranging from 36
consumption of high-protein diets. to 49%. Losses occur not only during the heat
Vitamin B6 occurs in animal tissues in the treatment but also during subsequent storage of
form of pyridoxal and pyridoxamine or as their milk. These storage losses have been attributed to
phosphates. Pyridoxine occurs in plant products. a conversion of pyridoxal to pyridoxamine and
Pyridoxine is stable to heat and strong alkali or then to a different form of the vitamin. Wendt and
acid; it is sensitive to light, especially ultraviolet Bernhart (1960) have identified this compound as
light and when present in alkaline solutions. bis-4-pyridoxal disulfide (Fig. 9.18). This com-
Pyridoxal and pyridoxamine are rapidly destroyed pound is formed by reaction of pyridoxal and
when exposed to air, heat, or light. Pyridoxamine is active sulfhydryl groups. The latter are formed
readily destroyed in food processing operations. during heat treatment of milk proteins. Exposure
Because it is difficult to determine this vita- of milk to daylight in clear glass bottles for 8 h
min in foods, there is a scarcity of information on resulted in a vitamin B6 loss of 21%.
its occurrence. Recent data establish the level in Food canning results in losses of vitamin B6 of
20–30%. Milling of wheat may result in losses of
up to 80–90%. Baking of bread may result in
Table 9.21  Vitamin B6 content of some foods losses of up to 17%.
Product Vitamin B6 (μg/g) A review of some stability data of vitamin B6
Wheat 3.2–6.1 as prepared by deRitter (1976) is given in
Whole wheat bread 4.2 Table 9.22.
White bread 1.0
Orange juice 0.52–0.60
Apple juice 0.35 Niacin
Tomatoes 1.51
Beans, canned 0.42–0.81 The term niacin is used in a generic sense for
Peas, canned 0.44–0.53 both nicotinic acid and nicotinamide (Fig. 9.19).
Beef muscle 0.8–4.0 Nicotinamide acts as a component of two impor-
Pork muscle 1.23–6.8 tant enzymes, NAD and NADP, which are
Milk, pasteurized 0.5–0.6 involved in glycolysis, fat synthesis, and tissue
Yeast 50 respiration. Niacin is also known as the pellagra

Fig. 9.18 Structural
formula of bis-4-­
pyridoxal disulfide
Water-Soluble Vitamins 387

Table 9.22  Vitamin B6 stability in foods


Product Treatment Retention (%)
Bread (added B6) Baking 100
Enriched corn meal 12 months @ 38 °C + 50% relative humidity 90–95
Enriched macaroni 12 months @ 38 °C + 50% relative humidity 100
Saccharomyces Carlsbergensis Chick Rat
Whole milk Evaporation and sterilization 30 55 65
Evaporation and sterilization +6 months @ 18 44 41
room temperature
Infant formula, Processing and sterilization 33–50 (natural)
liquid 84 (added)
Infant formula, dry Spray drying 69–83
Boned chicken Canning 57
Irradiation (2.79 megarads) 68
Source: From E. deRitter, Stability Characteristics of Vitamins in Processed Foods, Food Technol., Vol. 30, pp. 48–51,
54, 1976

Table 9.23  Niacin content of some foods


Niacin (mg/100 g)
Product edible portion
Barley (pearled) 3.1
Beans (green, snap) 0.5
Beans (white) 2.4
Beef (total edible) 4.4
Fig. 9.19  Structural formulas of (a) nicotinic acid and Beef kidney 6.4
(b) nicotinamide
Beef liver 13.6
Chicken (dark meat) 5.2
preventive factor. The incidence of pellagra has Chicken (light meat) 10.7
declined but is still a serious problem in parts of Corn (field) 2.2
the Near East, Africa, southeastern Europe, and Haddock 3.0
in North American populations that subsist on Milk 0.1
com diets. When com is treated with alkali or Mushrooms 4.2
lime, as for the tortilla preparation in Central Peanuts 17.2
America, the amount of available niacin can be Peas 2.9
greatly increased. Tryptophan can be converted Potatoes 1.5
by the body into niacin. Many diets causing pel- Spinach 0.6
lagra are low in good quality protein as well as in Wheat 4.3
vitamins. Corn protein is low in tryptophan. The Yeast (dry) 36.7
niacin of corn and other cereals may occur in a
bound form, called niacytin, that can be con- niacin equivalent of 16–33 mg. The RDA for
verted into niacin by alkali treatment. adults, expressed as niacin, is 6.6 mg per
The human requirement of niacin is related to 1000 kcal, and not less than 13 mg when caloric
the intake of tryptophan. Animal proteins contain intake is less than 2000 kcal.
approximately 1.4% of tryptophan, vegetable Good dietary sources of this vitamin are liver,
proteins about 1%. A dietary intake of 60 mg of kidney, lean meat, chicken, fish, wheat, barley,
tryptophan is considered equivalent to 1 mg of rye, green peas, yeast, peanuts, and leafy vegeta-
niacin. When this is taken into account, average bles. In animal tissues, the predominant form of
diets in the United States supply 500–1000 mg niacin is the amide. Niacin content of some foods
tryptophan per day and 8–17 mg niacin for a total are listed in Table 9.23.
388 9 Vitamins

Niacin is probably the most stable of the B Vitamin B12 (Cyanocobalamine)


vitamins. It is unaffected by heat, light, oxygen,
acid, or alkali. The main loss resulting from pro- This vitamin possesses the most complex structure
cessing involves leaching into the process water. of any of the vitamins and is unique in that it
Blanching of vegetables may cause a loss of has a metallic element, cobalt, in the molecule
about 15%. Processes in which brines are used (Fig.  9.20). The molecule is a coordination
may cause losses of up to 30%. Processing of ­complex built around a central tervalent cobalt
milk, such as pasteurization, sterilization, evapo- atom and consists of two major parts—a complex
ration, and drying have little or no effect on nico- cyclic structure that closely resembles the porphy-
tinic acid level. Virtually all the niacin in milk rins and a nucleotide-like portion, 5,6-dimethyl-l-
occurs in the form of nicotinamide. In many (α-D-ribofuranosyl) benzimidazole-3′-phosphate.
foods, application of heat, such as roasting or The phosphate of the nucleotide is esterified with
baking, increases the amount of available niacin. l-amino-2-­propanol; this, in turn, is joined by
This results from the change of bound niacin to means of an amide bond with the ­propionic acid
the free form. side chain of the large cyclic structure. A second

Fig. 9.20  Structural formula of cyanocobalamine


Water-Soluble Vitamins 389

linkage with the large structure is through the In milk, the vitamin occurs as cobalamine bound
coordinate bond between the cobalt atom and one to protein.
of the nitrogen atoms of the benzimidazole. The Vitamin B12 is not destroyed to a great extent
cyanide group can be split off relatively easily, for by cooking, unless the food is boiled in alkaline
example, by daylight. This reaction can be reversed solution. When liver is boiled in water for 5 min,
by removing the light source. The cyano group can only 8% of the vitamin B12 is lost. Broiling of
also be replaced by other groups such as hydroxo, meat may result in higher losses. Pasteurization
aquo, and nitroto. Treatment with cyanide will causes only a slight destruction of vitamin B12 in
convert these groups back to the cyano form. The milk; losses range from 7 to 10% depending on
different forms all have biological activity. pasteurization method. More drastic heat treat-
Cyanocobalamine is a component of several ment results in higher losses. Boiling milk for
coenzymes and has an effect on nucleic acid for- two to 5 min causes a 30% loss, evaporation
mation through its action in cycling 5-methyl-­ about 50%, and sterilization up to 87%. The loss
tetrahydrofolate back into the folate pool. The in drying of milk is smaller; in the production of
most important dietary sources of the vitamin are dried skim milk, the vitamin B12 loss is about
animal products. Vitamin B12 is also produced by 30%. Ultra-high-temperature sterilization of milk
many microorganisms. It is not surprising that does not cause more vitamin B12 destruction than
vitamin B12 deficiency of dietary origin only does pasteurization.
occurs in vegetarians.
The average diet in the United States is con-
sidered to supply between 5 and 15 μg/day. In Folic Acid (Folacin)
foods, the vitamin is bound to proteins via pep-
tide linkages but can be readily absorbed in the Folic acid is the main representative of a series of
intestinal tract. The RDA is 3 μg for adults and related compounds that contain three moieties:
adolescents. pterin, p-aminobenzoic acid, and glutamic acid
Few natural sources are rich in vitamin B12. (Fig.  9.21). The commercially available form
However, only very small amounts are required contains one glutamic acid residue and is named
in the diet. Good sources are lean meat, liver, pteroylglutamic acid (PGA). The naturally occur-
kidney, fish, shellfish, and milk (Table 9.24). ring forms are either PGA or conjugates with
varying numbers of glutamic acid residues, such
Table 9.24  Vitamin B12 content of some foods as tri- and heptaglutamates. It has been suggested
Product Vitamin B12 that folic acid deficiency is the most common
Beef muscle 0.25–3.4 μg/100 g vitamin deficiency in North America and Europe.
Beef liver 14–152 μg/100 g Deficiency is especially likely to occur in preg-
Milk 3.2–12.4 μg/L nant women.
Shellfish 600–970 μg/100 g (dry wt)
Egg yolk 0.28–1.556 μg/100 g

Fig. 9.21  Structural formula of folic acid


390 9 Vitamins

The vitamin occurs in a variety of foods, espe- mainly in the conjugated form; the folate in liver
cially in liver, fruit, leafy vegetables, and yeast occurs in the free form.
(Table  9.25) (Hurdle et al. 1968; Streiff 1971). The RDA for folacin is 400 μg for adults.
The usual form of the vitamin in these products is There is an additional requirement of 400 μg/day
a polyglutamate. The action of an enzyme (con- during pregnancy and 200 μg/day during
jugase) is required to liberate the folic acid for breastfeeding.
metabolic activity; this takes place in the intesti- Many of the naturally occurring folates are
nal mucosa. The folacin of foods can be divided extremely labile and easily destroyed by cook-
into two main groups on the basis of its availabil- ing. Folic acid itself is stable to heat in an acid
ity to L. casei: (1) the so-called free folate, which medium but is rapidly destroyed under neutral
is available to L. casei without conjugase treat- and alkaline conditions. In solution, the vitamin
ment; and (2) the total folate, which also includes is easily destroyed by light. Folate may occur in
the conjugates that are not normally available to a form more active than PGA; this is called
L. casei. About 25% of the dietary folacin occurs folinic acid or citrovorum factor, which is
in free form. The folate in vegetables occurs N5-formyl-5, 6, 7, 8-tetrahydro PGA (Fig. 9.22).
The folate of milk consists of up to 20% of
folinic acid. It has been reported that pasteuriza-
Table 9.25  Folate content of some foods tion and sterilization of milk involve only small
Product Folate (μg/g) losses or no loss. Hurdle et al. (1968) reported
Beef, boiled 0.03 that boiling of milk causes no loss in folate;
Chicken, roasted 0.07 however, boiling of potato results in a 90% loss
Cod, fried 0.16 and boiling of cabbage a 98% loss. Reconstitution
Eggs, boiled 0.30 of dried milk followed by sterilization as can
Brussels sprouts, boiled 0.20 occur with baby formulas may lead to signifi-
Cabbage, boiled 0.11 cant folacin losses. Fermentation of milk and
Lettuce 2.00 milk products may result in greatly increased
Potato, boiled 0.12 folate levels. Blanching of vegetables and cook-
Spinach, boiled 0.29 ing of meat do not appear to cause folic acid
Tomato 0.18 losses. Table 9.26 contains a summary of folate
Orange 0.45 stability data prepared by deRitter (1976). Citrus
Milk 0.0028 fruit and juices are relatively good sources of
Bread, white 0.17 folic acid, which is present mostly as the reduced
Bread, brown 0.38 5-methyl tetrahydro folate (monoglutamate
Orange juice, frozen reconstituted 0.50 form). There are also polyglutamate derivatives
Tomato juice, canned 0.10 present White (1991).

Fig. 9.22  Structural formula of folinic acid


Water-Soluble Vitamins 391

Table 9.26  Folic acid stability in foods Manufacture of cheese involves large losses dur-
Retention of folic acid ing processing, but during ripening the panto-
Product Treatment Free (%) Total (%) thenic acid content increases, due to synthesis by
Cabbage Boiled 5 min 32 54 microorganisms. Blanching of vegetables may
Potatoes Boiled 5 min 50 92 involve losses of up to 30%. Boiling in water
Rice Boiled 15 min – 10 involves losses that depend on the amount of
Beef, pork, Boiled 15 min <50 <50 water used.
and chicken
Various foods Cooked 27 55
Source: From E. deRitter, Stability Characteristics of Biotin
Vitamins in Processed Foods, Food Technol., Vol. 30,
pp. 48–51, 54, 1976
The structural formula (Fig. 9.24) contains three
asymmetric carbon atoms, and eight different ste-
reoisomers are possible. Only the dextrorotatory
Pantothenic Acid D-biotin occurs in nature and has biological
activity. Biotin occurs in some products in free
The free acid (Fig. 9.23) is very unstable and has form (vegetables, milk, and fruits) and in other
the appearance of a hygroscopic oil. The calcium products is bound to protein (organ meats, seeds,
and sodium salts are more stable. The alcohol and yeast). Good sources of the vitamin are meat,
(panthenol) has the same biological activity as liver, kidney, milk, egg yolk, yeast, vegetables,
the acid. Only the dextrorotatory or D form of and mushrooms (Table 9.28).
these compounds has biological activity. Biotin is important in a number of metabolic
Pantothenic acid plays an important role as a reactions, especially in fatty acid synthesis. The
component of coenzyme A, and this is the form in biotin supply of the human organism is only
which it occurs in most foods. partly derived from the diet.
Pantothenic acid occurs in all living cells and An important factor in biotin’s availability
tissues and is, therefore, found in most food prod- is that some of the vitamin is derived from
ucts. Good dietary sources include meats, liver, ­synthesis by intestinal microorganisms; this is
kidney, fruits, vegetables, milk, egg yolk, yeast, demonstrated by the fact that three to six times
whole cereal grains, and nuts (Table 9.27). In ani- more biotin is excreted in the urine than is
mal products, most of the pantothenic acid is ingested with the food. The daily intake of
present in the bound form, but in milk only about biotin is between 100 and 300 μg. No recom-
one-fourth of the vitamin is bound. mended dietary allowance has been established.
There is no recommended dietary allowance Biotin is deactivated by raw egg white. This is
for this vitamin because of insufficient evidence caused by the glycoprotein avidin. Heating
to base one on. It is estimated that adult dietary of avidin will destroy the inactivator capacity
intake in the United States ranges from 5 to for biotin.
20 mg/day, and 5 to 10 mg/day probably repre- Data on the stability of biotin are limited. The
sents an adequate intake. vitamin appears to be quite stable. Heat treat-
The vitamin is stable to air, and labile to dry ment results in relatively small losses. The vita-
heat. It is stable in solution in the pH range of 5–7 min is stable to air and is stable at neutral and
and less stable outside this range. Pasteurization acid pH. Pasteurization and sterilization of milk
and sterilization of milk result in very little or no result in losses of less than 10%. In the produc-
loss. The production and storage of dried milk tion of evaporated and dried milk, losses do not
involves little or no loss of pantothenic acid. exceed 15%.
392 9 Vitamins

Fig. 9.23  Structural formula of pantothenic acid. Pantothenic acid: R=COOH; Panthenol: R=CH2OH

Table 9.27  Pantothenic acid content of some foods


Product Pantothenic acid (μg/g)
Beef, lean 10
Wheat 11
Potatoes 6.5
Split peas 20–22
Tomatoes 1
Orange 0.7
Walnuts 8
Milk 1.3–4.2
Beef liver 25–60 Fig. 9.24  Structural formula of biotin
Eggs 8–48
Broccoli 46 Table 9.28  Biotin content of some foods
Product Biotin (μg/100 g)
Milk 1.1–3.7
Tomatoes 1
Vitamins as Food Ingredients Broad beans 3
Cheese 1.1–7.6
In addition to their role as essential micronutri- Wheat 5.2
ents, vitamins may serve as food ingredients for Beef 2.6
their varied functional properties (Institute of Beef liver 96
Food Technologists 1987). Vitamin C and vita- Lettuce 3.1
min E have found widespread use as antioxidants. Mushrooms 16
In lipid systems, vitamin E may be used as an Potatoes 0.6
antioxidant in fats that have little or no natural Spinach 6.9
tocopherol content. Ascorbic acid in the form of Apples 0.9
its palmitic acid ester, ascorbyl palmitate, is an Oranges 1.9
effective antioxidant in lipid systems. Ascorbyl Peanuts 34
palmitate prevents the formation of lipid free rad-
icals (Fig. 9.25) and thereby delays the initiation
of the chain reaction that leads to the deteriora- loxidase to quinones. The quinones rapidly
tion of the fat (Liao and Seib 1987). Ascorbyl pal- polymerize to form brown pigments. This reac-
mitate is used in vegetable oils because it acts tion is easily reversed by ascorbic acid (Fig. 9.27).
synergistically with naturally occurring tocopher- The carotenoids β-carotene and β-apo-8-­
ols. The tocopherols are fat-soluble antioxidants carotenal are used as colorants in fat-based as
that are used in animal fats. Ascorbic acid reduces well as water-based foods.
nitrous acid to nitric oxide and prevents the for- Other functions of ascorbic acid are inhibition
mation of N-nitrosamine. The reaction of nitrous of can corrosion in canned soft drinks, protection
acid and ascorbic acid is given in Fig. 9.26 (Liao of flavor and color of wine, prevention of black
and Seib 1987). Ascorbic acid is also widely used spot formation in shrimp, stabilization of cured
to prevent enzymic browning in fruit products. meat color, and dough improvement in baked
Phenolic compounds are oxidized by polypheno- goods (Institute of Food Technologists 1987).
Vitamins as Food Ingredients 393

Fig. 9.25  Prevention of lipid free radical formation in linoleic acid by ascorbyl palmitate

Fig. 9.26 Reaction
between nitrous acid and
ascorbic acid. Source:
From M.L. Liao and
P.A. Seib, Selected
Reactions of L-Ascorbic
Acid Related to Foods,
Food Technol., Vol. 41,
no. 11, pp. 104–107,
1987
394 9 Vitamins

Fig. 9.27  Reduction of ortho-quinone by ascorbic acid during enzymatic browning

deRitter, E. (1976). Stability characteristics of vitamins in


References processed foods. Food Technology, 30, 48–51.
Eitenmiller, R. R. (1997). Vitamin E content of fats and
Baltes, J. (1967). Tocopherols as fat stabilizers. In oils: Nutritional implications. Food Technology, 51(5),
K. Lang (Ed.), Tocopherol. Darmstadt: Steinkopff 78–81.
Verlag. Gross, J. (1987). Pigments in fruits. London: Harcourt
Bender, A. E. (1971). The fate of vitamins in food process- Brace Jovanovich.
ing operations. In M. Stein (Ed.), Vitamins. London: Hartman, A. M., & Dryden, L. P. (1978). The vitamins in
Churchill Livingstone. milk and milk products. In B. H. Webb, A. H. Johnson,
Bullock, D. H. (1968). Stability of vitamin C in enriched & J. A. Alford (Eds.), Fundamentals of dairy chemis-
commercial evaporated milk. Journal of Dairy try (pp. 325–401). Westport, CT: The Avi Publishing
Science, 51, 921–923. Company Inc.
Cook, J. D., & Reddy, M. B. (2001). Effect of ascorbic Hurdle, A. D., Barton, D., & Searles, I. H. (1968). A
acid intake on nonheme-iron absorption from a com- method for measuring folate in food and its appli-
plete diet. The American Journal of Clinical Nutrition, cation to a hospital diet. The American Journal of
73, 93–98. Clinical Nutrition, 21(10), 1202–1207.
deMan, J. M. (1981). Light-induced destruction of Institute of Food Technologists. (1987). Use of vitamins
vitamin A in milk. Journal of Dairy Science, 64, as additives in processed foods. Food Technology,
2031–2032. 41(9), 163–168.
deMan, J. M. (1986). Stability of vitamin A bead lets in Liao, M. L., & Seib, P. A. (1987). Selected reactions of
nonfat dry milk. Milchwissenschaft, 41, 468–469. l-ascorbic acid related to foods. Food Technology, 41,
104–107. 111.
References 395

Menger, A. (1957). Investigation of the stability of vita- Sattar, A., & deMan, J. M. (1973). Effect of packaging
min E in cereal milling products and baked goods. material on light-induced quality deterioration of milk.
Brot Gebck German, 11, 167–173. Canadian Institute of Food Science and Technology
Morton, R. A. (1967). The chemistry of tocopherols. In Journal, 6(3), 170–174.
K. Lang (Ed.), Tocopherol. Darmstadt: Steinkopff Stocker, R., & Kearney, J. F., Jr. (2004). Role of oxida-
Verlag. tive modifications in atherosclerosis. Physiological
National Academy of Sciences. (1974a). Recommended Reviews, 84(4), 1381–1478. https://doi.org/10.1152/
dietary allowances (8th ed.). Washington, DC: physrev.00047.2003.
National Academy Press. Streiff, R. R. (1971). Folate levels in citrus and other
National Academy of Sciences. (1974b). Proposed forti- juices. The American Journal of Clinical Nutrition,
fication policy for cereal-grain products. Washington, 24(12), 1390–1392.
DC: National Academy Press. Thaler, H. (1967). Concentration and stability of tocopher-
Odland, D., & Eheart, M. S. (1975). Ascorbic acid, min- ols in foods. In K. Lang (Ed.), Tocopherol. Darmstadt:
eral and quality retention in frozen broccoli blanched Steinkopff Verlag.
in water, steam and ammonia-steam. Journal of Food Wendt, G., & Bernhart, E. W. (1960). The structure of a
Science, 49, 1004–1007. sulfur-containing compound with vitamin B6 activ-
Ong, A. S. H. (1993). Natural sources of tocotrienols. In ity. Archives of Biochemistry and Biophysics, 88,
L. Packer & J. Fuchs (Eds.), Vitamin E in health and 270–272.
disease. New York: Marcel Dekker. White, D. R. (1991). Reverse phase HPLC/EC determina-
Rouseff, R. L., & Nagy, S. (1994). Health and nutritional tion of folate in citrus juice by direct injection with
benefits of citrus fruit components. Food Technology, column switching. Journal of Agricultural and Food
48(11), 126–132. Chemistry, 39, 714–717.
Enzymes
10
Chang Yong Lee and John M. deMan

The research on enzymes has immense practi-


Introduction cal importance in agriculture and food process-
ing. There is not a single food system that does
Enzymes, although minor constituents of many not involve enzyme reactions. Enzymes that are
foods, play a major and manifold role in foods. naturally present in foods may change the com-
They are highly specialized proteins with the spe- position of those foods; in some cases, such
cial ability to catalyze specific chemical reactions changes are desirable but in most instances are
in biological systems. Although they may undesirable, so the enzymes must be deactivated.
undergo change during the catalysis, they are The blanching of vegetables is an example of
unchanged at the end of the reaction. They are enzyme deactivation to prevent undesirable
highly selective catalysts, greatly accelerating change. Some enzymes are used as indicators in
both the rate and specificity of metabolic reac- analytical methods; phosphatase, for instance, is
tions from digestion of food to the synthesis of used in the phosphatase test of pasteurization of
DNA. An enzyme can catalyze only a single reac- milk. Enzymes are also used as processing aids in
tion of a single compound, called the enzyme’s food manufacturing. For example, rennin, con-
substrate. For example, amylase found in the tained in extract of calves’ stomachs, is used as a
human digestive tract catalyzes only the hydroly- coagulant for milk in the production of cheese.
sis of starch to yield glucose; cellulose and other Recent advances in biotechnology offer a new
polysaccharides are untouched by amylase. Other promise for tailoring enzymes for specific func-
enzymes have different specificities. Papain, for tions of particular applications and their produc-
example, a globular protein isolated from papaya tion in quantities for industrial uses. By
fruit, catalyzes the hydrolysis of many kinds of manipulating the genetic code of an enzyme, it is
peptide bonds, which makes papain useful as a now possible to target a specific amino acid resi-
meat tenderizer and a cleaner for contact lenses. due at any location for modification. During the
Enzymes act only to lower the activation energy last three decades, the use of cellulases, hemicel-
for a reaction, thereby making the reaction takes lulases and pectinase which account for approxi-
place more rapidly. Starch and water, for exam- mately 20% of the world enzyme market for
ple, react very slowly in the absence of a catalyst food, brewery, wine, and textile industries has
because the activation energy is too high. When increased considerably and many commercial
amylase is present, however, the energy barrier is enzyme producers are marketing tailor-made
lowered, and the hydrolysis reaction occurs enzyme preparations suitable for specific pur-
rapidly. poses (Wong 1995; Bhat 2000).

© Springer International Publishing AG 2018 397


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_10
398 10 Enzymes

Food science’s emphasis in the study of Whitaker (1972) has prepared an extensive
enzymes differs from that in biochemistry. The ­listing of the uses of enzymes in food process-
former deals mostly with decomposition reac- ing (Table 10.1) and this gives a good summary
tions, hydrolysis, and oxidation; the latter is of the many and varied possible applications
more concerned with synthetic mechanisms. of enzymes.

Table 10.1  Some uses and suggested uses of enzymes in foods and food processing
Enzyme Food Purpose or action
Amylases Baked goods Increase sugar content for yeast fermentation
Brewing Conversion of starch to maltose for fermentation; removal of starch
turbidities
Cereals Conversion of starch to dextrins, sugar; increase water absorption
Chocolate-­cocoa Liquidification of starches for free flow
Confectionery Recovery of sugar from candy scraps
Fruit juices Remove starches to increase sparkling properties
Jellies Remove starches to increase sparkling properties
Pectin An aid in preparation of pectin from apple pomace
Syrups and sugars Conversion of starches to low molecular weight dextrins (corn syrup)
Vegetables Hydrolysis of starch as in tenderization of peas
Cellulase Brewing Hydrolysis of complex carbohydrate cell walls
Coffee Hydrolysis of cellulose during drying of beans
Fruits Removal of graininess of pears; peeling of apricots, tomatoes
Dextran-­ Sugar syrups Thickening of syrup
sucrase Ice cream Thickening agent, body
Invertase Artificial honey Conversion of sucrose to glucose and fructose
Candy Manufacture of chocolate-coated, soft, cream candies
Lactase Ice cream Prevent crystallization of lactose, which results in grainy, sandy texture
Feeds Conversion of lactose to galactose and glucose
Milk Stabilization of milk proteins in frozen milk by removal of lactose
Tannase Brewing Removal of polyphenolic compounds
Pentosanase Milling Recovery of starch from wheat flour
Naringinase Citrus Debittering citrus pectin juice by hydrolysis of the glucoside, naringin
Pectic Chocolate-­cocoa Hydrolytic activity during fermentation of cocoa
enzymes
(useful) Coffee Hydrolysis of gelatinous coating during fermentation of beans
Fruits Softening
Fruit juices Improve yield of press juices, prevent cloudiness, improve concentration
processes
Olives Extraction of oil
Wines Clarification
Pectic Citrus juice Destruction and separation of pectic substances of juices
enzymes Fruits Excessive softening action
(deteriorative)
(continued)
Table 10.1 (continued)
Enzyme Food Purpose or action
Proteases Baked goods Softening action in doughs; cut mixing time, increase extensibility of doughs;
(useful) improvement in grain, texture, loaf volume; liberate β-amylase
Brewing Body, flavor and nutrients development during fermentation; aid in filtration
and clarification, chill-proofing
Cereals Modify proteins to increase drying rate, improve product handling
characteristics; manufacture of miso and tofu
Cheese Casein coagulation; characteristic flavors during aging
Chocolate-­cocoa Action on beans during fermentation
Eggs, egg products Improve drying properties
Feeds Use in treatment of waste products for conversion to feeds
Meats and fish Tenderization; recovery of protein from bones, trash fish; liberation of oils
Milk In preparation of soybean milk
Protein Condiments such as soy sauce and tamar sauce; specific diets; bouillon,
hydrolysates dehydrated soups, gravy powders, processed meats
Wines Clarification
Proteases Eggs Shelf life of fresh and dried whole eggs
(deteriorative) Crab, lobster Overtenderization if not inactivated rapidly
Flour Influence on loaf volume, texture if too active
Lipase Cheese Aging, ripening, and general flavor characteristics
(useful) Oils Conversion of lipids to glycerol and fatty acids
Milk Production of milk with slightly cured flavor for use in milk chocolate
Lipase Cereals Overbrowning of oat cakes; brown discoloration of wheat bran
(deteriorative) Milk and dairy Hydrolytic rancidity
products
Oils Hydrolytic rancidity
Phosphatases Baby foods Increase available phosphate
Brewing Hydrolysis of phosphate compounds
Milk Detection of effectiveness of pasteurization
Nucleases Flavor enhancers Production of nucleotides and nucleosides
Peroxidases Vegetables Detection of effectiveness of blanching
(useful) Glucose In combination with glucose oxidase
determinations
Peroxidases Vegetables Off-flavors
(deteriorative) Fruits Contribution to browning action
Catalase Milk Destruction of H2O2 in cold pasteurization
Variety of products To remove glucose and/or oxygen to prevent browning and/or oxidation; used
in conjunction with glucose oxidase
Glucose Variety of products Removal of oxygen and/or glucose from products such as beer, cheese,
oxidase carbonated beverages, dried eggs, fruit juices, meat and fish, milk powder,
wine to prevent oxidation and/or browning; used in conjunction with catalase
Glucose Specific determination of glucose; used in conjunction with peroxidase
determination
Polyphenol Tea, coffee, Development of browning during ripening, fermentation, and/or aging process
oxidase tobacco
(useful)
Polyphenol Fruits, vegetables Browning, off-flavor development, loss of vitamins
oxidase
(deteriorative)
Lipoxygenase Vegetables Destruction of essential fatty acids and vitamin A; development of off-flavors
Ascorbic acid Vegetables, fruits Destruction of vitamin C (ascorbic acid)
oxidase
Thiaminase Meats, fish Destruction of thiamine
Source: Reprinted with permission from Whitaker, J. R. (1972). Principles of enzymology for the food sciences, by
courtesy of Marcel Dekker, Inc
400 10 Enzymes

Nature and Kinetics of Enzymes proportional to the active enzyme concentration,


and dependent on substrate, inhibitor, and cofactor
Nature of Enzymes concentration, and on temperature and pH.

The catalytic properties of enzymes are quite Enzyme Concentration: The relationship between
specific, which makes enzymes useful in analyti- reaction velocity and enzyme concentration can
cal studies. Some enzymes consist only of pro- be illustrated as the following Fig. 10.2.
tein, but most enzymes contain additional In the first part of the curve, a straight line plot
nonprotein components such as carbohydrates, results when the amount of enzyme is increased in
lipids, metals, phosphates, or some other organic the presence of an excess of substrate that shows the
moiety. The complete enzyme is called holoen- concentration of an enzyme is directly proportional
zyme; the protein part, apoenzyme; and the non- to the rate of the reaction. In these conditions, dou-
protein part, cofactor. The compound that is bling the amount of enzyme will double the reaction
being converted in an enzymic reaction is called velocity. In the second part of the line, at higher lev-
substrate. In an enzyme reaction, the substrate els of enzyme concentration, the amount of sub-
combines with the holoenzyme and is released in strate becomes the limiting factor and the linear
a modified form, as indicated in Fig. 10.1. relationship cannot be maintained, causing the line
to flatten out. Under these conditions, adding more
enzyme has no effect on reaction rate.
Kinetics of Enzymes
Substrate Concentration: For a given amount of
Enzyme activity can be controlled in a number of enzyme under standard conditions, the initial
ways that are very important to food chemists. The reaction velocity (V0) varies with an increase of
velocity of an enzyme-catalyzed reaction is usually initial substrate concentration (S). Figure 10.3

Fig. 10.1  The nature of


enzymes—substrate
reactions
Nature and Kinetics of Enzymes 401

Fig. 10.2  Effect of


enzyme concentration
on reaction velocity

Fig. 10.3  Effect of


substrate concentration
on the initial velocity of
an enzyme catalyzed
reaction

shows the effect on V0 of varying (S) when the (E-S) explained by German scientists, Leonor
enzyme concentration is held constant. When Michaelis and Maud Menten in 1913. They pos-
substrate concentrations are low compare to tulated that enzyme first combines reversibly
available enzyme, V0 is nearly linearly related to with its substrate to form an enzyme-substrate
(S) because not all of the enzyme molecules are complex in a fast reversible step, then breaks
combined with substrate. Therefore, substrate down in a slower second step to yield the free
concentration is the limiting factor accounting enzyme (E) and the reaction product (P). The sec-
for the lower reaction rate. At high substrate con- ond step is slower and limits the rate of the over-
centration, V0 increases by smaller amounts in all reaction.
response to increases in (S) and almost unaf-
k1
fected by change in substrate concentration. This
plateau is called the maximum velocity (Vmax), as E+S⇔ E−S (10.1)
shown in Fig. 10.3. k2
This hyperbolic shape of the curve may be
k3
explained in terms of the following equation with (10.2)
the formation of an enzyme-substrate complex E−S→ E+P
402 10 Enzymes

At any given instant in an enzyme-catalyzed This Michaelis-Menten eq. (10.3) can be


reaction, the enzyme exists in two forms, the free a­lgebraically transformed into forms that are
form, E and the combined form, ES. At low (S), useful in the practical determination of
­ Km
most of the enzyme will be in the E form and the and Vmax.
rate will be proportional to (S) because of the By taking the reciprocal of both sides of the
equilibrium will be pushed toward formation of Michaelis-Menten equation:
more E−S as (S) is increased. The maximum ini-
1 K + (S) 1 Km (S)
tial rate of the catalyzed reaction (Vmax) is observed = m and = +
when all of enzyme is present as E−S. At this V0 Vmax (S) V0 Vmax (S) Vmax (S)

point the enzyme is saturated with its substrate, so
that further increases in (S) have no effect on rate. and simplifies to
The saturation effect is a distinguishing character-
1 K 1 1
istic of enzyme catalysis and responsible for the = m +
plateau observed in Fig. 10.3. After the E−S V0 Vmax (S) Vmax

breaks down to yield the product P, the enzyme is
free to catalyze another reaction (Eq. 10.2). This equation is a transform of the Michaelis-­
The above curve is described by the Michaelis-­ Menten equation called the Lineweaver-Burk
Menten equation: equation. For enzymes obeying the Michaelis-­
Menten relationship, a plot of 1/V0 versus 1/(S)
Vmax (S)
V0 = (10.3) yields a straight line as shown in Fig. 10.4.
K m + (S) This double-reciprocal presentation is also

called a Lineweaver-Burk plot. The straight line
where will have a slop of Km/Vmax, an intercept of 1/Vmax
V0 = initial reaction velocity, on the 1/V0 axis, and an intercept of −1/Km on the
(S) = initial substrate concentration, 1/(S) axis. The Km value which is sometimes used
Vmax = maximum reaction velocity, attained as an indication of the affinity of an enzyme for
when E−S is at its maximum value, its substrate can vary greatly from enzyme to
Km = Michaelis-Menten constant, the sub- enzyme, therefore, useful for the study and
strate concentration where one-half of the maxi- ­comparison of different enzymes. Vmax also varies
mum reaction velocity is attained. greatly from one enzyme to the next.
This equation shows the quantitative relation-
ship between the initial velocity V0, the maxi-
mum initial velocity Vmax, and the initial substrate
concentration (S), all related through the
Michaelis-Menten constant Km. The effects of
enzyme concentration and of substrate concen-
tration on the reaction velocity are often described
in terms of the order of the reaction that expresses
the reaction rate as a function of the concentra-
tion of one or more of the substances present. An
enzymatic reaction follows first-order kinetics
whenever the substrate concentration is much
less than the Km value for that particular reaction.
When the substrate concentration is much greater
than the Km value, the reaction is zero-order with
respect to substrate concentration, since all the
enzyme molecules are fully saturated with sub-
strate molecules. Fig. 10.4  Lineweaver-Burk (or double-reciprocal) plot
Nature and Kinetics of Enzymes 403

Effect of Temperature: Changes in temperature combination required for the inactivation of unde-
affect the rate of enzyme reaction. It is well sirable enzymes in foods. Some enzymes are
known that enzyme-catalyzed reaction occur known to regenerate when they are cooled follow-
more slowly when a food is placed in a refrigera- ing denaturation by heat. Peroxidase in vegetables
tor, but the reactions do not stop at 0–4 °C. Most which has been inactivated by scalding (blanch-
enzyme-catalyzed reactions decrease 1.4–2 time ing) can recover at least part of its enzyme activity
per 10 °C decrease in temperature. The usual during frozen storage.
effect of temperature on enzymatic reactions is in
two stages: (1) the rate of the reaction increases Effect of pH: The hydrogen ion concentration of
with increasing temperature up to maximum (see the medium in which the enzyme works affects
the following graph); (2) above this temperature greatly the activity of the enzyme because each
followed by decreasing activity at higher tem- enzyme works within a small pH range.
peratures, due to denaturation of the enzyme. Therefore, changing the pH of the system by 1 or
2 pH units from the optimum pH, can decrease
enzyme velocity to 0.5 or 0.1, respectively, of
that at the pH optimum. The greatest reaction
velocity is attained at the optimal pH of the
enzyme. Most enzymes pH optima lie often
within the pH range of 4.5–8.0: β-amylase, 4.8;
invertase, 5.0, and pectin methylesterase, 6.5–
8.0. Denaturation of the enzyme by high or low
pH can result in inactivation. Thus, it is important
that pH must be controlled for two main reasons:
(1) the enzyme reaction proceeds at a maximum
rate at a specific pH and (2) the range of maxi-
mum stability of an enzyme also occurs at a defi-
nite pH. In food industry, the pH is controlled
either to inhibit the enzyme activity or to produce
the maximum activity. In fruit and vegetable
The increasing temperature at the start pro- products, the pH is lowered by the addition of
duces greater molecular activity, which increases compounds such as citric or phosphoric acids. As
the rate of reaction. The enzyme activity increases shown in the following figure, when the activity
with temperature increase such that the reaction of an enzyme is plotted against varying pH values
velocity is approximately doubled for every 10 °C the result is ordinarily a bell-shaped.
rise up to 45–50 °C for most enzymes. The tem-
perature coefficient, Q10, is used as an expression
of the change in rate of reaction for a 10 °C change
in temperature. The value of Q10 is determined by
dividing the reaction rate at a given temperature
plus 10 °C by the reaction rate at that given tem-
perature. At higher temperature, the enzyme
begins to denature (loss of activity), followed by
rapid thermal inactivation at above 60–75 °C. Some
enzymes, such as peroxidase, are much more ther-
mostable than others. The criterion of heat stabil- Effect of Inhibitors: Some substances reduce or
ity is very valuable in characterizing an enzyme even stop the catalytic activity of enzymes in bio-
and is more pertinent in food processing. Heat- chemical reactions. They block or distort the
stability studies determine the temperature-time active site. Inhibitors that occupy the active site
404 10 Enzymes

and prevent a substrate molecule from binding to the active site of the enzyme, as well as the sub-
the enzyme are said to be active site-directed. It is strate, are important. But this complementarity
called competitive inhibitor, as they ‘compete’ may be even further expanded to cover amino
with the substrate for the active site. Competitive acid residues in the vicinity of the active site,
inhibitors are often compounds that resemble the hydrophobic areas near the active site, or the
substrate and combine with the enzyme. presence of a positive electrical charge near the
Inhibitors that attach to other parts of the enzyme active site (Parkin 1993). Types of specificity
molecule, perhaps distorting its shape, are said to may include group, bond, stereo, and absolute
be non-active site-directed are called noncom- specificity, or some combination of these. An
petitive inhibitors. example of the specificity of enzymes is given in
Practical methods of enzyme inhibition that Fig.  10.5, which illustrates the specificity of
may be used in the food industry are very limited proline-­specific peptidases (Habibi-Najafi and
due to problems associated with sensory quality Lee 1996). The amino acid composition of casein
and economic feasibility. In practice, the food is high in proline, and the location of this amino
industry is limited in enzyme inhibition or con- acid in the protein chain is inaccessible to com-
trol to inactivation by heat, pH, dehydration, high mon aminopeptidases and the di- and tripepti-
pressure processing or the use of chemicals such dases with broad specificity. Hydrolysis of the
as sulfur dioxide and phenolic antioxidants. proline bonds requires proline-specific pepti-
dases, including several exopeptidases and an
endopeptidase. Figure 10.5 illustrates that this
Specificity type of specificity is related to the type of amino
acid in a protein as well as its location in the
The nature of the enzyme-substrate reaction as chain. Neighboring amino acids also determine
explained in Fig. 10.1 requires that each enzyme the type of peptidase required to hydrolyze a par-
reaction is highly specific. The shape and size of ticular peptide bond.

Fig. 10.5  Mode of action of proline-specific peptidases. Adopted from from Habibi-Najafi, M. B., & Lee, B. H.
Bitterness in cheese: A review. Critical Reviews in Food Science and Nutrition, 36(5), 408
Hydrolases 405

Classification The hydrolases are classified on the basis of


the type of bond hydrolyzed. The most important
Enzymes are classified by the Commission on are those that act on ester bonds, glycosyl bonds,
Enzymes of the International Union of peptide bonds, and C–N bonds other than
Biochemistry. The basis for the classification is peptides.
the division of enzymes into groups according to
the type of reaction catalyzed. This, together with
the name or names of substrate(s), is used to Esterases
name individual enzymes. Each well-defined
enzyme can be described in three ways—by a The esterases are involved in the hydrolysis of
systematic name, by a trivial name, and by a ester linkages of various types. The products
number of the Enzyme Commission (EC). Thus, formed are acid and alcohol. These enzymes may
the enzyme α-amylase (trivial name) has the sys- hydrolyze triglycerides and include several
tematic name α-l,4-glucan-4-glucanohydrolase, lipases; for instance, phospholipids are hydro-
and the number EC 3.2.1.1. The system of lyzed by phospholipases, and cholesterol esters
nomenclature has been described by Whitaker are hydrolyzed by cholesterol esterase. The car-
(1972, 1974) and Parkin (1993). boxylesterases are enzymes that hydrolyze tri-
glycerides such as tributyrin. They can be
distinguished from lipases because they hydro-
Enzyme Production lyze soluble substrates, whereas lipases only act
at the water-lipid interfaces of emulsions.
Some of the traditionally used industrial Therefore, any condition that results in increased
enzymes (e.g., rennet and papain) are prepared surface area of the water-lipid interface will
from animal and plant sources. Recent develop- increase the activity of the enzyme. This is the
ments in industrial enzyme production have reason that lipase activity is much greater in
emphasized the microbial enzymes (Frost 1986). homogenized (not pasteurized) milk than in the
Microbial enzymes are very heat stable and non-homogenized product. Most of the lipolytic
have a broader pH optimum. Most of these enzymes are specific for either the acid or the
enzymes are made by submerged cultivation of alcohol moiety of the substrate, and, in the case
highly developed strains of microorganisms. of esters of polyhydric alcohols, there may also
Developments in biotechnology will make it be a positional specificity.
possible to transfer genes for the elaboration of Lipases are produced by microorganisms
specific enzymes to different organisms. The such as bacteria and molds; are produced by
major industrial enzyme processes are listed in plants; are present in animals, especially in the
Table 10.2. pancreas; and are present in milk. Lipases may
cause spoilage of food because the free fatty
acids formed cause rancidity. In other cases, the
Hydrolases action of lipases is desirable and is produced
intentionally. The boundary between flavor and
The hydrolases as a group include all enzymes off-flavor is often a very narrow range. For
that involve water in the formation of their prod- instance, hydrolysis of milk fat in milk leads to
ucts. For a substrate AB, the reaction can be rep- very unpleasant off-flavors at very low free fatty
resented as follows: acid concentration. The hydrolysis of milk fat
AB + HOH → HA + BOH in cheese contributes to the desirable flavor.
These differences are probably related to the
Table 10.2  Major industrial enzymes and the process used for their production
Submerged Surface Further Solid Solution Immobilized
Enzyme Source fermentation fermentation Intracellular Extracellular Concentration Precipitation Drying Pelleting purification product product product
Proteases
Rennet Calf stomach – – – ✓ – – ✓ – – ✓ ✓ –
Trypsin Animal pancreas – – – ✓ ✓ ✓ ✓ – ✓ ✓ – –
Papain Carica papaya fruit – – – ✓✓ – – ✓ – – ✓ ✓ –
Fungal Aspergillus oryzae ✓ ✓ – ✓ ✓ ✓ ✓ – – ✓ – –
Fungal (rennins) Mucor spp. ✓ ✓ – ✓ ✓ – ✓ – – ✓ ✓ –
Bacterial Bacillus spp. ✓ – – ✓ ✓ ✓ ✓ ✓ – ✓ ✓ –
Glycosidases
Bacterial Bacillus spp. ✓ – – ✓ ✓ – ✓ – – ✓ ✓ –
α-amylase
Fungal α-amylase Aspergillus oryzae ✓ – – ✓ ✓ – ✓ – – ✓ ✓ –
β-amylase Barley – – – ✓ ✓ ✓ ✓ – – ✓ – –
Amyloglucosidase Aspergillus niger ✓ – – ✓ ✓ – – – – – ✓ –
Pectinase Aspergillus niger – ✓ – ✓ ✓ – ✓ – – ✓ ✓ –
Cellulase Molds ✓ – – ✓ ✓ ✓ – – ✓ ✓ – –
Yeast lactase Kluyveromyces spp. ✓ – ✓ – – ✓ ✓ – – ✓ ✓ –
Mold lactase Aspergillus spp. ✓ ✓ – ✓ – ✓ ✓ – – ✓ – ✓
Others
Glucose isomerase Various microbial ✓ – ✓ – ✓ ✓ ✓ – – – – ✓
sources
Glucose oxidase Aspergillus niger ✓ – ✓ – – ✓ – – ✓ ✓ ✓ –
Mold catalase Aspergillus niger ✓ – ✓ – – ✓ – – – – ✓ –
Animal catalase Liver – – ✓ – ✓ ✓ ✓ – ✓ ✓ ✓ –
Lipase Molds ✓ – – ✓ ✓ ✓ ✓ – – ✓ – –
Source: From Frost, G. M. (1986). Commercial production of enzymes. In B. J. F. Hudson (Ed.), Developments in food proteins. Elsevier Applied Science Publishers Ltd
Hydrolases 407

background upon which these fatty acids are


superimposed and to the specificity for particular
groups of fatty acids of each enzyme. In seeds,
lipases may cause fat hydrolysis unless the
enzymes are destroyed by heat. Palm oil pro-
duced by primitive methods in Africa used to
consist of more than 10% of free fatty acids.
Such spoilage problems are also encountered in
grains and flour. The activity of lipase in wheat
and other grains is highly dependent on water
content. In wheat, for example, the activity of
lipase is five times higher at 15.1% than at 8.8%
moisture. The lipolytic activity of oats is higher
than that of most other grains.
Lipases can be divided into those that have a
positional specificity and those that do not. The
former preferentially hydrolyze the ester bonds
of the primary ester positions. This results in the
formation of mono- and diglycerides, as repre-
sented by the following reaction:

During the progress of the reaction, the con- increases with age, as is demonstrated by the
centration of diglycerides and monoglycerides increasing content of partial glycerides during
increases, as is shown in Fig. 10.6. The the aging of cheese (Table 10.4). In many cases,
β-monoglycerides formed are resistant to further lipolysis is induced by the addition of lipolytic
hydrolysis. This pattem is characteristic of pan- enzymes. In the North American chocolate indus-
creatic lipase and has been used to study the tri- try, it is customary to induce some lipolysis in
glyceride structure of many fats and oils. chocolate by means of lipase. In the production
The hydrolysis of triglycerides in cheese is an of Italian cheeses, lipolysis is induced by the use
example of a desirable flavor-producing process. of pregastric esterases. These are lipolytic
The extent of free fatty acid formation is much enzymes obtained from the oral glands located at
higher in blue cheese than in Cheddar cheese, as the base of the tongue in calves, lambs, or kids.
is shown in Table 10.3. This is most likely the Specificity for certain fatty acids by some
result of lipases elaborated by organisms growing lipolytic enzymes has been demonstrated.
in the blue cheese, such as P. roqueforti, P. cam- Pancreatic lipase and milk lipase are broad-­
emberti, and others. The extent of lipolysis spectrum enzymes and show no specificity for
408 10 Enzymes

Fig. 10.6  The course of


pancreatic lipase
hydrolysis of tricaprylin.
MG monoglycerides,
DG diglycerides, TG
triglycerides. Source:
From Boudreau, A., &
deMan, J. M. (1965).
The mode of action of
pancreatic lipase on
milkfat glycerides.
Canadian Journal of
Biochemistry, 43,
1799–1805

Table 10.3  Free fatty acids in some dairy products any of the fatty acids found in fats. Instead, the
Product Free fatty acids (mg/kg) fatty acids that are released from the glycerides
Fresh milk 415 occur in about the same ratio as they are present
Moderately rancid cream 1027 in the original fat. Specificity was shown by
Butter 2733 Nelson (1972) in calf esterase and in a mixed
Cheddar cheese 1793 (avg. of 12 samples) pancreatin-esterase preparation (Table 10.5).
Blue cheese 23,500–66,700 Pregastric esterases and lipase from Aspergillus
(range 3 samples) species primarily hydrolyze shorter chain-length
Source: From Day, E. A. (1966). Role of milk lipids in fatty acids (Arnold et al. 1975).
flavors of dairy products. In R. F. Gould (Ed.), Flavor
chemistry. American Chemical Society
Specificity of lipases may be expressed in a
number of different ways—substrate specific,
regiospecific, nonspecific, fatty acyl specific, and
stereospecific. Examples of these specificities
Table 10.4  Formation of partial glycerides in cheddar have been presented by Villeneuve and Foglia
cheese (1997) (Table 10.6).
Diglycerides Mono-glycerides Substrate specificity is the ability to hydro-
Product type (wt %) (wt %) lyze a particular glycerol ester, such as when a
Mild 7.4–7.6 1.0–2.0 lipase can rapidly hydrolyze a triacylglycerol,
Medium 7.6–9.7 0.5–1.4 but acts on a monoacylglycerol only slowly.
Old 11.9–15.6 1.1–3.2 Regiospecificity involves a specific action on
Hydrolases 409

Table 10.5  Free fatty acids released from milkfat by several lipolytic enzymes
Fatty acid Milk lipase Steapsin Pancreatic lipase Calf esterase Esterase pancreatin
4:0 13.9 10.7 14.4 35.00 15.85
6:0 2.1 2.9 2.1 2.5 3.6
8:0 1.8 1.5 1.4 1.3 3.0
10:0 3.0 3.7 3.3 3.1 5.5
12:0 2.7 4.0 3.8 5.1 4.4
14:0 7.7 10.7 10.1 13.2 8.5
16:0 21.6 21.6 24.0 15.9 19.3
18:1 and 18:2 29.2 24.3 25.5 14.2 21.1
18:0 10.5 13.4 9.7 3.2 10.1
Source: From Nelson, J. H. (1972). Enzymatically produced flavors for fatty systems. Journal of the American Oil
Chemists’ Society, 49, 559–562

Table 10.6  Examples of lipase specificities has opened up the possibility of tailor-making
Specificity Lipase triacylglycerols with a specific structure, and this
Substrate specific is especially important for producing high-value
Monoacylglyercols Rat adipose tissue fats such as cocoa butter equivalents. The cata-
Mono- and diacylglycerols Penicillium camembertii lytic activity of lipases is reversible and depends
Triacylglycerols Penicillium sp. on the water content of the reaction mixture. At
Regiospecific high water levels, the hydrolytic reaction prevails,
1,3-regioselective Aspergilllus niger whereas at low water levels the synthetic reaction
Rhizopus arrhizus is favored. A number of lipase catalyzed reactions
Mucor miehei are possible, and these have been summarized in
sn-2-regioselective Candida antarctica A Fig. 10.7 (Villeneuve and Foglia 1997). Most of
Nonspecific Penicillium expansum the lipases used for industrial processes have
Aspergillus sp. been developed from microbes because these
Pseudomonas cepacia usually exhibit high temperature tolerance.
Fatty acylspecific Lipases from Mucor miehei and Candida antarc-
Short-chain fatty acid (FA) Penicillium roqueforti tica have been cloned and expressed in industry-­
Premature infant gastric friendly organisms. Lipases from genetically
cis-9 unsaturated FA Geotrichum candidum engineered strains will likely be of major indus-
Long-chain unsaturated FA Botrytis cinerea
trial importance in the future (Godtfredsen 1993).
Stereospecific
Fatty acid–specific lipases react with either short-­
sn-1 stereospecific Humicola lanuginosa
chain fatty acids (Penicillium roqueforti) or some
Pseudomonas aeruginosa
long-chain fatty acids such as cis-9-unsaturated
sn-3 stereospecific Fusarium solani cutinase
fatty acids (Geotrichum candidum). Stereospecific
Rabbit gastric
lipases react with only fatty acids at the sn-1 or
Source: Reprinted with permission from Villeneuve, R., &
Foglia, T. A. Lipase specificities: Potential application in
sn-3 position.
lipid bioconversions. Journal of the American Oil The applications of microbial lipases in the
Chemists’ Society, 8, 641, © 1997, AOCS Press food industry involve the hydrolytic as well as
the synthetic capabilities of these enzymes and
either the sn-1 and sn-3 positions or reaction have been summarized by Godtfredsen (1993) in
with only the sn-2 position. The 1,3-specific Table 10.7.
enzymes have been researched extensively, The lipase-catalyzed interesterification pro-
because it is now recognized that lipases in cess can be used for the production of triacylg-
­addition to hydrolysis can catalyze the reverse lycerols with specific physical properties, and it
reaction, esterification or transesterification. This also opens up possibilities for making so-called
410 10 Enzymes

Fig. 10.7  Lipase catalyzed reactions used in oil and fat Potential application in lipid bioconversions. Journal of
modification. Source: Reprinted with permission from the American Oil Chemists’ Society, 8, 642, © 1997,
Villeneuve, R., & T. A. Foglia. Lipase specificities: AOCS Press

Table 10.7  Application of microbial lipases in the food industry


Industry Effect Product
Dairy Hydrolysis of milk fat Flavor agents
Cheese ripening Cheese
Modification of butter fat Butter
Bakery Flavor improvement and shelf-life prolongation Bakery products
Beverage Improved aroma Beverages
Food dressing Quality improvement Mayonnaise, dressing, and whipped toppings
Health food Transesterification Health foods
Meat and fish Flavor development and fat removal Meat and fish products
Fat and oil Transesterification Cocoa butter, margarine
Hydrolysis Fatty acids, glycerol, mono- and diglycerides
Source: Reprinted with permission from Godtfredsen, S. E. Lipases, enzymes in food processing. T. Nagodawithana
and G. Reed (Eds.), p. 210, © 1993, Academic Press
Hydrolases 411

structured lipids (Akoh 1997). An example is a specifically hydrolyze the 1,6-linkages between
triacylglycerol that carries an essential fatty acid chains, and the enzymes that split the 1,4-­linkages
(e.g., DHA-­docosahexaenoic acid) in the sn-2 between glucose units of the straight chains. The
position and short-chain fatty acids in the sn-1 latter group consists of endoenzymes that cleave
and sn-3 positions. Such a structural triacylglyc- the bonds at random points along the chains and
erol would rapidly be hydrolyzed in the diges- exoenzymes that cleave at specific points near the
tive tract and provide an easily absorbed chain ends. This behavior has been represented
monoacylglycerol that carries the essential fatty by Marshall (1975) as a diagram of the struc-
acid (Godtfredsen 1993). ture of amylopectin (Fig. 10.8). In this molecule,
The lipases that have received attention for the 1,4-α-glucan chains are interlinked by
their ability to synthesize ester bonds have been 1,6-α-glucosidic linkages resulting in a highly
obtained from yeasts, bacteria, and fungi. Lipases branched molecule. The molecule is com posed
can be classified into three groups according of three types of chains; the A chains carry no
to their specificity (Macrae 1983). The first substituent, the B chains carry other chains linked
group contains nonspecific lipases. These show to a primary hydroxyl group, and the molecule
no specificity regarding the position of the contains only one C chain with a free reducing
ester bond in the glycerol molecule, or the nature glucose unit. The chains are 25–30 units in length
of the fatty acid. Examples of enzymes in this in starch and only 10 units in glycogen.
group are lipases of Candida cylindracae,
Corynebacterium acnes, and Staphylococcus Alpha-Amylase (α-1,4-Glucan
aureus. The second group contains lipases with 4-Glucanohydrolase)
position specificity for the 1- and 3-positions of This enzyme is distributed widely in the animal
the glycerides. This is common among microbial and plant kingdoms. The enzyme contains 1
lipases and is the result of the sterically hindered gram-atom of calcium per mole. Alpha-amylase
ester bond of the 2-position’s inability to enter (α-1,4-glucan-4-glucanohydrolase) is an endoen-
the active site of the enzyme. Lipases in this zyme that hydrolyzes the α-l,4-glucosidic bonds
group are obtained from Aspergillus niger, Mucor in a random fashion along the chain. It hydro-
javanicus, and Rhizopus arrhizus. The third lyzes amylopectin to oligosaccharides that con-
group of lipases show specificity for particular tain two to six glucose units. This action,
fatty acids. An example is the lipase from therefore, leads to a rapid decrease in viscosity,
Geotrichum candidum, which has a marked spec- but little monosaccharide formation. A mixture
ificity for long-chain fatty acids that contain a cis of amylose and amylopectin will be hydrolyzed
double bond in the 2-position. The knowledge of into a mixture of dextrins, maltose, glucose, and
the synthetic ability of lipases has opened a whole oligosaccharides. Amylose is completely
new area of study in the modification of fats. The ­hydrolyzed to maltose, although there usually is
possibility of modifying fats and oils by immobi- some maltotriose formed, which hydrolyzes only
lized lipase technology may result in the produc- slowly.
tion of food fats that have a higher essential fatty
acid content and lower trans levels than is possi- Beta-Amylase (α-1,4-Glucan
ble with current methods of hydrogenation. Maltohydrolase)
This is an exoenzyme and removes successive
maltose units from the nonreducing end of the
Amylases glucosidic chains. The action is stopped at the
branch point where the α-1,6 glucosidic linkage
The amylases are the most important enzymes of cannot be broken by α-amylase. The resulting
the group of glycoside hydrolases. These starch-­ compound is named limit dextrin. Beta-amylase
degrading enzymes can be divided into two is found only in higher plants. Barley malt, wheat,
groups, the so-called debranching enzymes that sweet potatoes, and soybeans are good sources.
412 10 Enzymes

Fig. 10.8  Diagrammatic representation of amylopectin 1,4-bonds. The branch points are 1,6-α glucosidic bonds.
structure. Lines represent α-d-glucan chains linked by Source: From Marshall, J. J. (1975). Starch degrading
enzymes, old and new. Starke, 27, 377–383

Beta-amylase is technologically important in the be completely degraded to glucose. The enzyme


baking, brewing, and distilling industries, where is present in bacteria and molds and is used
starch is converted into the fermentable sugar industrially in the production of corn syrup and
maltose. Yeast ferments maltose, sucrose, invert glucose.
sugar, and glucose but does not ferment dextrins A problem in the enzymic conversion of corn
or oligosaccharides containing more than two starch to glucose is the presence of t­ ransglucosidase
hexose units. enzyme in preparations of α-amylase and glu-
coamylase. The transglucosidase catalyzes the
Glucoamylase (α-1,4-Glucan formation of oligosaccharides from glucose, thus
Glucohydrolase) reducing the yield of glucose.
This is an exoenzyme that removes glucose units Nondamaged grains such as wheat and barley
in a consecutive manner from the nonreducing contain very little α-amylase but relatively high
end of the substrate chain. The product formed is levels of β-amylase. When these grains germi-
glucose only, and this differentiates this enzyme nate, the β-amylase level hardly changes, but the
from α- and β-amylase. In addition to hydrolyz- α-amylase content may increase by a factor of
ing the α-1,4 linkages, this enzyme can also 1000. The combined action of α- and β-amylase
attack the α-1,6 linkages at the branch point, in the germinated grain greatly increases the pro-
albeit at a slower rate. This means that starch can duction of fermentable sugars. The development
Hydrolases 413

Table 10.8  Development of α-amylase during malting Pectic Enzymes


of barley at 20 °C
Days of steeping α-Amylase The pectic enzymes are capable of degrading
and germination (20° dextrose units) pectic substances and occur in higher plants and
0   0 in microorganisms. They are not found in higher
3  55
animals, with the exception of the snail. These
5 110
enzymes are commercially important for the
7 130
treatment of fruit juices and beverages to aid in
8 135
filtration and clarification and increasing yields.
Source: From Green, S. R. (1969). New use of enzymes in
The enzymes can also be used for the production
the brewing industry. MBAA Technical Quarterly, 6, 33–39
of low methoxyl pectins and galacturonic acids.
The presence of pectic enzymes in fruits and veg-
of α-amylase activity during malting of barley is etables can result in excessive softening. In
shown in Table 10.8. In wheat flour, high tomato and fruit juices, pectic enzymes may
α-amylase activity is undesirable, because too cause “cloud” separation.
much carbon dioxide is formed during baking. There are several groups of pectic enzymes,
Raw, nondamaged, and ungelatinized starch is including pectinesterase, the enzyme that hydro-
not susceptible to β-amylase activity. In contrast, lyzes methoxyl groups, and the depolymerizing
α-amylase can slowly attack intact starch gran- enzymes polygalacturonase and pectate lyase.
ules. This differs with the type of starch; for
example, waxy corn starch is more easily attacked  ectinesterase (Pectin Pectyl-Hydrolase)
P
than potato starch. In general, extensive hydroly- This enzyme removes methoxyl groups from
sis of starch requires gelatinization. Damaged pectin. The enzyme is referred to by several other
starch granules are more easily attacked by amy- names, including pectase, pectin methoxylase,
lases, which is important in bread making pectin methyl esterase, and pectin demethylase.
Alphaamylase can be obtained from malt, from Pectinesterases are found in bacteria, fungi, and
fungi (Aspergillus oryzae), or from bacteria (B. higher plants, with very large amounts occurring
subtilis). The bacterial amylases have a higher in citrus fruits and tomatoes. The enzyme is spe-
temperature tolerance than the malt amylases. cific for galacturonide esters and will not attack
non-galacturonide methyl esters to any large
Beta-Galactosidase (β-d-Galactoside extent. The reaction catalyzed by pectin esterase
Galactohydrolase) is presented in Fig. 10.9. It has been suggested
This enzyme catalyzes the hydrolysis of β-d-­ that the distribution of methoxyl groups along the
galactosides and α-l-arabinosides. It is best chain affects the reaction velocity of the enzyme
known for its action in hydrolyzing lactose and (MacMillan and Sheiman 1974). Apparently,
is, therefore, also known as lactase. The enzyme pectinesterase requires a free carboxyl group
is widely distributed and occurs in higher ani- next to an esterified group on the galacturonide
mals, bacteria, yeasts, and plants. Beta-­ chain to act, with the pectinesterase moving down
galactosidase or lactase is found in humans in the the chain linearly until an obstruction is reached.
cells of the intestinal mucous membrane. A con- To maintain cloud stability in fruit juices,
dition that is widespread in non-Caucasian adults high-temperature–short-time (HTST) pasteuriza-
is characterized by an absence of lactase. Such tion is used to deactivate pectolytic enzymes.
individuals are said to have lactose intolerance, Pectin is a protective colloid that helps to keep
which is an inability to digest milk properly. insoluble particles in suspension. Cloudiness is
The presence of galactose inhibits lactose required in commercial products to provide a
hydrolysis by lactase. Glucose does not have this desirable appearance. The destruction of the high
effect. levels of pectinesterase during the production of
414 10 Enzymes

Fig. 10.9  Reaction catalyzed by pectinesterase

Fig. 10.10  Reaction catalyzed by polygalacturonase

tomato juice and puree is of vital importance. The Table 10.9  Action of polygalacturonases
pectinesterase will act quite rapidly once the Type Preferred
tomato is broken. In the so-called hot-break of attack Enzyme substrate
method, the tomatoes are broken up at high tem- Random Endo-­polymethylgalacturonase Pectin
perature so that the pectic enzymes are destroyed Random Endo-polygalacturonase Pectic acid
instantaneously. Terminal Exo-­polymethylgalacturonase Pectin
Terminal Exo-polygalacturonase Pectic acid
Polygalacturonase (Poly-α-1,4-­
Galacturonide Glycanohydrolase)
This enzyme is also known as pectinase, and it substrates (pectins), whereas others act on sub-
hydrolyzes the glycosidic linkages in pectic sub- strates with free carboxylic acid groups (pectic
stances according to the reaction pattern shown in acids). These enzymes are named polymethyl
Fig. 10.10. The polygalacturonases can be divided galacturonases and polygalacturonases, respec-
into endoenzymes that act within the molecule on tively. The preferential mode of hydrolysis and the
α-1,4 linkages and exoenzymes that catalyze the preferred substrates are listed in Table 10.9.
stepwise hydrolysis of galacturonic acid molecules Endopolygalacturonases occur in fruits and in fila-
from the nonreducing end of the chain. A further mentous fungi, but not in yeast or bacteria.
division can be made by the fact that some Exopolygalacturonases occur in plants (for exam-
­polygalacturonases act principally on methylated ple, in carrots and peaches), fungi, and bacteria.
Hydrolases 415

 ectate Lyase (Poly-α-l,4-d-­


P Bacterial degradation of pectin in plant tissues is
Galacturonide Lyase) responsible for the spoilage known as “soft rot” in
This enzyme is also known as trans-eliminase; it fruits and vegetables. Commercial food grade pec-
splits the glycosidic bonds of a glucuronide chain tic enzyme preparations may contain several differ-
by trans elimination of hydrogen from the 4- and ent pectic enzymes. Usually, one type predominates;
5-positions of the glucuronide moiety. The reac- this depends on the intended use of the enzyme
tion pattern is presented in Fig. 10.11. The glyco- preparation.
sidic bonds in pectin are highly susceptible to this
reaction. The pectin lyases are of the endotype
and are obtained exclusively from filamentous Proteases
fungi, such as Aspergillus niger. The purified
enzyme has an optimum pH of 5.1 to 5.2 and iso- Proteolytic enzymes are important in many indus-
electric point between 3 and 4 (Albersheim and trial food processing procedures. The reaction
Kilias 1962). catalyzed by proteolytic enzymes is the hydroly-
sis of peptide bonds of proteins; this reaction is
Commercial Use shown in Fig. 10.12. Whitaker (1972) has listed
Pectic enzymes are used commercially in the the specificity requirements for the hydrolysis of
clarification of fruit juices and wines and for aid- peptide bonds by proteolytic enzymes. These
ing the disintegration of fruit pulps. By reducing include the nature of R1, and R2 groups, configura-
the large pectin molecules into smaller units and tion of the amino acid, size of substrate molecule,
eventually into galacturonic acid, the compounds and the nature of the X and Y groups. A major
become water soluble and lose their suspending distinguishing factor of proteolytic enzymes is
power; also, their viscosity is reduced and the the effect of R1, and R2 groups. The enzyme
insoluble pulp particles rapidly settle out. α-chymotrypsin hydrolyzes peptide bonds readily
Most microorganisms produce at least one but only when R1 is part of a tyrosyl, phenylalanyl, or
usually several pectic enzymes. Almost all fungi tryptophanyl residue. Trypsin requires R1 to
and many bacteria produce these enzymes, which belong to an arginyl or lysyl residue. Specific
readily degrade the pectin layers holding plant cells requirement for the R2 groups is exhibited by pep-
together. This leads to separation and degradation sin and the carboxypeptidases; both require R2 to
of the cells, and the plant tissue becomes soft. belong to a phenylalanyl residue. The enzymes

Fig. 10.11  Reaction catalyzed by pectin lyase

Fig. 10.12  Reaction catalyzed by proteases


416 10 Enzymes

require the amino acids of proteins to be in the conversion to rennin, the disulfide bridges remain
L-configuration but frequently do not have a strict intact. As the calves grow older and start to eat
requirement for molecular size. The nature of X other feeds as well as milk, the stomach starts to
and Y permits the division of proteases into endo- produce pepsin instead of rennin. The optimum
peptidases and exopeptidases. The former split activity of rennin is at pH 3.5, but it is most stable
peptide bonds in a random way in the interior of at pH 5; the clotting of cheese milk is carried out
the substrate molecule and show maximum activ- at pH values of 5.5–6.5.
ity when X and Y are derived. The carboxypepti- The coagulation or clotting of milk by rennin
dases require that Y be a hydroxyl group, the occurs in two stages. In the first, the enzymic
aminopeptidases require that X be a hydrogen, stage, the enzyme acts on κ-casein so that it can
and the dipeptidases require that X and Y both be no longer stabilize the casein micelle. The sec-
underived. ond, or nonenzymic stage, involves the clotting
Proteolytic enzymes can be divided into the of the modified casein micelles by calcium ions.
following four groups: the acid proteases, the ser- The enzymic stage involves a limited and specific
ine proteases, the sulfhydryl proteases, and the action on the κ-casein, resulting in the formation
metal-containing proteases. of insoluble para-κ-casein and a soluble macro-
peptide. The latter has a molecular weight of
Acid Proteases 6000–8000, is extremely hydrophilic, and
This is a group of enzymes with pH optima at low ­contains about 30 percent carbohydrate. The gly-
values. Included in this group are pepsin, rennin comacropeptide contains galactosamine, galac-
(chymosin), and a large number of microbial and tose, and N-acetyl neuraminic acid (sialic acid).
fungal proteases. Rennin, the pure enzyme con- The splitting of the glycomacropeptide from
tained in rennet, is an extract of calves’ stomachs κ-casein involves the breaking of a phenylalanine-­
that has been used for thousands of years as a methionine bond in the peptide chain. Other clot-
coagulating agent in cheese making. Because of ting enzymes—including pepsin, chymotrypsin,
the scarcity of calves’ stomachs, rennet substi- and microbial proteases—break the same bond
tutes are now widely used, and the coagulants and produce the same glycomacropeptide.
used in cheese making usually contain mixtures Pepsin is elaborated in the mucosa of the stom-
of rennin and pepsin and/or microbial proteases. ach lining in the form of pepsinogen. The high
Some of the microbial proteases have been used acidity of the stomach aids in the autocatalytic
for centuries in the Far East in the production of conversion into pepsin. This conversion involves
fermented foods such as soy sauce. splitting several peptide fragments from the
Rennin is present in the fourth stomach of the N-terminal end of pepsinogen. The fragments
suckling calf. It is secreted in an inactive form, a consist of one large peptide and several small
zymogen, named prorennin. The crude extract ones. The large peptide remains associated with
obtained from the dried stomachs (vells) contains pepsinogen by noncovalent bonds and acts as an
both rennin and prorennin. The conversion of inhibitor. The inhibitor dissociates from pepsin at
prorennin to rennin can be speeded up by addi- a pH of 1–2. In the initial stages of the conversion
tion of acid. This conversion involves an auto- of pepsinogen to pepsin, six peptide bonds are
catalytic process, in which a limited proteolysis broken, and continued action on the large peptide
of the prorennin occurs, thus reducing the molec- (Fig.  10.13) results in three more bonds being
ular weight about 14 percent. The conversion can hydrolyzed. In this process, the molecular weight
also be catalyzed by pepsin. The process involves changes from 43,000 to 35,000 and the isoelectric
the release of peptides from the N-terminal end point changes from 3.7 to less than 1. The pepsin
of prorennin, which reduces the molecular weight molecule consists of a single polypeptide chain
from about 36,000 to about 31,000. The molecule that contains 321 amino acids. The tertiary struc-
of prorennin consists of a single peptide chain ture is stabilized by three disulfide bridges and a
joined internally by three disulfide bridges. After phosphate linkage. The phosphate group is
Hydrolases 417

Fig. 10.13  Structure of


pepsinogen and its
conversion to pepsin.
Source: From Bovey,
F. A., & Yanari, S. S.
(1960). Pepsin. In P. D.
Boyer et al. (Eds.) The
enzymes (vol. 4).
Academic Press

attached to a seryl residue and is not essential for The molecular mass of the two enzymes is simi-
enzyme activity. The pH optimum of pepsin is lar, 35 kDa, but chymosin has a higher pI. Much
pH 2 and the enzyme is stable from pH 2–5. At of the chymosin used in cheese making is now
higher pH values, the enzyme is rapidly denatured obtained by genetic engineering processes. In the
and loses its activity. The primary specificity of production of soy sauce and other eastern food
pepsin is toward the R2 group (see the equation products, such as miso (an oriental fermented
shown in Fig. 10.12), and it prefers this to be a food) and ketjap (Indonesian type soy sauce), the
phenylalanyl, tyrosyl, or tryptophanyl group. acid proteases of Aspergillus oryzae are used.
The use of other acid proteases as substitutes Other products involve the use of the fungus
for rennin in cheese making is determined by Rhizopus oligosporus. Acid proteases also play a
whether bitter peptides are formed during ripen- role in the ripening process of a variety of soft
ing of the cheese and by whether initial rapid cheeses. This includes the Penicillia used in the
hydrolysis causes excessive protein losses in the blue cheeses, such as Roquefort, Stilton, and
whey. Some of the acid proteases used in cheese Danish blue, and in Camembert and Brie. The
making include preparations obtained from the molds producing the acid proteases may grow
organisms Endothia parasitica, Mucor miehei, either on the surface of the cheese or throughout
and Mucor pusillus. Rennin contains the enzyme the body of the cheese.
chymosin, and the scarcity of this natural enzyme
preparation for cheese making resulted in the use Serine Proteases
of pepsin for this purpose. Pepsin and chymosin This group includes the chymotrypsins, trypsin,
have primary structures that have about 50% elastase, thrombin, and subtilisin. The name of
homology and quite similar tertiary structures. this group of enzymes refers to the seryl residue
418 10 Enzymes

that is involved in the active site. As a conse- does not. The molecular weights of the enzymes
quence, all of these enzymes are inhibited by are quite similar; that of ficin is 25,500 and that
diisopropylphosphorofluoridate, which reacts of bromelain, 20,000–33,200. These enzymes
with the hydroxyl group of the seryl residue. catalyze the hydrolysis of many different com-
They also have an imidazole group as part of the pounds, including peptide, ester, and amide
active site and they are all endopeptides. The chy- bonds. The variety of peptide bonds split by
motrypsins, trypsin and elastase, are pancreatic papain appears to indicate a low specificity. This
enzymes that carry out their function in the intes- has been attributed (Liener 1974) to the fact that
tinal tract. They are produced as inactive zymo- papain has an active site consisting of seven sub-
gens and are converted into the active form by sites that can accommodate a variety of amino
limited proteolysis. acid sequences in the substrate. The specificity in
this case is not determined by the nature of the
Sulfhydryl Proteases side chain of the amino acid involved in the sus-
These enzymes obtain their name from the fact ceptible bond but rather by the nature of the adja-
that a sulfhydryl group in the molecule is essen- cent amino acids.
tial for their activity. Most of these enzymes are Commercial use of the sulfhydryl proteases
of plant origin and have found widespread use in includes stabilizing and chill proofing of beer.
the food industry. The only sulfhydryl proteases Relatively large protein fragments remaining
of animal origin are two of the cathepsins, which after the malting of barley may cause haze in beer
are present in the tissues as intracellular enzymes. when the product is stored at low temperatures.
The most important enzymes of this group are Controlled proteolysis sufficiently decreases the
papain, ficin, and bromelain. Papain is an enzyme molecular weight of these compounds so that
present in the fruit, leaves, and trunk of the they will remain in solution. Another important
papaya tree (Carica papaya). The commercial use is in the tenderizing of meat. This can be
enzyme is obtained by purification of the exudate achieved by injecting an enzyme solution into the
of full-grown but unripe papaya fruits. The puri- carcass or by applying the enzyme to smaller cuts
fication involves use of affinity chromatography of meat. The former method suffers from the dif-
on a column containing an inhibitor (Liener ficulty of uneven proteolysis in different parts of
1974). This process leads to the full activation of the carcass with the risk of overtenderizing some
the enzyme, which then contains 1 mole of sulf- parts of the carcass.
hydryl per mole of protein. The crude papain is
not fully active and contains only 0.5 mole of Metal-Containing Proteases
sulfhydryl per mole of protein. Bromelain is These enzymes require a metal for activity and
obtained from the fruit or stems of the pineapple are inhibited by metal-chelating compounds.
plant (Ananas comosus). The stems are pressed They are exopeptidases and include carboxypep-
and the enzyme precipitated from the juice by tidase A (peptidyl-l-amino-acid hydrolase) and B
acetone. Ficin is obtained from the latex of tropi- (peptidyl-l-lysine hydrolase), which remove
cal fig trees (Ficus glabrata). The enzyme is not amino acids from the end of peptide chains that
homogeneous and contains at least three different carry a free α-carboxyl group. The aminopepti-
proteolytic components. dases remove amino acids from the free α-amino
The active sites of these plant enzymes con- end of the peptide chain. The metalloexopepti-
tain a cysteine and a histidine group that are dases require a divalent metal as a cofactor; the
essential for enzyme activity. The pH optimum is carboxypeptidases contain zinc. These enzymes
fairly broad and ranges from 6 to 7.5 The enzymes are quite specific in the action; for example, car-
are heat stable up to temperatures in the range of boxypeptidase B requires the C-terminal amino
60–80 °C. The papain molecule consists of a acid to be either arginine or lysine; the require-
­single polypeptide chain of 212 amino acids. The ment for carboxypeptidase A is phenylalanine,
molecular weight is 23,900. Ficin and bromelain tryptophan, or isoleucine. These specificities are
contain carbohydrate in the molecule; papain compared with those of some other proteolytic
Hydrolases 419

Fig. 10.14  Specificity of some proteolytic enzymes

enzymes in Fig. 10.14. The carboxypeptidases Table 10.10 Protein hydrolysate products produced


are relatively small molecules; molecular weight from casein and whey protein concentrate (WPC)
of carboxypeptidase A is 34,600. The amino pep- Protein Average
tidases have molecular weights around 300,000. Hydrolysatea source molecular weightb ΑΝ/TΝc
Although many of the aminopeptidases are found Intact protein Casein 28,500 0.07
in animal tissues, several are present in microor- WPC 25,000 0.06
ganisms (Riordan 1974). Proteose Casein 6000 0.13
WPC 6800 0.11
Peptone Casein 2000 0.24
Protein Hydrolysates WPC 1400 0.24
Peptides Casein 400 0.48
WPC 375 0.43
Protein hydrolysates is the name given to a fam-
Peptides and Casein 260 0.55
ily of protein breakdown products obtained by
free amino acids
the action of enzymes. It is also possible to WPC 275 0.58
hydrolyze proteins by chemical means, acids, or
Source: Reprinted with permission from Lahl, W. J., &
alkali, but the enzymatic method is preferred. S. D. Braun. Enzymatic production of protein hydroly-
Many food products such as cheese and soy sauce sates for food use. Food Technology, 48(10), 69, © 1994,
are obtained by enzymatic hydrolysis. The pur- Institute of Food Technologists
a
Commercial hydrolysates produced by Deltown
pose of the production of protein hydrolysates is
Specialties, Fraser, NY
to improve nutritional value, cost, taste, antige- b
Determined by reverse-phase HPLC
nicity, solubility, and functionality. The proteins c
Ratio of amino nitrogen present in the hydrolysate to the
most commonly selected for producing hydroly- total amount of nitrogen present in the substrate
sates are casein, whey protein, and soy protein
(Lahl and Braun 1994). Proteins can be broad specificity and some preference for terminal
­hydrolyzed in steps to yield a series of proteoses, hydrophobic amino acids. Peptides containing ter-
peptones, peptides, and finally amino acids minal hydrophobic amino acids cause bitterness.
(Table 10.10). These products should not be con- Usually a mixture of different proteases is
fused with hydrolyzed vegetable proteins, which employed. The hydrolysis reaction is terminated
are intended as flavoring substances. by adjusting the pH and increasing the temperature
The extent of hydrolysis of protein hydrolysates to inactivate the enzymes. The process for produc-
is measured by the ratio of the amount of amino ing hydrolysates is shown in Fig. 10.15 (Lahl and
nitrogen to the total amount of nitrogen present in Braun 1994). Protein hydrolysates can be used as
the raw material (AN/TN ratio). Highly hydro- food ingredients with specific functional proper-
lyzed materials have AN/TN ratios of 0.50–0.60. ties or for physiological or medical reasons. For
To obtain the desired level of hydrolysis in a pro- example, hydrolyzed proteins may lose allergenic
tein, a combination of proteases is selected. Serine properties by suitably arranged patterns of hydro-
protease prepared from Bacillus licheniformis has lysis (Cordle 1994).
420 10 Enzymes

Fig. 10.15  Process for the production of protein hydroly- hydrolysates for food use. Food Technology, 48(10), 70, ©
sates. Source: Reprinted with permission from Lahl, 1994, Institute of Food Technologists
W. J., & Braun, S. D. Enzymatic production of protein
Oxidoreductases 421

Oxidoreductases or catechol oxidase. It is generally agreed


(Mathew and Parpia 1971) that these terms
Phenolases include all enzymes that have the capacity to oxi-
dize phenolic compounds to o-quinones. This can
The enzymes involved in enzymic browning are be represented by the conversion of o-­
known by the name polyphenoloxidase and are dihydroxyphenol to o-quinone,
also called polyphenolase, phenolase, tyrosinase,

The action of polyphenolases is detrimental peaches, bananas, avocados, tea leaves, and
when it leads to browning in bruised and ­coffee beans.
­broken plant tissue but is beneficial in the pro- In addition to changing o-diphenols into o-qui-
cessing of tea and coffee. The enzyme occurs nones, the enzymes also catalyze the conversion
in almost all plants, but relatively high levels of monophenols into o-diphenols (called mono-
are found in potatoes, mushrooms, apples, phenol monooxygenase or cresolase), as follows:

where BH2 stands for an o-diphenolic compound. involvement of copper is essential. Copper has
To distinguish this type of activity from the been found as a component of all polypheno-
one mentioned earlier, it is described as cresolase lases. The activity of cresolase involves three
activity, whereas the other is referred to as cate- steps, which can be represented by the following
cholase activity. For both types of activity, the overall equation (Mason 1956):

Protein − Cu 2 + − O2 + monophenol → Protein − Cu 2 + + o − quinone + H 2 O


422 10 Enzymes

Fig. 10.16  Phenolase catalyzed reactions. (a) Activation monophenols. Source: From Mason, H. S. Mechanisms of
of phenolase. (b–d) Two-step four-electron reduction of oxygen metabolism. Advances in Enzymology, 19,
oxycuprophenolase, and the associated hydroxylation of 79–233, © 1957

The protein copper-oxygen complex is formed dehydroxylating intermediate, and (Cu)n repre-
by combining one molecule of oxygen with the sents the actual charge designation of the cop-
protein to which two adjacent cuprous atoms are per at the active site. In preparations high in
attached. cresolase activity, n = 2, and in preparations
Catecholase activity involves oxidizing two high in catecholase activity, n = 1. The overall
molecules of o-diphenols to two molecules of reaction involves the use of one molecule of
o-quinones, resulting in the reduction of one mol- ­oxygen, one atom of which goes into the forma-
ecule of oxygen to two molecules of water. The tion of the diphenol, and the other, which is
action sequence as presented in Fig. 10.16 has reduced to water. This can be expressed in the
been proposed by Mason (1957). The enzyme-­ following equation given by Mathew and Parpia
oxygen complex serves as the hydroxylating or (1971):

enzyme

Monophenol + O2 + o − diphenol → o − diphenol + quinone + H 2 O

The substrates of the polyphenol oxidase possible. The first and obvious one, although
enzymes are phenolic compounds present in rarely practical, involves the exclusion of molec-
plant tissues, mainly flavonoids. These include ular oxygen. Another approach is the addition of
catechins, anthocyanidins, leucoanthocyanidins, reducing agents that can prevent the accumula-
flavonols, and cinnamic acid derivatives. tion of o-quinones. Heat treatment is effective in
Polyphenol oxidases from different sources show deactivating the enzymes. Metal complexing
distinct differences in their activity for different agents may deactivate the enzyme by making the
substrates. Some specific examples of polyphe- copper unavailable.
nolase substrates are chlorogenic acid, caffeic One of the most useful methods involves the
acid, dicatechol, protocatechuic acid, tyrosine, use of L-ascorbic acid as a reducing agent. This is
catechol, dihydroxyphenylalanine, pyrogallol, practiced extensively in the commercial produc-
and catechins. tion of fruit juices and purees. The ascorbic acid
To prevent or minimize enzymic browning of reacts with the o-quinones and changes them
damaged plant tissue, several approaches are back into o-diphenols (Fig. 10.17).
Oxidoreductases 423

Fig. 10.17  Reaction of


l-ascorbic acid with
o-quinone in the
prevention of enzymic
browning

 lucose Oxidase (β-d-Glucose:


G the oxygen by the use of a mixture of glucose oxi-
Oxygen Oxidoreductase) dase and catalase will prevent these peroxides
from forming. The glucose oxidase promotes the
This enzyme catalyzes the oxidation of d-glucose formation of gluconic acid with uptake of one
to δ-d-gluconolactone and hydrogen peroxide in molecule of oxygen. The catalase decomposes the
the presence of molecular oxygen, as follows: hydrogen peroxide formed into water and one
enzyme
half-molecule of oxygen. The net result is the
C6 H12 O6 → C6 H10 O6 + H 2 O2 uptake of one half-molecule of oxygen. The over-

all reaction can be written as follows:
The enzyme is present in many fungi and is
highly specific for β-d-glucopyranose. It has been Glucose + ½ O2 glucose
 oxidase
→ gluconic acid
catalase
established that the enzyme does not oxidize glu-
cose by direct combination with molecular oxygen. Recent information suggests that this applica-
The mechanism as described by Whitaker (1972) tion is not effective because of reversible inhibi-
involves the oxidized form of the enzyme, flavin tion of the glucose oxidase by the dyes used in
adenine dinucleotide (FAD), which serves as a soft drinks below pH 3 (Hammer 1993).
dehydrogenase. Two hydrogen atoms are removed This enzyme mixture can also remove glucose
from the glucose to form the reduced state of the from eggs before drying to prevent Maillard type
enzyme, FADH2, and δ-d-­gluconolactone. The browning reactions in the dried product.
enzyme is then reoxidized by molecular oxygen.
The gluconolactone is hydrolyzed in the presence
of water to form d-gluconic acid.  atalase (Hydrogen Peroxide:
C
In food processing, glucose oxidase is used to Hydrogen Peroxide Oxidoreductase)
remove residual oxygen in the head space of bot-
tled or canned products or to remove glucose. Catalase catalyzes the conversion of two mole-
Light has a deteriorative effect on citrus beverages. cules of hydrogen peroxide into water and molec-
Through the catalytic action of light, peroxides are ular oxygen as follows:
formed that lead to oxidation of other components, catalase
resulting in very unpleasant off-­flavors. Removing 2 H 2 O2 → 2 H 2 O + O2
424 10 Enzymes

This enzyme occurs in plants, animals, and  eroxidase (Donor: Hydrogen


P
microorganisms. The molecule has four subunits; Peroxide Oxidoreductase)
each of these contains a protohemin group, which
forms part of four independent active sites. The The reaction type catalyzed by peroxidase
molecular weight is 240,000. Catalase is less involves hydrogen peroxide as an acceptor, and a
stable to heat than is peroxidase. At neutral pH, compound AH2 as a donor of hydrogen atoms, as
catalase will rapidly lose activity at 35 °C. In shown:
addition to catalyzing the reaction shown above peroxidase
(catalatic activity), catalase can also have peroxi- H 2 O2 + AH 2 → 2 H2 O + A

datic activity. This occurs at low concentrations
of hydrogen peroxide and in the presence of In contrast to the action of catalase, no molec-
hydrogen donors (e.g., alcohols). ular oxygen is formed.
In plants, catalase appears to have two func- The peroxidases can be classified into the two
tions. First is the ability to dispose of the excess groups, iron-containing peroxidases and flavopro-
H2O2 produced in oxidative metabolism, and sec- tein peroxidases. The former can be further subdi-
ond is the ability to use H2O2 in the oxidation of vided into ferriprotoporphyrin peroxidases and
phenols, alcohols, and other hydrogen donors. verdoperoxidases. The first group contains ferri-
The difference in heat stability of catalase and protoporphyrin III (protohemin) as the prosthetic
peroxidase was demonstrated by Lopez et al. group (Fig. 10.18). The common plant peroxidases
(1959). They found that blanching of southern (horseradish, fig, and turnip) are in this group and
peas for 1 min in boiling water destroys 70–90% the enzymes are brown when highly purified. The
of the peroxidase activity and 80–100% of the second group includes the peroxidases of animal
catalase activity. tissue and milk (lactoperoxidase). In these
The combination of glucose oxidase and cata- enzymes, the prosthetic group is an iron porphyrin
lase is used in a number of food processing appli- nucleus but not protohemin. When highly purified,
cations, including the removal of trace glucose or these enzymes are green in color. Flavoprotein
oxygen from foods and in the production of glu- peroxidases occur in microorganisms and animal
conic acid from glucose. Greenfield and Lawrence tissues. The prosthetic group is FAD.
(1975) have studied the use of these enzymes in The linkage between the iron-containing
their immobilized form on an inorganic support. prosthetic group and the protein can be stabilized

Fig. 10.18 Structural
formula of
ferriprotoporphyrin III
(protohemin). Source:
From Whitaker, J. R.
(1974). Food related
enzymes. American
Chemical Society
Oxidoreductases 425

by bisulfite (Embs and Markakis 1969). It is The peroxidase test is used as an indicator of
suggested that the bisulfite forms a complex
­ satisfactory blanching of fruits and vegetables.
with the peroxidase iron, which stabilizes the However, it has been found that the enzymes
enzyme. causing off-flavors during frozen storage can,
Because of the widespread occurrence of per- under some conditions, be regenerated.
oxidase in plant tissues, Nagle and Haard (1975) Regeneration of enzymes is a relatively common
have suggested that it plays an important role in phenomenon and is more likely to occur the
the development and senescence of plant tissues. faster the temperature is raised to a given point in
It plays a role in biogenesis of ethylene; in regu- the blanching process. The deactivation and 3
lating ripening and senescence; in the degrada- reactivation of peroxidase by heat was studied by
tion of chlorophyll; and in the oxidation of Lu and Whitaker (1974). The rate of reactivation
indole-3-acetic acid. was at a maximum at pH 9 and the extent of reac-
The enzyme can occur in a variety of multiple tivation was increased by addition of hematin.
molecular forms, named isoenzymes or iso- The deactivation of peroxidase is a function of
zymes. Such isoenzymes have the same enzy- heating time and temperature. Lactoperoxidase is
matic activity but can be separated by completely deactivated by heating at 85 °C for
electrophoresis. Nagle and Haard (1975) sepa- 13 s. The effect of heating time at 76 °C on the
rated the isoperoxidases of bananas into six deactivation of lactoperoxidase is represented in
anionic and one cationic component by gel elec- Fig. 10.19, and the effect of heating temperature
trophoresis. By using other methods of separa- on the deactivation constant is shown in
tion, an even greater number of isoenzymes was Fig.  10.20. Lactoperoxidase can be regenerated
demonstrated. under conditions of high temperature short time
Peroxidase has been implicated in the forma- (HTST) pasteurization. Figure 10.21 shows the
tion of the “grit cells” or “stone cells” of pears regeneration of lactoperoxidase activity in milk
(Ranadive and Haard 1972). Bound peroxidase that is pasteurized for 10 s at 85 °C. The regen-
but not total peroxidase activity was higher in the eration effect depends greatly on storage temper-
fruit that contained excessive stone cells. The ature; the lower the storage temperature, the
stone cells or sclereids are lignocellulosic in smaller the regeneration effect.
nature. The presence of calcium ions causes the Lactoperoxidase is associated with the serum
release of wall bound peroxidase and a conse- proteins of milk. It has an optimum pH of 6.8 and
quent decrease in the deposition of lignin. a molecular weight of 82,000.

Fig. 10.19 Deactivation
of lactoperoxidase as a
function of heating time.
Source: Adapted from
Kiermeier, F., & Kayser,
C. (1960). Heat
inactivation of
lactoperoxidase (in
German). Zeitschrift für
Lebensmittel-­
Untersuchung und
-Forschung, 113
426 10 Enzymes

Fig. 10.20 Dependence
of the rate constant of
heat deactivation of
lactoperoxidase on the
heating temperature.
Adapted from Kiermeier,
F., & Kayser, C. (1960).
Heat inactivation of
lactoperoxidase (in
German). Zeitschrift für
Lebensmittel-­
Untersuchung und
-Forschung, 113

Fig. 10.21 
Regeneration in ultra
high temperature treated
milk as a function of
storage temperature. (a)
2 °C; (b) 20 °C; (c)
37 °C. Adapted from
Kiermeier, F., & Kayser,
C. (1960). Heat
inactivation of
lactoperoxidase (in
German). Zeitschrift für
Lebensmittel-­
Untersuchung und
-Forschung, 113

 ipoxygenase (Linoleate: Oxygen


L acids with a high stereo- and regioselectivity;
Oxidoreductase) type II lipoxygenase is less specific for free lin-
oleic acid and acts as a general catalyst for autox-
This enzyme, formerly named lipoxidase, is pres- idation. Type I reacts with fats in a food only after
ent in plants and catalyzes the oxidation of unsat- free fatty acids have been formed by lipase
urated fats. The major source of lipoxygenase is action; type II acts directly on triacylglycerols.
legumes, soybeans, and other beans and peas. Lipoxygenase is highly specific and attacks
Smaller amounts are present in peanuts, wheat, the cis-cis-1,4-pentadiene group contained in the
potatoes, and radishes. Lipoxygenase is a fatty acids linoleic, linolenic, and arachidonic, as
metallo-protein with an iron atom in its active follows:
center. In plants two types of lipoxygenase exist:
−CH = CH − CH 2 − CH = CH −
type I lipoxygenase peroxidizes only free fatty
Oxidoreductases 427

Most manifestations of lipoxygenase in foods


are undesirable. However, it is used in baking to
bring about desirable changes. Addition of soy-
bean flour to wheat flour dough results in a
bleaching effect, because of oxidation of the xan-
thophyll pigments. In addition, there is an effect
on the rheological and baking properties of the
dough. It has been suggested that lipoxygenase
acts indirectly in the oxidation of sulfhydryl
groups in the gluten proteins to produce disulfide
bonds. When raw soybeans are ground with water
to produce soy milk, a strong and unpleasant fla-
vor develops that is called painty, green, or beany.
Carrying out the grinding in boiling water
instantly deactivates the enzyme, and no off-­
flavor is formed. Blanching of peas and beans is
Fig. 10.22  Essential steps in the mechanism of the
essential in preventing the lipoxygenase-­catalyzed
lipoxygenase-­catalyzed oxidation of the 1,4-pentadiene
group development of off-flavor. In addition to the
development of off-flavors, the enzyme may be
responsible for destruction of carotene and vita-
min A, chlorophyll, bixin, and other pigments.
The specificity of this enzyme requires that In some cases the action of lipoxygenase leads
both double bonds are in the cis configuration; in to development of a characteristic aroma. Galliard
addition, there is a requirement that the central et al. (1976) found that the main aroma com-
methylene group of the 1,4-pentadiene group pounds of cucumber, 2-trans hexenol and 2-trans,
occupies the ω-8 position on the fatty acid chain 6-cis-nonadienal, are produced by reaction of
and also that the hydrogen to be removed from linolenic acid and lipoxygenase to form hydro-
the central methylene group be in the l-position. peroxide (Fig. 10.23); these are changed into cis
Although the exact mechanism of the reaction is unsaturated aldehydes by hydroperoxide lyase.
still in some doubt, there is agreement that the The cis unsaturated aldehydes are transformed by
essential steps are as represented in Fig. 10.22. isomerase into the corresponding trans isomers.
Initially, a hydrogen atom is abstracted from the These same substances in another matrix would
ω-8 methylene group to produce a free radical. be experienced as off-flavors. The use of lipoxy-
The free radical isomerizes, causing conjugation genase as a versatile biocatalyst has been
of the double bond and isomerization to the trans described by Gardner (1996).
configuration. The free radical then reacts to
form the ω-6 hydroperoxide.
Lipoxygenase is reported to have a pH opti-  anthine Oxidase (Xanthine: Oxygen
X
mum of about 9. However, these values are Oxidoreductase)
determined with linoleic acid as substrate, and
in natural systems the substrate is usually pres- This enzyme catalyzes the conversion of xan-
ent in the form of triglycerides. The enzyme has thine and hypoxanthine to uric acid. The reaction
a molecular weight of 102,000 and an isoelec- equation is given in Fig. 10.24; heavy arrows
tric point of 5.4. The peroxide formation by indicate the reactions catalyzed by the enzyme
lipoxygenase is inhibited by the common lipid and the dashed arrows represent the net result of
antioxidants. The antioxidants are thought to the catalytic process (Whitaker 1972). Although
react with the free radicals and interrupt the oxi- xanthine oxidase is a nonspecific enzyme and
dation mechanism. many substances can serve as substrate, the rate
428 10 Enzymes

Fig. 10.23  Lipoxygenase catalyzed formation of aroma oleic acid hydroperoxide isomers by a hydroperoxide
compounds in cucumber. Source: Reprinted from cleavage enzyme system in cucumber (Cucumis Sativus)
Galliard, T., Phillips, D. R., & Reynolds, J. The formation fruits. Biochimica et Biophysica Acta, 441, 184, Copyright
of cis-3-nonenal, trans-2-nonenal and hexanol from lin- 1976, with permission from Elsevier Science

Fig. 10.24  Oxidation of hypoxanthine and xanthine to uric acid by xanthine oxidase. Source: From Whitaker, J. R.
(1972). Principles of enzymology for the food sciences. Marcel Dekker, Inc
Immobilized Enzymes 429

of oxidation of xanthine and hypoxanthine is Immobilized Enzymes


many times greater than that of other substrates.
Xanthine oxidase has been isolated from milk One of the most important recent developments
and obtained in the crystalline state. The molecu- in the use of enzymes for industrial food process-
lar weight is 275,000. One mole of the protein ing is the fixing of enzymes on water-insoluble
contains 2 moles of FAD, 2 gram-atoms of inert supports. The fixed enzymes retain their
molybdenum, 8 gram-atoms of nonheme iron, activity and can be easily added to or removed
and eight labile sulfide groups. The eight labile from the reaction mixture. The use of immobi-
sulfide groups are liberated in the form of H2S lized enzymes permits continuous processing and
upon acidification or boiling at pH 7. The opti- greatly increased use of the enzyme. Various pos-
mum pH for activity is 8.3. The xanthine oxidase sible methods of immobilizing enzymes have
in milk is associated with the fat globules and, been listed by Weetall (1975) and Hultin (1983).
therefore, follows the fat into the cream when A schematic representation of the available meth-
milk is separated. It seems to be located in small ods is given in Fig. 10.25. The immobilizing
particles (microsomes) that are attached to the fat methods include adsorption on organic polymers,
globules. The microsomes also contain the glass, metal oxides, and siliceous materials such
enzyme alkaline phosphatase. The microsomes as bentonite and silica; entrapment in natural or
can be dislodged from the fat globules by synthetic polymers, usually polyacrylamide;
mechanical treatment such as pumping and agita- microencapsulation in polymer membranes;
tion and by heating and cooling. The enzyme is ion exchange; cross-linking; adsorption and
moderately stable to heat but no less so than cross-­linking combined; copolymerization; and
peroxidase. covalent attachment to organic polymers.

Fig. 10.25  Methods of immobilizing enzymes


430 10 Enzymes

The chemistry of immobilizing enzymes has When a high molecular weight substrate is used,
been covered in detail by Stanley and Olson the immobilizing should not be done by entrap-
(1974). ment, microencapsulation, or copolymerization,
A summary of immobilization methods has because enzyme and substrate cannot easily get
been provided by Adlercreutz (1993) and is pre- in contact. One of the promising methods
sented in Exhibit 10.1. In membrane reactors, the appears to be covalent coupling of enzymes to
reaction product is separated from the reaction inorganic carriers such as porous silica glass par-
mixture by a semipermeable membrane. In two-­ ticles. Not all of the immobilized enzyme is
phase systems, a hydrophobic reaction product can active, due to either inactivation or steric hin-
be separated from the aqueous reaction mixture by drance. Usually, only about 30–50% of the
transfer to the organic solvent phase. In aqueous- bound enzyme is active.
aqueous systems, two incompatible polymers in Immobilized enzymes can be used in one of
aqueous solution form a two-phase system. two basic types of reactor systems. The first is the
stirred tank reactor where the immobilized
enzyme is stirred with the substrate solution. This
Exhibit 10.1Summary of Enzyme is a batch system and, after the reaction is com-
Immobilization Methods plete, the immobilized enzyme is separated from
the product. The other system employs continu-
1. chemical methods ous flow columns in which the substrate flows
• covalent binding through the immobilized enzyme contained in a
• cross-linking column or similar device. A simplified flow dia-
2. physical methods gram of such a system is given in Fig. 10.26.
• adsorption The characteristics of immobilized enzymes
• physical deposition are likely to be somewhat different than those of
• entrapment the original enzyme. The pH optimum can be
• in polymer gels shifted; this depends on the surface charge of the
• in microcapsules carrier (Fig. 10.27). Another property that can be
• membranes changed is the Michaelis constant, Km. This value
3. two-phase systems can become either larger or smaller. Immobilizing
• organic-aqueous may result in increased thermal stability
• aqueous-aqueous (Fig. 10.28), but in some cases the thermal stabil-
ity is actually decreased.
Many examples of the use of immobilized
Immobilizing enzymes is likely to change enzymes in food processing have been reported.
their stability, and the method of attachment to One of the most important of these is the use of
the carrier also affects the degree of stability. immobilized glucose isomerase obtained from

Fig. 10.26  Flow diagram of an immobilized enzyme sys- Stanley, W. L., & Olson, A. C. (1974). The chemistry of
tem (column operation of lactase immobilized on phenol-­ immobilizing enzymes. Journal of Food Science, 39,
formaldehyde resin with glutaraldehyde). Source: From 660–666
Immobilized Enzymes 431

Fig. 10.27  Effect of immobilizing on the pH optimum of papain. Source: From Weetall, H. H. (1975). Immobilized
enzymes and their application in the food and beverage industry. Process Biochemistry, 10, 3–6

Fig. 10.28  Effect of immobilizing of the thermal stability of papain. Source: From Weetall, H. H. (1975). Immobilized
enzymes and their application in the food and beverage industry. Process Biochemistry, 10, 3–6

Streptomyces for the production of high-fructose through which the com syrup flows. The product
com syrup (Mermelstein 1975). In this process, obtained by this process is a syrup with 71% sol-
the enzyme is bound to an insoluble carrier such ids that contains about 42% fructose and 50%
as diethyl amino ethyl cellulose or a slurry of the glucose; it has high sweetening power, high fer-
fixed enzyme coated onto a pressure-leaf filter. mentability, high humectancy, reduced tendency
The filter then serves as the continuous reactor to crystallize, low viscosity, and good flavor.
432 10 Enzymes

Examples of the use of immobilized enzymes Carasik, W., & Carroll, J. O. (1983). Development of
immobilized enzymes for production of high-fructose
in food processing and analysis have been listed
com syrup. Food Technology, 37(10), 85–91.
by Olson and Richardson (1974) and Hultin Cordle, C. T. (1994). Control of food allergies using pro-
(1983). l-aspartic acid and l-malic acid are pro- tein hydrolysates. Food Technology, 48(10), 72–76.
duced by using enzymes contained in whole Embs, R. J., & Markakis, P. (1969). Bisulfite effect on
the chemistry and activity of horseradish peroxidase.
microorganisms that are immobilized in a poly-
Journal of Food Science, 34, 500–501.
acrylamide gel. The enzyme aspartase from Frost, G. M. (1986). Commercial production of enzymes.
Escherichia coli is used for the production of In B. J. F. Hudson (Ed.), Developments in food proteins
aspartic acid. Fumarase from Brevibacterium (Vol. 4). New York: Elsevier Applied Science Publishers.
Galliard, T., et al. (1976). The formation of cis-3-­nonenal,
ammoniagenes is used for l-malic acid
trans-2-nonenal and hexanal from linoleic acid
production. hydroperoxide isomers by a hydroperoxide cleavage
The most widely used immobilized enzyme enzyme system in cucumber (Cucumis sativus) fruits.
process involves the use of the enzyme glucose Biochimica et Biophysica Acta, 441, 181–192.
Gardner, H. W. (1996). Lipoxygenase as a versatile
isomerase for the conversion of glucose to fruc-
biocatalyst. Journal of the American Oil Chemists'
tose in com syrup (Carasik and Carroll 1983). Society, 73, 1347–1357.
The organism Bacillus coagulans has been Godtfredsen, S. E. (1993). Lipases. In T. Nagodawithana
selected for the production of glucose isomerase. & G. Reed (Eds.), Enzymes in food processing. San
Diego, CA: Academic Press.
The development of the immobilized cell slurry
Greenfield, P. F., & Lawrence, R. L. (1975).
has not proceeded to the point where half-lives of Characterization of glucose oxidase and catalase
the enzyme are more than 75 days. A half-life is on inorganic supports. Journal of Food Science, 40,
defined as the time taken for a 50% decrease in 906–910.
Habibi-Najafi, M. B., & Lee, B. H. (1996). Bitterness in
activity. Such immobilized enzyme columns can
cheese: A review. Critical Reviews in Food Science
be operated for periods of over three half-lives. and Nutrition, 36, 397–411.
The second important application of immobi- Hammer, F. E. (1993). Oxidoreductases. In
lized enzymes is the hydrolysis of lactose to glu- T. Nagodawithana & G. Reed (Eds.), Enzymes in food
processing. San Diego, CA: Academic Press.
cose and galactose in milk and milk products by
Hultin, H. O. (1983). Current and potential uses of immo-
lactase (Sprössler and Plainer 1983). Several lac- bilized enzymes. Food Technology, 37(10), 66–82, 176.
tase sources are available; from yeast, Lahl, W. J., & Braun, S. D. (1994). Enzymatic production
Saccharomyces lactis and S. fragilis, or from of protein hydrolysates for food use. Food Technology,
48(10), 68–71.
fungi, Aspergillus oryzae or A. niger. The
Liener, I. E. (1974). The sulfhydryl proteases. In J. R.
enzymes vary in their optimum pH and optimum Whitaker (Ed.), Food related enzymes, Advances in
temperature, as well as other conditions. chemistry series 136. Washington, DC: American
It is to be expected that the use of immobilized Chemical Society.
Lopez, A., et al. (1959). Catalase and peroxidase activ-
enzymes in food processing will continue to
ity in raw and blanched southern peas, Vigna senensis.
grow rapidly in the near future. Food Research, 24, 548–551.
Lu, A. T., & Whitaker, J. R. (1974). Some factors affect-
ing rates of heat inactivation and reactivation of
horseradish peroxidase. Journal of Food Science, 39,
References 1173–1178.
MacMillan, J. D., & Sheiman, M. I. (1974). Pectic
Adlercreutz, P. (1993). Immobilized enzymes. In enzymes. In J. R. Whitaker (Ed.), Food related enzymes,
T. Nagodaurithana & G. Reed (Eds.), Enzymes in food Advances in chemistry series 136. Washington, DC:
processing. San Diego, CA: Academic Press. American Chemical Society.
Albersheim, P., & Kilias, U. (1962). Studies relating to Macrae, A. R. (1983). Lipase catalyzed interesterification
the purification and properties of pectin transelimi- of oils and fats. Journal of the American Oil Chemists'
nase. Archives of Biochemistry and Biophysics, 97, Society, 60, 291–294.
107–115. Marshall, J. J. (1975). Starch degrading enzymes, old and
Arnold, R. G., et al. (1975). Application of lipolytic new. The Star, 27, 377–383.
enzymes to flavor development in dairy products. Mason, H. S. (1956). Structures and functions of the phe-
Journal of Dairy Science, 58, 1127–1143. nolase complex. Nature, 177, 79–81.
Bhat, M. K. (2000). Cellulases and related enzymes in Mason, H. S. (1957). Mechanisms of oxygen metabolism.
biotechnology. Biotechnology Advances, 18, 355–383. Advances in Enzymology, 19, 79–233.
References 433

Mathew, A. G., & Parpia, H. A. B. (1971). Food browning in chemistry series 136. Washington, DC: American
as a polyphenol reaction. Advances in Food Research, Chemical Society.
19, 75–145. Sprössler, B., & Plainer, H. (1983). Immobilized lac-
Mermelstein, N. H. (1975). Immobilized enzymes pro- tase for processing whey. Food Technology, 37(10),
duce high-fructose corn syrup. Food Technology, 93–95.
29(6), 20–26. Stanley, W. L., & Olson, A. C. (1974). The chemistry of
Nagle, N. E., & Haard, N. F. (1975). Fractionation and immobilizing enzymes. Journal of Food Science, 39,
characterization of peroxidase from ripe banana fruit. 660–666.
Journal of Food Science, 40, 576–579. Villeneuve, P., & Foglia, T. A. (1997). Lipase speci-
Nelson, J. H. (1972). Enzymatically produced flavors for ficities: Potential application in lipid bioconversions.
fatty systems. Journal of the American Oil Chemists’ Journal of the American Oil Chemists’ Society, 8,
Society, 49, 559–562. 640–650.
Olson, N. F., & Richardson, T. (1974). Immobilized Weetall, H. H. (1975). Immobilized enzymes and their
enzymes in food processing and analysis. Journal of application in the food and beverage industry. Process
Food Science, 39, 653–659. Biochemistry, 10, 3–6.
Parkin, K. L. (1993). General characteristics of enzymes. Whitaker, J. R. (1972). Principles of enzymology for the
In T. Nagodawithana & G. Reed (Eds.), Enzymes in food sciences. New York: Marcel Dekker.
food processing. San Diego, CA: Academic Press. Whitaker, J. R. (1974). Food related enzymes. In
Ranadive, A. S., & Haard, N. F. (1972). Peroxidase local- Advances in chemistry series 136. Washington, DC:
ization and lignin formation in developing pear fruit. American Chemical Society.
Journal of Food Science, 37, 381–383. Wong, D. W. S. (1995). Food enzymes: Structure and mech-
Riordan, J. F. (1974). Metal-containing exopeptidases. In anism (pp. 17–36). New York, NY: Chapman & Hill.
J. R. Whitaker (Ed.), Food related enzymes, Advances
Fruits and Vegetables
11
Chang Yong Lee

solid matter of fruits and vegetables is made up of


Major and Minor Components carbohydrates along with small amounts of pro-
tein and fat (Table 11.1). Also represented, usu-
Chemical composition of fruits and vegetables is ally only in relatively small amounts, are many
highly dependent on photosynthesis and absorp- other classes of organic compounds and a wide
tion of water and minerals during the period of range of mineral elements drawn from the soil.
growth and maturation on the parent plant. Many of these “minor” constituents can have a
However, following harvest, the sources of nutri- most important influence on the properties of
ents and water that supplied the growing entities fruits and vegetables—on their color, flavor,
are no longer available and they become indepen- nutritive value and biological activity, and in
dent units in which respiratory processes now play some cases on their texture.
a proportionately major role, with the uptake of
oxygen and loss of carbon dioxide and water.
Deterioration accompanies the use of the reserves Water
of reparable materials (mainly carbohydrates)
accumulated during photosynthesis that are no lon- Water is the most abundant single constituent of
ger replenished. Therefore, chemical constituents fruits and vegetables. Most fresh produce con-
of fresh fruits and vegetables are decided by these tains more than 80% water, with some tissues,
growth, maturation and respiration processes. such as cucumber, lettuce, and melons, contain-
Fruits and vegetables contain a very wide ing about 95% of the total weight. The starchy
range of different chemical compounds and show tubers and seeds, for example yam, cassava and
considerable variations in composition and in corn, contain less water, but even they usually
structure. Apart from the obvious inter-specific comprise more than 50% water. It was suggested
differences, no two entities are exactly the same, that water plays four important functions in the
e.g. two fruits from the same plant. Moreover, an plant cell walls as a part of the matrix gel: it is a
individual fruit or vegetable, being largely com- structural component; it can act as wetting agent
posed of living tissues which are metabolically interrupting direct hydrogen bonding between
active, is constantly changing in composition, the polymers; it can help in stabilizing conforma-
rate and extent of such changes depending on the tions of polymers; and it acts as a solvent for the
physiological role and stage of maturity. presence and transport of salts and low molecu-
The most ubiquitous of all biological com- lar weight organic compounds (Northcote 1972).
pounds is water ranging 80–90%. Most of the Especially, its contribution to texture of fresh

© Springer International Publishing AG 2018 435


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_11
436 11  Fruits and Vegetables

Table 11.1  Ranges of proximate composition in fresh tion to simple sugars, the structural framework of
fruits and vegetables
plant tissues is largely composed of complex mol-
Water Carbohydrate Lipid Protein ecules built up from monosaccharides. The total
(% Fresh weight) carbohydrate content of fruits and vegetables can
Fruits 80–90 5–20 0.1–0.5 0.5–3 range from as little as 2% of the fresh weight in
Vegetables 70–90 2–25 0.1–0.3 5–7 some cucurbitaceous fruits (melons, cucumbers,
squashes) to over 30% in vegetables containing
fruits and vegetables can be very significant. storage starch. The total carbohydrate includes
Given an unlimited supply of available mois- polysaccharides, which apart from starch are
ture, the water content of a living plant tissue largely confined to the cell walls, and sugars,
assumes a characteristic maximum value which mainly sucrose, glucose and fructose, which accu-
is associated with a state of complete turgor of mulate mainly in the cell sap of fruits.
the component cells. The internal pressure (up In general, fruits and vegetables contain more
to 9 atm) developed by osmotic forces is exactly reducing sugars than sucrose (Table 11.2),
balanced by the inward pressure of the fully although in many cases the reverse is true. Other
extended cell wall in which the tissue is physi- sugars, such as xylose, mannose, arabinose,
cally incapable of absorbing any further water. galactose, maltose, sorbose, and cellobiose, may
Significant variations in water content can occur also be present. Plant tissues may also contain
within a species, since the water content of indi- sugar alcohols, such as sorbitol, and sugar acids,
vidual cells varies considerably. The actual such as ascorbic acid. Sucrose is the dominant
water content is dependent on the availability of oligosaccharide. Maltose occurs in small amounts
water to the tissue at the time of harvest. It may in grapes, bananas and guavas. d-Sorbitol is
also be markedly affected by cultural conditions abundant in Rosaceae fruits (pomme fruits, stone
which influence structural differentiation. fruits), at the concentration range in 300–
Interspecific differences in moisture content are 800 mg/100 mL in apple juice. Tropical and sub-­
generally smaller than differences between dif- tropical fruits tend to have the higher level of
ferent types of tissue. glucose and fructose with persimmon, litchi,
The susceptibility to wilting of harvested banana and pomegranate having combined levels
fruits and vegetables varies according to the of the two sugars of more than 10%, while grape
extent to which their external surfaces are struc- is probably the only temperate fruit with more
turally modified to reduce water loss. Leaves are than 10%. Sucrose occurs at 8–10% in the tropi-
especially liable to postharvest wilting. Since cal fruits, banana, mango, guava, and others
water supports various chemical reactions, there- (Wills et al. 1989). Some vegetables, such as
fore, removal of water, such as dehydration, broccoli and onions contain insignificant amounts
retards many reactions and inhibits growth of of raffinose and stachyose that cause flatulence
microorganisms, thus improving shelf lives of when ingested because humans do not produce
processed fruit and vegetable products. the α-galactosidase needed to hydrolyze the
galactose-galactose bond.
The principal carbohydrate of plant tissues,
Carbohydrates which is not associated with cell walls, is starch,
a linear (α 1,4) or branched (α 1,4;1,6) polymer
During the growth and maturation of plants, car- of d-glucose. Starch is localized in intracellular
bohydrates, mainly in the forms of sugars and plastids that have species-specific shape, size,
transient starch, are elaborated as a result of pho- and optical properties. Industrially, starch is
tosynthesis. Starch, having been formed in the obtained from such crops as potatoes, sweet pota-
storage cells and tissues, may become transformed toes, or cereals that may contain up to 40% starch
into sugars, particularly sucrose, glucose, and on a fresh weight basis. Since starch can be uti-
fructose, during the postharvest period. In addi- lized as energy source, some vegetables with
Carbohydrates 437

Table 11.2  Distribution of free sugars in some fresh fruits and vegetables
Name Variety Glucose Fructose Sucrose Maltose Raffinose Stachyose
Apple G. Delicious 0.98 7.10 3.80 0.38
Apricot Alfred 1.80 0.92 5.82
Blueberry Blue Crop 3.92 4.04 0.24
Grape Risling 6.39 6.34 1.90 3.08
Peach Elberta 1.26 1.40 7.12
Pear Bartlett 1.00 7.88 1.28 0.48
Sweet Cherry Schmidt 6.32 5.94 0.64
Strawberry Sparkle 2.20 2.52 1.56 0.14
Asparagus M. Wash. 1.02 1.40 0.30
Broccoli Primo 0.60 0.52 0.40 0.10 0.18
Cabbage Early Head 1.40 1.14 0.26
Carrot Nantes 0.92 0.82 3.64
Celery Early Fortune 0.36 0.36 0.24
Lettuce Grand Rapids 0.17 0.24 0.10
Onion Premier 1.76 1.18 0.84 0.66 0.24
Spinach Virg. Savory 0.05 0.03
Sweet corn Seneca Chief 0.52 0.56 3.10
Tomato New Yorker 1.20 1.34 0.02
Lee et al. (1970)

high starch content such as cassava, yam, and group of polysaccharides that contain various
sweet potato are the major energy source in many kinds of hexose and pentose sugars. The middle
developing countries. Although starch contrib- lamella, composed mainly with pectic substances,
utes more calories to the normal human diet than is the outermost portion of the cell wall that plays
any other single substance, it is absent or negli- the primary role in intercellular adhesion. More
gible in most ripe fruits and many vegetables. than one-third of the dry material of the primary
Other polysaccharides are cellulose, hemicellu- cell walls and the middle lamella of fruits and veg-
lose and pectins. etables are made up with pectic substances.
A substantial proportion of carbohydrates in Chemically they are bound by covalent bonding,
fruits and vegetables is made of the cell wall and hydrogen bonding, and ionic bonding. Therefore,
middle lamella that have important effects on the canning, cooking, storage, or ripening processes
texture of the products and human health. that bring about texture changes are accompanied
Cellulose, hemicellulose, six substances and lig- by significant changes in the characteristics of the
nins are found in cell wall; the cellulose gives pectic substances.
rigidity as well as resistance to tearing; pectic sub- Fruits and vegetables are important sources of
stance and hemicellulose give plasticity together dietary fiber in our diet. During the last 30 years,
with ability to stretch. Cellulose is one of the most results from extensive research have demon-
abundant substances in the biosphere, is largely strated that fiber is important for gastrointestinal
insoluble and indigestible by human beings due to tract function and that foods rich in fiber help pre-
lack of enzymes. Cellulose is a β-(1→4) linkages vent and manage a variety of diseases:
with about 8000–12,000 glucose units per chain. ­cardiovascular disease, diabetes mellitus, obesity
In the primary cell walls, it occurs as linear asso- and weight control, and others, including lower-
ciations of the polymer molecules called fibrils and ing the risk of death. Major fruits and vegetables
up to 70% of the fibril is crystalline form (Preston that provide significant amounts of dietary fibers
1974). The hemicelluloses are a heterogeneous are listed in Table 11.3.
438 11  Fruits and Vegetables

Table 11.3  Total dietary fiber content of selected fruits A large number of simple nitrogenous sub-
and vegetables (g/100 g fresh)
stances have been found to occur in the tissues of
Total Total fruits and vegetables. Free amino acids and
Fruit fiber Vegetables fiber
related amines such as asparagine and glutamine,
Apple 2.4 Asparagus 2.1
normally those which are also present in the pro-
Apricot 2.0 Beans, Lima 4.9
teins of the tissue, appear to make up the bulk (up
Banana 2.6 Beet 2.8
to 80%) of this soluble fraction of the total nitro-
Blueberry 2.4 Broccoli 2.6
gen. Asparagine and glutamine or the related
Cherry, sweet 2.1 Brussels sprout 3.8
Mango 1.6 Cabbage 2.5
acids aspartic and glutamic appear to be espe-
Grapes, American 0.9 Carrot 2.8 cially abundant in many species—citrus fruits,
Cantaloupe 0.9 Cauliflower 2.0 potato, tomato, strawberry, gooseberry and black-
Orange 2.4 Celery 1.6 berry, and together these compounds often repre-
Peach 1.5 Cucumber 0.5 sent more than half of the non-protein nitrogen.
Pear 3.1 Kale 3.6
Plum 1.4 Lettuce, Romaine 2.1
Pineapple 1.4 Lettuce Green 1.3 Lipids
Leaf
Raspberry 6.5 Onion 1.7 The lipid content of fresh fruits and vegetables is
Strawberry 2.0 Peas 2.6 small (0.1–1%) but significant in terms of storage
Watermelon 0.4 Pepper, sweet 1.7 and quality, due to undesirable flavor changes
Potato 2.2 resulting from oxidation. In fruits, only seeds and
Source: USDA (2006) nuts contain significantly higher levels of lipids.
The lipids of fruits and vegetables (with the excep-
tion of the avocado and the olive), are, like the pro-
teins, largely confined to the cytoplasmic layers in
 roteins and Nitrogenous
P which they are especially associated with the sur-
Compounds face membranes. In addition to triacylglycerides,
glyco- and phospholipids, carotenoids, triterpe-
Proteins, though commonly representing less noids and wax are present. Lipid and lipid-like
than 1% of the fresh weight of fruit and vegetable materials are particularly prominent in the protec-
tissue, must be considered as structural constitu- tive tissues at the surfaces of plant organs—in the
ents since they are the major solid components of cuticle, epidermis and corky layers. These include
the cytoplasm of living cells. Leguminous seeds wax-like substances which are soluble in fat sol-
are richest in protein, containing up to about 8%. vents and contain mixtures of fatty acids, hydroxy-
Some leafy vegetables and sweet corn can con- acids, alcohols, esters, ketones, ethers and
tain up to about 4% of protein, but in most other hydrocarbons. In climacteric fruits, such as toma-
products the level is below 3%. The protein con- toes, the increase in respiration rate is known to
tent of fruits is usually particularly low, seldom accompany a large increase in phosphatidyl cho-
rising above about 1%, but is mostly functional line and a breakdown of total lipid during the post-
as enzymes. Enzyme systems, which are of such climacteric stage (Guclu et al. 1989).
primary importance in the physiology and post-
mortem behavior of fruits and vegetables, always
contain a protein moiety, and traces of protein, Minor Composition
probably enzymatic, are found in parts of the cell
other than the cytoplasmic layer, e.g. in the cell Organic Acids
wall. Enzymes may have either a beneficial or
detrimental effect on processing. Major enzymes Small quantities of various organic acids of the
associated with quality of fruit and vegetable Krebs’ cycle or shikimic acid pathway intermedi-
products will be discussed later. ates are produced and accumulate in vacuoles of
Minor Composition 439

COOH
COOH COOH
O
CH2
H C OH CH2 C OH

HO C COOH H C OH
HO C H
C OH

CH2 COOH
COOH O

COOH
Citric acid Tartaric Acid Malic Acid Oxalic Acid

Fig. 11.1  Chemical structures of common aliphatic organic acids in fruits and vegetables

fruits and vegetables. Some of these acids and Table 11.4  Major organic acids in some fresh fruits and
various others such as oxalic and tartaric acids, vegetables (mg/100 g fresh weight)
which have not thus far been linked with particu- Fruits and
lar metabolic cycles, can accumulate in the tissues vegetables Malic Citric Others
in considerable amounts. As a result, fruits and Grape 1095 61 Tartaric 1061
Lemon 228 5149
vegetables are normally acidic; the acid content
Melon 14 1005
ranging widely from very low levels in some veg-
Orange 131 2049
etables, such as sweet corn and leguminous seeds,
Peach 2183 673
to high level in certain fruits such as lemon, black
current, grape, and loganberry. Among vegeta- Broccoli 120 210
bles, spinach and rhubarb show an unusually high Cabbage, white 100 140
Carrot 240 90 Oxalic 0–60
level of acidity due to high content of oxalic acid.
Cauliflower 190 10
Green beans 112 34 Oxalic 20–45
Aliphatic Acids: The most widely occurring and
Green pepper 190 77
abundant acids in fruit and vegetable tissues are
Lettuce 575 118
citric and malic, each of which can, in particular
Rhubarb 910 137 Oxalic 230–500
examples, constitute over 2% of the fresh weight. Tomato 81 1064
Lemons generally contain over 3% of citric acid.
Belitz and Grosch 1999; Flores et al. 2012
In most species either citric acid or malic acid is
the predominant individual acid constituent but in lettuce, cauliflower, carrot, artichoke, cucur-
there are one or two notable exceptions. The bits, okra, onion, celery, parsnip, turnip, green
blackberry produces mainly isocitric in place of pepper, and green beans. Others are succinic, gly-
citric acid, while the avocado is exceptional in colic, gluoxylic, oxalo-acetic, benzoic (antifungal
being deficient in both of the major plant acids activity in cranberry), fumaric, α-ketoglutaric,
citric and malic. Citric acid is the principal acid pyruvic, aconitic, lacto-isocitric, malonic, quinic,
of citrus fruits, of black and red currant, raspber- and shikimic. In addition, various free uronic,
ries, loganberries, strawberries, cranberries, pine- glutamic, and short-chain fatty acids are often
apples, pomegranates and pears. Malic acid present in small amounts.
predominates in apples, most drupe fruits (plums,
cherries, apricots, etc.), and cucurbitaceous fruits, Aromatic (cyclic) acids: Some phenolic acids
banana and rhubarb (Fig. 11.1 and Table 11.4). contribute to color and taste of products.
Vegetables also differ in the relative abundance Hydroxycinnamic acids and their derivatives
of citric and malic acids. In tomato, along with p-coumaric, ferulic, caffeic and sinapic acids
potato, many leafy vegetables, and beet root, cit- are widespread in fruits and vegetables. These
ric acid is the main acid. Malic acid predominates acids are rarely free but are present mainly as
440 11  Fruits and Vegetables

Table 11.5  Chlorogenic acid content in common fruits


Chlorogenic acid
Fruit (mg/100 g fresh wt.) References
Apples
 Empire 2.56 Coseteng and Lee (1987)
 McIntosh 10.40
  Golden Delicious 7.71
  Granny Smith 3.2 Mattila and Kumpulainen (2002)
Pear
  d’Anjou, Bosc 3–7 Spanos and Wrolstad (1992)
Red raspberry 1.5 Mattila and Kumpulainen (2002)
Strawberry 2.9

acid derivatives. For example, caffeic acid is weight and caffeic acid dominated in most of
esterified with quinic acid, giving rise to chlo- fruit samples (Table 11.6). Chlorogenic and
rogenic acid. Esters of other hydroxycinnamic caffeic acids have also been implicated in the
acids with quinic acids are also widespread. non-enzymatic blackening of potato tissue after
Hydroxycinnamic acid biosynthesis starts with cooking, due to the formation of complexes
phenylalanine. Chlorogenic acid, the ester of with ferrous iron.
caffeic acid and quinic acid, is the most widely
occurring member of phenolic compounds. It is
the major phenolic compounds in apples and
pears (Table 11.5). It is the main substrate
involved in the enzymatic oxidative discolor-
ation of products such as apple, pear, peach,
potato and sweet potato. A recent report
(Mattila et al. 2006) on phenolic acids in fruits
determined by HPLC after hydrolyses showed
that the content of total phenolic acids as agly-
cones ranged from 0 to 103 mg/100 g fresh Chlorogenic acid
Minor Composition 441

Table 11.6  Phenolic acid content in some fruits (mg/100 g fresh weight)
Fruit CAF FER SIN VAN P-COUM GAL TOT
Apple, Lobo 4.30 0.27 0.66 0.09 0.66 7.2 13
Apple, Granny Smith 1.3 n.d. n.d. n.d. 0.41 5.4 7
Apple, Red Delicious 5.4 0.27 0.10 n.d. 1.0 6.5 14
Banana 0.20 5.4 n.d. 0.44 0.46 n.d. 7
Grape, red 3.4 0.43 n.d. 1.07 3.8 3.1 19
Grape, green 3.4 n.d. n.d. n.d. 1.17 2.8 7
Plum, dark 23.4 1.47 0.14 1.27 2.1 n.d. 28
Cherry 17.1 0.46 n.d. 1.17 5.1 n.d. 28
Pear 6.5 0.29 0.10 0.27 0.7 n.d. 8
Orange 3.3 9.4 2.2 0.44 1.78 n.d. 18
Grapefruit 5.5 11.6 0.99 1.66 1.35 n.d. 21
Kiwi fruit 1.5 n.d. n.d. 0.19 0.25 n.d. 3
Watermelon 0.12 0.35 n.d. 0.23 0.37 n.d. 2
CAF caffeic acid, FER ferulic acid, SIN sinapic acid, VAN vanillic acid, P-COUM p-coumaric acid, GAL gallic acid,
TOT total of phenolic acids after hydrolysis (Mattila et al. 2006)

Phenolic Compounds and minerals. In addition, the astringency of


many fruits and beverages is due to the precipita-
Plants consumed by humans contain thousands of tion of salivary proteins with plant polyphenols.
phenolic compounds because phenolic com- Therefore, polyphenol compounds are directly
pounds comprise one of the largest and most related to their nutritional and sensory character-
ubiquitous groups of plant metabolites. They are istics of foods. However, current interest stems
formed to protect the plant from photosynthetic from the observations that dietary phenolic com-
stress, reactive oxygen species, wounds, and her- pounds have antioxidative, anti-inflammatory,
bivores. The term “phenolic compounds” and anticarcinogenic activities. Numerous epide-
embraces a wide range of compounds that pos- miological data have shown a very strong inverse
sess an aromatic ring bearing a hydroxyl substitu- relationship between consumption of fruits and
ent, including their functional derivatives. vegetables and the risk of heart disease and can-
Phenolic compounds are commonly present in cer (Joshipura et al. 2001) and many investigators
most fruits and vegetables (Table 11.7). Many have attributed benefits to various antioxidants in
phenolic compounds participate in both enzy- fruits and vegetables, including phenolic com-
matic and nonenzymatic browning reactions. In pounds (Willcox et al. 2004).
addition to color, polyphenol compounds also In broad sense, polyphenol compounds can
contribute to food flavor and other qualities. For be divided into two main groups, monomer and
example, astringency of polyphenols and its ratio polymer. Monomer can be divided to flavonoid
with sugar and acid are important and useful cri- subgroup and phenolic acid subgroup: flavo-
teria for determining the overall quality of fresh noid subgroup includes flavone, flavonol, flavo-
fruits, fruit beverages, and wines. Some polyphe- none, flavononol, isoflavone, anthocyanine,
nols, such as chalcones and related compounds chalcone, and aurone; phenolic acid subgroup
found in citrus fruits, are exceedingly sweet or includes chlorogenic acid, gallic acid and
bitter. Both bitterness and astringency of wine are ellagic acid. Polymer (tannin group) can be
due to phenolic compounds present in grapes. divided into two sub groups, condensed tannin
Traditionally, they were related to antinutritional and hydrolysable tannin. Hydrolysable tannin
effects, by decreasing absorption and digestibility sub group can be divided to gallotannin and
of food because of their ability to bind proteins ellagtannin. Among these p­ henolic compounds,
442 11  Fruits and Vegetables

Table 11.7  Flavonoid content of selected fruits and vegetables (mg/100 g edible portion)
Anthocyanidina Flavan-3-ols Flavonols
Produce Cyanidin Epicatechin Catechin Kaempferol Quercetin
Apple, with skin 2.44 6.07 0.89 0.02 4.27
Apricot 5.47 4.79 2.08
Banana 7.39(Del) 0.02 6.10
Beans, Kidney, canned 0.35 1.66
Blackberry 90.31 4.66 37.06 0.06 1.76
Blueberry 61.35(Mal) 13.69 37.24 1.81 5.05
Cabbage, red 72.86 0.38
Carrot 0.10 0.31
Chard, Swiss 2.15 4.30 2.63
Cherry, sour 6.64 3.83 0.30 2.92
Cherry, sweet 75.18 6.97 1.31 2.64
Cranberry 41.81 4.37 0.39 0.09 15.09
Grape, red 34.71(Mal) 1.20 0.82 1.38
Onion 6.16 1.10 33.43
Peach 1.61 2.34 4.92 – 0.68
Pear 12.18 3.76 0.27 4.51
Plum 39.68 2.44 17.55 0.01 12.45
Strawberry 31.27(Pel) 0.12 3.32 0.46 1.14
Source: USDA (2006)
a
Anthocyanidin: Del Delphinidin, Mal Malvidin, Pel Pelargonidin

of which approximately 8000 are known to that are very water soluble (e.g. Quercetin
occur in plants the flavonoids form the largest 3-sulfate). The size of molecule varies greatly,
group with more than 5000 known structures. ranging from monomer, catechol with molecu-
Phenolic compounds range from structures that lar weight of 110 to the complex polymers
are very lipophilic (e.g. Tangeretin) to those which has molecular weight of over 1500.
Minor Composition 443

Flavonoids, diphenylpropanes (C6-C3-C6) bound usually at the C3 position. Some flavonoids


constitute one of the most distinctive polyphenolic in common fruits and vegetable are known to have
groups and occur ubiquitously in plant foods and various biological effects in vitro and in vivo and
are common component in fruits and vegetables. their significance of health beneficial effects has
They occur generally as O-glycosides with sugars been an important issue in the recent years.

Minerals potatoes, broccoli and cauliflowers are good


sources of potassium among vegetables and
The total amount of mineral matter in fruits and bananas among fruits. Peas, beans, and dark green
vegetables varies in different products from as leafy vegetables (spinach and collard) are good
little as 0.1% in some varieties of yam to as much sources of iron. Vegetables in general are richer in
as 0.4% in kohlrabi. The most common mineral minerals than are fruits (Table 11.8).
constituents in fruits and vegetables are ­potassium, The mineral elements can have an important
calcium, magnesium, iron, phosphorus, sulfur influence on the quality of fruit and vegetable
and nitrogen, together with certain other elements products: calcium can have a marked effect on tex-
such as sodium, aluminum and silicon which, ture. Metallic constituents also strongly influence
though not essential to the plant, are often well- color by their combination with organic compo-
represented in the soil. Copper, manganese, zinc, nents, while the trace metals are all constituents of
boron, and molybdenum, all essential micro- prosthetic groups of tissue enzymes which control
nutrients, are also consistently present. The most the metabolic activity of harvested products and
abundant individual mineral element in fruits and can cause marked changes in quality during and
vegetables is potassium, 60–600 mg. Spinach, subsequent to various processing procedures.
444 11  Fruits and Vegetables

Table 11.8  Minerals in some fruits and vegetables (mg/100 g FW)


Fruit or vegetable Ca Fe Mg P K Na Zn
Apple 6 0.12 5 11 107 1 0.04
Banana 5 0.26 27 22 358 1 0.15
Grape, muscadine 37 0.26 14 24 203 1 0.11
Orange, Navel 43 0.13 11 23 166 1 0.08
Peach 6 0.25 9 20 190 0 0.17
Plum 6 0.17 7 16 157 0 0.10
Watermelon 7 0.24 10 11 112 1 0.10

Broccoli 47 0.73 21 66 316 33 0.41


Cabbage 40 0.47 12 26 170 18 0.18
Cauliflower 22 0.42 15 44 299 30 0.27
Green Bean 37 1.03 25 38 211 6 0.24
Pea 43 2.08 24 53 200 4 0.27
Potato 12 0.78 23 57 421 6 0.29
Spinach 99 2.71 79 49 558 79 0.53
Tomato 10 0.27 11 24 237 5 0.17
Source: USDA., ARS (2011)

Many fruits and vegetables are important Carotenoids: The carotenoids are a group of
sources of vitamins (Table 11.9). They are the yellow, orange, and orange-red fat soluble pig-
major source of vitamin A and C in our diet. ments composed of isoprene units and widely
Provitamin A carotenoids are found in many distributed in fruits and vegetables. They are
fruits and vegetables and one of the most abun- present in the lipid material along with the chlo-
dant carotenoids is β-carotene, which exhibits rophylls. The green color of the chlorophyll
the greatest amount of provitamin A activity. In masks the yellow to red color of the carotenes
general, yellow, orange, and red fruits and vege- except in very young leaves.
tables such as papaya, apricot, peaches, canta- Chemically, carotenoids are polyene hydro-
loupes, tomatoes, carrots, sweet potatoes, winter carbons biosynthesized by only plants that
squash, pumpkins, contain significant amounts formed from eight isoprene units (tetraterpenes):
of β-carotene (provitamin A carotenoid).
Ascorbic acid occurs in large amounts in aspara- CH2 = C -CH = CH2
gus, papaya, oranges, cantaloupe, cauliflower, / Isoprene unit
broccoli, Brussels sprouts, green pepper, grape CH3
fruit, lemon and strawberries. The vitamin con-
tent of a given species can vary considerably Some carotenoids are derived by hydrogena-
with variety, growing conditions, maturity, post- tion, dehydrogenation, and/or cyclization of the
harvest storage, and processing conditions. basic structure of the 40-C carotenoids.
Carotenoids are divided into two main classes:
carotenes and xanthophylls. Carotenes are pure
Pigments polyene hydrocarbons and xanthophylls contain
oxygen in the form of hydroxy, epoxy or oxo
Although the total quantity of pigments in fruits groups. The basic carbon structure for lycopene
and vegetables is very limited, but they are excep- shows the symmetrical arrangement around the
tionally important to consumers’ pleasure. The central pair of carbons. This structure can be
main pigments of fruits and vegetables can be clas- cyclized to form β-carotene and the 15-15′ car-
sified as: (1) the carotenoids, (2) the chlorophylls, bon pair forms the center of the molecule. Other
(3) the anthocyanins, and (4) the anthoxanthins. carotenoids have different end groups with the
Postharvest Deterioration 445

Table 11.9  Major vitamins in some fruits and vegetables (fresh weight)
Vitamin A Vitamin C Vitamin B6 Thiamin Niacin
Fruit or vegetable (RAE, μg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g)
Apple 4.6 0.041 0.17 0.091
Apricot 3 10.0 0.054 0.030 0.600
Banana 96 8.7 0.367 0.031 0.665
Bluberry 3 9.7 0.052 0.037 0.418
Cherry, sweet 3 7.0 0.049 0.027 0.154
Grapefruit 3 31.2 0.053 0.043 0.204
Grape, American 58 4.0 0.110 0.092 0.300
Melon, honeydew 3 18.0 0.088 0.038 0.418
Orange 11 53.2 0.060 0.087 0.282
Peach 16 6.6 0.025 0.024 0.806
Pear 1 4.3 0.029 0.012 0.161
Pineapple 3 47.8 0.112 0.079 0.500
Strawberry 1 58.8 0.047 0.024 0.386
Watermelon 28 8.1 0.045 0.033 0.178

Asparagus 38 5.6 0.091 0.143 0.978


Beans, Snap 35 12.2 0.141 0.082 0.743
Broccoli 31 89.2 0.175 0.071 0.639
Asparagus 38 5.6 0.091 0.143 0.978
Beans, Snap 35 12.2 0.141 0.082 0.743
Broccoli 31 89.2 0.175 0.071 0.639
Brussels sprout 38 85.0 0.219 0.139 0.745
Cabbage 5 36.6 0.124 0.061 0.234
Carrot 835 5.9 0.138 0.066 0.983
Cauliflower 0 48.2 0.184 0.05 0.507
Corn, sweet 9 6.8 0.093 0.155 1.770
Onion 0 7.4 0.12 0.046 0.116
Pea, green 38 40.0 0.169 0.266 2.090
Potato, russet 0 5.7 0.345 0.082 1.036
Spinach 469 28.1 0.195 0.078 0.724
Squash, winter 68 11 0.154 0.07 0.5
Sweet potato 709 2.4 0.209 0.078 0.557
Tomato 42 13.7 0.08 0.037 0.594
Cabbage 5 36.6 0.124 0.061 0.234
USDA., ARS (2011)

same central structure. About 60 different end occur in fruits and vegetables during the ripening
groups are known, comprising about 500 known process and after harvest influence their use as
carotenoids. foods. Following harvest, the sources of nutrients
and water that supplied the growing fruits and
vegetables are no longer available. In order to sus-
Postharvest Deterioration tain life processes, respiration continues to pro-
duce metabolic energy with the uptake of oxygen
Today’s consumers have unlimited access to and loss of carbon dioxide and water. Consequently,
an amazing range of fresh produce throughout the postharvest deterioration accompanies with the
year. A major problem, however, has been main- use of the reserved materials accumulated during
taining the produce at high quality. Changes that photosynthesis that are no longer replenished.
446 11  Fruits and Vegetables

Therefore, in order to increase shelf-life of fruits small communication channels, called plas-
and vegetables and reduce deterioration, increased madesmata linking their cytoplasmic masses.
attention has been given in recent years to the The wall is semi-permeable to water and solutes.
postharvest physiology of fresh fruits and vegeta- The material within the cell is protoplasm com-
bles. In order to conserve high quality of fruits and posed of a very large number of different mole-
vegetables, we have to understand the basic bio- cules. An important organelle found in plant cells
chemical reactions that are taking place in plant but not in animal cells is granular green plastids
cells before and after harvest. Therefore, the called chloroplast, where photosynthesis occurs.
nature and causes of losses, due to both wastage Another unique to plant cells is a large central
and reduced quality, between harvest and con- vacuole, a compartment that stores water and a
sumption should be studied. variety of chemicals. The cytoplasm contains
The magnitude of postharvest losses in fresh several important organelles, such as nucleus,
fruits and vegetables is estimated 5–25% in mitochondria, chloroplasts (chlorophylls), chro-
developed countries and 25–80% in developing moplasts (carotenoids), and amyloplasts which
countries, depending upon the commodity, culti- are membrane-bound bodies with specialized
var, and handling conditions. In tropical regions, functions. Important processes which occur in
which include a large proportion of the develop- the cytoplasm include the breakdown of storage
ing countries, these losses can assume consider- reserves of carbohydrate by glycolysis and pro-
able economic and social importance. To reduce tein synthesis. Other types of plant cells besides
these losses, producers and handlers must under- parenchyma cells are the conducting cells, the
stand the biological and environmental factors supporting cells, and the protective cells. The
involved in deterioration and use postharvest walls of these cells are composed primarily of
techniques that delay senescence and maintain cellulose and lignin.
the best possible quality. Thus the main role of The most important fact in the postharvest
food scientists or postharvest physiologists is to handling of fruits and vegetables is that harvested
devise methods by which deterioration of pro- fruits and vegetables are living structures. One
duce is restricted as much as possible during the readily accepts that produce is a living, biological
period between harvest and consumption. entity when it is attached to the growing parent
plant in its agricultural environment. But even
after harvest, the produce is still in living state as
Cellular Components it continues to perform the metabolic reactions
and Physiology of Fruits and maintain the physiological systems which
and Vegetables were present when it was attached to the plant.
An important feature of plants is that they respire
The cells of fruits and vegetables are typical plant by taking up oxygen and giving off carbon diox-
cells. The main type of cell in the edible portion ide and heat. They also transpire, that is, lose
of most fruits and vegetables is the parenchyma water. Therefore, the produce is dependent
cell. The walls of parenchyma cells in young entirely on its own food reserves, moisture con-
plants are composed almost entirely bounded by tent, and its environmental conditions. The intact
a more or less rigid cell wall composed of cellu- plant cells must respond to various physical and
lose, fiber and other polymers such as pectic sub- chemical challenges from the environment. It is
stances, hemicelluloses and lignins. A layer of important to recognize that different commodi-
pectic substances forms the middle lamella and ties, species and cultivars differ markedly in
acts to bind adjacent cells together. As the plant response to the environmental conditions.
grows older the nature of these cementing sub- Therefore, no single condition or treatment is
stances often changes, lignins and other com- ideal for extending the shelf-life of all fruits and
pounds are deposited, and the cellulose layer of vegetables. Some of the major factors that influ-
the cell wall thickens. Adjacent cells often have ence the respiration rate and deterioration will be
Cellular Components and Physiology of Fruits and Vegetables 447

discussed. The texture of fruits and vegetables Table 11.10  Fruits classified according to respiratory
behavior during ripening
depends on the turgor of living cells as well as on
the occurrence of supporting tissues and the Climacteric fruits Nonclimacteric fruits
cohesiveness of the cells. Turgor pressure is cre- Apple Muskmelon Blackberry Olive
ated by the cell contents and the partially elastic Apricot Nectarine Cherry Orange
cell wall in a delicately balanced force. It main- Avocado Papaya Cranberry Pea
tains the cell at a normal volume yet allows the Banana Passion fruit Cucumber Pepper
Blueberry Peach Date Pineapple
exchange of substances. When the balanced force
Guava Pear Grape Raspberry
is disrupted by wilt, cooking or processing, the
Kiwifruit Plum Lemon Strawberry
cell walls dies and the vacuoles are no longer
Mango Tomato Lime Watermelon
covered by a living membrane of protoplasm and
Kader (2002)
eventually the texture of food becomes soft.

citrus, pineapple and strawberry, that do not


Respiration exhibit a respiratory climacteric, are known as
the non-climacteric class of fruits. Non-­
Respiration is the major process in living tissues climacteric fruit exhibit most of the ripening
and can be described as the oxidative breakdown changes at the plant, although these usually occur
of the more complex materials normally present more slowly than those of the climacteric fruits.
in cells, such as starch, sugars, and organic acids, All vegetables are considered to have a non-­
into simpler molecules, such as carbon dioxide climacteric type of respiratory pattern. A clear
and water, with the concurrent production of understanding of the control mechanism of fruit
energy and other molecules which can be used by ripening has important commercial benefits.
the cell for synthetic reactions. Numerous Climacteric and non-climacteric fruits may be
enzymes are associated with these reactions. further differentiated by their response to applied
ethylene and by their pattern of ethylene
C6H12O6 + 6O2 → 6CO2 + 6H2O + Energy ­production during ripening. It has been estab-
lished that all fruits produce minute quantities of
Respiration rate is an excellent indicator of ethylene during development. However, coinci-
metabolic activity of the tissue and thus is a use- dent with ripening, climacteric fruits produce
ful guide to the potential storage life of the pro- much larger amounts of ethylene than non-cli-
duce. The loss of stored reserves during macteric fruits. This difference between the two
respiration means the hastening of senescence as classes of fruits is further exemplified by the
they are exhausted. Therefore, the rate of deterio- internal ethylene concentration found at several
ration of post-harvested commodities is generally stages of development and ripening. The internal
proportional to the respiration rate. A large group ethylene concentration of climacteric fruits var-
of fruits that include tomato, mango, banana, and ies widely, but that of non-climacteric fruits
apple, show a pronounced increase in respiration changes little during development and ripening.
coincident with ripening. Such an increase in res- Ethylene, applied at concentration as low as 0.1–
piration is known as a “respiratory climacteric,” 1.0 μL/L for 1 day, is normally sufficient to has-
and this group of fruits is known as the climac- ten full ripening of climacteric fruits, but the
teric class of fruits (Table 11.10). The intensity magnitude of the climacteric is relatively inde-
and duration of the respiratory climacteric varies pendent of the concentration of applied ethylene.
widely amongst fruit. The respiratory climac- In contrast, applied ethylene merely increases the
teric, as well as the complete ripening process, respiration of non-­climacteric fruits, the magni-
may proceed while the fruit is either attached to tude of the increase being dependent on the con-
or detached from the plant. Those fruits, such as centration of ethylene.
448 11  Fruits and Vegetables

 nvironmental Factors that Influence


E Atmospheric condition: Since oxygen and carbon
Deterioration dioxide are directly involved with respiration of
fruits and vegetables, the composition of gases in
Temperature: Temperature is the environmental the storage atmosphere can affect the deteriora-
factor that most influences the deterioration rates tion rates. Therefore, reduction of oxygen or ele-
of postharvest commodities. The rate of respira- vation of carbon dioxide can either delay or
tion reactions, within the physiological tempera- accelerate the deterioration of fresh produce. The
ture range, increases exponentially with increase magnitude of these effects depends on the com-
in temperature. For each increase of 10 °C above modity, cultivar, physiological age, temperature,
optimum, the rate of deterioration increases by concentrations of oxygen and carbon dioxide,
two- to threefold. Therefore, exposure to undesir- and duration of storage time. Today, commercial
able temperatures results in many physiological controlled atmosphere (CA) storage and modi-
disorders. Many enzymes are inactive beyond fied atmosphere (MA) storage are frequently
35 °C and inactivated at above 40 °C. At above used to extend the shelf life of fruits and vegeta-
35 °C, metabolism becomes abnormal and results bles. Commercial use of CA storage is greatest
in a breakdown of membrane integrity and struc- worldwide on apples and pears.
ture, with rapid deterioration of the produce. The Other factors: Ethylene (C2H4) is a plant hor-
lower limit for normal metabolism is the freezing mone that regulates many aspects of growth,
point of the tissue which is usually between 0 °C development, ripening and senescence, espe-
and −2 °C. Within these temperature limits, cially in climacteric fruits. Therefore, its effects
respiratory changes associated with ripening and can be desirable or undesirable. Ethylene can be
senescence proceed, but at different rates. used to promote faster and more uniform ripen-
Therefore, temperature management is the most ing of fruits picked at the early mature stage,
effective tool for extending the shelf life of fresh while exposure to ethylene can be detrimental to
commodities. In general, lowering of tempera- the quality of vegetables. Exposure of some com-
ture can retard the rate of respiration and undesir- modity, such as potato or endive, to light results
able physiological changes. However, exposure in greening due to formation of chlorophylls.
of the commodity to too low temperature results
in chilling and freezing injury. Chilling injury
occurs in some commodities (especially those of  ffects of Processing on Fruits
E
tropical or subtropical fruits) held at temperature and Vegetables
above their freezing point and below 15 °C,
depending on commodity. The basic purpose of processing of fruits and
vegetables is to extend the shelf life and to main-
Relative humidity: The rate of water loss from tain the edible quality of the products by curtail-
fruit and vegetables depends on the vapor pres- ing the activity of microorganisms and the
sure deficit between the commodity and the sur- chemical changes that would otherwise adversely
rounding ambient air, which is influenced by affect. However, the severity of common com-
temperature and relative humidity. Therefore, to mercial processing methods for fruits and vegeta-
control the rate of water loss from the produce, bles, such as dehydration, canning, or freezing,
one should lower the temperature to reduce the results in extensive changes on the quality char-
vapor pressure difference between the produce acteristics of a given commodity. Some alteration
and the air, or provide a barrier to water loss. An may be desirable for inactivation of anti-­
effective method for reducing water loss from nutritional factors or softening of hard tissue or
fruits and vegetables is to increase the relative creation of new flavor and may also be undesir-
humidity of the air. The use of very high relative able by loss of nutrients, color, texture, or flavor.
humidity, however, favors the growth of Undesirable chemical changes include oxidative
microorganisms. rancidity of fats, loss of ascorbic acid and other
Effects of Processing on Fruits and Vegetables 449

vitamins through oxidation, degradation of pec- Carbohydrate Reactions


tin, and discoloration. The chemical changes that
occur when fruits and vegetables are cooked, Thermal processing softens plant tissues through
steamed, canned, dried or frozen are very com- the action of heat on pectic substances. The large
plex. Numerous chemical reactions that associ- molecules of insoluble protopectins are hydro-
ated with the profound changes in sensory and lyzed during heat processing to smaller soluble
nutritional qualities have been studied, but many pectin and pectic acid. Total pectic substances as
of the exact reaction mechanisms are still well as protopectin in the raw vegetables were
unknown. Various enzyme activities in fruits and decreased while the amounts of pectins and pec-
vegetables are also important factors for the sen- tates were increased after steaming. This was due
sory and nutritional qualities. Cellular disruption to solubilization of protopectin to pectins
during peeling, dicing, size reduction, etc. before (Table 11.11). On the other hand mild heat treat-
the enzymes are inactivated contributes signifi- ments can activate endogenous pectin methyles-
cantly to the chemical changes that occur. terase and this will partially demethylate pectic
Blanching is the most common types of heat pro- substances allowing internal calcium to effect
cesses used for fruits and vegetables. Blanching crosslinking of pectic polymers with a resulting
is a brief heat treatment involving exposure of the increase in tissue firmness. Starch granules in the
tissues to hot water or steam for a few minutes at presence of water gelatinize, resulting in changes
about 100 °C. Prior to freezing, or dehydration, in texture and water-holding capacity of the
blanching is done primarily to inactivate starch. Aqueous suspensions of starch can
enzymes, whereas prior to heat sterilization undergo hydrolysis in the presence of acid or
blanching is done for several reasons, the most enzymes and this alters texture and sweetness.
important is to remove air from tissue. Under cer- Sugars can undergo caramelization reactions pro-
tain conditions, blanching may serve to activate ducing a brown color and a typical burnt flavor,
enzymes in such a way as to promote desirable and can degrade into a variety of compounds that
changes. For example, blanching of some green influence flavor (e.g. furfural and 5-­hydroxymethyl
vegetables can stabilize their bright color (activa- furfural). Sugar in the presence of amino groups
tion of chlorophyllase). Similarly, the texture-­ can engage in the Maillard reaction resulting in a
firming effect of blanching on certain vegetables brown color, distinctive flavor, textural changes,
has been attributed to activation of pectin methy- loss of protein nutritive value and perhaps the
lesterase, which in turn catalyzes the conversion development of toxic compounds.
of pectin to pectic acids. Thus, in any processing
method, both the growth of undesirable organ-
isms and chemical, physical, and biochemical Protein Reactions
reactions of the food itself must be considered.
Some of the major chemical reactions associated Moderate heating results in protein denaturation.
with the sensory and nutritional qualities during The consequences can include inactivation of
processing are: enzymes, inactivation of proteinaceous inhibitors
of digestive enzymes (e.g. trypsin inhibitors in

Table 11.11  Effect of steaming on pectic substances of carrots and parsnips


Pectic material Raw Steamed 45 min
Carrots (%) Parsnips (%) Carrots (%) Parsnips (%)
Total pectic substance 18.6 16.4 13.2 15.0
Protopectin 14.1 10.2 3.6 5.7
Pectin 3.7 4.7 8.8 7.9
Peptic acid/pectates 0.8 1.6 1.3 2.1
Simpson and Halliday (1941)
450 11  Fruits and Vegetables

soy products), decreasing protein solubility, value of foods. In some cases the breakdown of
alteration of protein functionality (water—0.8— fatty acids is initiated by such simple processing
holding capacity and emulsifying capabilities) as cutting. This enables enzymes present in one
and alterations in texture. Severe heating of pro- area of vegetable to act on compounds of their
teins in the presence (6) of water can result in areas of the same tissue. The intact tissue of
protein crosslinking (formation of isopeptide cucumbers and tomatoes has very little flavor. By
0.7), alterations in flavor and texture, and a cutting the tissue, the enzyme (lipoxygenase)
decrease in protein digestibility. In acidic solu- almost instantaneously develops flavoring com-
tion, some (7.9) protein hydrolysis can occur pounds by breaking down primarily linoleic and
with effects on texture and flavor. Severe heating linolenic acids present in the triacylglycerols.
in the presence (2.1) of water and the absence of Secondary reactions, such as isomerization,
air can result in desulfuration with accompanying hydrogenation to alcohol, oxidation to acids,
alterations in color and flavor. Oxidation of pro- esterification, aldol condensation and many oth-
teins can result in protein crosslinking, thio-­ ers will follow. When heated in the presence of
disulfide interchange reactions and oxidative pigments, peroxidizing lipids result in decolor-
degradation products that influence the texture of ization; when heated in the presence of flavor,
products. In the presence of alkali, heating can flavor changes occur; and when heated in the
result in denaturation, racemization and forma- presence of vitamins (A, C, D, E, folate), loss of
tion of lysinoalanine-type compounds. Changes vitamin activity can occur.
in protein solubility, texture, and nutritive value
can result. Heating in the presence of active car-
bonyls will result in the Maillard reaction and Color Change
Strecker degradation which have pronounced
effects in color (browning), flavor, protein nutri- Several major natural pigments in fruits and veg-
tive value, and texture. Heating in the presence of etables degrade significantly when exposed to
lipids can result in the formation of lipid-protein heat. One of the most important chemical reac-
complexes and an alteration of protein texture tions that influence the color quality of green
and functionality. vegetables during heat processing is a degrada-
tion of chlorophylls. Color changes are encoun-
tered most visibly in processing of green peas,
Lipid Reactions green beans, kale, Brussels sprouts and spinach.
Green vegetables contain volatile acids that are
Heated lipids can undergo several types of chem- partially given off during heat processing due to
ical reactions that influence the sensory proper- cellular disruption and lower the pH of vegetable
ties, functionality, and nutritive value. tissue. Since chlorophylls are very sensitive to
Triacylglycerols can be hydrogenated in the pres- any pH below 7, they lose magnesium in the
ence of hydrogen at elevated temperatures and acidic condition, changing color from green to
pressures. In the presence of enzymes or acid, olive drab (pheophytin). Conversion of chloro-
water and heat, triacylglycerols can be hydro- phylls to pheophytins, which is accompanied by
lyzed that affects flavor and increases their sus- a color change in spinach, beans, asparagus and
ceptibility to other degradative reactions. Heat peas is shown in Table 11.12. Chlorophyllase is
and oxidizing conditions (oxygen and inorganic mostly inactivated when vegetables are blanched,
or organic catalysts) can result in oxidation of hence, chlorophyllides and pheophorbides are
unsaturated lipids. This is a very common reac- rarely detected. However, in the fermentation of
tion for lipids leading to the formation of cucumbers chlorophyllase is active. The result is
­aldehydes, ketones, acids, hydrocarbons, dimers a color change from dark green to olive-green,
and polymers which in turn, have profound caused by large amount of pheophorbides. Okra
effects on the flavor, color, texture, and nutritive and turnip greens showed considerable formation
Effects of Processing on Fruits and Vegetables 451

Table 11.12 Pheophytin a, b and pyropheophytin a, b in temperature short-time processing or a combina-


commercial canned vegetables
tion of HTST processing and pH control. Some
Pheophytin Pyropheophytin improvement in color retention has been achieved
(µg/g DW) (µg/g DW) by producing the more heat stable phytol-free
a b a b pigments. However, results were inconsistent and
Spinach 830 200 4000 1400
were influenced by biological variability attribut-
Beans 340 120 260 95
able to differences in growing conditions.
Asparagus 180 51 110 30
Anthyocyanins can undergo color changes
Peas 34 13 33 12
during heating especially if the pH is altered or
Von Elbe and Schwartz (1996)
tin is present. Anthocyanin losses also occur
enzymatically. Enzyme systems involved include
of chlorophylides and pheophorbides when glucosidases (anthocyanases), peroxidases and
blanched at 82 °C, but considerable less of these phenolases. Since these enzyme systems are
pytol-free derivatives when blanched at present in many fruits and vegetables, inactiva-
100 °C. The difference indicates deactivation of tion is essential. Colorless proanthocyanidins
chlorophyllase at elevated temperature. In snap with metal ions can result in pink discoloration of
beans, the pigments were unaffected by blanch- fruits, such as pear. Also nonenzymatic or
ing temperature, because of the low initial Maillard browning may contribute to discolor-
enzyme activity. Considerable formation of ation of fruits and vegetables during heat pro-
phytol-­free derivatives in unblanched frozen peas cessing and subsequent storage when conditions
was observed, whereas none was detected in peas are favorable for these reactions.
blanched before freezing. A change in color
occurs during drying of vegetables, its extent
being dependent on water activity. The conver- Flavor Change
sion of chlorophylls to pheophytins continues in
blanched vegetables even during frozen storage. Heating fruits and vegetables can cause dramatic
In beans and Brussels sprouts, immediately after changes in flavor. Most flavor changes can be
blanching (2 min at 100 °C), the pheophytin con- attributed to the loss of certain compounds by
tent amounts to 8–9%, while after storage for 12 volatilization or by formation of new substances
months at −18 °C it increases to 68–83%. via degradation or addition reactions. For exam-
Pheophytin content rises from 0% to only 4–6% ple, the major carbonyl component of the fresh
in paprika peppers and peas under the same con- banana, 2-hexanal, is not detected in the heat pro-
ditions. Chlorophylls are further subject to both cessed puree. Volatile losses are also important to
enzymatic and nonenzymatic oxidative changes. flavor change in other canned foods, including
A number of studies contribute the bleaching of pear, cabbage, and onion. Volatile losses may
chlorophylls to the enzyme activities of lipase also continue through chemical mechanisms dur-
and lipoxygenase. Regreening of processed veg- ing post-processing storage. For example, in
etable has been observed and attributed to the plain tin cans this may occur at the tin/product
formation of metallochlorophyll complexes interface by reduction of aldehydes and ketones
involving copper or zinc. A number of attempts to primary and secondary alcohols. New volatiles
have been made to preserve chlorophyll during may also arise in canned foods. Formation of
green vegetable preservation, but none has been hydrogen sulfide and other volatile sulfur com-
fully successful. Since acidic conditions favor pounds in heated products, such as cabbage and
pheophytin formation, many studies have corn, are examples of this kind. Methyl sulfide is
involved pH control. The improved color generated during heat processing of tomatoes,
­retention obtained at higher pHs is soon lost dur- and off-flavor in canned apple sauce has been
ing storage. Other investigators have attempted to attributed to the formation of caproic acid.
minimize heat exposure by applying high-­ Nonvolatile substances may also contribute to
452 11  Fruits and Vegetables

flavor changes in heated foods. Off-flavors in be firmer than those heated in the absence of this
some canned fruits and vegetables are attributed solute. Sucrose firms the texture of canned fruit
to the formation of ammonium pyrrolidonecar- by osmotic dehydration of the tissue, which in
boxylate from the cyclization of glutamine. turn appears to facilitate cohesion of cell wall
Moreover, failure to completely inactivate polysaccharides. While severe heat processing
enzymes may also result in flavor changes during tends to soften plant tissues through the action of
processing and subsequent storage. In some heat on pectic substances, mild heat treatments
instances, enzymes are naturally heat resistant can be used to activate endogenous pectin methy-
(peroxidase), and in others the enzyme acquires lesterase. This will partially demethylate pectic
heat resistance because of the manner in which it substances allowing internal calcium to effect
is physically situated in the tissues. Examples of crosslinking of pectic polymers with a resulting
the latter are β-glucosidase in plum kernels and increase in tissue firmness.
lipoxygenase in the cob of sweet corn. Peroxidase,
which can catalyze production of off-flavors in
vegetables, can partially “regenerate” of “reacti- Nutrient Loss
vate” following HTST thermal processing.
The destruction of nutrients during the thermal
process is dependent on time/temperature treat-
Texture Change ment used as the basis of the process, rate of heat
transfer into the product, and thermal stability of
As discussed previously, texture of fruits and nutrients and their solubility in water (Tables
vegetables is likely to soften as a result of heat 11.13 and 11.14). Increasing the rate of heat
treatment. This is due to various factors including transfer into the product has been one of the pri-
the loss of turgor pressure, loss of extracellular mary research in commercial development.
and vascular air, and the denaturation or degrada- Leaching losses of water soluble vitamins and
tion of cell wall constituents and other polysac- minerals can be substantial if plant products are
charides. Failure to inactivate pectin-hydrolyzing exposed to hot water. Therefore, optimizing
enzymes early in the process can result in unde- nutrient retention in commercial processing has
sirable texture changes in certain products, such been studied extensively. The use of high-­
as canned tomatoes. Starch granules gelatinize temperature short-time (HTST) processes is par-
during heating in an aqueous environment, and ticularly adaptable to aseptic processing to retain
this reaction undoubtedly influences the texture high percent of vitamins (Table 11.15). In the
of certain commodities. The deterioration of tex- processing of fruits and vegetables, the HTST
ture can be minimized by choosing less mature processes are most likely to retain more of the
produce, lessening the severity of thermal pro- water soluble nutrients and to minimize the loss
cesses, choosing cultivars that are less suscepti- of heat labile vitamins such as C. The blanching
ble to softening, and by adding acids, pectic of vegetables prior to canning or dehydration is
enzyme inhibitors, or calcium salts to the canning important if losses of vitamin C and vitamin A
medium. Fruits heated in sucrose syrups tend to through enzymatic activity are to be avoided.

Table 11.13  Losses (%) of nutrients in the canning process


Vitamin
Product A B1 B2 Niacin B6 C
Asparagus 43.3 66.7 55.0 46.6 64.0 54.5
Green beans 51.7 62.5 63.6 40.0 50.0 78.9
Carrots 9.1 66.7 60.0 33.3 80.0 75.0
Spinach 32.1 80.0 50.0 50.0 75.0 72.5
Harris and Karmas (1975)
Dehydration of Fruits and Vegetables 453

Table 11.14  Retention of ascorbic acid in peas at vari- destruction. Destruction of carotenoids has been
ous stages of processing and after cooking
associated with lipoxygenase activity during the
Fresh Frozen Canned Air-dried direct oxidation of fatty acids. Nonenzymatic
Retention of ascorbic acid (%) oxidation of carotenoids with concurrent color
Blanching – 75 70 75 loss is of major concern in dehydrated fruit and
Processing – 75 63 45 vegetable products. The relative stability of carot-
Thawing – 71 – – enoids against oxidation when present in water-­
Cooking 44 39 36 25
containing foods suggests a protective effect of
Harris and von Loesecke (1960) water, and therefore, water activity is related to
carotenoid stability in dehydrated products.
Table 11.15  Effect of HTST processing on nutrient
losses
Pyridoxin loss
Product Thiamin loss (%) (%)
 ehydration of Fruits
D
HTST Conv. HTST Conv.
and Vegetables
Strained lime 15.8 40.3 9.5 10.1
beans Commercial dehydration is carried out by using
Tomato juice 0 2.8 0 0 conveyor or tunnel dryer, spray or drum dryer, flu-
concentrate idized bed dryer, or freeze-dryer depending on the
Harris and Karmas (1975) state of the commodity to a residual moisture con-
tent of 4–8%. Temperature of drying chambers
Table 11.16  Changes in concentration of β-carotene in varies depending on the dryers and commodities.
cooked dehydrated carrots Solar drying is still a common process in southern
β-Carotene content (µg/g and tropical countries for dates, fig or raisins.
Carrot solids) Dehydration reduces the natural water content of
Fresh 980–1860 fruits and vegetables below the level of critical for
Explosive puff-dried 805–1060 the growth of microorganisms (12–15%) without
Freeze-dried 870–1125 being detrimental to important nutrients, flavor and
Conventional air-dried 636–987 appearance. Freeze-drying is known to provide
Dellamonica and McDowell (1965) high quality products with good shape retention
and porous structure that is readily rehydrated.
One of the two most important vitamins in our Since the rate of a chemical reaction in a food is
diet derived from in fruits and vegetables is a a function of many factors, mainly reactant con-
group of provitamin A carotenoids and they pres- centration, temperature, pH, oxidation-­ reduction
ent in the stable trans-configuration. However, potential and inhibitors and catalysts along with
processing and storage under acidic conditions, migration of solutes, the dehydration process is
elevated temperatures, and/or light would favor usually accompanied by significant changes. The
isomerization and isomers are far less in biologi- concentration of important components such as
cal activity. Therefore, extended holding at ele- protein and carbohydrates leads to accelerate their
vated temperatures or heat sterilization can have chemical reactions, like Maillard browning, result-
some effect on the total carotenoid content and ing in a darker color and development of new flavor
vitamin A value. Changes in the b-carotene con- compounds. Oxidative reactions degrade proteins,
tent of cooked dehydrated carrots (Table 11.16) lipids, vitamins, and original aroma compounds.
illustrate typical extents of degradation during Sulfite treatment is used to prevent both enzymatic
processing and typical exposure to oxygen. and nonenzymatic browning reactions, stabilizes
Other losses of carotenoids can occur because vitamin C and prevents microbial contamination
of enzymatic and nonenzymatic oxidation. during processing and storage. Dehydrated fruits
Losses have been reduced in blanched, compared and vegetables are light, air and moisture sensitive
to unblanched tissue, suggesting enzymatic and therefore require careful packaging.
454 11  Fruits and Vegetables

Canning of Fruits and Vegetables (below −30 °C) for 2–3 h to avoid migration of


soluble solids, formation of large ice crystals and
Stone fruits, pears, pineapple are major canned microbial growth. Some fruits are covered, prior
fruits and strawberries are canned to a lesser to freezing, with a 30–50% sugar solution (and
extent. First, all fruits are sorted and washed. citric acid and ascorbic acid) to improve aroma,
Apples and pears are peeled and sliced, cherries texture, and color of the fruits. Frozen fruit is
are freed from seeds and stems, plums, prunes, stored at −18–25 °C which has shelf life of 2–3
apricots and peaches are halved and the seeds are years. Asparagus, beans, peas, Brussels sprout,
removed, strawberry calix is removed and red broccoli, carrots, cauliflower, spinach, sweet
current stems are removed. Specialized equip- corns, and potatoes are very suitable for freezing.
ment has been developed for these procedures. In They are blanched (boiling water or steam) for
order to prevent discoloration during processing, 2–5 min as in the case of canning. Immediately
ascorbic acid is added. To enhance aroma and after blanching, the vegetables are cooled and
taste, sugar and citric acid solutions are added frozen at −40 °C. Freezing is usually carried out
and the can is vacuum sealed at 77–95 °C for using conventional freezing method in plate or
4–6 min and heat sterilized under the required air freezers then stored at −18–20 °C. Major
conditions for each commodity. losses of sensory and nutritional qualities of fro-
Blanching is the most common operation in the zen fruits and vegetables loss sensory and nutri-
canning process of vegetables. It serves not only tional qualities are usually taking place at the
to inactivate the enzymes, but to remove the air primary processing steps of washing and blanch-
present in plant tissue and to induce shrinkage or ing, but they are preserved during freezing and
softening of the product (leafy vegetables, such as thawing. However, freezing method, among dif-
spinach). Brine (1–3% NaCl solution) often serves ferent processing methods, is the most favorable
as filling liquid with citric acid (up to 0.05%), cal- method in retaining the original quality of fresh
cium slats for firming the plant tissue. Sterilization fruits and vegetables.
is carried out in rotation retorts and the time and
temperature vary depending on the commodity,
size of cans, and the state of the product. After the Lactic Acid Fermentation
required sterilization, the cans are quickly cooled.
In order to retain better quality, high temperatures Pickled vegetables by the lactic acid fermentation
and shorter times (HTST) sterilization process is have a long history as one of the oldest food pres-
commonly used today. As indicated before, the ervation methods. Cucumber, cabbage, green
losses in sensory and nutritional qualities are beans and others, are produced by spontaneous
expected. Nutrient losses in fruits and vegetables lactic acid fermentation at lower pH that inhibits
during canning vary greatly (from 0 to 91%) the growth undesirable microorgnisms and stabi-
depending on the nutrient and product. Minimizing lizes some nutrients and sensory qualities. Today,
losses of sensory and nutritional qualities due to sauerkraut and kimichi manufactured by fer-
canning process have been the major tasks for mented cabbages with probiotic lactic acid bacte-
food scientists for the last 6–70 years. ria are considered as probiotic foods. Vegetables
are treated with 2.5–3% salt at about 20 °C for
lactic acid fermentation. For sauerkraut, white
Freezing of Fruits and Vegetables cabbage heads are shredded, mixed with salt (at
1.8–2.5%), and packed in layers in barrels or
Fruits and vegetables are often frozen and stored large tanks. The lactic acid fermentation initiated
as an end product or for further processing. by starter cultures at 18–24 °C and lasts for 3–6
Among fruits, apples, apricots, cherries, pineap- weeks. During the first 48 h of anaerobic fermen-
ples and strawberries are commonly frozen. tation, the pH falls from around 6.2 to 3.7–4.2.
Properly sized fruits pieces are rapidly chilled The acid formed promotes the growth of
References 455

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Mattila, P., & Kumpulainen, J. (2002). Determination of
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CoSeteng, M. Y., & Lee, C. Y. (1987). Changes in apple histological studies of disintegration of cell mem-
polyphenoloxidase and polyphenol concentrations brane materials in vegetables during cooking. Food
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Dellamonica, E. S., & McDowell, P. E. (1965). apple, pear, and white grape juices and their changes
Comparison of beta-carotene content of dried car- with processing and storage—A review. Journal of
rots prepared from three dehydrated processes. Food Agricultural and Food Chemistry, 40, 1478–1487.
Technology, 19, 1597–1599. Von Elbe, J. H., & Schwartz, S. (1996). Colorants.
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Herbs and Spices
12
Zhuohong Xie and John W. Finley

Herbs and spices are important food ingredients. compounds which are basic to the aromas.
The human consumption of herbs and spices can Terpenes represent a very large family of mole-
date back to 5000 BC. The world production of cules which are frequently utilized for defense by
spices is estimated to be 8,730,271 tons in 2013 the plants. Terpenes are the sources of typical
(FAOSTAT). The major producer countries are aromas from bark and needles of conifers, citrus
India, China, Thailand and USA. Table 12.1 sum- products and flowers. The aromas have been
marizes some of the more common spices, the described as pine-like, citrus, fresh, and floral.
portion of the plant and their region of cultiva- These compounds contribute to the overall flavor
tion. Herbs and spices are consumed as is or for- of a broad range of herbs and spices. They are
mulated into various food, beverage and dietary frequently associated with fresh flavors. They
supplement products. Due to their characteristic tend to be volatile so they can be lost during
chemical compounds, herbs and spices are used cooking. The phenolic compounds tend to be
to flavor foods and beverages, to inhibit micro- more dominant in the flavor of the herb or spice.
bial growth and preserve food quality. Increasing Examples of phenolics contributing dominant
evidence also suggest consumption of certain flavors are vanillin in vanilla, cinnamaldehyde in
herbs and spices bring in potential health bene- cinnamon and Eugenol in cloves.
fits. Although the definitions sometimes overlap, Figure 12.1 illustrates some typical structures
generally herbs are plant leaves or flowering parts of phenolics and terpenes found in spices and
either fresh or dried and spices are small pieces herbs. It can be seen that the phenolic compounds
from roots, bark or seeds of plants. Most spices contain a benzene ring appended with at least
also contain essential oils which are normally one hydroxyl group. Terpenes all contain a block
recovered by steam distillation. or at least five carbons in a zigzag formation.
The flavor contribution of spices and herbs Both phenolic compounds and terpenes include a
comes primarily from the aroma of the volatile wide range of compounds with many aroma
compounds in the product. Most spices are fairly characteristics.
complex mixtures of compounds that define the Spices contain volatile oils, usually referred to
distinctive character of the product. In some cases as essential oils, which are isolated by steam dis-
one compound may dominate the aroma profile tillation. The primary constituents in essential
for example cloves, cinnamon, anise and thyme. oils are mono- and sesquiterpenes, phenolics or
The aroma compounds in herbs and spices are phenol ethers (Fig. 12.2).
either phenolics or terpene based compounds. The volatile components in spices are fre-
Table 12.2 summarizes the terpene and phenolic quently complex mixtures, however in some

© Springer International Publishing AG 2018 457


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_12
458 12  Herbs and Spices

Table 12.1  Common spices and region of origin


Common name Latin name Region of cultivation
Fruits
Black Pepper Piper nigrum Tropical and sub-tropical regions
Vanilla Vanilla plantifolia, Vanilla fragans, Madigascar, Comore Island, Mexico, Uganda,
Vanillathaitensis, Vanilla pompon Indonesia
Allspice Pimenta diocia Central America, Caribbean Islands
Paprika (Bell Pepper) Capsicum annuum Widely distributed throughout the world
Chili (Tobasco) Capsicum frutescens
Bay tree fruits and leaves Crocus spinosa Laurus nobilis Mediterranean region
Juniper berries Juniperus communis Temperate climate
Aniseed Pimpinella anisum Mediterranean and southwest/Asia
Caraway Carum carvi Temperate climate
Coriander Coriandrum sativum Southern Europe and northern Africa to
southwestern Asia
Dill Anethum graveolens Tolerates temperatures slightly below freezing
Seeds
Fenugreek Trigonfoenum greacum Temperate and Mediterranean
Mustard Sinapsis alba Temperate regions
   White Brassica nigra
   Black
Nutmeg Myristica fragrans India, Indonesia, Siri Lanka
Cardamom Elettaria cardamomum India, Siri Lanks
Flowers
Cloves Syzygium aromatiicum
Saffron Crocus sativus Mediterranean, India, Australia
Caper Capparis spinosa Mediterranean
Rhizomes
Ginger Zingeiber Officianale South China, India, Japan. Caribbean, Africa
Turmeric Curcuma longa India, China, Indonesia
Bark
Cinnamon Cinnamomum zeylanicum, C. China, Siri Lanka, Indonesia, Caribbean
aromaticum, C. burmanti
Roots
Horseradish Armoracia rusticana Temperate regions
Leaves
Basil Ocimum basilcum Mediterranean, India
Parsley Petroselinum crispum Temperate regions
Savory Satureia hortensis Temperate regions
Tarragon Artemisia dracunculus Temperate regions
Marjorum Origanum majorana Mediterranean, temperate regions
Oregano Origanum heracleoticum, O. onites Temperate regions
Rosemary Rosmarinus officialis Mediterranean
Sage Salvia officialis Mediterranean
Chives Allium schoenoprasum Temperate regions
Thyme Thymus vulgaris Temperate regions
12  Herbs and Spices 459

Table 12.2  Aromas contributed by phenolic and terpenes


Chemical compounds Aroma descriptor
Phenolics
Eugenol Clove
Cinnamaldehyde Cinnamon
Anethole Anise
Vanillin Vanilla
Thymol Thyme
Cavacrol Oregano
Estragole Caraway
Terpenes
Pinenes Pine needles and bark
Limonene, terpinene, citral Citrus fruits
Geraniol Roses
Linalool Lily of the valley
Cineole Eucalyptus
Menthol and Menthone Peppermint
l-Carvone Spearmint
d-Carvone Caraway

Fig. 12.1  Structures of


phenolics and terpenes
found in herbs and
spices

cases the dominant volatiles define the aroma. The antimicrobial effects of herbs and spices
For example eugenol in cloves, cinnamaldehyde are well recognized. Foodborne bacteria are sen-
in cinnamon and carvone in caraway. Table 12.3 sitive to extracts from herbs and spices such as
summarizes the principal aroma compounds oregano, clove, cinnamon, citral, garlic, corian-
found in some of the major spices. der, rosemary, parsley, lemongrass, sage, vanil-
The aroma of many spices is the result of the lin and mustard (Tajkarimi et al. 2010; Ceylan
major aroma producing compounds such as cin- and Fung 2004). The essential oil fraction of
namaldehyde in cinnamon, eugenol in cloves and herbs and spices are rich in antimicrobial com-
(E)-anethole in caraway. Other spice aromas are pounds (Ceylan and Fung 2004). Phenols, alco-
much more complex. hols, aldehydes, ketones, terpenes, ethers,
460 12  Herbs and Spices

Fig. 12.2 Aromatic
compounds found in
essential oils

Table 12.3  Major aroma compounds found in spices and herbs


Spice/herb Major aroma components
Black pepper Germacrene d (11.01%), limonene (10.26%), β-pinene (10.02%), α-phellandrene (8.56%),
β-caryophyllene, (7.29%), α-pinene (6.40%) and cis-β-ocimene (3.19%); P. guineense (black)—β-
caryophyllene (57.59%), β-elemene (5.10%), bicyclogermacrene (5.05%) and α-humulene (4.86%)
White pepper β-Caryophyllene (51.75%), cis-β-­ocimene (6.61%), limonene (5.88%), β-pinene (4.56%), linalool
(3.97%) and α-humulene (3.29%)
Vanilla Vanillin, (r) (+)-trans—α-ionone, p-hydroxybenzylmethylether
Allspice Eugenol, β-caryophyllene, methyleugenol, 1,8-cineole,α-phellandrene
Bay leaf Cineole, α-pinene, β-pinene, α-phellandrene, linalool
Juniper berries α-Pinene, myrcene, β-pinene, δ3-carene, (e)-anethole
Aniseed (e)-Anethole
Caraway (s)(+)-Carvone, limonene
Coriander (s)(+)-Linalool, (r)(−)-linalool, linalyl acetate, citral, 2-alkeales c10–c14
Dill (herb) (s)-Carvone, (s)(+)-phellandrene, (3r,4s,8s)(+)-epoxy-p-menth-1-ene, myristican, limonene
Fenugreek Linalool, 3-isobutyl-2-­methoxypyrazine, 2-methoxy-3-­isopropylpyrazine,
3-hydroxy-4,5-dimethyl-2(5H)-furanone
Nutmeg α-Pinene, β-pinene, sabinene, limonene, safrole, myristicin, 1,8-cineole
Cardamom 1,8-Cineole, α-terpinyl acetate, limonene, sabinene
Clove Eugenol, β-caryophyllene, eugenol acetate
Saffron Safranal, 2,6,6-trimethyl-4-hydroxy-1-­cyclohexene-1-formaldehyde
Ginger (−)Zingiberene, β-bisabolene, (−)-sesquiphellandrone, (+)-ar-cucumene, citral, citronellyl acetate
Turmeric Turmerone, ar-turmerone, zngiberene
Cinnamon Cinnamaldehyde, eugenol, safrole, linaloor, camphor
Parsley p-Mentha-1,3,8-triene, 2-sec-butyl-3-methoxypyrazine, 2-isopropyl-3-­methoxypyrazine,
(z)-6-decanal, (e,e)-2,4-decadienal, myrcene
Marjoram cis-Sabinehydrate, trans-­sabaninehydrate, 1-terpinen-4-ol
Rosemary 1,8-Cineol, camphor, α-pinene, camphene
Oregano Carvacrol, thymol
Sage 1,8-Cineol, camphor, α-pinene, thujone
Thyme Thymol, p-cymene, carvacol, linalool
Adapted from Belitz et al. (2009)
12  Herbs and Spices 461

Table 12.4  Major antimicrobial compounds found in spices and herbs


Spice/herb Major antimicrobial compounds
Rosemary Anethole, apigenin, ascorbic acid, borneol, bornyl acetate, caffeic acid, camphor, delta-3-carene,
carveol, caryophyllene, chlorogenic acid, 1,8-cineole, p-cymene, genkwanin, geraniol, glycolic
acid, limonene, linalool, methyl eugenol, niacin, pinene, rosmarinic acid, safrole, terpinene,
thujone, ursolic acid
Cinnamon Benzaldehyde, camphene, camphor, caryophyllene, 1,8-cineole, cinnamaldehyde, cuminaldehyde,
p-cymene, eugenol, farnesol, furfural, guaiacol, limonene, linalool, methyl eugenol, methyl
salicylate, myrcene, niacin, pinene, piperitone, safrole, terpineol
Thyme Borneol, bornyl acetate, caffeic acid, camphene, delta-3-carene, beta-­carotene, carvacrol,
chlorogenic acid, 1,8-cineole, p-cymene, geraniaol, limonene, linalool, methionine, myrcene,
niacin, pinene, rosmarinic acid, terpinen-4-ol, terpineol, thymol, tryptophan, ursolic acid
Oregano Anethole, apigenin, borneol, cadinene, caffeic acid, camphor, delta-3-carene, beta-carotene,
carvacrol, carvone, caryophyllene, 1,8-cineole, p-cymene, geraniol, kaempferol, limonene, linalool,
luteoline, niacin, pinene, rosmarinic acid, terpinene, thymol, ursolic acid
Bay Acetic acid, benzaldehyde, borneol, bornyl acetate, cadinene, delta-3-­carene, camphene,
caryophyllene, 1,8-cineole, costulonide, cubebin, p-cymene, eugenol, geraniol, limonene, linalool,
methyl eugenol, myrcene, pinene, terpinen-4-ol, terpineol
Garlic Ajoene, allicin, alliin, allistatin-I, allistatin-II, arginine, ascorbic acid, choline, citral, diallyl
disulfide, diallyl disulfide, geraniaol, glutamic acid, linalool, niacin, trypthophan
Cloves Anethole, benzaldehyde, carvone, caryophyllene, chavicol, cinnamaldehyde, ellagic acid, eugenol,
eugenyl acetate, furfural, gallic acid, kaempferol, linalool, methyl eugenol
Allspice Anethole, ascorbic acid, cadinene, delta-3-carene, beta-carotene, caryophyllene, copaene,
p-cymene, eugenol, linalool, methyl eugenol, myrcene, pinene, selinene, terpinen-­4-­ol, terpinene
Sage Apigenin, ascorbic acid, borneol, bornyl acetate, camphor, beta-carotene, caryophyllene,
chlorogenic acid, 1,8-cineole, limonene, linalool, myrcene, niacin, pinene, rosmarinic acid,
terpinen-4-ol, terpinene, thujone, ursolic acid
Vanilla Acetaldehyde, acetic acid, anisaldehyde, benzaldehyde, benzoic acid, cresol, eugenol, furfural,
guaiacol, vanillic acid, vanillin, vanillyl alcohol
Adapted from Duke (1994)

hydrocarbons, alkaloids and peptides in herbs can be used as a stabilizer of fatty oil and
and spices are the effective antimicrobial com- ­antioxidant in sausages. The antioxidant com-
ponents (Ciocan and Bara 2007; Ceylan and pounds in common herbs and spices are listed in
Fung 2004). The antimicrobial compounds in Table 12.5. It is noteworthy that the category of
common herbs and spices are summarized in aroma compounds and antimicrobial compounds
Table 12.4. sometimes overlap, meaning that the same com-
Oxidation is one of the reasons for deteriora- pound may have both properties.
tion of food quality. Free radicals form chain Recent scientific data support that chemical
reactions in the food matrix, causing rancidity compounds in herbs and spices may provide
and unfavorably change in color and flavor. The health benefits including antioxidant and anti-­
chained reaction can be stopped by radical scav- inflammatory effects, reducing risks of cardio-
enging agents, also known as antioxidants. Strong vascular diseases and diabetes, improving the
antioxidants, including certain vitamins, phenols overall health, cognition, GI health etc. (Butt
(flavonoids), terpenoids, carotenoids, phytoestro- et al. 2013; Suhaj 2006; Gruenwald et al. 2010;
gens, minerals, etc. are found in many herbs and Jamal et al. 2006; Singletary 2010). The health
spices (Suhaj 2006). For example, rosemary benefits will be elaborated in the following indi-
extract containing diterpenes and carnosic acid vidual herb and spice section.
462 12  Herbs and Spices

Table 12.5  Major antioxidant compounds found in spices and herbs


Spice/herb Major antioxidant compounds
Black Ascorbic acid, beta-carotene, camphene, carvacrol, eugenol, gamma-terpinene, lauric acid, linalyl
pepper acetate, methyl eugenol, myrcene, myristic acid, myristicin, palmitic acid, piperine, terpinen-4-ol,
ubiquinone
Caraway Beta-carotene, camphene, camphene, carvacrol, gamma-terpinene, gamma-­terpinene, lauric acid,
myrcene, myrcene, myristic acid, myristicin, myristicin, palmitic acid, quercetin, tannin, terpinen-4-ol
Chili Alanine, ascorbic acid, beta carotene, caffeic acid, campesterol, capsaicin, capsanthin, chlorogenic
pepper acid, hesperidin, histidine, kaempferol, lauric acid, lutein, methionine, myrcene, myristic acid, myristic
acid, p-coumaric acid, palmitic acid, pentadecanoic acid, quercetin, scopoletin, stigmasterol, terpinen-
4-ol, tocopherol, tryptophan
Coriander Apigenin, ascorbic acid, beta-carotene, beta-carotene, beta-sitosterol, caffeic acid, camphene,
chlorogenic acid, gamma-terpinene, isoquercitrin, myrcene, myristic acid, myristicin,
p-hydroxybenzoic acid, p-hydroxybenzoic acid, palmitic acid, protocatechuic acid, protocatechuic
acid, quercetin, rhamnetin, rutin, scopoletin, tannin, terpinen-4-ol, trans-anethole, vanillic acid,
vanillic acid
Dill Alpha-tocopherol, anethole, ascorbic acid, beta-sitosterol, caffeic acid, camphene, carvacrol,
chlorogenic acid, eugenol, ferulic acid, gamma-­terpinene, histidine, isoeugenol, isorhamnetin,
kaempferol, lauric acid, methionine, myrcene, myristic acid, myristicin, palmitic acid, quercetin,
scopoletin, selenium, stigmasterol, terpinen-4-ol, trans-anethole, vicenin
Ginger 6-Gingerol, 6-shogaol, alanine, ascorbic acid, beta-carotene, beta-­sitosterol, caffeic acid, camphene,
capsaicin, chlorogenic acid, curcumin, delphinidin, ferulic acid, gamma-­terpinene, kaempferol, lauric
acid, methionine, myrcene, myricetin, myristic acid, p-coumaric acid p-hydroxybenzoic acid, palmitic
acid, quercetin, selenium, shikimic acid, sucrose, terpinen-4-ol, tryptophan, vanillic acid, vanillin
Nutmeg Camphene, cyanidin, eugenol, gamma-terpinene, isoeugenol, kaempferol, lauric acid, methyl eugenol,
myrcene, myristic acid, myristicin, myristicin, oleanolic acid, palmitic acid, quercetin, terpinen-4-ol
Oregano camphene, carvacrol, gamma-­terpinene, linalyl acetate, myrcene, terpinen-4-ol, thymol
Red Alanine, alpha-tocopherol, ascorbic acid, beta-carotene, beta-sitosterol, caffeic acid, campesterol,
(sweet) camphene, capsaicin, capsanthin, chlorogenic acid, eugenol, gamma-terpinene, hesperidin, histidine,
pepper lupeol, lutein, methionine, myrcene, myristic acid, p-coumaric acid, palmitic acid, pentadecanoic acid,
scopoletin, selenium, stigmasterol, terpinen-4-ol, tocopherol, tryptophan
Rosemary apigenin, ascorbic acid, beta-carotene, beta-sitosterol, caffeic acid, camphene, camphene, camphene,
carnosic acid, carnosol, carvacrol, chlorogenic acid, gamma-terpinene, hesperidin, hispidulin,
isorosmanol, labiatic acid, luteolin, luteolin-3′-o-(3″-o-acetyl)-beta-d-glucuronide, luteolin-3′-o-(4″-o-
acetyl)-beta-d-glucuronide, methyl eugenol, myrcene, oleanolic acid, rosmadial, rosmanol,
rosmaridiphenol, rosmarinic acid, rosmariquinone, squalene, tannin, terpinen-4-ol, thymol, trans-
anethole, ursolic acid
Sage Alanine, apigenin, ascorbic acid, beta-carotene, beta-sitosterol, caffeic acid, campesterol, camphene,
carnosic acid, carnosol, carnosol, carnosolic acid, catechin, chlorogenic acid, cholesterol, chrysoeriol,
ferulic acid, fumaric acid, gallic acid, gamma-­terpinene, hispidulin, labiatic acid, linalyl acetate,
luteolin, myrcene, oleanolic acid, p-coumaric acid, palmitic acid, rosmanol, rosmarinic acid, salicylic
acid, selenium, stigmasterol, terpinen-4-ol, thymol essential oil, ursolic acid, uvaol, vanillic acid
Thyme 4-Terpineol, alanine, anethole essential oil, apigenin, ascorbic acid, beta-­carotene, caffeic acid,
camphene, carvacrol, chlorogenic acid, chrysoeriol, eriodictyol, eugenol, ferulic acid, gallic acid,
gamma-­terpinene, isochlorogenic acid, isoeugenol, isothymonin, kaempferol, labiatic acid, lauric acid,
linalyl acetate, luteolin, methionine, myrcene, myristic acid, naringenin, oleanolic acid, p-coumaric
acid, p-hydroxybenzoic acid, palmitic acid, rosmarinic acid, selenium, tannin, thymol, tryptophan,
ursolic acid, vanillic acid
Turmeric Ascorbic acid, beta-carotene, caffeic acid, curcumin, eugenol essential oil, p-coumaric acid,
protocatechuic acid, syringic acid, vanillic acid
Adapted from Suhaj (2006)
Black Pepper 463

Black Pepper Piperine is the most abundant constituent of


pepper oleoresin (Borges and Pino 1993). The
Both black and white pepper are widely con- pungency of black pepper (P. nigrum L.) was
sumed. Black pepper is harvested before it is attributed to the presence of piperine, the struc-
fully ripe the flesh is removed and the seed is ture is shown in Fig. 12.4. After removal of piper-
dried. White pepper, which has a milder flavor, is ine form the pepper resin the oleoresin was
from the seed or ripe fruit. The major aromatic named chavicine (Govindarajan 1977). Chavicine
compound in black pepper is (−)-rotundone. The was thought to possess greater bite the tongue
main differences between black and white pepper than crystalline piperine, however it was later
is the concentrations in the product. The losses of shown that piperine in solution was very pungent.
aroma compounds account for the instability of Later research demonstrated that piperine was
pepper after it is ground. Therefore, fresh ground the major pungent principle and chavicine is a
pepper provides better flavor in most food mixture of piperine and several minor alkaloids.
applications. Five additional pungent alkaloids have been
Blackening of fresh pepper is an oxidation of identified in pepper extracts. They are piperet-
(3,4-dihydroxy phenyl) ethanol glycoside by an tine, piperylin, piperolein, A and B and pipera-
o-diphenol oxidase (PPO) which is present in the nine (Zachariah et al. 2008).
fruit. Bandyopadhyay et al. (1990) reported dur- Structures of some of the major components in
ing the conversion of green pepper to black pep- black pepper oil are found in Figs. 12.3 and 12.4.
per 75% of the total pheolic content is lost. The primary and most important in pepper
Included is a complete loss of o-diphenol oxidase oleoresin is piperine. Black pepper oil contrib-
oxidizable phenolic compounds. The major sub- utes the aroma, oleoresin constitutes the compo-
strate for o-diphenol oxidase is 3,4-dihydroxy-­6- nents that complete the overall taste with the
(N-ethylamino) benzamide. Early work alkaloidpiperine imparts pungency.
established the presence of α-pinene, β-pinene, β-Carophyllene, is the most abundant sesquiter-
1-β-phellandrene, dllimonene, piperonal, and pene hydrocarbon present in pepper oil. Other
dihydrocarveol. sesquiterpene hydrocarbons are β-bisabolene, α-
The volatile oils in black pepper constitute and β-cadinenes, calamenene, α-copaene, α and
2–5% of the dry berries. Pepper oil is traditionally β-cubebenes, ar-curcumene, β- and δ-elemenes,
prepared by steam distillation of the black pep- β-farnesene, α-guaiene, α- and γ-humulenes, iso-
percorns. Liquid carbon dioxide is now emerging caryophyllene, γ-muurolene, α-santalene, α- and
as a useful technology to prepare essential oils β-selinenes, ledene, sesquisabinene and zingibe-
including pepper oil. Liquid carbon dioxide was rene (Zachariah and Parthasarathy 2008).
described by Ferreira et al. (1999). In studies on Beyond contribution to flavor, piperine is also
black pepper berries from India and Malaysia, the the major ingredient related to black pepper’s
presence of optically active monoterpenes, indi- potential health benefits. Piperine in black pepper
cated (±)-linalool, (+)-α-phellandrene, (−)-limo- may be responsible for antioxidant, antimicro-
nene, myrcene, (−)-α-pinene, 3-methylbutanal bial, anti-inflammatory, gastro-protective, antide-
and methylpropanal were the most important pressant and anti-cancer activities of the spice (Li
aroma compounds in black pepper. Additionally, 2006; Butt et al. 2013). Most of these, however,
2-isopropyl-3-methoxypyrazine and 2,3-diethyl-­ are results from in vitro and animal studies; thus,
5-methylpyrazine identified as causing a moldy further clinical research is warranted to explore
or musty off aroma in a black pepper sample from the efficacy and safety on the health promoting
Malaysia (Jagella and Grosch 1999a). use of black pepper.
464 12  Herbs and Spices

Fig. 12.3  Major aroma


compounds in black
pepper

Fig. 12.4  Piperine and


chavicine are the
primary pungent
compounds in black and
white pepper

Table 12.6  World country vanilla production


Vanilla Country Production (tons) %
Madagascar 3719 48
Like Cocoa, vanilla originated in Central Indonesia 2000 26
America, where the Aztecs used it to flavor cocoa Papua New Guinea 510 6.6
well before the discovery of the New World. The Mexico 420 5.4
vanilla plant is a member of the orchidaceae fam- China 286 3.7
ily. The cultivated species of vanilla include Turkey 280 3.6
Vanilla fragans, Vanilla pompona and Vanilla Uganda 218 2.8
tahitensis. The vanilla bush is a tropical plant that Tonga 186 2.4
requires a warm and humid climate. The temper- 2014 Top vanilla producers (Source: FAOSTAT 2017)
ature must be as even as possible. The major
vanilla-producing countries are Madagascar,
Indonesia, China, and Turkey. Madagascar which (Table 12.6). Vanilla is especially popular in ice
cultivates 64,000 ha, is the largest producer and creams, beverages, desserts, dairy products,
some argue has the highest quality vanilla chocolate, confectionery products and pastry.
Vanilla 465

Fig. 12.5 Enzymatic
hydrolysis of vanillin
glucoside to produce
vanillin

Fig. 12.6  Some major


aroma compounds in
vanilla

Vanilla is unique among spices because the pro- as it begins to split on the end. Overripe fruits are
cessing is considerably more involved to produce likely to split, causing a reduction in market
the culinary form. Fresh vanilla pods have little or value. The commercial value of vanilla beans is
no taste. Vanillin which is the primary aroma/fla- fixed based on the length and appearance of
vor constituent is bound as a glycoside which is the pod.
enzymatically hydrolyzed to release the vanillin. Fruit is more than 15 cm (5.9 in) in length, it
The enzymatic reaction is induced by a sequence belongs to first-quality product. The largest fruits
of blanching or steaming. During the curing pro- (greater than 16–21 cm) are reserved for the
cess, the vanillin glycoside is hydrolyzed to yield gourmet vanilla market. Fruits between 10 and
vanillin and glucose and some other minor aro- 15 cm long, pods are under the second-quality
matic substances as shown in Fig. 12.5 category, and fruits less than 10 cm in length are
Vanilla fruit grows quickly on the vine and under the third-quality category.
matures for harvest in about 6 months. Vanilla There are four basic steps in curing
fruits ripen at widely differing rates, thus daily vanilla beans killing, sweating, slow-drying, and
harvest is required to ensure the optimum flavor conditioning of the beans (Havkin-Frenkel et al.
from every fruit. Individual pods are hand-picked 2004, 2005).
466 12  Herbs and Spices

In order to initiate the enzymatic hydrolysis of Vanilla is one of the more complex aro-
the vanillin glycosides the vegetative cells of the mas containing over 170 compounds. Vanillin
vanilla bean must be disrupted. This process is the primary aroma material thus “imita-
called killing disrupts the cells and tissue of the tion” vanilla produced enzymatically is used
fruits, imitating the enzymatic reactions respon- in many food applications. Natural vanillin is
sible for releasing the aromatic compounds. produced from the enzymatic hydrolysis of a
Killing can be accomplished by heating in hot glycoside during fermentation of the fruit. Two
water, freezing, or scratching, heating in an oven other important contributors to the aroma of
or exposing the beans to direct sunlight. The dif- fresh vanilla are (R) (+)-trans-α-ionone and
ferent methods give different profiles of enzy- p-hydroxybenzylmethylether.
matic activity allowing the glycosidase enzymes Other aroma compounds in vanilla have been
to act on the glucovanillin to release the free van- reported including aromatic carbonyls, aromatic
illin (Frenkel et al. 2010; Arana 1944). alcohols, aromatic acids, aromatic esters, phenols
Hot-water killing is accomplished by dipping and phenol ethers, aliphatic alcohols, carbonyls,
the beans in hot water 63–65 °C (145–149 °F) for acids, esters and lactones. The levels of vanillin,
3 min, or at 80 °C (176 °F) for 10 s. Fruits can be p-hydroxybenzaldehyde and their respective
scratched lengthwise to disrupt the cells structure acids (vanillic acid and p-hydroxybenzoic acid),
of the fruits (Arana 1944). Fruits can be frozen are used as indicators of vanilla bean quality in
and thawed to initiate the release of enzyme and commercial operations (Klimes and Lamparsky
substrate however they must be thawed for the 1976; Adedeji et al. 1993; Ranadive 1994; Azeez
sweating stage. Frozen or quick-frozen fruits 2008). The major aroma compounds in vanilla
must be thawed again for the subsequent sweat- are shown in Fig. 12.6.
ing stage. Fruits can also be tied bundles and Chemical composition of processed vanilla
rolled in blankets and heated in and oven at 60 °C Composition of processed vanilla beans variable
for 36–48 h. The Aztecs exposed the fruits to and complex due to a number of variables such
sunlight until they turned dark brown to imitate as species, growth conditions, soil composition,
the enzymatic reaction (Frenkel et al. 2010). fruit maturity and mainly, the type of process-
Sweating is a combination of hydrolytic and ing. All these variables define the relative con-
oxidative processes. Sweating is the process of tent of the chemical constituents in the processed
holding the fruits, for 7–10 days at temperature beans, which makes it difficult to define their
of 45–65 °C with high humidity. At the end of the typical composition. Perez-Silva et al. (2006)
sweating process the fruits are brown and have studied the volatile compounds in vanilla beans
developed the characteristic vanilla flavor and combining GC/MS analysis with GC-olifactory
aroma. At the end of sweating they contain analysis to characterize the aroma contributors
60–70% moisture content by weight (Frenkel in vanilla beans. Table 12.7 summarizes the
et al. 2010). results of the study.
The drying process reduces the beans to Vanillin can be synthesized by a number of
25–30% moisture which is sufficient to prevent methods and is frequently used as a replacement
rotting and stabilizes the aroma compounds in the for the much more expensive natural vanilla. The
pods. Traditionally the drying is accomplished by ‘classical’ synthesis of vanillin from eugenol or
exposing the beans to intermittent shade and sun- isoeugenol was developed in 1896 and it remained
light. Drying is the most difficult stage to control the preferred method for about 50 years. Vanillin
because it can result in uneven drying which can is now prepared industrially in large amounts by
lead to loss is in vanillin (Frenkel et al. 2010). the Reimer–Tiemann reaction, starting with guai-
Conditioning is accomplished by storing the acol, from which it is formed along with
pods for 5–6 months in closed boxes. During this o-­vanillin. A newer and more common means to
stage the fragrance develops. The cured vanilla produce vanillin is from the reaction of guracol
fruits contain an average of 2.5% vanillin. and glyoxylic acid as shown in Fig. 12.7.
Vanilla 467

Table 12.7  Aroma compounds from vanilla beans and odor quality (from Perez-Silva et al. 2006)
Compounds ppm Odor quality Intensitya
Phenols
Guaiacol 9.3 Chemical, sweet spicy +++
4-Methylguaiacol 3.8 Sweet, woody +++
p-Cresol 2.6 Balsamic, woody, spicy ++
4-Vinylguaiacol 1.2 Chemical, phenolic +
4-Vinylphenol 1.8 Sweet, woody ++
Vanillin 19,118 Vanilla, sweet +++
Acetovanillone 13.7 Vanilla, sweet, honey +++
Vanillyl alcohol 83.8 Vanilla like +++
p-Hydroxybenzaldehyde 873 Vanilla like, biscuit ++
p-Hydroxybenzyl alcohol 65.1 Vanilla like, sweet ++
Aliphatic acids
Acetic acid 124 Sour, vinegar ++
Isobutyric acid 1.7 Buttery ++
Butyric acid <1 Buttery, oily +
Isovaleric acid 3.8 Buttery, oily ++
Valeric acid 1.5 Cheese +++
Alcohols
2,3-Buranediol (Isomer 2) 8.0 Floral, oily +
Anisyl alcohol 2.4 Herbal ++
Aldehydes
2-Heptenal 2.1 Green, oily +
E-2-decenal 1.8 Herblike, floral ++
(E,Z)-2,4-decadienol 1.4 Herb-like, fresh ++
(E,E)-2,4-decadienol 1.2 Fatty, wood ++
Esters
Methyl salicylate <1 Chalk +++
Methyl cinnamate 1.1 Sweet ++
Ethyl linoleate 13.5 Sweet ++
Ketone
3-Hydroxy-2-butanone 14.6 Buttery +
Unknownb 6.2 Vanilla-like, chemical +++
Aroma-active compounds detected by GC-O analysis of representative aroma extract from vanilla beans
a
(+) Weak, (++) medium, (+++) strong
b
Mass fragmentation 91(90), 74(37), 69(34),89(25), 57(24), and RI 2528

Vanillin is also recovered as a by-product of by adding vanilla beans to a liquid preparation.


paper pulp manufacture. Synthetic vanillin is Natural vanilla gives a brown or yellow color to
used in both food and non-food applications, in preparations, depending on the concentration.
fragrances and as a flavoring in pharmaceutical Good-quality vanilla has a strong aromatic flavor,
preparations. but food with small amounts of low-quality
In culinary applications vanilla flavoring in vanilla or artificial vanilla-like flavorings are far
food can be achieved by adding vanilla extract or more common.
468 12  Herbs and Spices

Fig. 12.7  Chemical approaches for the production of vanillin

most expensive spices after saffron and vanilla.


Cardamom The volatile oil components of cardamom first
reported in detail by Nigam et al. (1965) and fur-
Cardamom is a spice used in South and Southeast ther summarized by Guenther (1975). The oil
Asia, Middle East and Nordic countries. It is predominantly composed of oxygenated com-
made from seeds of two plants (Elettaria carda- pounds, all which contribute to the aroma. Many
momum and Amomum subulatum). The major of the alcohols, esters and aldehydes are found in
production countries are Guatemala, India and many spice oils. The predominant volatiles in
Sri Lanka. Cardamom is considered one of the cardamom are 1,8-cineole and the esters,
Ginger 469

Table 12.8  Main volatile components in cardamom oil (steam volatile) and non-­ volatile components
Component Total oil (%) responsible for the pungency.
α-Pinene 1.5 Ginger oleoresin can be prepared from dried
β-Pinene 0.2 ginger by extraction using a variety of organic
Sabinene 2.8 solvents. The oleoresin contains the major organ-
Myrcene 1.6 oleptically important volatile oil and pungent
α-Phellandrene 0.2 principles. In addition the oleoresin includes
Limonene 11.6 ­triglycerides and a small amount of free fatty
1,8-Cineole 36.3 acids. Preparation and storage of the dried spice
γ-Terpinene 0.7 and oleoresin influences the organoleptic proper-
p-Cymene 0.1 ties of these products. During storage of dried
Terpinolene 0.5 spice and particularly ground spice significant
Linalool 3.0 amounts of the volatile components in the oil are
Linalyl acetate 2.5 lost to evaporation. The oleoresin is prone to loss
Terpinen-4-ol 0.9 of pungency during as a result of degradation of
α-Terpineol 2.6 the pungent gingerols. Heating ginger or the
α-Terninyl acetate 31.3 oleoresin from ginger results in the loss of both
Citronellol 0.3
volatile and pungent factors.
Nerol 0.5
The fatty oil of ginger ranges from 2 to 12% in
Geraniol 0.5
dried gingers. Ginger fatty oil contain saturated
Methyl eugenol 0.2
and unsaturated fatty acids in a ratio of 46:53.
Trans-Nerolidol 2.7
The major fatty acids in ginger lipids are pal-
Source: Lawrence (1978); Govindarajan et al. (1982)
mitic, oleic and linoleic (Zachariah 2008). The
total lipid content varies widely (5.8–15%)
α-terpinyl and linalyl acetates (Lewis et al. 1966; among ginger varieties (Govindarajan 1982). The
Salzer 1975; Korikanthimath et al. 2002). The typical fatty acid profile of ginger lipid is shown
major components in cardamom oil are listed in in Table 12.9.
Table  12.8. Structures of the main components The composition of ginger oleoresin has a
are found in Fig. 12.8. Preliminary studies have wide range depending on the variety and extrac-
found consumption of cardamom may have anti- tion conditions. Most commercial dried gingers
oxidant (Kikuzaki et al. 2001), gut stimulatory contain 3.5–10% oleoresins and 15–30% volatile
and inhibitory, gastroprotective (Jamal et al. oils (Govindarajan 1982).
2006), diuretic and sedative effects (Gilani et al. The non-volatile portion referred to as gin-
2008), as well as alleviation of diarrhea, consti- gerols contain a 1-(4′-hydroxy-3′-methoxyphenyl)-
pation, and high blood pressure (Gilani et al. 5-hydroxyzlkan-3-one structure. The non-volatile
2008). pungent constituents in ginger include gingerols,
shogaols, zingerone and paradols. All of these
groups of compounds except zingerone contain
Ginger hydrocarbon chains of 4–8 -CH2- groups. The pun-
gent non-volatile components in ginger are illus-
Ginger is the rhizome of Zingiber officinale Roscoe trated in Fig. 12.9.
and is one of the most widely used spices in a wide The volatile composition of ginger oil is very
range of condiment for various foods and bever- complex. Miyazawa and Kameoka (1988) identi-
ages. Z. officinale Roscoe, which is used for com- fied 72 components in the volatile oil extracted
mercial ginger, and is grown extensively in many from the air-dried rhizomes.
tropical countries. Like pepper (Piper nigrum) and The main components were zingiberene
the fruits of the Capsicum species, ginger is char- (21.8%), geranial (9.9%), geraniol (9.4%),
acterized by two classes of constituents: the odor β-bisabolene (7.9%), nerol (7.1%), 1,8-cineol
470 12  Herbs and Spices

Fig. 12.8 Major
components of
cardamon oil
(Chempakam and
Sindhu 2008)

Table 12.9  Fatty acid composition of ginger lipids (1982). It was proposed that Citral and citronellyl
Fatty acid Fatty acid (%) acetate were the most important attributes of
Caprylic acid 1.4 aroma. Freshly prepared oil is rich in Zingiberene
Capric acid 4.1 and β-sesquiphellandrene, and ar-Curcumene,
Lauric acid 7.6 increases during storage, and can be used as an
Myristic acid 3.5 indicator of aoil age and processing conditions.
Pentadecanoic acid 0.4 The ratio of zingiberene and β-sesquiphellandrene
Palmitic acid 23.2 to ar-curcumene = 2:3 as a stable marker for
Heptadecanoic acid 1.3 identifying pure ginger oil. The lemony note
Stearic acid 3.3 from the citrals combined with the α-terpineol,
Oleic acid 22.9 β-sesquiphellandrene and ar-curcumene com-
Linoleic acid 23.2 bine to form the basis of the characteristic ginger
Linolenic acid 6.6 flavor.
Arachidic acid 1.1 Nerolidol contributes to the woody note; and
cis- and trans-β-sesquiphellandrol were also con-
(6.2%), terpineol (5.6%), borneol (5.4%), phel- tributors to the ginger flavor (Govindarajan
landrene (3.1%), linalool (1.7%), methyl nonyl 1982). Sesquiterpenes, particularly zingiberene,
ketone (1.6%) and camphene (1.4%); the other was identified as a major characteristic of
components accounted for ∼1% each of the vola- ginger.
tile oil. The structures of the primary volatile Ginger has been used as folk medicine since
compounds in ginger are shown in Fig. 12.10. ancient time. With help from recent advances in
The impact of composition of ginger on gin- analytical chemistry and nutrition, the bioactive
ger oil quality was summarized by Govindarajan compounds in ginger including gingerols,
Turmeric 471

Fig. 12.9  Non volatile components of ginger oil

Fig. 12.10 Primary
volatile compounds in
ginger

β-carotene, capsaicin, caffeic acid, curcumin and


salicylate have been identified (Tapsell et al. Turmeric
2006). In vitro and animal studies suggest ginger
has antioxidant, anti-inflammatory, and anti-­ Turmeric, Curcuma longa L. (Zingiberaceae) is a
platelet properties, as well as lipid and blood spice cultivated in warm, rainy regions like India,
pressure lowering activities (Singletary 2010). In China, Indonesia, Jamaica and Peru (Govindarajan
addition, a systematic review summarized 12 ran- 1980). The spice belongs to the genus Curcuma,
dom clinical trials and concluded that ginger con- which includes several plant species with
sumption is able to reduce pregnancy-­associated underground rhizomes and roots. About 40
nausea symptoms (Viljoen et al. 2014). species of the genus are indigenous to India
­
472 12  Herbs and Spices

(Velayudhan et al. 1999). The use of turmeric


dates back nearly 4000 years to the Vedic culture
in India, where it was used as a culinary spice but
also had some religious significance.
Turmeric is used as a food additive to improve
the palatability, storage and preservation of food.
For example, turmeric is used in a wide range of
curry powders and mustards. In Asian cuisines,
dry or fresh turmeric, or ground turmeric, are
used for vegetable and meat dishes and soup-like
dishes (Govindarajan 1980). Turmeric oleoresin
extract is used in brine pickle (Eiserlie 1966;
Cripps 1967) and mayonnaise and relish formu-
lations. It can be included in non-alcoholic bever-
ages, or for garnishing and in some ice creams
(Perotti 1975). Turmeric has in intense red orange
color and it is commonly used as a natural food
color (Govindarajan 1980).
The processing and storage condition can
affect the quality of turmeric. Both dried rhi-
zomes and leaves are used as raw materials to
extract the volatile oil. Dried rhizomes contain
5–6% and leaves contain about 1.0–1.5% oil
which is extracted by steam distillation. A large
portion of volatile oil can be lost during process-
ing (Chempakam et al. 2008a). The curcuminoids
(a general terms for curcumin, demethoxycur-
cumin and bisdemethoxycurcumin) deteriorate
on exposure to light and are also prone to oxida-
tion which caused discoloration (Buescher and
Fig. 12.11  Curcuminoids found in turmeric
Yang 2000).
The primary component contributing to the
unique aroma of turmeric is ar-turmerone. The volatile oil and delivers a fresh, clean, mildly
primary compounds responsible for the color in pungent, woody-pungent, woody-spicy aroma of
the rhizomes are curcumin (1,7-bis (4-hydroxy-­ turmeric (Chempakam et al. 2008b).
3-methoxy prenyl)-1, 6-heptadiene-3, 5-dione) Turmeric has gained lots of popularity recently
and two related demethoxy compounds, deme- due to increasing evidence on health benefits as
thoxy curcumin and bis-demethoxycurcumin well as promotion from celebrities and top chefs.
(Fig. 12.11). Curcumin is insoluble in water but In most studies, the active component of turmeric
highly soluble in ethanol and acetone. is considered to be curcumin. Consumption of
The oleoresin from turmeric is primarily used curcumin showed lipid lowering effects in
as a food color, and secondarily, to contribute a patients with type-2 diabetes and metabolic syn-
characteristic mild spicy aroma compatible with drome (Neerati et al. 2014; Yang et al. 2014).
mustard, pickles, relish formulae, etc. Curcuminoids may reduce inflammation in
Commercial oleoresin from turmeric has a cur- patients with cardiovascular diseases indicated
cuminoid content of 4.5–5.0%. It is highly vis- by lowering C-reactive protein in blood (Sahebkar
cous with a dark brownish-orange color. The 2014). Curcumin was found to have anticancer
product contains 30–40% curcumin, 15–20% activities in numerous studies (Hallman et al.
Cinnamon 473

2017; Yue et al. 2016). A review also summarized Table 12.10  Botanical sources of cinnamon and cassia
that turmeric/curcumin products both oral and Cinnamon variety Region of origin/production
topical may improve skin health (Vaughn et al. Cinnamomum verum Presl True or Ceylon cinnamon
2016). Conclusive evidence on these health ben- (syn. C. zeylanicum Nees)
efits are warranted. C. cassia Presl Cassia, Chinese cinnamon,
“Cassia lignea”
C. burmannii Blume Indonesian cassia
C. loureirii Nees Vietnamese cassia
Cinnamon
C. tamala (Buch.-Ham.) Indian cassia
Nees & Eberm
Cinnamon and its close relative, cassia, are
From Leela (2008)
among the earliest, most popular spices used by
mankind. The genus Cinnamomum (family:
Lauraceae) consists of 250 species of trees and food seasonings, sauces and pickles, baked
shrubs distributed in Southeast Asia, China and goods, confectionery, cola-type drinks, tobacco
Australia. True cinnamon, Cinnamomum verum flavors and in dental and pharmaceutical prepara-
syn. C. zeylanicum, is a native of Sri Lanka and tions. Perfumery applications are limited because
South India. Cassia cinnamon is derived from the oil has some skin-sensitizing properties.
different sources, including Chinese cassia (C. Cinnamon leaf oil has a warm, spicy aroma, but
cassia syn. C. aromatica) from China and the aroma can be harsh and it lacks the body of
Vietnam, Indonesian cassia (C. burmannii) from bark oil. Leaf oils major constituent is eugenol
Sumatra and the Java region and Indian cassia (C. rather than cinnamaldehyde. Leaf oil is used in
tamala) from the north-eastern region of India seasonings and savory snacks, as a low cost fra-
and Myanmar (Baruah and Nath 2004; Leela grance in soaps and insecticides. Table 12.11 lists
2008). Sri Lanka is the major cinnamon produc- the profiles of the volatile compounds found in C.
ing country with 60% of the world cinnamon zeylanicum leaves and bark.
trade. The dried inner bark of the cinnamon tree Cinnamomum verum (C. zeylanicum) is used
is used as a spice. Cinnamon oleoresin is obtained to produce both leaf and bark oils for flavoring
by solvent extraction of the bark, is used mainly and perfumes. The major component of bark oil
for flavoring food products such as cakes and is cinnamaldehyde and of leaf oil is eugenol. In
confectionary products. The volatile oil and oleo- cinnamon the volatile components are found in
resin from cassia are also used for flavoring soft other parts of the plant including root bark, fruits,
drinks and other beverages (Leela 2008). The flowers, twigs and branches. The most important
genus Cinnamomum comprises several hundred source is the bark which varies from 0.4 to 2.8%
species which occur naturally in Asia and (Angmor et al. 1972; Wijesekera 1978;
Australia. They are evergreen trees and shrubs Krishnamoorthy et al. 1996). The oil from the
and most species are aromatic. C. verum, the stem bark of a commercial cinnamon sample
source of cinnamon bark and leaf oils, is a tree contained 75% cinnamaldehyde, 5% cinnamyl
indigenous to Sri Lanka, although most oil now acetate (Gruenwald et al. 2010), 3.3% caryophyl-
comes from cultivated areas (Leela 2008). The lene, 2.4% linalool and 2.2% eugenol
major varieties and growing regions are listed in (Senanayake et al. 1978).
Table 12.10. Cassia oil is distilled from a mixture of leaves,
Cinnamon bark oil delivers a delicate aroma twigs and fragments of bark. Cinnamaldehyde is
of the spice and a sweet and pungent taste. Its the major constituent and it is used mainly for
major constituent in cinnamon bark is cinnamal- flavoring cola-type drinks, with smaller amounts
dehyde, however it is the minor components used in bakery products, sauces, confectionery
impart the characteristic odor and flavor that and liqueurs. Like cinnamon bark oil, its use as a
makes C. zeylanicum unique. The cinnamon bark fragrance is limited by its skin sensitizing
oil is used in food flavoring such as meat and fast properties.
474 12  Herbs and Spices

Table 12.11  The primary volatile components in C. zeylanicum leaf and bark extracts
Leaf Bark
Volatile oil Oleoresin Volatile oil Oleoresin
% % % %
α-Thujene 0.1 α-Pinene tr
α-Pinene 0.5 Camphene tr
β-Pinene tr Sabinene tr
Myrcene tr β-Pinene tr
α-Phellandrene 1.9 Limonene tr
p-Mentha-1(7),8-diene tr 1,8-Cineole tr
p-Cymene 0.7 Camphor tr
1,8-Cineole 0.7 Z-cinnamaldehyde tr 1.5
Terpinolene tr E-cinnamaldehyde 97.7 50.0
α-Terpineol tr α-Copaene  0.8 4.6
α-Cubebene tr α-Amorphene  0.5
Eugenol 87.3 87.2 δ-Cadinene  0.9 7.8
β-Caryophyllene 1.9 1.4 Terpinen-4-ol 0.1
Aromadendrene 1.1 0.8 β-Caryophyllene 1.0
α-Amorphene tr 0.4 Coumarin 16.6
Germacrene-d 0.6 0.2 α-Muurolene 4.4
Bicyclogermacrene 3.6 1.7 β-Bisabolene 1.4
δ-Cadinene 0.4 0.6 Cadina-1(2), 4-diene 1.8
Spathulenol 0.5 1.7 Ortho-methoxy cinnamaldehyde 1.5
Sabinene tr Cubenol 0.5
γ-Terpinene tr 1-Heptadecene 0.2
Terpinen-4-ol tr 1-Nonadecene 0.4
δ-Elemene 1.0 Tetracosane 0.1
Viridiflorol 0.3 Octacosane 0.1
Methoxy-eugenol 0.1 Nonacosane 0.2
Isospathulenol 0.3
Neophytadiene 0.3
Docosane 0.1
Nonacosane 0.1
Vitamin-E 0.2
Total 99.4 97.1 Total 100 92.3
Adapted from Singh et al. (2007)

The volatile distribution of C. Cassia differs The primary components influencing cinna-
significantly form C. zeylanicum. Table 12.11 mon quality are associated with the volatile com-
compares the bark and leaf volatiles from C. zey- ponents, cinnamon also contains several
lanicum. The dried inner bark of cinnamon and diterpenes. The diterpenes include cinncassiols
cassia contains volatile oil, fixed oil, tannin, resin A, B, C1 and their glucosides, cinncassiols C2
proteins, cellulose, pentosans, mucilage, starch, and C3, cinncassiols D1, D2 and D3 and their
calcium oxalate and mineral elements. Cinnamon glucosides, cinncassiol E, cinnzeylanol, cinnzey-
delivers a spicy aroma from its volatile oil which lanin, anhydrocinnzeylanol and anhydrocinnzey-
is composed of a mixture of monoterpenes, ses- lanin. Cinnamon also contains several benzyl
quiterpenes and phenylpropenes. The major vola- isoquinoline alkaloids, flavanol glucosides, cou-
tile components in cinnamon are illustrated in marin, b-sitosterol, cinnamic acid, protocatechuic
Fig. 12.12. acid, vanillic acid and syringic acid.
Cinnamon 475

Fig. 12.12  The major


volatile components in
cinnamon (Leela 2008)

Some important non-volatile components The in vitro investigation of cinnamon has


reported in Cinnamomum and Cassia are listed in revealed that its extract mimics the function of
Table  12.12 and the structures are shown in insulin, which potentiates insulin action in iso-
Fig. 12.13. lated adipocytes (Broadhurst et al. 2000).
Cinnamon is one of the oldest herbal medi- Cinnamon extract has been shown to improve the
cines, which has been recorded in Chinese publi- insulin receptor function (Jarvill-Taylor et al.
cations 4000 years before (Qin et al. 2003). 2001; Leela 2008). Current in vitro and in vivo
Cinnamon has been used to treat dyspepsia, gas- evidence suggests that cinnamon has
tritis, blood circulation disturbance and inflam- anti-inflammatory, antimicrobial, antioxidant,
­
matory diseases in many countries since ancient antitumor, cardiovascular, cholesterol lowering,
times (Yu et al. 2007). The significant anti-aller- and immunomodulatory effects (Scaglione et al.
gic, anti-ulcerogenic, antipyretic, anaesthetic and 1989). In vitro studies have demonstrated that
analgesic activities have been confirmed previ- cinnamon acts as an insulin mimetic, to potenti-
ously (Kurokawa et al. 1998; Lee and Ahn 1998). ate insulin activity or to stimulate cellular glucose
476 12  Herbs and Spices

Table 12.12  Non-volatile components in cinnamon and cassia


Compound Plant part References
Lyoniresinol 3 α-O-β-glucopyranoside C. cassia stem bark Miyamura et al. (1983)
2,4,5-Trimethoxy phenol β-d apiofuranosyl-(1-6)-β-­glucopyranoside C. cassia stem bark Miyamura et al. (1983)
Syringaresinol C. cassia stem bark Miyamura et al. (1983)
5,7,3′-Trimetyl (−) epicatechin C. cassia stem bark Miyamura et al. (1983)
5,7-Dimethyl-3′-,4′-di-O-­methylene (±) epicatechin C. cassia stem bark Miyamura et al. (1983)
Cinnamic aldehyde cyclic glycerol-1,3-acetol (9,2′ trans) C. cassia stem bark Miyamura et al. (1983)
Cinnamic aldehyde cyclic glycerol-1,3-acetol (9,2′ cis) C. cassia stem bark Miyamura et al. (1983)
Cimmacassiol D4 C. cassia stem bark Nohara et al. (1982)
Cimmacassiol D4 -glucoside C. cassia stem bark Nohara et al. (1982)
2′-Hydroxy cinnamaldehyde C. cassia stem bark Kwon et al. (1996)
3-(2-Hydroxy phenyl)-propanoic acid C. cassia stem bark Tanaka et al. (1989)
Cinncassiol E C. cassia Nohara et al. (1985)
From Leela (2008)

Fig. 12.13  Major non-volatile components in cinnamon


Ginseng 477

metabolism. Animal studies have demonstrated been added into tea, energy drink, soup, candies,
strong hypoglycemic properties. The use of cin- as well as dietary supplements in food and
namon as an adjunct to the treatment of type 2 beverage.
diabetes mellitus is the most promising area, but The major bioactives in ginseng are ginsen-
further research is needed before definitive rec- osides, belonging to triterpene saponins. Most
ommendations can be made. ginsenosides consist of a dammarane aglycone
with different sugar moieties. The common gin-
senosides are shown in Fig. 12.14. The composi-
Ginseng tion of ginsenosides are affected by species, age,
growing condition, harvesting method, as well as
Ginseng is a traditional herb that has been used storage condition (Leung and Wong 2010).
for a 1000 years by ancient East Asian. It was The therapeutic potentials of ginseng and gin-
considered to be a panacea thus having the genus senosides have been studied extensively.
name Panax. There are 11 species in the genus Consumption of ginseng has positive effects on
Panax considered as ginseng. The edible part is cardiovascular diseases (Lee and Kim 2014),
the fleshy root of ginseng plant. Ginseng is immune modulation (Scaglione et al. 1990),
grown in Asia and North America with China, enhancement on Cognitive function (Vogler et al.
South Korea, Canada, and the US being the four 1999). The mechanism on efficacy of ginseng on
major producing countries, accounting for 99% cardiovascular diseases may include: inhibition
of the annual 80,080 tons’ production (Baeg and of free radical production, stimulation of NO pro-
So 2013). As a functional food ingredient, gin- duction, improvement in blood circulation,
seng has been found in over 11,000 retail prod- adjustment of vasomotor function, and improve-
ucts (Innova Market Insights 2017). Ginseng has ment in the lipid profile (Lee and Kim 2014).

Fig. 12.14 Structures
of common
ginsenosides.
Abbreviation: Glc
glucopyranoside,
Ara(pyr)
arabinopyranoside, Rha
rhamnopyranoside.
Adapted from Choi et al.
(2002)

Ginsenoside R1 R2 R3
Rb1 -O-Glc-Glc -H -O-Glc-Glc
Rc -O-Glc-Glc -H -O-Glc-Ara (pyr)
Re -OH -O-Glc-Rha -O-Glc
Rf -OH -O-Glc-Glc -OH
Rg1 -OH -O-Glc -O-Glc
478 12  Herbs and Spices

Ginkgo Food Fraud Risks

Ginkgo (Ginkgo biloba) is a woody plant that has Herbs and spices are usually high value crops
been grown widely in the world. Similar to gin- that require delicate care and intensive labor.
seng, ginkgo leaf is considered a medicinal herb They are mostly sold in powder, minced, oleo-
that has a rich history of consumption in China. resin and essential oil forms. The combining
Ginkgo has a variety of bioactives categorized factors above have made herbs and spices
­
into flavonoids (glycosides of kaempferol, ­products susceptible to Economically Motivated
quercetin, and isorhamnetin) and terpenoids
­ Adulteration (EMA). Fraudulent adulteration of
(ginkgolide A, B, C, and J) (Kleijnen and spices and herbs with added colors have been
Knipschild 1992). Figure 12.15 showed the struc- seen in international trade and retail markets
ture of common ginkgolides. Ginkgo is usually (USP 2017). Market surveillance has showed
made into extracts and is mainly added into 25% of the oreganos in the markets are adulter-
dietary supplements or as prescribed herbal ated with other plants (BBC 2015). Tested by
drugs. Standardized extracts from the leaves of Taiwan FDA, lead chromate has been found in
ginkgo have a set ratio of bioactives. For exam- one turmeric product at lead level as high as
ple, EGb761 contains 24% of flavonoids and 6% 1600 ppm (Liao 2016). Sudan dyes have been
of terpenoids. The extract EGb761 are commonly identified in paprika powders and oleoresins
prescribed for dementia and cognitive impair- (USP 2017). With the enforcement of FSMA act,
ment (O’Hara et al. 1998). A review of nine clini- preventive controls have to be taken to ensure
cal trial found a 240 mg dose of EGb761 was able product quality and safety against EMA. The
to improve cognition for the whole group of mitigation of risks would require a comprehen-
patients with Alzheimer’s disease, vascular or sive supply chain control, a good QA testing sys-
mixed dementia, as well as for the Alzheimer’s tem, as well as information tools such as database
disease subgroup (Weinmann et al. 2010). capturing fraudulent activities to be in place.

References
Adedeji, J., Hartman, T. G., & Ho, C. (1993). Flavor
characterization of different varieties of vanilla beans.
Perfumer and Flavors, 18, 115–133.
Angmor, J. E., Dicks, D. M., Evans, W. C., & Sandra,
D. K. (1972). Studies on Cinnamomum zeylanicum.
Part 1. The essential oil components of C. zeylanicum
Nees grown in Ghana. Planta Medica, 21, 3.
Arana, F.E. (1944, October). Vanilla curing and its chemis-
try. Bulletin (Federal Experiment Station of the United
States Department of Agriculture in Mayaguez, Puerto
Rico) (42): 1–17.
Azeez, S. (2008). Vanilla. In V. A. Parthasarathy,
C. Bhageerathy, & T. J. Zachariah (Eds.), Chemistry of
Ginkgolide R1 R2 spices. Cambridge, MA: CABI.
A -H -H Baeg, I. H., & So, S. H. (2013). The world ginseng market
B -OH -H and the ginseng (Korea). Journal of Ginseng Research,
C -OH -OH 37(1), 1–7.
J -H -OH Bandyopadhyay, C., Narayan, V. S., & Variyar, P. S. (1990).
Phenolics of green pepper berries (Piper nigrum L).
Fig. 12.15 Structures of terpenoids (ginkgolides) in Journal of Agricultural and Food Chemistry, 38(8),
Ginkgo 1696–1699.
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Beer and Wine
13
John W. Finley

Alcoholic beverages are produced by fermenta- duced in 7000–6650 BC. A Sumerian poem hon-
tion of sugars to ethanol. Starting materials range oring Ninkasi, the patron goddess of brewing,
from simple sugars to complex carbohydrates contains the oldest surviving beer recipe describ-
that are reduced to simple sugars by hydrolytic ing the production of beer from barley 3900 years
cleavage of starches and dextrins. Beer and wind ago. Beer became vital to all the grain-growing
represent direct products from fermentation civilizations of classical Western antiquity,
whereas vodka, rum, whiskey and other distilled including Egypt. Knowledge of brewing was
spirits and a distillation step. passed on to the Greeks who then passed the
Fermented beverages have been produced by knowledge on to the Romans. Early Romans con-
many cultures since some of the earliest days of sumed beer, but during Republican times wine
civilization. Mead, was produced in Asia from displaced beer as the preferred alcoholic bever-
honey during the Vedic period (around 1700– age (Alba-Lois and Segal-Kischinevzky 2010).
1100 BC), and later mead was produced by the Sumerian and Egyptian texts dating from
Greeks, Celts, Saxons, and Vikings. In Egypt, about 2100 BC described medical uses of alco-
Babylon, Rome, and China, wine was made from holic beverages. The Hebrew Bible recommends
grapes and beer from malted barley. In Godoy giving alcoholic drinks to those who are dying or
et al. (2003). depressed, so that they can forget their misery
Most substances that contain fermentable sug- (Proverbs 31:6–7).
ars can undergo spontaneous fermentation by During the Middle Ages, in Europe beer, often
wild yeasts in the air. Beer-like beverages were of very low strength, was an everyday drink for
independently invented among various cultures all classes and ages of people.
throughout the world. Analysis of residues in In South America produced a beer-like bever-
ancient pottery jars suggests that both beer and age was produced from cassava or maize. The
wine were produced about 7000 years ago in maize or cassava had to be chewed before fer-
what is today Iran. In Mesopotamia, the oldest mentation to provide amylase which converted
evidence of beer is believed to be a 6000-year-­ the starches to fermentable sugars. In ancient
old. Wine’s first appearance is from around 6000 Japan rice was chewed to hydrolyze thee rice
BC in Georgia. The earliest firm evidence of starch in sake production.
wine production dates back to 5400 BC in Iran. Louis Pasteur determined that the fermenta-
In China chemical analysis of Neolithic jars con- tion process result from the action of living yeast
firmed that a fermented drink made of grapes, converting transforming glucose into ethanol. He
hawthorn berries, honey, and rice was being pro- also demonstrated that only microorganisms are

© Springer International Publishing AG 2018 483


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_13
484 13  Beer and Wine

capable of converting sugars into alcohol from Rum played a crucial part of the triangular
grape juice, and that the process occurs in the trade between Britain, Africa, and the North
absence of oxygen (Barnett 2000; Pasteur 1876). American colonies that once dominated the
Today, beer brewing and wine making are Atlantic economy.
agricultural industries which evolved from
ancient and empirical knowledge which emerged
from many different cultures around the world. Alcoholic Fermentation
The ancient art of fermentation has been com-
bined with basic scientific knowledge and applied Ethanol contained in alcoholic beverages is pro-
toward modern production processes (Hornsey duced by yeast fermentation. Wine is produced
2003). by fermentation of the natural sugars present in
Fermentation has been used to preserve and grapes; cider and perry are produced by similar
produce food and beverages since the Neolithic fermentation of natural sugar in apples and pears,
age. Lactic acid fermentation has been used to respectively; and other fruit wines are produced
preserve many foods including cheeses, yogurt, from the fermentation of the sugars in any other
pickles and kimchi as well as for producing alco- kinds of fruit. Mead is produced by fermentation
holic beverages such as beer and wine and beer. of the natural sugars present in honey. Beer,
Wine was consumed in Classical Greece at whiskey, and vodka are produced by fermenta-
breakfast or at symposia, and in the 1st century tion of grain starches that have been converted to
BC it was part of the diet of most Roman citizens. sugar by the enzyme amylase, which is present in
Both the Greeks and the Romans generally drank grain kernels that have been malted. Other
diluted wine (the strength varying from one part sources of starch (e.g. potatoes and unmalted
wine and one part water, to one part wine and grain) may be added to the mixture where amy-
four parts water). lase is present to hydrolyze the starches to sugars.
In Europe during the Middle Ages, beer, often Whiskey and vodka are also distilled; gin and
of very low strength, was an everyday drink for related beverages are produced by the addition of
all classes and ages of people. A document from flavoring agents to a vodka-like feedstock during
that time mentions nuns having an allowance of distillation.
six pints of ale each day. Cider and pomace wine Rice wines (including sake) are produced by
were also widely available; grape wine was the the fermentation of grain starches converted to
prerogative of the higher classes. sugar by the mold Aspergillus oryzae. Baijiu,
Hard liquor, particularly brandy and rum, soju, and shōchū are distilled from the product of
were provided for sailors during the long sea voy- the fermentation. Rum and some other beverages
ages of the Age of Exploration, when European are produced by fermentation and distillation of
powers plied the seas during the fifteenth, six- sugarcane. Rum is usually produced from the
teenth, and early seventeenth centuries. Great sugarcane product molasses.
Britain’s longtime superiority at sea may also Fermentation is the metabolic process that
owe a debt to its navy’s drink of rum-based converts sugar to acids, gases or alcohol. The
choice, grog, which was made a compulsory bev- simplest representation summarizing the fermen-
erage for sailors in the late eighteenth century. tation process is shown in Fig. 13.1.

Fig. 13.1  Glucose is fermented to pyruvic acid and then converted to ethanol and carbon dioxide
Alcoholic Fermentation 485

The fermentation by yeast takes place in an Glycolysis occurs in cytosol of the cell in
anaerobic environment (when the electron trans- most organisms. The most common type of
port chain is unusable) and becomes the cell’s glycolysis is the Embden–Meyerhof–Parnas
­
primary means of ATP (energy) production. The (EMP pathway).
process converts the pyruvate produced by gly- Pyruvate is metabolized in processes like the
colysis and NADH and pyruvate to yield ethanol. Krebs cycle to produce energy under aerobic
In a process called oxidative phosphorylation conditions. The products of this type of metabo-
NADH and pyruvate are used to generate ATP. lism are ATP, H2O, and CO2. When abundant
The overall process is called the Embden– supplies of sugars present under anaerobic condi-
Meyerhoff–Parnas pathway which describes the tions which occur in bread fermentation, beer
conversion of glycose or xylose for energy pro- wort and wine production yeast undergo alco-
duction and production of ethanol and CO2 as holic fermentation. This type of metabolism
illustrated in Fig. 13.2. yields much smaller amounts of energy com-
Glycolysis is the first step which converts glu- pared to aerobic respiration.
cose to pyruvate and results production of 2 The alcoholic fermentation utilizes the two
NADH and 2 ATP molecules in the process. pyruvates produced from glycolysis of glucose.
C6H12O6 + 2 NAD+ + 2 ADP + 2 Pi  → 2 The pyruvate molecules are decarboxylated by
CH3COCOO− + 2 NADH + 2 ATP + 2 H2O + 2 pyruvate decarboxylase to form acetaldehydes
(Pyruvate) and CO2. At the active sites on pyruvate
Glycolysis is a sequence of ten enzyme-cata- decarboxylase pyruvate reacts with cofactors
­
lyzed reactions. Intermediates enter the pathway thiamine pyrophosphate (TPP) and magnesium
at various points in the pathway. For example, to release carbon dioxide. The final step to form
most monosaccharides, such as fructose and ethanol is the addition of a hydrogen ion to the
galactose, can be converted to one of the glycoly- acetaldehyde. The hydrogen ion comes from the
sis intermediates. Glycolysis is an oxygen inde- NADH produced during glycolysis. The ethanol
pendent metabolic pathway that does not use in beer and wine production prevents the growth
molecular oxygen for any of its reactions. of other microbes.

Fig. 13.2  Embden–Meyerhoff–Parnas pathway for production of pyruvate which enters the TCA cycle and ethanol
486 13  Beer and Wine

Saccharomyces cerevisiae is the yeast gener- Malting commences with steeping of barley in
ally used in the beer and wine making process. water at 14–18 °C for up to 48 h, until it reaches
The strains of yeast used in the beer making pro- a moisture content of 42–46%. This is usually
cess ferments the different types of sugars found achieved in a three-stage process, with the steeps
in the wort, pre-fermented beer, to produce etha- being interspersed with ‘air rests’ that allow the
nol. Saccharomyces cerevisiae has been has been barley to get some oxygen (to ‘breathe’).
in use for thousands of years. Beer technology Raising the moisture content allows the grain to
was utilized by from Germanic and Celtic tribes germinate, a process that usually takes 3–5 days at
around the first century AD. The oldest known 16–20 °C. In germination, the enzymes break
example of yeast used in fermentation of a bever- down the cell walls and some of the protein in the
age was found in China around 7000 BC. starchy endosperm, which is the grain’s food
Saccharomyces cerevisiae can be found all reserve, rendering the grain friable. Amylases are
around the world on the surface of fruits and produced in germination and these are important
plants, in the soil, the gastrointestinal tract of ani- for the mashing process in the brewery.
mals, and the skin surface of animals. Progressively increasing the temperature dur-
ing kilning arrests germination, and regimes with
progressively increasing temperatures over the
Beer range 50 to perhaps 110 °C are used to allow dry-
ing to <5% moisture, whilst preserving heat-­
Beer is produce by fermentation of malted barley sensitive enzymes. The more intense the kilning
(with or without other added starch or sugar process, the darker the malt and the more roasted
sources), water hops yeast and water. Additions and burnt are its flavor characteristics.
to the malt, called adjuncts are barley, wheat, rice Brewing is typically divided into seven steps:
or corn starches, their hydrolysis products result- Mashing, Lautering, Boiling, Fermenting,
ing in fermentable sugars. Additional enzymes Conditioning, Filtering, and Packaging.
from microbial sources can be added to enhance Mashing is the process of mixing malted and
carbohydrate hydrolysis or to clarify beer. He milled grain with water, and heating this mixture
aroma and flavor of beer comes from the bitter up with holding or rests at certain temperatures to
taste of hops, kiln dried products from the barley allow enzymes in the malt to break down the
malt and aroma compounds resulting from the starch in the grain into sugars, typically maltose.
fermentation. The fermentation process is based The malt is crushed in a “malt mill” to reduce
on the Embden–Meyerhoff–Parnas pathway for the particle size of the grain kernels, increase
glycolysis and alcoholic fermentation. their surface area, and separate the fermentable
The production of beer includes the malting of portions from the husks. The resulting grist is
barley, the preparation of wort fermentation and mixed with heated water in a vat called a “brew-
delivery. The process is outlined in Fig. 13.3. ing kettle” for a process known as “mashing”.
All beers are brewed using a process based the During this process, natural enzymes within the
fermentation of malted grain. Many types of malt break down much of the starch into sugars
beers are produced resulting from variation in which play a vital part in the fermentation pro-
starting materials and regional preferences. cess. Mashing usually takes 1–2 h, and during
Barley, wheat or sometimes rye are the preferred this time various temperature profiles activate
starting materials. different enzymes depending upon the type of
Malt is produced by germinating the grain to malt being used. Hydrolysis of the starchy raw
breakdown starches to fermentable sugars, after material by the amylase enzymes convert the
which the grain is dried in a kiln and sometimes starches into fermentable sugars such as glucose
roasted. The germination process activates sev- and maltose.
eral enzymes, particularly a-amylase and Lautering is the separation of the fermentable
b-­
amylase, which hydrolyze the starch in the extracts from the grain and removing the spent
grain into sugar. grain. The process is conducted in either a lauter
Beer 487

malting milling
water steeping germination kilning

barley roller mill

cooling mashing grist


brewing

trub “spent”
grain water
hops

wort

plate heat exchanger whirlpool kettle lauter tun mash mixer


separator
fermentation maturation filtration flash pasteurization packaging

yeast excess yeast

green beer

maturation tank filtration unit plate heat exchanger keg


fermentation yessel
filling and tunnel pasteurization finished product
crowning

© 2009 Enoyclopaedia Britannica, Inc. bottles conveyor belt labeled bottles cans

Fig. 13.3  Production of beer

tun, a wide vessel with a false bottom, or a mash non-malted grains such as corn and rice, which
filter, a plate-and-frame filter designed the sepa- are widely used as adjuncts in North American
ration. Lautering has two stages: first wort run-­ beers. Finally, a mash rest temperature of 149–
off, during which the extract is separated in an 160 °F (65–71 °C) is used to convert the starches
undiluted state from the spent grains, and sparg- in the malt to sugar. Finally the mash temperature
ing, in which extract which remains with the may be raised to 165–170 °F (about 75 °C)
grains is rinsed off with hot water. A mash rest (known as a mashout) to deactivate enzymes.
temperature of 104 °F or 40 °C activates beta-­ After the mashing, the mash is pumped to a
glucanase, which breaks down gummy beta-­ lauter tun where the resulting liquid is strained
glucans in the mash, reducing the viscosity of the from the grains in a process known as lautering.
mash. A mash rest from 120 to 130 °F (49 °C to The lauter tun generally contains a slotted “false
55 °C) activates various proteinases, which bottom” or other form of manifold which acts as
hydrolyze proteins reducing the chance of haze in a strainer allowing for the separation of the liquid
the beer. This rest is generally used only with from the grain. A lauter tun is the traditional ves-
undermodified (i.e. undermalted) malts which are sel used for separation of the extracted wort.
popular in Germany and the Czech Republic, or While the basic principle of its operation has
488 13  Beer and Wine

remained the same since its first use, technologi- After the hops are removed by filtration the
cal advances have led to better designed lauter wort is cooled to fermentation temperatures
tuns capable of quicker and more complete before yeast is added. The wort is pumped into
extraction of the sugars from the grain. the heat exchanger, and goes through every other
The false bottom in a lauter tun has thin slits to gap between the plates. The cooling medium,
hold back the solids and allow liquids to pass usually water, goes through the other gaps. The
through. The solids, not the false bottom, form a ridges in the plates ensure turbulent flow. A good
filtration medium and hold back small solids, heat exchanger can drop 95 °C wort to 20 °C
allowing the otherwise cloudy mash to run out of while warming the cooling medium from about
the lauter tun as a clear liquid. The false bottom 10 °C to 80 °C. After cooling the yeast is added
of a lauter tun is today made of wedge wire, for fermentation.
which can provide a free-flow surface in the bot- Fermentation, as a step in the brewing pro-
tom of the tun. cess, starts as soon as yeast is added to the cooled
Sparge water is sprayed on top of the spent wort. The yeast fermentation converts the sugars
grains in the tun to help extract remaining fer- in the wort to alcohol and carbon dioxide.
mentable sugars. http://www.sterkensbrew.be/ Fermentation tanks come in all sorts of forms,
sbm/beer from enormous tanks which can look like storage
A mash filter is a plate-and-frame filter. The silos, to five gallon glass carboys in a homebrew-
empty frames contain the mash, including the er’s closet.
spent grains, and have a capacity of around one When the sugars in the fermenting beer have
hectoliter. The plates contain a support structure been almost completely digested, the fermenta-
for the filter cloth The plates, frames, and filter tion slows down and the yeast starts to settle to
cloths are arranged in a carrier frame like so: the bottom of the tank. The beer is cooled to
frame, cloth, plate, cloth, with plates at each end almost freezing, which encourages settling of the
of the structure. Newer mash filters have bladders yeast, and causes proteins to coagulate and settle
that can press the liquid out of the grains between out with the yeast. Phenolic compounds which
spargings. The grain does not act like a filtration can cause unpleasant flavors become insoluble in
medium in a mash filter. the cold beer and their removal results in a
At this point the liquid is known as wort. The smoother flavor for the beer.
wort is moved into a large tank known as a “cook- Some beers undergo a fermentation in the bot-
ing tun” or kettle where it is boiled with hops and tle, giving natural carbonation. This may be a
sometimes other ingredients such as herbs or sug- second or third fermentation. They are bottled
ars. The boiling process serves to terminate enzy- with a viable yeast population in suspension. If
matic processes, precipitate proteins, isomerize there is no residual fermentable sugar left, sugar
hop resins, concentrate and sterilize the wort. may be added. The resulting fermentation
Hops add flavor, aroma and bitterness to the beer. ­generates CO2 which is trapped in the bottle,
At the end of the boil, the hopped wort settles remaining in solution and providing natural car-
to clarify using hop filters. SBM does not use the bonation. http://www.sterkensbrew.be/sbm/beer_
whirlpool system for hop separation. http://www.
sterkensbrew.be/sbm/beer_
Boiling the wort, ensures its sterility, and thus Raw Materials
prevents the growth of undesired microorgan-
isms. During the boil hops are added, which con- Barley is the most central ingredient for beer
tribute bitterness, flavor, and aroma compounds brewing. Different barley strains impart unique
to the beer. The heat of the boil, causes proteins characteristic taste and body in different beers.
in the wort to coagulate and the pH of the wort to Malted barley is barley that has been allowed to
drop slightly. germinate (sprout) to a degree and is then dried.
Hops 489

This is accomplished industrially by increasing Hops are the cone-like flowers of the female
the water content of the seed to 40–45% by soak- hop vine (Humulus Lupulus). At the base of hop
ing it for a period close to 40 h. The seed is then flower, there is a soft resin called Lupulin Oil
drained and held at a constant temperature (60 °F) which contains the compounds that deliver the
for close to 5 days until it starts to sprout. The bitter flavor and “hoppy” aroma. Hops are grown
barley is slowly dried in a kiln at temperatures in Germany, southern England, southern
gradually rising to 122 °F for lighter malts and Australia, Tasmania, Oregon and Washington
220 °F for darker malts. This kiln drying takes State. The 50 plus varieties of hops deliver unique
about 30 h. Finally, the rootlets from the partially bitterness in flavor or aroma.
germinated seeds are removed. Buttery and Ling (1966) identified over 100
The germination process converts starch into hop oil components. Among the 110 components
simpler sugars used by the plant in its initial in beer, Tressl et al. (1978) was able to identify
growing stage. The conversion of starch to sugar 47 that were derived from hops. Figure 13.4 illus-
is accomplished by amylase enzymes that the trates the hop oil constituents as defined by
seed produces during germination process. The Sharpe and Laws 1981, where 50–80% of the hop
germination and drying stages capture ferment- oil is comprised of hydrocarbons.
able sugars, soluble starch, and the diastase Steinhaus and Schieberle (2000) found 23
enzymes for beer brewing. Malted barley is the potent aroma compounds in the hop variety
primary source of the fermentable sugar con- Spalter Select. The compounds were subse-
sumed by the yeast. However many beers add quently categorized by flavor dilution factors.
other carbohydrate sources called adjuncts, such The most potent aroma constituents were trans-­
as corn, wheat or rice. 4,5-epoxy-(E)-2-decenal, (which confers a
metallic note), linalool (flowery) and myrcene
(geranium like). Kishimoto et al. (2006) discov-
Hops ered 4-mercapto-4-methylpentan-2-one (4MMP)
added a fruity aroma in American, Australian and
Hops may be the most important ingredient in New Zealand hops cultivars.
determining the flavor beer. The current revival Steinhaus and Schieberle (2000) utilized aroma
craft beer many of which a strong hops flavor as extract dilution analysis (AEDA) to assess the
resulted in more studies of hop chemistry. contributions of various compounds to hoppy

Fig. 13.4  Groups of hop oil constituents (Sharpe and Laws 1981)
490 13  Beer and Wine

aroma in beer. They observed that only a few of Hanke et al. (2010) studied the effect on
the volatiles in hops contributed to the aroma of threshold values of binary mixtures of hop aroma
the beer. Linalool was confirmed in various works compounds.
to significantly contribute to the hoppy aroma in They reported that a mixture of caryophyllene
beer (Sakuma et al. 1991). Takoi et al. (2010) and nerol had a flavor threshold of 170 μg/L
found that linalool acts synergistically with gera- whereas the individual compounds had thresholds
niol and citronellol. They also discovered that the of 210 mg/L and 1200 μg/L, respectively. Addition
presence of β-citronellol (lime aroma) depends on of a combination of linalool and farnesene resulted
geraniol metabolism by the yeast. Kishimoto et al. in a combined threshold of 500 μ/L, however,
(2006) identified 19 hop derived components in farnesene alone had threshold of about 2000 μg/L.
hopped beers using GC olfactometry. Within The contribution of hops to the final aroma of
these, 4-­mercapto-4-methylpentan-2-one (4MMP, beer is most noticeable when late-hopping or
blackcurrant-like) and 3-mercaptohexan-1-ol dry-hopping is applied. Late-hoping is the
(muscat-, blackcurrant-like) proved to contribute ­addition at the end of boil. This approach reduces
to the hoppy aroma. Nielsen 43 was also able to the exposure of the bee with hops to heat which
identify important contributors to the aroma from causes unnecessary evaporation of many valu-
hops compounds (Table 13.1). able aroma compounds.
Schönberger and Kostelecky (2011) summa- Dry-hopping is mainly applied during the
rized the contribution of hops to the aroma of lagering step. Addition at this stage is based on
beer: cold extraction of hop material into an alcoholic
solution.
• Green and grassy flavors can be attributed to The hop oils are water soluble and are highly
aldehydes, e.g., hexanal volatile. The hops are added to the wort at the end
• Citrus flavours can be attributed to esters, of boiling to minimize the loss of volatile compo-
nerol and linalool nents. The choice of hops alters the composition
• Floral and fruity flavors can be attributed to of the hop oils which account for the bitter flavor
linalool, geraniol, β-ionone, citronellol, to and hop taste of beer.
-mercapto-4-methylpentan-2-one and Hop oil contains (0.5–3% volatile compo-
3-­mercaptohexan-1-ol and other ketones, nents, and 3–6% non-volatiles) which are present
epoxides and Esters (Marriott 2001) in the hop polyphenolic fraction. In addition to
• Herbal flavors can be attributed to oxidized the aroma and flavor the polyphenolic fraction
sesquiterpenes (Kowaka et al. 1983). also contributes to the mouthfeel of the beer.

Table 13.1  Identified aroma compounds in strong hopped beer by Kishimoto et al. (2009), Nielsen (2006), and
Lermusieau and Collin (2003)
Sensory character compound Compound Threshold in beer
Black currant 4-Mercapto-4-methylpentan-2-one 10–50 ppb
Black currant 3-Mercaptohexan-1-ol 55 ppb
Resinous Myrcene 30–1000 ppm
Floral, citrus Linalool 8–80 ppm
Floral, rose like Geraniol 4–40 ppm
Ethyl 2-methyl-butanoate 1 ppm
Ethyl 4-methyl-pentanoate
Fruity, herbal cis-Rose oxide 5–50 ppb
Hoppy Pineapple (e,z)1,3,5-Undecatriene
Cheesy 2-Methylbutyric acid
Grape tobacco, black tea Beta damascenone/phenyl ethyl alcohol
Cedarwood Caryophylla-3,8-dien-(13)-dien-5-beta-ol
Cheesy/onion/ garlic Various sulfur compounds
Hops 491

The complex mixtures of hop components in beer the bitter components have been carefully studied
are present in very low but they are very important and are well understood because there are only a
to the flavor, aroma and mouth feel of the final few precursors are present in hops (Verzele and
product. In addition to the complex nature of the De Keukeleire 1991; Benitez et al. 1997). The
hops going into the beer many volatile materials most important hop compounds are the hop acids,
are lost during processing and other components which are defined as alpha-acids or humulones
are oxidized. The complex composition of the hop and beta-acids or lupulones (Fig. 13.5). The two
oils becomes even more complex during wort boil- groups of compounds each comprise three con-
ing. Many brewers conserve part of the aroma stituents which differ in the side chains. The three
compounds in hops by adding the hops near the different side chains are derived from the
end of the wort boiling step. Hops also can account branched chain amino acids, leucine, valine and
for up to one third of the total polyphenols in beer. isoleucine. These hop acids can constitute up to
The low-molecular-weight proanthocyanidins 25% of the dry weight of the hop cones. The pro-
determine the colloidal stability of beer. Boiling portions of the individual compounds very
results in major changes in the complex polyphe- greatly among hop varieties variety and, and are
nol composition of wort (Forster et al. 1995). also influenced by growing conditions. The hop
The barley and hops contribute complex mix- acids occur as pale yellowish solids in the pure
tures of polyphenolics which are further altered by state, are weak acids, exhibit very poor solubility
oxidation during processing. Hop polyphenols are in water and have almost no bitter taste.
found as monomers, dimers, trimers. In beer they The most abundant component in alpha-acids
are also found bound to protein. The polyphenols is humulone. Depending on the hop variety the
combine slowly with proteins to form chill haze relative amounts of humulone range from 20 to
when the beer is cooled after brewing, but the haze 50%, adhumulone is a consistent 15% of the mix-
re-dissolves when the beer is warmed up. Upon ture in most hops. Cohumulone is often associ-
aging the polyphenols polymerize and grow larger, ated with a poor hop quality (Pollach et al. 1996).
becoming insoluble at room temperature resulting The beta-acids are very sensitive to oxidation
in an irreversible haze (De Keukeleire 2000). and most products result in off flavors. In order to
One of the primary contributions of hops in avoid the problem many brewers select hop vari-
beer is the bitter flavor. Bitter profiles in beer and eties with low concentrations of beta-acids.

Fig. 13.5  Alpha- and beta acids form hops


492 13  Beer and Wine

Humulone is the most abundant alpha-acid in The concentrations vary widely, from 15 ppm in
most hops. The relative amounts of humulone typical American lager beers to nearly 100 ppm
and cohumulone vary widely in different variet- in bitter English ales. The perceived taste of beer
ies of hops. Adhumulone constitutes invariably is also influenced complexes formed with the
approximately 15% of the mixture. humalones and the sugars in the beer.
During the boiling of wort, the humalones To standardize discussions of beer flavors an
form the hops undergo major chemical changes. International Beer Flavor Terminology was
The most significant change is the thermal developed by the American Society of Brewing
­isomerization of the alpha-acids or humulones to Chemists in collaboration with the European
the iso-alpha-acids or isohumcontraction Brewery Convention and the Master Brewers
(Fig. 13.6). Each humulone is converted to two Association of the Americas. These organiza-
epimeric forms (cis-isohumulones and trans-­ tions felt that by providing brewers, researchers,
isohumulones). The difference is based on the judges, and marketing people with a simple and
spatial arrangement of the tertiary alcohol func- easily understandable terminology system, com-
tion at and the prenyl side chain at. The trans and munication among beer industry professionals
cis designations refer to the fact that indicate that would improve.
these groups point in opposite faces and to the In Fig. 13.7 the descriptive terms are arranged
same face of the five-membered ring, respec- in three tiers. The center circle allows you to clas-
tively. Six major iso-alpha-acids (cis-­ sify the terms as they relate to odor and taste. You
isohumulone and trans-isohumulone, start with the odor and determine which term best
cis-isocohumulone and trans-isocohumulone, describes the first smell you experience. Then
cis-isoadhumulone and trans-isoadhumulone) move on to the next level which contains com-
are present in beer as a result of the thermal mon descriptive terms that allow you to expand
isomerization of the three major alpha-acids, on your initial reaction. The third tier makes it
humulone, cohumulone and adhumulone, possible for you to pinpoint the flavor more spe-
respectively. The ratio of the isohumulones cifically. If you have decided that the odor is ini-
depends on the reaction conditions. In wort, the tially fruity, then you might also be able to specify
cis-/trans- ratio is approximately 68:32. The cis- whether it is more apple-like, banana-like, or
isomers are more stable with a half-life of 5 both. You can then move on to secondary odors or
years than the trans-isomers which have a half- work on the tastes.
life of about 1 year. During storage of beer, the There a wide range of beers and each brewery
changes in the cis:trans ratio results in changes and beer has unique character. Table 13.2 sum-
in the in taste of the beer. Thus, during process- marizes the general characteristics of some
ing it is desirable to maximize the levels of cis- American beer types. Beers range from very light
isohumulones. The iso-alpha-acids constitute in color and body to very heavy stouts and brown
the are the most significant fraction of hops in ales. There is also a wide range of bitterness con-
beer and account for the bitter taste of beer. tributed by the hops.

Fig. 13.6  Humalone structures from hops


Yeast 493

Fig. 13.7  The ASBC Beer Flavor Wheel design combines the original flavor wheel developed by M. C. Meilgaard,
C. E. Dalgliesh, and J. F. Clapperton in 1979 with the updated, detailed terminology provided by experts

strains of yeast. Carefully cultivated strains of


Yeast yeast are used in the brewing of beer. Selection of
the most desirable yeast for the specific beer is
Yeast is responsible for the converting of sugar to critical. The two types of yeast used for beer
alcohol and carbon dioxide in the fermentation brewing are top-fermenting yeast (Saccharomyces
stage. It also contributes to the fermentation of cerevisiae) and bottom-fermenting yeast
the beer. There are thousands of varieties and (Saccharomyces uvarum). The top-fermenting
Table 13.2  Characteristics of some American type beers
494

American beer type Description Alcohol by weight Bitterness SRM Color


Lager Light fruity-ester aroma, low hop aroma and low malt sweetness 3.20–4.00 4 10 1.5–4
Hop bitterness is not perceived to very low
Corn, rice, or other grain or sugar adjuncts often used
Body is light
Light lager Pale to medium-amber. “light” refers to relatively low body and reduced calories 2.8–3.5 4–10 1.5–4.0
Hop aroma is absent or low. Malt sweetness is very low but evident
Hop flavor is absent or very low. Hop bitterness is very low to low. Corn, rice, or other grain or sugar
adjuncts or all-malt formulations are also made. High in carbonation
Pilsner Medium-low to medium malt aroma is present. Hop aroma is medium to high. Up to 25% corn and/or 3.8–5.0 7–20 2–8
rice in the grist should be used. Medium-low to medium malt flavor. Hop flavor is medium to high. Hop
bitterness is medium to high
Body is light-medium to medium
Malt liquor Fruity-ester and complex alcohol aromas at low levels. Hop aroma is not perceived. Some residual 5.0–6.0 18–30 6–14
sweetness is perceived. Hop bitterness is very low. High alcoholic strength. Some malt liquors are just
slightly stronger than American lagers, while others approach bock strength. Fruity-ester and complex
alcohol at low levels
Dark lager Dark Lagers are light brown to very dark. Low malt aroma contains contributions from caramel and 3.2–4.4 14–24 14–25
roasted malts
Hop aroma is very low to low. Low malt flavor contains discreet contributions from caramel and roasted
malts. Non-malt adjuncts are often used. Hop flavor is very low to low. Hop bitterness is very low to
low
Golden or Blonde Golden or Blonde Ales are straw to light amber. Hop aroma is low to medium-­low. Light malt 3.2–4.0 30–45 3–7
Ales sweetness is present. Hop flavor is low to medium-low. Hop bitterness is low to medium. Fruity esters
may be perceived
Pale Ale Pale Ales are deep golden to copper or light brown. Low caramel malt aroma. Moderate fruity-ester 3.5–4.3 30–50 6–14
aroma Hop aroma is medium to medium-high, exhibiting floral, fruity, sulfur/diesel-like, citrus-like,
piney resinous characters. Hop flavor is medium to medium-high
Hop bitterness is medium to medium-high. Fruity-­ester flavor is moderate to strong
India Pale Ales IPAs are gold to copper or red/light brown 5.0–6.0 50–70 6–15
Fruity-ester aroma is moderate to very high. Hop aroma is high
Exhibits floral, fruity, sulfur/diesel-like, citrus-like, piney, resinous characters. Medium maltiness is
present. Hop flavor is high
Brown Ales Brown Ales are deep copper to very dark brown. Fruity-ester aromas should be subdued. Roasted malt 5.0–6.0 50–70 25–45
caramel-like and chocolate-like aromas are medium. Hop aroma is low to medium. Roasted malt caramel-like
and chocolate-like flavors should be medium. Hop flavor is low to medium. Hop bitterness is medium to high
Stout Stouts are black. Head retention is excellent. Fruity-ester aroma is low. Coffee-like roasted barley and 4.5–7.0 35–60 40+
roasted malt aromas are prominent. Hop aroma is medium to high. Low to medium malt sweetness with
low to medium caramel, chocolate, and/or roasted coffee flavor. Roasted barley and roasted malt
13  Beer and Wine

contribution to astringency is low. Hop bitterness is medium to high. Fruity-ester flavor is low
Wine 495

Mitochondrion
residues in pottery provides chemical evidence of
ancient wine making (McGovern et al. 1997,
Bud vacuole
2004; Jackson 2008). Wine production first began
Secretory
vesicles in what is now Iran and spread throughout the
Bud
Fertile Crescent. The technology was further
Nucleus
Golgi complex developed by the ancient Mesopotamian,
Pore in nuclear Egyptian, Greek, and Roman civilizations. Once
membrane
the basic technology was established there was
Vacuole
Endoplasmic reticulum limited progress in wine processing. With the
Vacuolar membrance advent of modern chemistry by Dalton and the
subsequent investigations of Pasteur brought
Lipid granule
wine chemistry forward as a modern discipline.
Bud scar
Cell membrane
The establishment of the role of microorganisms
Cell wall
in fermentation in the late 1800s led to an
Vacuolar
improved understanding of the chemistry pio-
granules neered by Pasteur. The focus shifted to analysis
Storage granule of the major components that effect wine quality,
Thread-like mitochondrion
including ethanol, sugars, and organic acids.
Methods of analysis for ethanol and the other
Fig. 13.8  Main components of a typical yeast cell components, including some aroma compounds
were described by Mulder in 1857. This was one
yeast is similar to the yeast for baking bread. It is of the earliest treatises to focus on wine chemis-
applied for making ales and stouts. The bottom-­ try as opposed to wine production (Ebeler and
fermenting yeast is used in the production of Thorngate 2009).
lagers and steam beer. Wine is produced in many places around the
Most top fermenting yeast have tendency to world. Table 13.3 contains the 2014 wine produc-
flocculate on the surface of the beer during the tion form some of the largest wine producing
first few days of fermentation. Some of this yeast countries and the per capita consumption of some
settles to the bottom of the fermenter, however of the largest uses. It is interesting to note that the
the largest portion of the yeast remain dispersed highest per capita wine consumption is the
in the fermentation. Top Fermenting yeast, used Vatican City State (54.26 L/year), Andorra
in ale production, grows best in the temperature (46.26 L/year) and Croatia (44.2 L/year). There
range of 55–75 °F.
Bottom fermenting yeast, used in lager pro- Table 13.3  Leading wine producing countries and wine
duction performs better in the temperature range consumption
55–32 °F. The fermentation is much slower using Per capita
lager yeast, this time is often referred to as Production consumption
Country in liters (L/year)
Lagering (Fig. 13.8).
France 4,670,100 42.66
Italy 4,473,900 33.30
Spain 3,820,400 21.26
Wine United States 3,021,400 10.25
Argentina 1,519,700 24.46
Archeological evidence has traced winemaking Australia 1,200,000 24.53
to the Neolithic period (8500–4000 BC) in the South Africa 1,131,600  7.38
Trans-Caucasus and ancient Near East (Michel China 1,117,800  1.18
et al. 1993). Tartaric acid is an excellent marker Chile 1,050,000 17.46
for grape wine because few other fruits produce http://www.wineinstitute.org/files/World_Per_Capita_
tartaric acid in significant quantities. Measuring Wine_Consumption_Revised_Nov_2015.pdf
496 13  Beer and Wine

are hundreds of commercial varieties of wine older vines. If grapes were planted from seed,
grapes. In Tables 13.4 and 13.5 the flavor/aroma they would no longer be genetically identical to
characteristics and the regions of production for their parent, and would represent a different spe-
major white and red wines are summarized. cies. The only way to make more of a desired
There are many wine varieties cultivated but the grape variety (Riesling) is to clonally propagate it,
Vitis vinifera, L.ssp. is the most prevalent for wine and grafting is a common approach.
production. There are over 800 cultivars of Vitis During the second half of the nineteenth cen-
vinifera in production world. There is a wide varia- tury phylloxera, a devastating louse was brought
tion in cluster size, size, shape, sugar content and into Europe from American vines. Rootstocks
color of the grapes. Grapes are either table grapes were developed from North American species,
for fresh consumption or wine grapes which can be including V. riparia, V. rupestris, and V. berland-
used for Red, rose or white wine production. ieri which result in more robust vines. Most com-
mon are Vitis riparia based rootstocks.
Grapevines are trained to grow along a trellis
Wine Grape Production supports it which enables the grower to deter-
mine the amount and positioning of both leaves
In order to maintain identity of grape varieties and fruit. Newly planted grapevines require 4–5
new grape vines are produced from cuttings of years before reaching full productivity. Grape
existing varietal vines which can be started vines can continue producing viable crops for
directly in soil or they can be grafted onto existing 50–100 years.

Table 13.4  Regions of production and tastes of some white wines


Variety Regions of production Flavor descriptors
Chardonnay Chardonnay is the principle white wine of Chardonnay wines are often full-bodied (and
Burgundy (Bourgogne, France), where it more velvety) than other white wines. Notes
originated. Areas of production include: include raspberry, vanilla, tropical fruit,
California, Oregon, Washington, France, butterscotch peaches, tea tomato, tobacco with
Australia, Italy, South Africa, Chili, rich citrus (lemon, grapefruit) flavors. Frequently
Argentina aged in oak which results in buttery notes
Sauvignon blanc Sauvignon blanc is grown in the Bordeaux Piercing aroma, fruity, grassy, herbal, green fruit,
region where it is blended with Semillon. musky
The Loire valley and New Zealand
produce some excellent sauvignon blanc
varietals
Moscato/Muscat Grows in most vine-friendly climates, Orange blossom rose petal, peach, apricot
including Italy, the Rhône Valley (where it
is called muscat blanc à petits grains) and
Austria (where it is called Muskateller)
Also produced in California, Australia,
and South America
Pinot grigio Italy, France Pinot grigio is also grown in Italy it is light bodied, with pear, citrus and
Pino Gris the western coastal regions of the mineral flavors French Alsace are richer rounder
USA. Loire Valley and Pinot gris in the wines with spicy, honey, peach and pear flavor
rest of France. In Germany and Austria Oregon Alsace Pinot Gris providing aromatic,
Pinot grigio is known as the Ruländer or fruity flavors
Grauer Burgunder
Gewürztraminer Gewürztraminer is from Alsace, Germany, Typical taste is fruity flavors with floral aromas
the US West Coast, and New York. Also of rose petals, peaches, lychees, and allspice
New Zealand
Riesling Riesling is the classic German grape of the Riesling wines are much lighter than Chardonnay
Rhine and Mosel. Alsace and the Eastern wines. Green apple, citrus and peach flavors
US is also excellent but typically drier.
Does well in New Zealand and Australia
Wine 497

Table 13.5  Regions of production and tastes of some red wines


Variety Regions of production Taste descriptors
Petite Sirah Syrah excels in California, Australia, Very dark, big tannins, plum, blackberry fruit flavors
Argentina and Chile Burgandy, Oregon
and New Zealand
Merlot A major variety in Bordeaux blends, merlot Typical scents include black cherry, plums and
is now grown in Italy, Romania, California, jammy flavors. When aged on oak secondary flavors
Washington State, Chile, Australia of vanilla, leather tobacco and earthy
Cabernet Bordeayx left, Margaux, Pauillac, St.-Julien In Bordeaux the wine it is full bodied with black
Sauvignon and Graves current high acidity and tannins
It is part of the great red Médoc wines of In warmer climates exhibit black cherry, blueberry,
France, and among the finest reds in menthol and medium body
Australia, California and Chile
Malbec Malbec has its origins in the French Malbec’s characteristics vary greatly depending on
Bordeaux region. Now the signature grape where it is grown and how it is transformed. Ripe
for Argentina vineyards near Mendoza tannins and black fruit flavors Malbec is often
blended with other varieties such as Cabernet Franc,
Cabernet Sauvignon, Merlot
Pinot noir Produces very high quality red wines of Sour cherry and strawberry spicy with medium to low
Burgundy, and good wines from Austria, acidity and light tannins
California, Oregon, and New Zealand
Zinfandel Primarily found in California, Zinfandel Black fruit and spice flavors
originates from Italy (where it is called
primitivo)
Barbera A classic red of Italian origin. Widespread Juicy black cherry and plum fruit, a silky texture and
in California excellent acidity

Sugar, pH and acid levels are the major fac- replace the chlorophyll resulting in the color
tors for determining when grapes are ready for changes in the berries. Malic acid is consumed as
harvesting, however, tannins in the grapes also the primary metabolic substrate. Tartaric acid
need to reach a certain level of maturity. stays unchanged as a result, tartaric acid, becomes
Determining the optimum time of harvest is the predominant acid in the grape berries Liang
a critical viticultural decision. It is difficult et al. (2011). Concomitant with the decrease
to assess grape maturity in the vineyard and in acidity hexose sugars accumulate in the
therefore predict the ultimate wine quality. berries. Sucrose produced by photosynthesis
In vineyards now, ripeness evaluation includes is transferred from the leaves to the berries
more than an analysis of Brix (sugar content), where it is hydrolyzed to glucose and fructose.
titratable acidity and pH. Winemakers use flavor/ Physiologically, the sugar concentration increases
aroma assessment in addition to the routine to levels above 18% occasionally reaching 25%.
standards. Most grapes are harvested at between 18 and
In viticulture the onset of ripening is called 25% sugar content. The greater the concentration
veraison. Veraison represents the transition from of sugars in the grape juice, the greater the poten-
berry growth to berry ripening, and many changes tial for alcohol production. Grapes for sweet
occur at veraison. Prior to veraison the grapes are wines, such as dessert wines are allowed to ripen
hard and green with low sugar levels and very longer, producing more sugar than will be
high acid levels (primarily malic acids). Veraison ­fermented leaving a sweet wine.
lasts a week or less. During this period the grapes Also: titratable acidity also decreases because
go through several changes which impact their (i) the berry expands, approximately doubling in
sugar, acid, tannin and mineral composition. The size, and (ii) there is exchange of H+ for K+ and
concentration of phenolic compounds in the skin, other cations by the berry Veraison occurs from
primarily anthocyanins in red wine grapes which 30 to 70 days depending on the climate.
498 13  Beer and Wine

The decrease in free acids, combined with Wine Production


the buildup of potassium, results in a rise in the
pH level of the grape juice. The general rela- After the harvest, the grapes are taken into a win-
tionship between the concentrations of the cat- ery and prepared for the primary fermentation.
ions, potassium and sodium, and acidity is Figure 13.9 outlines the divergent processes for
expressed as a function of the titratable acidity. the production of red or white table wines. Red
The balance among potassium and sodium wine is produced from the must (pulp) of red or
­contents and the tartrate to malate ratio deter- black grapes and fermentation occurs together
mine the pH and titratable acidity of the grape with the grape skins, which give the wine its
juice (Boulton 1980). color. White wine is produced by fermenting
The color of the grape berries change with the juice from crushed grapes where the skins are
development of phenolic compounds including removed.
anthocyanins in the skins. Vitis vivifera grapes Crushing is the process where the berries are
contain non-volatile aroma precursors including squeezed breaking the skins releasing the juice
unsaturated lipids, carotenoids, S-cysteine conju- from the berries. The presence of stems in the
gates, glucoconjugates and S-methylmethionine, mix facilitates pressing by allowing juice to flow
These non-volatile components are transformed past flattened skins. More common would be to
into volatile compounds providing the character- add only the crushed fruit, sometimes enzyme
istic aromas of wine varieties during the progres- treated. In red wine production stems of the
sion of the grapes from crushing through aging grapes are usually removed before fermentation
and storage. Flavor precursors are primarily because the stems have relatively high tannin
glycosides and S-amino acid conjugates.
­ content; in addition to tannin they can also give
Additionally, the concentration of tannins in the the wine a vegetal aroma (due to extraction of
grape increases in several areas of the grape 2-methoxy-3-isobutylpyrazine which has an
including the skin, seeds and stem. aroma reminiscent of green bell peppers). This is
Most strains of winemaking yeast have diffi- one of many undesirable compounds; the stems
culties surviving in an alcohol solution above take up room in the fermenter that could other-
15% alcohol This leaves a certain amount of wise hold grapes.
residual sugar which influences the sweetness of Crushed grapes are often treated with sulfites
the wine. or sulfur dioxide which is often added as SO2 gas,
The presence of alcohol (particularly etha- to protect constituents in the juice that are sensi-
nol) in the wine contributes much more than tive to oxidation. The treatment prevents enzy-
just intoxication. It has an immense impact of matic browning caused by the polyphenoloxidase
the weight and mouthfeel of the wine as well as enzyme. The treatment also suppresses the
the balance of sweetness, tannins and acids. growth of undesirable microbes including acetic
In wine tasting, ethanol diminishes perception acid producing bacteria, wild yeasts and molds.
of acids and tannins, and increases perception Wine fermentation can occur as a result of the
of body. natural yeasts found on the surface of the grapes
As sugar levels in the grape rise, acid levels or after inoculation of pasteurized wine must.
fall. Wines need some degree of acidity in order Wild yeasts include Saccharomyces apiculatus
to have a balanced flavor. The major acids in and exiguous. Commercial wine yeasts used for
wine are tartaric and malic acid with citric and inoculation are derived from Saccharomyces
succinic acids playing a minor role. Titratable cerevisiae var. ellipodes or pastorianus. The pure
acidity is not a measure of tartaric acid. Rather, wine yeast strains use for inoculation are chosen
wine acidity is expressed in units of tartaric acid for their desirable fermenting characteristics.
equivalents. A wine can have 0 g/L tartaric acid, These yeast strains can produce high alcohol
but still have 5 g/L “as tartaric acid”. wines containing up to 15.5% alcohol. Typically,
Wine 499

Fig. 13.9  Flow diagram White wines grapes Red wines


for the production of red
and white wine from
crushing to bottling Crushing /
Destemming

Fermentation

Pressing
White grape pomace Red grape pomace
Juice lees Lees
Settling /
Racking

Lees
Fermentation Aging Lees

Aging

Clarification

Lees
Stabilization

Bottling

these strains which are used for red wine fermen- Red wines are produced by destemming and
tation are resistant to alcohol and tannins. Some crushing the grapes into a tank and leaving the
strains of yeast are resistant to sulfites thus they skins in contact with the juice throughout the fer-
are more useful in sulfite treated wines. mentation (maceration). White wines can be pro-
After inoculation of the wine must with yeast, duced from red grapes by pressing of uncrushed
white wines are typically fermented up to 21 days fruit minimizing the contact between grape juice
at below 20 °C and red wines are fermented at and skins. One example is the production of
20–24 °C. At the end of the primary fermentation blanc de noirs a sparkling wine, manufactured
(5–7 days) most of the sugar has been converted from Pinot noir, a red vinifera grape. Almost all
to alcohol. During this period tannins, protein, white wines are produced with crushing and
tartrate and cell debris settle to the bottom of the destemming They are pressed directly as picked
fermentation vessel. Yeast is normally already to avoid extraction of tannins from the skins or
present on the grapes. The primary, or alcoholic grape seeds. Some winemakers crush white
fermentation can be done with the natural yeast, grapes allowing a short period of skin contact,
however results can be unpredictable depending usually for 3–24 h.
on the exact types of yeast that are present. For After alcoholic fermentation, some wines
this reason specific cultured yeast is often added can undergo a secondary fermentation known as
to the must. malolactic fermentation. This process is particu-
Red wines derive most of their color from larly desirable for high-acid wine because it
grape skins, therefore contact between the juice reduces the acidity of the wine by the conver-
and skins are essential for color extraction. sion of the dicarboxylic l-malic acid to the
500 13  Beer and Wine

monocarboxylic l-lactic acid and carbon are crushed there is a considerable amount of
dioxide. This process is conducted by the action juice immediately liberated (called free-run
of lactic acid bacteria, including Oenococcus juice) that can be used for vilification. Typically,
oeni, Leuconostoc spp., and Pediococcus spp. this free-run juice is of a higher quality than the
(Liu 2002). press juice, however most wineries use presses to
During or after the alcoholic fermentation, a increase yield of juice by 15–30%. Increasing the
secondary, or malolactic fermentation can also pressure increases the amount of tannin extracted
take place where specific strains of bacteria (lac- from the skins, rendering the pressed juice exces-
tobacter) convert malic acid into the milder lactic sively tannic or harsh.
acid. The fermentation is carried out by the lactic In winemaking cold stabilization is used to
acid bacteria, Oenococcus oeni, and various spe- facilitate the precipitation of potassium tartrate
cies of Lactobacillus or Pediococcus. The pri- crystals from the wine. These tartrate crystals
mary function malolactic fermentation is to look like grains of clear sand. The cold stabiliz-
convert l-malic acid, to l-lactic acid. This con- ing process is done immediately after fermenta-
version is accompanied by the production of car- tion, the temperature of the wine is dropped to
bon dioxide. near freezing temperatures and is held for 1–2
weeks. Large wineries more commonly use ion
exchange, electrodialysis, and/or addition of sub-
stances that inhibit precipitation.
Bentonite addition is often used to remove
haze-causing proteins from white and rose wines.
Sulfur dioxide is added to wine to prevent oxi-
dation and to undesired fermentation or spoilage.
It is normally added either as liquid sulfur diox-
ide or as sodium or potassium metabisulfite. In
The sourness decreases because the titratable the production of white wine sulfur dioxide can
acidity decreases because lactic is monoprotic be added prior to or immediately after alcoholic
and malic is diprotic Lactic acid tastes less sour fermentation. When it is added after alcoholic
than malic acid. fermentation it will have the effect of preventing
Although acid reduction is the most obvious or stopping malolactic fermentation, bacterial
result of the growth of lactic acid bacteria in spoilage and help protect against the damaging
wine, their action can also significantly modify effects of oxygen.
the wine’s aroma, flavor and mouthfeel. Wine contains both volatile and non-volatile
Oenococcus oeni typically produces substances compounds that contribute to the flavor and
that have pleasant and O. oeni does not produce aroma of wine. During the fermentation and first
off-aromas like many other spoilage lactic acid months of aging, chemical reactions occur result-
bacteria in wine. The metabolism of sugars by ing in changes in the aroma. As a wine ages and
certain lactic acid strains can result in formation matures, changes and developments in aroma
of acetic acid and other undesirable compounds: will continue to take place but at a slower rate.
intentional malolactic fermentation can stabilize Wine grapes contain many non-volatile pre-
a wine by preventing growth of spoilage lactic cursors for aroma and flavor including unsatu-
acid bacteria. Lactic fermentation also produces rated lipids, carotenoids, S-cysteine, conjugates,
diacetyl is the most important of these substances, glycoconjugates and S-methylmethionine. These
it provides the buttery characteristic in the wine. procursors are non-volatile and odorless; how-
Pressing is the act of applying pressure to ever, they are susceptible to chemical and/or
grapes or pomace in order to separate juice or enzymatic reactions that occur during process-
wine from grapes and grape skins. Pressing is not ing and aging. These reactions are often driven
always a necessary act in winemaking; if grapes by the genetic makeup of the particular wine
Wine 501

variety as well as processing and storage condi- Table 13.6  Sensory modalities and selected chemical
components contributing to grape and wine flavor (from
tions (Baumes 2009).
Polaskova 2008)
Wine aroma is a compilation of many factors
Sensory Chemical compounds
including compounds derived from multiple bio-
modality Attribute in wine
chemical sources and the processes of wine mak-
Taste Sweet Glucose, fructose,
ing. The contribution of the grape derived aroma Sour glycerol, ethanol
compounds include monoterpenes, norisopren- Salty Tartaric acid
oids, aliphatics, phenylpropanoids, methoxypyr- Bitter Sodium chloride,
potassium chloride
azines and volatile sulfur compounds (Ebler and
Catechin
Thorngate 2009, González-Barreiro et al. 2013). Aroma Floral, Lily Linalool
Secondary metabolites from microbial metabo- of the valley Isoamyl acetate
lism of sugar, fatty acids, cinnamic acid, organic Banana like
nitrogen compounds (proteins, nucleic acids and Tactile Viscosity Polysaccharides,
pyrimidines) all can contribute to aroma com- Astringency mannoproteins
Pungency/heat Tannins
pounds found in wine (Chatonnet et al. 1992; Ethanol
Herraiz and Ough 1993; Bartowsky and Pretorius Color Red Anthocyanins
2009; Robinson et al. 2014). Terpenes are altered
by acid conditions (Skouromounis and Sefton
2002; Versini et al. 2002) and by enzymatic catal- Wine aroma is highly variable among wines
ysis modify both nonaroma and aroma active and many classes of compounds contribute.
products(Rapp 1998, Ugliano 2009). Oxidative Some of the significant contributors are monoter-
processes in wine take place during winery pro- penes, norisoprenoids, aliphatics, higher alco-
duction of wine, storage and packaging hols, esters, phenylpropanoids, methoxypyrazines,
(Karbowiak et al. 2009; Ghidossi et al. 2012). and volatile sulfur (Francis and Newton 2005;
Tasting wine is essentially the act of smelling Ebeler and Thorngate 2009).
these vaporized aroma compounds and sensing In white wine, monoterpenes are contributors
the taste factors on the tongue. Wines differ to the aroma of Muscat, Gewürztraminer and
widely in both volatile aroma compounds as well Riesling wines (Ribéreau-Gayon et al. 1975;
as non-volatile compounds that impact taste. Rapp 1998; Mateo and Jimeńez 2000). Linalool
Polaskova et al. (2008) have summarized some of and α-terpineol in contribute a floral note in the
the major interactions associated with grape and white wines. (Williams et al. 1981; Günata et al.
wine flavor. Table 13.6 summarizes the taste attri- 1985; De La Presa-Owens and Noble 1997; Lee
butes and chemical compounds responsible for and Noble 2003, 2006; Campo et al. 2005;
sensor responses in wine. It is also important to Robinson et al. 2014).
remember that the color comes from anthocya- Norisoprenoids are present in all wine grapes
nins in the red wine. and are at higher levels in highly aromatic
As mentioned different grapes and resulting grape cultivars (Winterhalter et al. 1990a;
wine result in the wide range of aromas we expe- Schneider et al. 2001). The norisoprenoids are
rience in wine varieties. In Table 13.7 some of the important contributors to the aroma of many
primary odorants and their characteristic aroma wine varieties including Semillon, Sauvignon
are described. blanc, Chardonnay, Merlot, Syrah, and Cabernet
Wine aromas typically contain hundreds of Sauvignon (Razungles et al. 1993; Sefton et al.
volatile compounds. The use of GCMS combined 1993, 1994, 1996; Sefton 1998). The norisopren-
with aroma ports on the gas chromatograph has oids are derived from carotenoids found in
allowed identification of the compounds that in grapes and wines. β-Carotene and lutein con-
contribute most to the aroma of the wine. stitute 85% of the carotenoids, with minor con-
Table 13.7 identifies some of the most important tents of neochrome, neoxanthin, violaxanthin,
providing the base of wine flavor. luteoxanthin, flavoxanthin, lutein-5,6-epoxide
502 13  Beer and Wine

Table 13.7  Impact odorants contributing to varietal aromas of selected wines (from Polaskova 2008)
Variety Odorant Odor quality Sensorya References
Muscat Linalool, Geraniol, nerol Floral 170 Stevens et al. (1966),
Wenzel and deVries (1968)
Riesling 1,1,6-Trimethyl-1,2-­ Kerosene, 20 Ohloff (1978), Simpson
dihydronaphthalene bottle age (1978)
Cabernet Sauvignon, 3-Isobutyl-2-methoxypyrazines Bell pepper 2 Lacey et al. (1991), Allen
Sauvignon blanc et al. (1991), Roujou et al.
Cabernet franc, (2000), Belancic and
Merlot Carmenere Agosin (2007)
Gerwurtzraminer Cis-Rose oxide Geranium oil, 200 Bauer et al. (1997), Guth
carrot leaves (1995, 1996, 1997a, b)
Vitis labrusca, Vitis o-Aminoacetophenone Foxy, sweet 0.2 (in air) Acree et al. (1990), Baek
rotundfolia et al. (1997)
Sauvignon blanc 4-Methyl-4-mercaptopentan-2- Black current 0.6 Guth (1997a, b, 1998)
Scheurebe one
Grenache rose, 3-Mercapto-hexanol Grapefruit, 50 Ferreira et al. (2002),
Sauvignon blanc, citrus peel, Tominaga et al. (2006)
Semillon Passion fruit
Shiraz Rotundone Black pepper 16 Siebert et al. (2008), Wood
et al. (2008)
Threshold in water ng/L
a

and zeaxanthin, and cis isomers of lutein and the overall aroma of the wines. It is important to
β-carotene the next most abundant (Mendes-­ recognize that low levels of methoxypyrazines
Pinto 2009). Carotenoids accumulate prior to contribute to varietal character, however, high
veraison in the grape exocarp (skin) (Razungles levels are generally considered undesirable.
et al. 1988; Robinson et al. 2014). Methoxypyrazines contribute to the characteris-
Volatile phenylpropanoids are derived from tic aroma of both grape juice and wines including
hydrolyzed of glycoside in juices and wines, in Sauvignon blanc (Allen et al. 1991; Lacey et al.
Chardonnay wine phenylpropanoids make up 10 1991), Cabernet Sauvignon (Allen et al. 1991,
to 20% of the total hydrolyzed volatile fraction in 1994), Cabernet franc (Roujou de Boubée et al.
Chardonnay juice (Sefton et al. 1993). Their flavor 2000), Merlot (Sala et al. 2004; Belancic and
attributes include the dried fig, tobacco, and choc- Agosin 2007; Robinson et al. 2014).
olate aromas in Cabernet Sauvignon and Merlot The basis for wine flavor crosses over many
musts (Francis et al. 1998). Methyl anthranilate, varieties of grape and wine. Ferreira (2010) iden-
which is responsible for the distinctive “foxy” tified the aroma compounds that for base back-
aroma and flavor of the Washington Concord ground for many wines. They are summarized in
grape (Vitis labrusca) (Wang and De Luca 2005) Table 13.8.
and may also contribute to the aroma of Pinot noir The conjugated thiol precursors are highest in
(Moio and Etievant 1995; Robinson et al. 2014). the skin (Roland et al. 2011). The glutathione can
The peppery, pea or asparagus aromas in be conjugated to the C6-aldehyde, (E)-2-hexenal,
wines come from the 3-alkyl-2-­methoxypyrazines, which is presumably reduced enzymatically to
including 3-isobutyl- 2-methoxypyrazine (IBMP), the alcohol by alcohol dehydrogenase (Capone
3-isopropyl-2-methoxypyrazine (IPMP), and and Jeffery 2011). It has also been discovered
sec-butyl-2-methoxypyrazine (SBMP) (Sala that grapes can contain 3-S-cysteinylglycinehexan-­
et al. 2004). These compounds have very strong 1-ol, an intermediate in the catabolism of the glu-
aromas and are at very low concentrations in tathione conjugate to the cysteine conjugate
wine (ng/L), however they are important to (Capone et al. 2011).
Wine

Table 13.8  Aroma compounds forming the base of wine flavor (Ferreira 2010)
Miscellaneous Fusel alcohols Organic acids Isoacids Organic acid ethyl esters Fusel alcohol acetates Ethyl esters of isoacids
Ethanol Isobutanol Acetic acid Isobutyric acid Ethyl acetate Isobutyl acetate Ethyl isobutyrate
Diacetyl Isoamyl alcohol Butyric acid 2-Methyl butyric acid Ethyl butyrate Isoamyl Ethyl 2-methyl butyrate
Acetaldehyde Hexanol β-Phenylethanol Hexanoic acid Isovaleric acid Ethyl hexanoate acetate β-Phenylethyl acetate Ethyl isovalerate
Methionol Octanoic acid Ethyl octanoate
Decanoic acid Ethyl decanoate
503
504 13  Beer and Wine

Thiols/Mercaptan-sulfur compound can contrib- et al. 2005). Yeast strains differ greatly in their
ute an aroma of garlic and onion that is ­considered ability to use nitrogen and amino acids (Carrau
a wine fault. They have also been found to contrib- et al. 2008).
ute to some of the varietal aromas associated with Many flavor and aroma compounds are formed
Cabernet Sauvignon, Gewürztraminer, Merlot, from amino acids are alcohols, associated esters
Muscat, Petit Manseng, Pinot blanc, Pinot griso, and volatile acids Amino acids can be catabo-
Riesling, Scheurebe, Semillon and Sylvaner. lized into higher alcohols by the pathway referred
Volatile thiols can result in pleasant herbaceous, to as the Ehrlich reaction. In Fig. 13.10 the
fruity, mineral, smoky, and toasty aromas in wine Ehrlich reactions and products marked with an
(Dubourdieu and Tominaga 2009). The predomi- asterisk. The Ehrlich pathway compounds are
nant volatile sulfur compounds in wines are H2S, generally of less importance than the compounds
methanethiol, dimethylmercaptans (dimethylsul- produced by the anabolic pathway. In addition to
fide, dimethyldisulfide, dimethyltrisulfide), methyl- the three nonpolar branched-­chain amino acids
thioesters (S-methyl thioacetate, S-methyl (valine, leucine, and isoleucine), other amino
thiopropanoate, and S-methyl thiobutanoate), and acids can also be broken down to other metabo-
liberated glutathione and cysteine polyfunctional lites via this reaction (Table 13.8) (Ardö 2006).
thiols (4-­mercapto-­4-methylpentan-2-one, 4MMP; The most important odor-related products (higher
3-­mercaptohexan-1-ol, 3MH; and 3-­mercaptohexyl alcohols and volatile fatty acids) shown in
acetate, 3MHA) (Swiegers and Pretorius 2007; Table 13.8 are produced from valine, leucine, and
Dubourdieu and Tominaga 2009; Roland et al. isoleucine. The first step is a transamination reac-
2010). Many other sulfur-­ containing compounds tion in which the amino group from the amino
have been identified in wines (Mestres et al. 2000, acid is transferred to α-ketoglutarate to form an
2002; Bailly et al. 2006; Sarrazin et al. 2007; α-keto acid and glutamate (Fig. 13.10) (Conway
Dubourdieu and Tominaga 2009). and Hutson 2000; Davoodi et al. 1998; Taylor
Some of the aromas perceived in wine are and Jenkins 1966).
from esters created by the reaction of acids and Many fermentation volatiles that are impor-
alcohol in the wine. Esters can develop during tant to wine aroma are metabolic products from
fermentation, with the influence of yeast, or dur- yeast metabolism of hexose sugars. Straight-­
ing aging. Yeast strain used during fermentation chain fatty acids and their corresponding ethyl
and temperature are the primary factors control- esters were derived almost exclusively from hex-
ling ester formation. Wines eventually approach oses. Most fusel alcohols and their acetate esters
an equilibrium where with alcohols, acids, and are derived from hexose metabolism in the yeast
esters. During processing and aging the bouquet indicating the importance of anabolic pathways
of the wine is constantly changing due to the con- in their formation Fig. 13.11 illustrates the ratios
centration, formulation and splitting of different of volatile compounds arising from the metabo-
esters. This is partly the reason why a wine will lism of hexoses (Nisbet et al. 2014).
have one set of aromas at one time and other aro- From this it can be seen that the chemical
mas later in its life (Bardi et al. 1999). reactions that influencing wine flavor and aroma
During alcoholic fermentation yeast can use are complex and varied. Wine flavor and aroma
amino acids in several ways, particularly for pro- continues to be an intense area of research.
tein synthesis, or for other metabolic processes. Professional wine tasters will often mentally
Because grapes are low in Nitrogen compounds, cycle through a list of potential aromas (and may
available Nitrogen often limits growth of yeast. use visual aids like the aroma wheel (Fig. 13.12)
The nitrogen composition of the grape must developed by Ann C. Noble of University of
affects not only the kinetics of alcoholic fermen- California, Davis) until one choice stands out and
tation, but also the production of aromatic com- can be identified in the wine (http://winearoma-
pounds, ethanol, and glycerol (Hernandez-Orte wheel.com/).
Wine 505

Fig. 13.10  A simplified metabolic map of yeast aroma responsible for the production of higher alcohols and vol-
compound production, indicating known metabolic link- atile acids. Cofactors and transition metabolites are shown
ages. Bold type indicates aroma compounds important to in italics. αKG is α-keto glutamate and Glu is glutamate
this study. Compounds marked with an asterisk constitute (Polášková et al. 2008)
a diagrammatical representation of the Ehrlich pathway,

Fig. 13.11  Percentage of carbon found in volatiles that originated from hexoses during yeast metabolism. The mean is
indicated by the circles, and the error bars represent the 95% confidence interval
506 13  Beer and Wine

Fig. 13.12  Wine aroma wheel

rendus du 4e Symposium International d’Oenologie,


References Bordeaux, 1989 (pp. 25–30).
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Acree, T. E., Lavin, E. H., Nishida, R., & Watanabe, S. W. V. (1991). Contribution of methoxypyrazines to
(1990). ο-Amino-acetophenone, a foxy smelling com- sauvignon blanc wine aroma. American Journal of
ponent of Labruscia. In Y. Bessiere & A. F. Thomas Enology and Viticulture, 42, 109–112.
(Eds.), Flavour science and technology (pp. 49–52). Allen, M. S., Lacey, M. J., & Boyd, S. (1994). Determination
Chichester: Wiley. of methoxypyrazines in red wines by stable isotope dilu-
Alba-Lois, L., & Segal-Kischinevzky, C. (2010). Yeast tion gas chromatography-mass spectrometry. Journal of
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Allen, M. S., Lacey, M. J., Brown, W. V., & Harris, R. lism. Biotechnology Advances, 24, 238–224.
L. N. (1990). Occurrence of methoxypyrazines in Baek, H. H., Cadwallader, K. R., Marroquin, E., & Silva,
grapes of Vitis vinifera cv. Cabernet Sauvignon J. L. (1997). Identification of predominant aroma
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Bailly, S., Jerkovic, V., Marchand-Brynaert, J., & Collin, Chatonnet, P., Dubourdieu, D., Boidron, J. N., & Pons,
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Genetically Modified Crops
14
W. Jeffrey Hurst and John W. Finley

When man evolved from a hunter gatherer they (known as the F1 generation) was purple-­
began to grow crops for food. They found that flowered. When Mendel self-fertilized the F1
selection of crops improved the quality and yield generation pea plants, he obtained a purple flower
of foods for food production. The selection of to white flower ratio in the F2 generation of 3 to 1.
seeds led to the evolution of new crops that are The results of this cross are tabulated in the
more productive and nutritious crops. Punnett square in Fig. 14.1. Mendel hypothesized
The domestication of plants by humans to pro- that allele pairs segregate, from each other during
duce plants with more desirable traits than wild the production of gametes. Because allele pairs
plants drove early man to become dependent on separate during gamete production, a sperm or
identification and propagation of plants with egg carries only one allele for each inherited trait.
desirable attributes. Seed selection for desired When sperm and egg unite at fertilization, each
traits began between 9000–11,000 years ago. contributes its allele, restoring the paired condi-
Initially early farmers simply selected food plants tion in the offspring. This is called the Law of
with particular desirable characteristics, and Segregation. The presence of an allele does not
employed these as progenitors for subsequent mean that the trait will be expressed in the indi-
generations, resulting in an accumulation of valu- vidual that possesses it. If the two alleles of an
able traits over time. inherited pair differ (the heterozygous condition),
1856 and 1863 Gregor Mendel investigated of one determines the organism’s appearance and is
plant hybridization led to his laws of inheritance. called the dominant allele; the other has no
This work became well known in the 1900s and noticeable effect on the organism’s appearance
formed the basis of the science of genetics. The and is called the recessive allele. Thus, in the
laws he developed stimulated research by many example above dominant purple flower allele will
plant scientists dedicated to improving crop pro- hide the phenotypic effects of the recessive white
duction through plant breeding. He conducted flower allele. This is known as the Law of
hybridization experiments with garden peas Dominance. These laws formed the basis of
(Pisum sativum) which led to two generalizations understanding inheritance and allowed more
which later became known as Mendel’s Principles effective selection in breeding of plants.
of Heredity or Mendelian inheritance. Mendel Gregor Mendel’s experiments with plant
discovered that, when he crossed purebred white hybridization led to his establishing laws of
flower and purple flower pea plants (the parental inheritance. Once this work became well known,
or P generation), the result was not a blend. it formed the basis of the new science of genetics,
Rather than being a mix of the two, the offspring which stimulated research by many plant

© Springer International Publishing AG 2018 511


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_14
512 14  Genetically Modified Crops

must evaluate the progeny sorting for desired


changes as well as undesired changes. In 1973,
Cohen et al. described recombinant-DNA
(rDNA) techniques that allowed scientists to cut
gene sequences from the DNA of one organism
and splice them into the DNA of another organ-
ism (Cohen et al. 1973), the path was paved for a
new approach to increase genetic diversity for
use in breeding organisms, including crops hence
genetic engineering.
Many present-day crops currently under culti-
vation are traceable to plant domestication in
Fig. 14.1  Representation of the Punnett square for inher- ancient times. Nearly all of the domesticated
ence of flower color in pea plants the https://commons. plants used today for food and agriculture were
wikimedia.org/w/index.php?curid=2063426 domesticated in the various centers of origin
around the world. In these centers there is still a
­scientists dedicated to improving crop production great diversity of closely related wild plants. For
through plant breeding. example wheat in its early form came from the
However, successful commercial plant breed- Fertile Crescent and corn as we know it came
ing concerns began to be founded from the late from Central America.
nineteenth century. Gartons Agricultural Plant One major technique of plant breeding is
Breeders in England was established in the 1890s selection, the process of selectively propagating
by John Garton, who was one of the first to cross-­ plants with desirable characteristics and eliminat-
pollinate agricultural plants and commercialize ing or “culling” those with less desirable
the newly created varieties. He began experi- characteristics.
menting with the artificial cross pollination first Deliberate interbreeding of closely or dis-
of cereal plants, then herbage species and root tantly related individuals produces new crop vari-
crops and developed far reaching techniques in eties or lines with desirable properties. Plants are
plant breeding (Graften and Ridley 2006; Ford crossbred to introduce traits/genes from one vari-
1960). ety or line into a new genetic background. One
In the late nineteenth and early twentieth cen- example is when a mildew-resistant pea is
turies the better understanding of genetics pro- crossed with a high-yielding but susceptible pea
vided plant breeders, with the tools to make the new cross bred peas with mildew resistance
modifications in plants with increasing precision. without losing the high-yield characteristics.
They changed the expression of traits in plants by Progeny from the first cross would then be
crossing specific parent plants to produce new crossed with the high-yielding parent to ensure
varieties with desired traits. They also developed that the progeny were most like the high-yielding
more rapid methods to generate and detect parent in a step referred to as backcrossing. Plants
genetic changes. The improved technology led to may also be crossed with themselves to produce
targeted and more efficient breeding of improved inbred varieties for breeding.
varieties (Mba2013). DNA mutation rare in Classical breeding relies largely on homolo-
nature (Ossowski et al. 2010), but scientists gous recombination between chromosomes to
applied mutagenic chemicals or radiation to generate genetic diversity. Conventional plant
induce mutations in DNA at increased rates breeders also make use in vitro techniques such
(Roychowdhury and Tah 2013) thus increasing as protoplast fusion, embryo rescue or mutagen-
the genetic variation in the species. Both natural esis to generate diversity and produce hybrid
and induced mutations are random so breeders plants that would not exist in nature.
Genetically Modified Crops 513

Breeders have attempted to incorporate the modern corn illustrates man’s direction of genetic
following traits into crops using conventional selection to yield enhanced genotypes of a crop. It
techniques: is important to realize that this is an early example
of a genetically enhanced organism which resulted
1. Improved quality, such as increased nutrition, from selection and cross breeding. Figure 14.2
improved flavor, or greater beauty illustrates the evolution both the tassel (male) and
2. Increased yield of the crop ear (female) portion of corn from the parent teo-
3. Increased tolerance of environmental pres-
sinte to modern hybrid corn.
sures (salinity, extreme temperature, drought) These breeding techniques resulted in large
4. Resistance to viruses, fungi and bacteria yield increase in the United States in the early
5. Increased tolerance to insect pests twentieth century. After World War II, the Green
6. Increased tolerance of herbicides Revolution increased crop production in the
7. Longer storage period for the harvested crop developing world in the 1960s. These break-
through were based on three essential crops;
The development of modern day corn serves as maize, wheat and rice. The development of
an example of the evolution of corn which began hybrid maize was followed by high-yielding and
about 10,000 years ago. Starting with Teosinte we input-responsive “semi-dwarf wheat” (for which
see the directed evolution of corn through selec- the CIMMYT breeder N.E. Borlaug received the
tive plant breeding. Figure TSC1 illustrates the Nobel prize for peace in 1970), and third came
evolution of corn from teosinte to modern hybrid high-yielding “short statured rice” cultivars.
corn. The farmers selected corn seeds from crops Dwarfing of wheat delivered a critical agro-
that were easier to grind, tasted better or had nomic quality of dwarf plants with thick stems
larger kernels. The evolution from teosinte to making them less prone to collapse under the

Fig. 14.2  Evolution of


maze from teosinte to
modern corn
514 14  Genetically Modified Crops

weight of the extra grain — a trait called lodging. It also opened the possibility of plants with many
Borlaug worked wheat that had tall, thin stalks. new and useful attributes.
The taller of wheat grasses better compete for These directed changes in plants which
sunlight, but tend to collapse under the weight of includes direct transfer of a desired DNA from a
the extra grain resulting from rapid growth spurts microorganism or another plant into a trans-genic
from Nitrogen fertilizer. In 1953, he acquired a species that has the desired characteristics.
Japanese dwarf variety of wheat called Norin 10 Figure 14.3 illustrates the principal of gene inser-
developed by Orville Vogel, that had been crossed tion in plants.
with a high-yielding American cultivar called Intensive research in molecular genetics has
Brevor Reitz (1970). Norin 10/Brevor is semi-­ led to the development of recombinant DNA
dwarf (one-half to two-thirds the height of stan- technology. Advancement in biotechnological
dard varieties) and produces more stalks and thus techniques has opened many possibilities for
more heads of grain per plant. Borlaug crossbred breeding crops. Mendelian genetics allowed
the semi-dwarf Norin 10/Brevor cultivar with his plant breeders to perform genetic transformations
disease-resistant cultivars to produce wheat culti- in a few crops, molecular genetics has provided
vars that were adapted to tropical and sub-­tropical the key to both the manipulation of the internal
climates (Hedden 2003). genetic structure, and the “crafting” of new culti-
In efforts to develop new varieties of grains vars with targeted attributes such as virus resis-
seeds were submitted to mutation breeding. tance, insect resistance and herbicide tolerance.
Mutation breeding is the process of exposing
seeds to chemicals or radiation in order to gener- Genetically modified organism
ate mutants with searching for desirable traits to insecticide gene created
be bred with other cultivars. The mutated seeds using recombinant
DNA technology
were grown to screen for positive characteristics plasmid
vectors
From 1930 to 2014 more than 3200 mutagenic
plant varietals have been released (Schouten and digestion with
restriction enzymes
Jacobsen (2007; FAO/IAEA 2014). The crops
released are either direct mutants (70%) or their
progeny (30%) (Maluszynsk et al. 2000), ot
cleaved
which food and agricultural crops account for cleaved
DNA vectors
75% of released mutagenic species (Ahloowali
2004). The FAO/IAEA reported in 2014 over
1000 mutant varietals of major staple crops were growing plant cells
being grown worldwide. take up insecticide gene
from plasmid vectors
It is important to realize that mutation breed-
ing produces completely random mutations
which ultimately result in new or modified pro-
teins in the new plant. The process is very tedious select for
insecticidal cells cells used
and it can take many years to identify a new and for plant
insects that feed on
useful plant. the plants will die
propagation
The accumulation of this previous work illus-
trates that plant scientists have been “engineer-
ing” plants for at least 10,000 years. In the early
1990s the next step in the evolution of the tech-
nology was to move genes from one species to
another by direct gene insertion. The process is © 2009 Encyclopaedia Britannica, Inc.
significantly faster and more precise than muta-
tion breeding of crossing and back crossing. Fig. 14.3  The principal of gene insertion in DNA
Genetically Modified Crops 515

Recombinant DNA technology is the joining tion that the enzyme polygalacturonidase was a
together of DNA molecules from two different key since it dissolved cell wall pectin. A group
species which are inserted into a host organism to from Celgene proposed to limit this enzyme by
produce new genetic combinations. The recombi- developing an antisense gene. The researchers
nant organisms have value and promise in medi- hoped that this would retard ripening allowing it
cine, agriculture, and industry. Recombinant DNA to remain firm longer. In 1987, Calgene identified
technology allows the isolation of a segment of and cloned the tomato fruit pg gene and in 1992
DNA or a gene for a specific protein. With that presented a petition to the FDA and in 1994
fragment the nucleotide sequence can be deter- approved the addition of a kanamycin resistance
mined, the transcripts, mutate the sequence in gene construct needed to create the PG-antisense
highly specific ways if needed, and reinsert the tomato. Work continued and in late 1994 the
modified sequence into a living organism. Both Flavr-Savr tomatoes was introduced. Demand
agriculture and medicine have benefited signifi- was high and remained high but production costs
cantly from this technology. It should be pointed were also high and the product was not profitable.
out that it is one more step in the timeline of plant While it may have been a technological success it
science and improving our crops. Currently crops was a commercial failure and did nothing for the
or foods modified by modern genic technology are cause of biotechnology so generally, the applica-
referred to as Genetically Modified Organisms or tion of biotechnology and transgenic foods has
GMOs. Simplistically GMO’s have their genetic become the purview of commercial agriculture.
composition altered hence they can code for a new Currently there is a substantial amount of food
property. The gene needs a mechanism to turn it grown using DNA recombinant technology with
on. This on switch is called the promoter segment. approx. 85% of the corn grown in the US being
One of the more widely used promoters is named GMO and almost 90% of the soybeans. This is
35S. When a new GMO has been developed with not all the GMO crops but encompass a substan-
a new trait, the resulting gene construct is called an tial percentage. The GMO crops have various
event with events being developed regularly. These traits. Examples of two of the more common
events undergo various regulatory and safety crops with their associated traits follow. Roundup
reviews before being approved for use. One area Ready Soybeans contain a proteins that interferes
that unfortunately is growing is the development most with the EPSPS pathway. Round Up known
of unapproved events. as glyphosate is a general purpose pesticide used
Genetically modified (GM) foods were first not only in agriculture but in homes to eliminate
approved for human consumption in the United weeds. While good to eliminate weeds, it also
States in 1994, and by 2014–2015 about 90% of eliminates healthy crops such as flowers, crops
the corn, cotton, and soybeans planted in the and ornamentals. In the case of Roundup Ready
United States were GM. By the end of 2010, GM Soy, the GMO trait allows the farmer the ability
crops covered more than ten million square kilo- to use Round Up to eliminate weeds while not
metres (3.86 million square miles) of land in 29 killing the soy. Furthermore, a farmer can be
countries worldwide—one-tenth of the world’s more productive eliminating tedious weeding.
farmland. The majority of GM crops were grown The second example is BT corn having been
in the Americas. In the agricultural arena, the encoded with a gene that eliminates the corn
technique was applied to soybeans in 1988, pav- borer allowing for more corn per acre. Based on
ing the way for one of the most successful crops, data from the end of 2012 there were 170 million
glyphosate tolerant soy. While of substantial hectares in production that includes 312 events in
importance to commercial agriculture, very few 29 species with 3497 approvals in 59 countries.
consumers were aware of this development. Table  14.1 provides a partial listing of some of
Likely, the introduction of the “Flavr Savr” the key proteins expressed by some GM crops.
tomato in 1994 was the first GMO crop many had Figure 14.4 gives a simplistic view of the steps in
seen. In the 1980s there was anecdotal informa- genetic modification.
Table 14.1  Genetically engineered traits deregulated and approved for field release in the United States as of 2015
Crop Scientific Name Trait Year approved Developer
Alfalfa Medicago sativa Glyphosate HRa,b 2005, 2010 Monsanto/Forage Genetics
Reduced Lignin 2015 Monsanto/Forage Genetics
Apple Malus domestica Non Browning 2015 Okanagan Specialty Fruits
Canola Brassica napus/ Oil Profile Alteredc 1994 Calgene
Brassica rapa Glufosinate HR 1995 Bayer
Phytase 1998 BASF
Glyphosate HR 1999 Monsanto
Maize, Zea mays Glufosinate HR 1995 AgrEvo
field Bt IR 1995 Monsanto
Glyphosate HR 1996 Monsanto
Increased Lysine 2006 Monsanto
Alpha-Amylase 2011 Syngenta
Drought Tolerance 2011 Monsanto
Male Sterility/Color 2011 DuPont
ACCased HR 2014 Dow
2,4-D HR 2014 Dow
Increased Ear Biomass 2015 Monsanto
Maize, Zea mays Bt IRe 1998 Novartis
sweet Glyphosate HR 2011 Monsanto
Papaya Carica papaya Ring Spot Virus VRf 1996 Cornell University, University of Hawaii
USDA Agricultural Research Service
Plum Prunus domestica Plum Pox VRc 2007 USDA Agricultural Research Service
Potato Solanum tuberosum Bt IR 1995 Monsanto
Potato Virus Y VRc 1999 Monsanto
Potato Leafroll VRc 2000 Monsanto
Low Acrylamide 2015 Simplot Plant Sciences
Nonbrowning 2015 Simplot Plant Sciences
Resistance to Late 2015 Simplot Plant Sciences
Blight Pathogen
Rice Oryza sativa Gulfosinate HR 1999 AgrEvo
Squash Cucurbita pepo Zucchini Yellow VR 1994 Upjohn
Watermelon Mosaic 1994 Upjohn
VR 1996
Cucumber Mosaic VR
Soybean Glycine max Glyphosate HR 1994 Monsanto
Glufosinate HR 1996 Bayer
Sulfonylurea HR 2007 DuPont
Modified Oil 2009 DuPont
High Oleic Oil 2010 DuPont
Isoxaflutole HRc 2013 Syngenta
Mesotrione HRc 2014 Syngenta
Imidazolinone HR 2014 BASF
2,4-D HR 2015 Dow
Dicamba HR 2015 Monsanto
Sugar Beta vulgaris Glyphosate HRg 2005 Monsanto
Beet Glufosinate HR 1998 AgrEvo
Tomato Solanum lycopersicum Fruit Ripening Alteredc 1992 Calgene
a
HR herbicide resistance
b
Returned to regulated status in 2007; returned to deregulated status in 2011
c
Not in production in 2015
d
Acetyl CoA Carboxylase inhibitor herbicide
e
IR insect resistance (different Bacillus thuringienis Cry genes inserted to encode proteins that kill specific species)
f
VR virus resistance
g
Returned to regulated status in 2010 because of litigation; Returned to deregulated status in 2011
DATA SOURCES: http://www.cera-gmc.org/GMCropDatabase; http://www.isaaa.org/gmapprovaldatabase/; http://
www.aphis. usda.gov/biotechnology/petitions_table_pending.shtml.
Adapted from NAS, 2016
Genetically Modified Crops 517

Agrobacterium tumefaciens
bacterium

Inserted T-DNA
carrying foreign
4 The plasmid gene
is reinserted
Restriction into a bacterium. 5 The bacterium is
cleavage used to insert the
site T-DNA carrying the
Ti foreign gene into the
6 The plant cells
T-DNA plasmid chromosome of a
are grown in
plant cell.
culture.
Recombinant
1 The plasmid is removed Ti plasmid
from the bacterium, and
the T-DNA is cut by a
restriction enzyme. 3 The foreign DNA is
inserted into the T-DNA
of the plasmid. 7 A plant is generated from a cell
clone. All of its cells carry the
2 Foreign DNA is cut
foreign gene and may express
by the same enzyme.
it as a new trait.
© BENJAMIN/CUMMINGS

Fig. 14.4  Gene insertion in plants via Agrobacterium tranfection

One of the earlier techniques used to insert delivery system for gene engineering in plants
genes into plants was the use of the Agrobacterium (Schell and Van Montagu 1977; Joos et al. 1983).
as a vector to insert the new DNA into a plant. This work then laid the groundwork for the inser-
Agrobacterium tumefaciens causes a condition tion of specific genes into a plant using the
known as crown-gall disease in plants. Crown-­ Agrobacterium. One can also argue that the gene
gall is characterized by a tumor-like growth or transfer has been going on for a very long time
gall on the infected plant. The tumors are initiated and we have learned to use it effectively for spe-
by the transfer of a DNA segment from the bacte- cific crop improvements.
rial tumor-inducing plasmid. The plasmid T-DNA The genes to be introduced into the plant are
is integrated semi-randomly into the genome of cloned into a plant transformation vector that
the host cell where the tumor morphology genes contains the T-DNA region of the bacterial plas-
on the T-DNA are expressed, causing the forma- mid, together with a selectable marker. Frequently
tion of a gall (Francis and Spiker 2004). The abil- an antibiotic marker gene was incorporated into
ity of Agrobacterium to transfer genes to plants the plasmid in conjunction with the other desired
and fungi is used in biotechnology, in particular, genes to enable selection for plants that have
genetic engineering for plant improvement. A been successfully transformed. Plants are grown
modified Ti or Ri plasmid can be used. The plas- on media containing antibiotic following
mid is ‘disarmed’ by deletion of the tumor induc- ­transformation, and those that do not have the
ing genes; the only essential parts of the T-DNA T-DNA integrated into their genome will die.
are its two small (25 base pair) border repeats, at Transformation with Agrobacterium can be
least one of which is needed for plant transforma- accomplished by incubating either protoplasts or
tion. Marc Van Montagu and Jozef Schell at the leaf discs with the Agrobacterium to cause the
University of Ghent (Belgium) discovered the plasmid insertion. From the callus that results,
gene transfer mechanism between Agrobacterium whole plants regenerated using plant tissue
and plants, which resulted in the development of ­culture. The transformation with Agrobacterium
methods to alter Agrobacterium into an efficient is illustrated in Fig. 14.4.
518 14  Genetically Modified Crops

Agrobacterium does not infect all plant spe-


cies0 but other techniques have been applied for
plant transformation one of which is the gene
gun. A gene gun is biolistic particle delivery sys-
tem, originally designed for plant transformation
by injecting cells with genetic material. The plas-
mid DNA is coated on elemental particle of a
heavy metal. The gene gun is able to transform
almost any type of cell, including plants, and is
not limited to genetic material of the nucleus: it
can also transform organelles, including plastids.
Gene insertions intended to transform prokary-
otic genomes generally have the gene or genes of
interest, at least one promoter and terminator
sequence, and a reporter gene; which is a gene
used to ease detection or removal of those cells
which didn’t integrate the construct into their
DNA.[5] These genes may each have their own pro- Fig. 14.5  Illustrates the principal of the Gene Gun shoot-
moter and terminator, or be grouped to produce ing DNA coated particles into plant callus cells
multiple gene products from one transcript, in
which case binding sites for translational machin- The term genetic modification and so-called
ery should be placed between each to ensure maxi- genetically modified organisms (GMOs) is fre-
mum translational efficiency. In any case the entire quently misused. All types (organic, conven-
construct is flanked by regions called border tional) of agriculture modify the genes of plants
sequences which are similar in sequence to loca- so that they will have desirable traits. Traditional
tions within the genome, this allows the construct forms of breeding change the plant’s genetics
to target itself to a specific point in the existing indirectly by selecting plants with specific traits,
genome (Slater et al. 2008). The target of a gene while genetic engineering changes the traits by
gun is often a callus of undifferentiated plant cells making changes directly to the DNA. In tradi-
growing on gel medium in a Petri dish. After the tional breeding, crosses are made in a relatively
gold particles have impacted the dish, the gel and uncontrolled manner. In conventional plant breed-
callus are largely disrupted. However, some cells ing, the breeder selects the parents to cross, the
are not killed in the impact, and have incorporated results are unpredictable because the DNA from
enveloped a DNA coated gold particle, which the parents recombines randomly. In contrast,
eventually migrates to and integrates into a plant genetic engineering allows highly precise transfer
chromosome. Figure 14.5 illustrates the principal of genes, quick and efficient tracking of genes in
of the gene gum. The propellant can be com- new varieties. This ultimately results in increased
pressed gas or 22 caliber blanks. efficiency in developing new crop varieties with
The modified cells from the callus are treated new and desirable traits (Popping 2010).
with a series of plant hormones, such as auxins
and gibberellins, and each may divide and differ-
entiate into the differentiated tissue cells of the  pplications of Genetically
A
plant. This capability of total re-generation is Modified Crops
called totipotency. The new plant that originated
from a successfully shot cell will express new The introduction of the “Flavr Savr” tomato in
genetic (heritable) traits as illustrated in Fig. 14.6. 1994 was the first GMO crop to be introduced
In the Figure the use of the gene gun is used to into the market. In the 1980s there was anecdotal
incorporate insect resistance into a tomato plant. information that the enzyme polygalacturonidase
Applications of Genetically Modified Crops 519

Fig. 14.6  Application of the Gene Gun to introduce insect resistance in a tomato plant

(PG) was key to softening of tomato fruit because the tomatoes to remain firm longer. In 1987,
it dissolved cell wall pectin. Calgene proposed to Calegene identified and cloned the tomato fruit
limit this enzyme by developing an antisense PG gene and in 1992 presented a petition to the
gene. The goal was to retard ripening allowing FDA. In 1994 FDA approved the addition of a
520 14  Genetically Modified Crops

kanamycin resistance gene construct needed to The seven other food crops of which GE vari-
create the PG-antisense tomato. Work continued eties were grown in 2015 were apple (Malus
and in late 1994 the Flavr-Savr tomatoes was domestica), canola (Brassica napus), sugar beet
introduced. While it may have been a technologi- (Beta vulgaris), papaya (Carica papaya), potato,
cal success it was a commercial failure and did squash (Cucurbita pepo), and eggplant (Solanum
nothing for the cause of biotechnology so gener- melongena) (James 2015). The contribution of
ally, the application of biotechnology and trans- GE varieties to the production of those crops was
genic foods has become a major point of small, except for canola; GE varieties of canola
discussion in agriculture. Currently there is a constituted 24% of the 36 million hectares
substantial amount of food grown using DNA planted in 2015 (James 2015) rd of all land
recombinant technology with approx. 85% of the planted to maize worldwide that year (James
corn grown in the US being GMO and almost 2006, 2015).
90% of the soybeans. This is not all the GMO The most economically important crop modi-
crops but encompass a substantial percentage. fications to date are herbicide resistance, Insect
The GMO crops have various traits. Examples of resistance and virus resistance.
two of the more common crops with their associ- Herbicide resistance introduces the ability of a
ated traits follow. Roundup Ready Soybeans con- crop to resist the application of certain herbicides
tain a protein that interferes most with the EPSPS that are used for weed control. Herbicide resis-
pathway. Round Up known as glyphosate is a tant traits have been developed for nine different
general purpose pesticide used not only in agri- herbicides and introduced into eight herbicide
culture but in homes to eliminate weeds. While resistant traits for soybeans, six for cotton, three
good to eliminate weeds, it also eliminates for canola, three for maize, two for sugar beet,
healthy crops such as flowers, crops and orna- and one for alfalfa. Some crop varieties that had
mentals. In the case of Roundup Ready Soy, the stacked traits for resistance to two herbicides (for
GMO trait allows the farmer the ability to use example, glyphosate and 2,4-D or glyphosate and
Round Up to eliminate weeds while not killing dicamba). Since it was first introduced in for soy-
the soy. Furthermore, a farmer can be more pro- beans in 1996, glyphosate resistance has been
ductive eliminating tedious weeding. The second introduced in alfalfa, canola, cotton, maize, and
example is BT corn having been encoded with a sugar beet by 2015.
gene that eliminates the corn borer allowing for Insect-resistant (IR) trait incorporates insecti-
more corn per acre. Based on data from the end cidal properties produced internally by a plant
of 2012 there were 170 million hectares in itself. An example of insect resistance is the
­production that includes 312 events in 29 species introduction of transfer of a gene coding for a
with 3497 approvals in 59 countries. crystalline (Cry) protein from the soil bacterium
About 12 of global cropland was used to pro- Bacillus thermogenesis. The Cry is toxic to the
duce genetically modified crops in 2015 (FAO target insect when the insect feeds on the plant.
2015; James 2015). Commercially available The Cry proteins can control many insect pests—
crops in production in 2015 included nine food primarily moths, beetles, and flies (Höfte and
crops, three non-food crops, and two types of Whiteley 1989). In 2015 insect resistant varieties
flowers. Maize and soybean were the most of cotton, eggplant, maize, poplar, and soybean
widely grown genetically modified crops. were in commercial production (NAS 2016).
Production of genetically modified genetically Virus resistance prevents a plant from being
modified maize has increased substantially since susceptible to specific viral diseases. The virus
its first commercial release in 1996 to include resistance In crops target the coat-protein gene of
53.7 million hectares by 2015. Genetically mod- the targeted virus. The transgene prevents the
ified soybean also increased rapidly from their virus from replicating successfully in the host
introduction in 1996 to over 92 million hectares plant. Commercially grown virus resistant variet-
in 2015 (James 2015). ies of papaya were first introduced in the state of
Testing 521

Hawaii in 1998. Virus resistant squash ws also proteins are expressed at very low levels. An
commercialized in the United States in the late example of an immunochromatography based
1990s NAS, 2016). method, also called “dipstick” is seen in Fig. 14.7.
A second approach is to test for the fragments
such as 34S, 35S and NOS using either PCR or
Testing RT-PCR with several commercial test protocols
with kits available to test for the specific insert.
The ability to determine whether a crop has been Obviously before any of these techniques, sam-
genetically modified is important since consum- ples need to be extracted and prepared for analy-
ers and regulators require that information. There sis using one of several techniques available. In
are two basic types of testing that is performed on qualitative PCR, the specifity of DNA poly-
selected commodities; protein and DNA. In the merase is used to allow for amplification of target
development of the gene sequence for a crop, the sequences. In standard PCD, two pairs of primers
new gene is sandwiched between two segments; are used with one being a sense sequence and the
a promoter and a terminator. There are a number other being antisense. These sequences are ampli-
of promoter and terminator segments which fied numbers of times approaching a million.
come from a novel source hence are readily iden- After the amplification, these segments can be
tified. Two of the most common promoter seg- separated by agarose gel electrophoresis but
ments are 34S and 35S which come from other techniques such as HPLC have been used.
Cauliflower Mosaic Virus (CaMV) and the The approach that is an alternative to the qualita-
Figwort Mosaic Virus (FMV). A relatively com- tive PCR is Quantitative Real time PCR in which
mon terminator marker is NOS from Nopaline the separation of fragments is performed auto-
Synthase. When testing for GM content there are matically. Should an organization not choose to
two approaches. In the first approach, one can perform testing, there are several contract labs
test for the expressed protein using an ELISA or that can perform this assay. (Ahmed 2002).
an Immunochromatography method. ELISA tests While there is interest in the various technol-
for a large number of compounds have been in ogy involved in GMO testing, in recent years a
use for decades. While these are useful, the num- new phenomenon has come into being which is
ber of possible proteins to test for is limited and GMO verification services with the most visible

Fig. 14.7 Sample
lateral flow device
522 14  Genetically Modified Crops

that this did not trigger labeling. Regulation


1998/1139/EC which regulated Round Up Ready
soy and BT137 maize and 1997/1813/EC required
labeling of biotech corn and soy based on the
presence of transgenic protein or DNA. These
regulations created additional confusion as no
thresholds were established and 2000/49/EC and
2000/50/EC introduced a 1% labeling threshold
Fig. 14.8  Non-GMO project verified symbol for adventitious material which is defined as
material that is contained in the food even after
all attempts to exclude it. Over the next 3 years
there was an evolution of the regulations with
2003/1829/EC/and 2003/1830/EC which added
additional biotech crops past Round Up Ready
Soy and BT maize and added feed in addition to
food. The current level that triggers labeling is
0.9% GMO content but there is still confusion on
this topic as there can be different interpretations
as what this means. Obviously unapproved events
are not allowed. In contrast to Europe, Japan has
focused on the GMO content of final food prod-
uct rather than ingredients with a 5% level
Fig. 14.9  USDA PVP symbol (European Union 2011).
The US does not have universal labeling
requirements but two bills have been introduced
being the non-GMO Verification Project. The in the US congress that would have standardized
project has numerous requirements with the final labeling with one of them recently defeated. In a
result being that a manufacturer then can add the parallel fashion a number of states have passed
symbol to their product (Fig. 14.8). laws requiring GMO labeling with Vermont
Additionally, in late 2015, the USDA being the farthest along with food manufacturers
announced a Process Verified Program (PVP) preparing for its implementation in 2017
which allows a supplier manufacturer to place (Figs. 14.10 and 14.11).
another type of symbol on their project
(Fig. 14.9).
Future Challenges

Regulation As this topic area evolves, there are going to be


challenges on a number of fronts. With the total
At the time this chapter is being written, the regu- number of approved events in the hundreds, a cat-
latory landscape is unclear since countries and egory of unapproved events continues to grow. A
now individual states in the US are developing paper published in the International Journal of
action levels that could trigger a requirement for Food Contamination on the GMO Contamination
labeling a product as containing GMO. In 1997 Register between the years of 1997 and 2013
the EC developed the novel food regulation indicated that it had recorded 396 incidents
1997/258/EC. For GMO containing foods it across 63 countries (Price and Cotter 2014). An
required evidence that foods were safe for human in-depth analysis revealed that rice had the high-
consumption and required labeling if foods were est number accounting for almost 1/3 of the inci-
not substantially equivalent. It was interesting dents even though there are is no commercial
Future Challenges 523

Fig. 14.10 Pro-GMO
Labeling Ad

selected pharmaceuticals. Finally this chapter


presents a snapshot of this topic area in early
2016 with changes occurring on a regular basis.

GMO Dictionary

0.9%: The level used in EU countries to deter-


mine labeling thresholds.
35S: Promoter DNA fragment from CaMV
used as marker to indicate GM content. Also used
as a marker when testing samples.
Fig. 14.11  Example of a GMO label
34S: Promoter DNA fragment from FMV
used as marker to indicate GM content. Also used
growing of gm rice anywhere in the world. The as a marker when testing samples.
majority of the rice incidents occurred with 17,025: A number indicating a lab has met
LLRICE in the US and BT63 Rice from China certain quality requirements. Many customers
with the conclusion that the detection of these require that 17,025 accredited labs perform the
unapproved events being dependent on both rou- analysis of their samples
tine and targeted monitoring. As a corollary to Adventitious Contamination The presence of
these developments, a review of test kits whether GMOs in traditional crops is difficult to avoid.
it be for expressed protein or genetic markers is Minute traces in food products are tolerated if
limited indicating the need for additional empha- their presence is accidental or the result of techni-
sis in the area. A final concern is the need to feed cally unavoidable contamination during growing,
an ever increasing world population making the harvesting, transport or processing.
need for more efficient production of food. In a Base Building blocks of the nucleic acids
recent article in Nature titled “India needs Home DNA and RNA. Four bases are present in DNA:
Grown GM Food to Stop Starvation”, the author adenine (A), cytosine (C), guanine (G) and thy-
of this commentary stresses the need for India to mine (T). In RNA, thymine is replaced by uracil
develop “homegrown” GM IP focusing on com- (U). These four bases encode the genetic infor-
modities that are critical to the country. mation; thus, the four letters A, C, G and T are
The area of transgenic crops is going to con- sometimes called “the alphabet of life”.
tinue to evolve with not only development in food Bt A protein that is toxic to chewing insects
for human consumption but also the continuing and is produced by the soil bacterium Bacillus
developments in Pharm animals where modifica- thuringiensis and has long been used as a bio-
tions are made providing for the development of logical pesticide.
524 14  Genetically Modified Crops

Chromosome: The self-replicating genetic herbicides by the incorporation of certain gene(s)


structure of cells, containing genes, which deter- either through genetic engineering or traditional
mines inheritance of traits. Chemically, each breeding methods. The genes allow the herbicides
chromosome is composed of proteins and a long to be applied to the crop to provide effective weed
molecule of DNA. control without damaging the crop itself.
Dipstick; Defined as immunochromatography Identity preservation: The segregation of one
and used on commodities in the field to determine crop type from another at every stage from pro-
GM content. Not suitable for processed foods. duction and processing to distribution. This pro-
Uses same technique as home pregnancy test kits. cess is usually performed through audits and site
Cross-pollination: Fertilization of a plant with visits and provides independent third-party veri-
pollen from another plant. Pollen may be trans- fication of the segregation.
ferred by wind, insects, other organisms, or Insecticide resistance: The development or
humans. selection of heritable traits (genes) in an insect
DNA (deoxyribonucleic acid): The chemical population that allow individuals expressing the
substance from which genes are made. DNA is a trait to survive in the presence of levels of an
long, double-stranded helical molecule made up insecticide (biological or chemical control agent)
of nucleotides which are themselves composed that would otherwise debilitate or kill this species
of sugars, phosphates, and derivatives of the four of insect. The presence of such resistant insects
bases adenine (A), guanine (G), cytosine (C), and makes the insecticide less useful for managing
thymine (T). The sequence order of the four pest populations.
bases in the DNA strands determines the genetic Insect-resistance management: A strategy for
information contained. delaying the development of pesticide resistance
EU Labeling: see 0.9%. by maintaining a portion of the pest population in
Event: A set of trait in a plant giving it unique a refuge that is free from contact with the insecti-
properties such as herbicide resistance with the cide. For Bt crops this allows the insects feeding
provider of the traits having IP. on the Bt toxin to mate with insects not exposed
GMO Analysis: A series of steps including to the toxin produced in the plants.
extraction, isolation, analysis and interpretation Insect-resistant crops: Plants with the ability
of data. to withstand, deter or repel insects and thereby
Genetic engineering: Manipulation of an prevent them from feeding on the plant. The traits
organism’s genes by introducing, eliminating or (genes) determining resistance may be selected
rearranging specific genes using the methods of by plant breeders through cross-pollination with
modern molecular biology, particularly those other varieties of this crop or through the intro-
techniques referred to as recombinant DNA duction of novel genes such as Bt genes through
techniques. genetic engineering.
Genetically engineered organism (GEO): An Intellectual property rights: The legal protec-
organism produced through genetic engineering. tion for inventions, including new technologies
Genetic modification: The production of heri- or new organisms (such as new plant varieties).
table improvements in plants or animals for spe- The owner of these rights can control their use
cific uses, via either genetic engineering or other and earn the rewards for their use. This encour-
more traditional methods. Some countries other ages further innovation and creativity for the ben-
than the United States use this term to refer spe- efit of us all. Intellectual property rights protection
cifically to genetic engineering. includes various types of patents, trademarks,
Genetics: The study of the patterns of inheri- and copyrights.
tance of specific traits. Molecular biology: The study of the structure
Genome: All the genetic material in all the and function of proteins and nucleic acids in bio-
chromosomes of a particular organism. logical systems.
Herbicide-tolerant crops: Crops that have been Nucleotide: A subunit of DNA or RNA con-
developed to survive application(s) of particular sisting of a nitrogenous base (adenine, guanine,
Future Challenges 525

thymine, or cytosine in DNA; adenine, guanine, Non-GMO: An internal term indicating a


uracil, or cytosine in RNA), a phosphate mole- product or ingredient does not require llabeling
cule, and a sugar molecule (deoxyribose in DNA of GM content
and ribose in RNA). Many of nucleotides are Non-GE: See Non-GMO
linked to form a DNA or RNA molecule. Test Free: Test results indicating the sample
Plant-incorporated protectants (PIPs): was Test negative or had detectable but no quan-
Pesticidal substances introduced into plants by tifiable GM content
genetic engineering that are produced and used Test Negative: Test results indicating no
by the plant to protect it from pests. The protein detectable GM content
toxins of Bt are often used as PIPs in the forma- Quantitative Result: A test result indicating
tion of Bt crops. the presence or absence of GM content. A result
Polymerase chain reaction (PCR): A tech- indication the presence of Gm content would like
nique used to create a large number of copies of a trigger the need for a quantitative test
target DNA sequence of interest. One use of PCR Qualitative Result; A test result resulting in
is in the detection of DNA sequences that indi- how much GM content in a sample
cate the presence of a particular genetically engi- RTPCR; The final step in GMO analysis when
neered organism (Vollenhofer et al. (1999)). isolate DNA are amplified 2,000,000 times and
Promoter: A region of DNA that regulates the are able to be detected
level of function of other genes. Also see 35S NOS: Terminator DNA fragment from
and 34S. Nopaline Synthase ad used as marker to indicate
Protein: A molecule composed of one or more GM content. Also used as a marker when testing
chains of amino acids in a specific order. Proteins samples
are required for the structure, function, and regu- Real Time PCR: See RTPCR
lation of the body’s cells, tissues, and organs, and Round up: A type of herbicide used to elimi-
each protein has a unique function. nate weeds. Chemically known as glyphosate
Recombinant DNA (rDNA): A molecule of Roundup Ready: A particular type of plant
DNA formed by joining different DNA segments resistant to Roundup allowing for increased
using recombinant DNA technology. productivity
Recombinant DNA technology: Procedures Ingredients
used to join together DNA segments in a cell-free
system (e.g. in a test tube outside living cells or
organisms). Under appropriate conditions, a Flour
recombinant DNA molecule can be introduced
into a cell and copy itself (replicate), either as an There are currently no approved non-GMO wheat
independent entity (autonomously) or as an inte- events. Additionally, based on analytical data and
gral part of a cellular chromosome. discussions with laboratories and vendors, it is
Ribonucleic Acid (RNA): A chemical sub- unlikely that one will be able to finds a test nega-
stance made up of nucleotides compound of sug- tive source of flour. There are vendors that sell a
ars, phosphates, and derivatives of the four bases non-GMO flour that tests positive. What this
adenine (A), guanine (G), cytosine (C), and uracil means is the samples likely have been tested with
(U). RNAs function in cells as messengers of result indicating a detectable but not quantifiable
information from DNA that are translated into result. One of the reasons for this phenomena is a
protein or as molecules that have certain structural concept called adventitious contamination. In the
or catalytic functions in the synthesis of proteins. case of flour, this concept mean that in each load of
RNA is also the carrier of genetic information for wheat there can be several percent GMO corn or
certain viruses. RNAs may be single or double soy resulting in a detectable result for the flour and
stranded. products made with it. A follow on quantitative
Terminator A segment of DNA indicating the determination will not be successful as the sample
end of a particular gene sequence. See NOS contains not quantifiable DNA. The final result is
526 14  Genetically Modified Crops

that almost 100% of flour will test positive. This Joos, H., Timmerman, B., Montagu, M. V., & Schell,
J. (1983). Genetic analysis of transfer and stabiliza-
should be taken into account as we move forward.
tion of Agrobacterium DNA in plant cells. The EMBO
Sunflower Lecithin: With increasing pressure Journal, 2(12), 2151–2160.
on IP soy lecithin supplies, manufacturers have Maluszynsk, M. K., Nichterlein, K., van Zanten, L., &
looked for alternatives such as sunflower lecithin. Ahloowalia, B. S. (2000). Officially released mutant
varieties – the FAO/IAEA Database. Mutation
There currently is no GMO sunflower.
Breeding Review, 12, 1–84.
Mba, C. (2013). Induced mutations unleash the potentials
of plant genetic resources for food and agriculture.
References Agronomy, 3, 200–231.
NAS (National Academy of Sciences). (2016). Genetically
engineered crops: Experiences and prospects. NAS
Ahmed, F. E. (2002). Detection of genetically modified
Press. Retrieved from http://www.nap.edu/23395.
organisms in food. TRENDS in Biotechnology, 20(5),
Ossowski, S. K., Schneeberger, J. I., Lucas-Lledo, N.,
215–223.
Warthmann, R. M., Clark, R. G., Shaw, D., Weigel, D.,
Ahloowali, B. S. (2004). Global impact of mutation-­
& Lynch, M. (2010). The rate and molecular spectrum
derived varieties (PDF). Euphytica, 135, 187–204.
of spontaneous mutations in Arabidopsis thaliana.
Cohen, S. N., Chang, A. C. Y., Boyer, H., & Helling, R. B.
Science, 327, 92–94.
(1973). Construction of biologically functional bac-
Popping, B. (2010). Genetically modified organisms.
terial plasmids in vitro. Proceedings of the National
In B. Popping, C. Diaz-Amigo, & Hoenicke (Eds.),
Academy of Sciences of the United States of America,
Molecular biological and immunological -techniques
70, 3240–3244.
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European Union. (2011). Overview on the detection,
New York: Wiley.
interpretation and reporting on the presence of unau-
Price, B., & Cotter, J. (2014). The GM contamination reg-
thorised genetically modified materials ENGL ad hoc
ister: A review of recorded contamination incidents
working group on “unauthorised GMOs”. Guidance
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Laboratories (ENGL) European Commission Joint
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Research Centre’ Institute for Health and Consumer
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Protection Luxembourg: Publications Office of the
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Roychowdhury, R., & Tah, J. (2013). Mutagenesis—A
FAO/IAEA. (2014). Plant Breeding and Genetics Joint
potential approach for crop improvement. In K. R.
FAO/IAEA Division of Nuclear Techniques in Food
Hakeem, P. Ahmad, & M. Ozturk (Eds.), Crop
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Arabidopsis thaliana transformants without selection
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the Acquisition of Agri-biotech Applications.
Additives and Contaminants
15
W. Jeffrey Hurst, John W. Finley,
and John M. deMan

Introduction Food Toxins

For centuries, ingredients man has utilized food In the 1500s Paracelsus expressed the classic
additives to improve and preserve foods. For cen- toxicology maxim “All things are poison and
turies, we have used salt to preserve meats and nothing is without poison; only the dose makes a
fish, added herbs and spices to improve the flavor thing not a poison.” This is often condensed to:
of foods, preserved fruit with sugar, and pickled “The dose makes the poison”.
vegetables like in a vinegar solution. Consumers There is also the possibility of harmful or toxic
now have the expectation of flavorful, nutritious, substances entering the food supply unintention-
safe, convenient, colorful and affordable. Food ally through direct contamination, through envi-
additives and processing techniques help =deliver ronmental pollution, as a result of processing or
foods with these attributes. There is also a strong through deliberate adulteration for economic ben-
consumer desire for clean labels and foods with- efit. In addition, many foods can contain toxic
out additives. This presents the food chemist with materials. It should be kept in mind that most
many conundrums about the use of additives. chemicals have a safe range of use but at very high
There are thousands of ingredients approved doses can be toxic. Most compounds that we
for use in foods. The Food and Drug would consider in food have a dose response
Administration (FDA) maintains a list of over where they are inconsequential at low doses then
3000 ingredients in its data base “Everything can become toxic and very toxic. This can be seen
Added to Food in the United States”, many of with ingredients ranging from pesticides to com-
which we use at home every day (e.g., sugar, bak- mon ingredients like salt. The concentration effects
ing soda, salt, vanilla, yeast, spices and colors) are very different but the typical dose response
partially sliced loaf of bread. curve is similar for most chemicals or food ingre-
All food additives are carefully regulated by dients. The key is to identify the safe level and
federal authorities and various international orga- typically we identify the “no adverse effect level”
nizations to ensure that foods are safe to eat and (NOAEL). This can be seen in Fig. 15.1.
are accurately labeled. The toxicity of some materials will differ
In addition to ingredients that are added greatly among individuals. There is a distribution
directly for benefits foods can contain toxic mate- what concentration of a material will affect indi-
rials that are produced by the plant, animal, fun- viduals. There are variations in sensitivity to even
gus or as a result of microbial contamination, the most common food ingredients. For example,

© Springer International Publishing AG 2018 527


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8_15
528 15  Additives and Contaminants

rials represent both toxins produced by the spe-


cies and contaminants resulting from
microbiological activity such as aflatoxins.
Fungal toxins, also called mycotoxins, are
produced by fungi or molds. Most of the interest
in fungal toxins is concerned with the so-called
storage fungi, molds that grow on relatively dry
cereals and oilseeds. These belong to two com-
mon genera, Aspergillus and Penicillium. The
most common of the fungal toxins are the afla-
toxins formed by members of the Aspergillus fla-
vus group. The aflatoxins were discovered as a
Fig. 15.1  Dose response curve for an ingredient or a drug result of widespread poisoning of turkeys in the
illustrating the NOAEL early 1960s in England through feeding of toxic
peanut meal. The aflatoxins belong to the most
some populations are sensitive to salt while the powerful toxins known and are highly carcino-
bulk of the population is unaffected. Figure 15.2 genic. A dose of 1 mg given to rats for short or
illustrates the distribution in sensitivity to food, long periods can result in liver cancer, and a diet
drugs or toxins for the same dose of a food ingre- containing 0.1 ppm of aflatoxin produces liver
dient or a drug.aflatoxin. tumors in 50% of male rats (Spensley 1970).
Many naturally occurring food compounds There are at least eight aflatoxins, of which the
may be toxic as naturally-occurring constituent, more important are designated B1, B2, G1, and
microbial activity handling or processing, the G2. The names result from the blue and green
incidence of adverse reactions from food is rela- fluorescence of these compounds when viewed
tively low. The low incidence of adverse effects is under ultraviolet light. Aflatoxin B1 is a very
the result solutions by the US Food and Drug powerful liver carcinogen; a level of 15 ppb in the
Administration (FDA) and other regulatory agen- diet of rats resulted in tumors in 100% of cases
cies. The application of specifications, action lev- after 68 weeks (Scott 1969). Ducklings are used
els, tolerances, warning labels and prohibitions as test animals because they are especially sensi-
have been effective in controlling the number of tive to aflatoxins. The aflatoxins, for which the
adverse events from food consumption. formulas are shown in Fig. 15.3, can occur in
Regardless of preventative measures to protect many foods but are particularly common in pea-
consumers from natural food toxins, consump- nuts. Roasting of peanuts reduces the level of
tion of small levels of these materials is inevita- aflatoxin; for example, roasting for a half hour at
ble. The risk for toxicity due to consumption of 150° may reduce aflatoxin B1 content by as much
food toxins is relatively low, however, there is as 80% (Scott 1969). However, aflatoxin may
always the possibility of toxicity due to contami- still be carried over into peanut butter. In addi-
nation, overconsumption, allergy or an unpre- tion, aflatoxins have been found in cottonseed
dictable idiosyncratic response. meal, rice, sweet potatoes, beans, nuts, and
Sources of food born toxins include certain wheat. Through ingestion of moldy feed by ani-
sea foods, microbial contamination and produc- mals, aflatoxins may end up as contaminants in
tion of mycotoxins and natural toxins present in milk and meat. Aflatoxins found in milk may be
plant fungal and sea foods. The increased use of M1 or M2, where M stands for metabolic; these
dietary supplements by consumers also present are also toxic. The development of aflatoxins
sources of natural toxins. Table 15.1 contains a depends very much on temperature and moisture
list of a few natural toxins found in the food sup- conditions. With peanuts, contamination occurs
ply. These materials cause a range of symptoms mostly during the drying period. Improper drying
from gastrointestinal upset to death. These mate- and storage are responsible for most of the
Food Toxins 529

Fig. 15.2 Typical
distribution of
sensitivities to food
ingredients or drugs in a
population

Table 15.1  Natural toxins found in foods, their food source and effects when consumed
Toxic material Food source Effects
Ciguatoxin from Dinoflagellates Consumption of subtropical and tropical Combination of gastrointestinal,
(marine algae) in the genus marine finfish that have accumulated neurological, and, occasionally,
Gambierdiscus ciguatoxins through their diets cardiovascular disorders
Shellfish poisoning is caused by Paralytic shellfish poisoning(PSP) PSP—can be deadly
a group of toxins produced by Diarrhetic shellfish poisoning(DSP) DSP—nausea, vomiting, diarrhea
planktonic algae consumed by Neurotoxic shellfish poisoning (NSP) GI effects plus NSP-Neurological loss
shellfish of short-term memory
Amnesic shellfish poisoning(ASP) (ASP) and AZP nausea, vomiting,
Azaspiracid shellfish poisoning(AZP) diarrhea
Scombrotoxin is a combination Scombrotoxin is a combination of Tingling or burning in or around the
of substances, histamine substances, histamine prominent among mouth or throat, rash or hives, drop in
prominent among them them. Histamine is produced during blood pressure, headache, dizziness,
decomposition of fish itching of the skin, nausea, vomiting,
diarrhea
Tetrodotoxin (TTX) and related Pufferfish Multiple species Muscle is Death is from respiratory-muscle
compounds (e.g. edible Toxin is in gonads (mainly paralysis and usually occurs within
4,9-anhydroTTX, 4-epiTTX, ovary), liver, intestines, and skin can 4–6 h, with a known range of about
11-deoxyTTX, tetrodonic acid) contain levels of tetrodotoxin sufficient 20 min to 8 h if victims survive the initial
to produce rapid death 24 h, they are expected to recover full
Mushroom toxins: Amanitin, Consumption of raw or cooked fruiting Protoplasmic poisons—life-­
Gyromitrin, Orellanine, bodies (mushrooms, toadstools) of a threatening poisons
Muscarine, Ibotenic Acid, number of species of higher fungi. T Neurotoxins—profuse sweating, coma,
Muscimol, Psilocybin, Coprine convulsions, hallucinations,
excitement, depression, spastic colon
Gastrointestinal irritants—transient
nausea, vomiting, abdominal
cramping, and diarrhea
Aflatoxins (AFs) are mycotoxins Produced mainly by certain strains of Cancer, impaired protein formation,
produced by certain fungi and Aspergillus flavus and A. parasiticus impaired blood coagulation, toxic
can cause serious illness in hepatitis, AFB1 is the most potent
animals and humans known natural carcinogen and is the
The four major aflatoxins are Occur in a broad range of agricultural most abundant of the AFs
AFB1, AFB2, AFG1, and AFG2 commodities, such as corn and nuts
Pyrrolizidine alkaloids are a Widely distributed in the plant pain, particularly in the right upper
large class of naturally occurring kingdom, particularly in the part of the abdomen; nausea; vomiting;
alkaloids containing Boraginaceae, Compositae, and swollen belly; swollen veins on the
pyrrolizidine rings Leguminosae families belly; puffiness from fluid; and fever
Phytohaemagglutinin (kidney Red kidney bean (Phaseolus vulgaris) Usually begins with extreme nausea
bean lectin) poisoning and kinkoti bean poisoning and vomiting within 1–3 h of ingestion
are examples of names for the illness of the product, with diarrhea
caused by phytohaemagglutinin developing later
Compiled from: www.fda.gov/food/foodborneillnesscontaminants/causesofillnessbadbugbook/ucm2006773.htm
530 15  Additives and Contaminants

Fig. 15.3 Chemical
structure of aflatoxins B1,
B2, G1, G2, M1 and M2

c­ ontamination. This has been found to apply for ation. Experiments on milling showed that the
rice. Optimum conditions for growth of vomitoxin was distributed throughout the milled
Aspergillus flavus are 25° to 40 °C with a relative products and was not destroyed by the bread-­
humidity greater than 85%. making process. Patulin is another Aspergillus
Sterigmatocystin, which is a carcinogenic metabolite and has been indicated as a food con-
metabolite of Aspergillus ochraceus, has been taminant, especially in fruits, as a result of stor-
found to be a natural contaminant of foods, espe- age rot. It has been found as a constituent of apple
cially com. Molds of the species Fusarium pro- juice (Harwig et al. 1973).
duce several mycotoxins in countries with Control measures for prevention of aflatoxin
moderate climates (Andrews et al. 1981). Two of production focus on reduction of water activity to
these are zearalenone and deoxynivalenol a point where the fungus is unable to grow and
(Fig.  15.4). Zearalenone, of F-2 toxin, is pro- maintenance of low water activity during storage.
duced by Fusarium molds that grow on com A moisture content of 18.0–19.5% in cereal
(Marasas et al. 1979) that is immature or high in grains is required for growth and toxin produc-
moisture at harvest. Deoxynivalenol, also known tion by A. flavus. Aflatoxin-contaminated com-
as vomitoxin, has been found in wheat and barley modities can be detoxified by treatment with
(Trenholm et al. 1981; Scott et al. 1983). During ammonia, calcium hydroxide, or a combination
the wet summer of 1980, wheat grown in Ontario of formaldehyde and calcium hydroxide
showed sprouting of kernels and pink discolor- (Palmgren and Hayes 1987).
Food Additives 531

Fig. 15.4 Chemical
structures of
sterigmatocystin,
ochratoxin A,
zearalenone,
deoxynivalenol
(vomitoxin), and patulin

Food additives can be divided into two major


Food Additives groups, intentional additives and incidental addi-
tives. Intentional additives are chemical sub-
The term food additive means any substance the stances that are added to food for specific
intended use of which results, or may reasonably purposes. All additives whether intentional, uun-
be expected to result, directly or indirectly in its intentional or incidental additives are regulated
becoming a component or otherwise affecting the by strict governmental controls. The U.S. law
characteristics of any food (including any sub- governing additives in foods is the Food Additives
stance intended for use in producing, manufac- Amendment to the Federal Food, Drug and
turing, packing, processing, preparing, treating, Cosmetic Act of 1958. According to this act, a
packaging, transporting, or holding food; and food additive is defined as follows:
including any source of radiation intended for Any substance that is reasonably expected to
any such use), if such a substance is not generally become a component of food is a food additive that
recognized, among experts qualified by scientific is subject to premarket approval by FDA, unless the
training and experience to evaluate its safety, as substance is generally recognized as safe (GRAS)
among experts qualified by scientific training and
having been adequately shown through scientific experience to evaluate its safety under the condi-
procedures (or, in the case of a substance used in tions of its intended use, or meets one of the other
food prior to January 1, 1958, through either sci- exclusions from the food additive definition in sec-
entific procedures or experience based on com- tion 201(s) of the Federal Food, Drug, and Cosmetic
Act (FFDCA). Any food additive that is intended to
mon use in food) to be safe under the condition of have a technical effect in the food is deemed unsafe
its intended use; except that such a term does not unless it either conforms to the terms of a regulation
include pesticides, color additives and substances prescribing its use or to an exemption for investiga-
for which prior sanction or approval was granted. tional use. Otherwise, in accordance with section
409 of the Act, the substance is deemed an unsafe
532 15  Additives and Contaminants

food additive. Any food that contains an unsafe ness, including life-threatening botulism. One
food additive is adulterated under section 402(a)(2) group of preservatives—antioxidants—prevents
(C) of the FFDCA. fats and oils and the foods containing them from
Similarly, any substance that is added to food becoming rancid or developing an off-flavor. They
and imparts color to the food is a color additive also prevent cut fresh fruits such as apples from
(see color additive definition in §201(t) of the turning brown when exposed to air.
FFDCA and 21 CFR 70.3(f)). Any color additive in To Improve or Maintain Nutritional Value:
food is deemed unsafe unless its use is either per- Vitamins and minerals (and fiber) are added to
mitted by regulation or exempt by regulation. many foods to make up for those lacking in a per-
Unlike the definition for food additive, there is no son’s diet or lost in processing, or to enhance the
GRAS exemption for color additives. Any food nutritional quality of a food. Such fortification and
that contains an unsafe color additive is adulterated enrichment has helped reduce malnutrition in the
under section 402(c) of the FFDCA. U.S. and worldwide. All products containing added
The decision tree below will help in determin- nutrients must be appropriately labeled.
ing the regulatory status of a food ingredient. It is Improve Taste, Texture and Appearance: Spices,
the responsibility of the manufacturer of any food natural and artificial flavors, and sweeteners are
to ensure that all ingredients used are of food- added to enhance the taste of food. Food colors
grade purity and comply with specifications and maintain or improve appearance. Emulsifiers, stabi-
limitations in all applicable authorizations. The lizers and thickeners give foods the texture and con-
overall regulatory status of a food is affected by the sistency consumers expect. Leavening agents allow
regulatory status of each individual food ingredi- baked goods to rise during baking. Some additives
ent. To determine compliance, consider each help control the acidity and alkalinity of foods,
authorization to be composed of three parts: while other ingredients help maintain the taste and
appeal of foods with reduced fat content.
• The identity of the substance,
• Specifications including purity and physical The use of food additives strictly regulated by
properties, and national and international laws. The National
• Limitations on the conditions of use.
Academy of Sciences (1973) has listed the pur-
To assure a customer that an ingredient that is poses of food additives as follows:
being shipped to them is not adulterated or mis-
branded, the ingredient manufacturer may want to
provide a letter of guaranty with the shipment (see • to improve or maintain nutritional value
21 CFR 7.13 for suggested forms of guaranty). • to enhance quality
• to reduce wastage
Food additives perform a variety of useful • to enhance consumer acceptability
functions in foods. Food ingredients that are • to improve keeping quality
intentionally introduced into foods to aid in pro- • to make the food more readily available
cessing, to act as preservatives, or to improve the • to facilitate preparation of the food
quality of the food are called intentional addi-
tives. Food ingredients have been used for many The use of food additives is in effect a food
years to preserve, flavor, blend, thicken and color processing method, because both have the same
foods, and have played an important role in objective—to preserve the food and/or make it
reducing serious nutritional deficiencies among more attractive. In many food processing tech-
consumers. These ingredients also help ensure niques, the use of additives is an integral part of
the availability of flavorful. the method, as is smoking, heating, and ferment-
The FDA defines the utility of food additives ing. The National Academy of Sciences (1973)
as follows: has listed the following situations in which addi-
To Maintain or Improve Safety and Freshness: tives should not be used:
Preservatives slow product spoilage caused by
mold, air, bacteria, fungi or yeast. In addition to • to disguise faulty or inferior processes
maintaining the quality of the food, they help con- • to conceal damage, spoilage, or other inferiority
trol contamination that can cause foodborne ill-
• to deceive the consumer
Food Additives 533

• if use entails substantial reduction in impor- cessed foods make the use of chemical food pre-
tant nutrients servatives imperative. Some of the commonly
• if the desired effect can be obtained by eco- used preservatives—such as sulfites, nitrate, and
nomical, good manufacturing practices salt—have been used for centuries in processed
• in amounts greater than the minimum neces- meats and wine. The choice of an antimicrobial
sary to achieve the desired effects agent has to be based on a knowledge of the
antimicrobial spectrum of the preservative, the
There are several ways of classifying inten- chemical and physical properties of both food
tional food additives. One such method lists the and preservative, the conditions of storage and
following three main types of additives: handling, and the assurance of a high initial
quality of the food to be preserved (Davidson
1. complex substances such as proteins or
and Juneja 1990).
starches that are extracted from other foods
(for example, the use of caseinate in sausages
and prepared meats) Benzoic Acid
2. naturally occurring, well-defined chemical

compounds such as salt, phosphates, acetic Benzoic acid occurs naturally in many types of
acid, and ascorbic acid berries, plums, prunes, and some spices. As an
3. substances produced by synthesis, which may additive, it is used as benzoic acid or as benzoate.
or may not occur in nature, such as coal tar The latter is used more often because benzoic
dyes, synthetic β-carotene, antioxidants, pre- acid is sparsely soluble in water (0.27% at 18 °C)
servatives, and emulsifiers and sodium benzoate is more soluble
(66.0 g/100 mL at 20 °C). The undissociated
Some of the more important groups of inten- form of benzoic acid is the most effective antimi-
tional food additives and their functions are crobial agent. With a pKa of 4.2, the optimum pH
described in Table 15.2. range is from 2.5 to 4.0. This makes it an effec-
Under the Food Additives Amendment, two tive antimicrobial agent in high-acid foods, fruit
groups of ingredients were exempted from the drinks, cider, carbonated beverages, and pickles.
regulation process. It is also used in margarines, salad dressings, soy
GROUP I—Prior-sanctioned substances—are sauce, and jams.
substances that FDA or USDA had determined
safe for use in food prior to the 1958 amendment.
Examples are sodium nitrite and potassium nitrite Parabens
used to preserve luncheon meats.
GROUP II—GRAS (generally recognized as Parabens are alkyl esters of p-hydroxybenzoic
safe) ingredients—are those that are generally acid. The alkyl groups may be one of the
recognized by experts as safe, based on their ­following: methyl, ethyl, propyl, butyl, or heptyl.
extensive history of use in food before 1958 or Parabens are colorless, tasteless, and odorless
based on published scientific evidence. Among (except the methyl paraben). They are nonvolatile
the several hundred GRAS substances are salt, and nonhygroscopic. Their solubility in water
sugar, spices, vitamins and monosodium gluta- depends on the nature of the alkyl group; the lon-
mate (MSG). Manufacturers may also request ger the alkyl chain length, the lower the solubil-
that FDA review the industry’s determination of ity. They differ from benzoic acid in that they
GRAS Status (Hall 1975). have antimicrobial activity in both acid and alka-
Preservatives or antimicrobial agents play an line pH regions.
important role in today’s supply of safe and sta- The antimicrobial activity of parabens is pro-
ble foods. Increasing demand for convenience portional to the chain length of the alkyl group.
foods and reasonably long shelf life of pro- Parabens are more active against molds and
534

Table 15.2  Common food additives and their functions


Types of ingredients Applications Examples of uses Products in foods
Preservatives Prevent food spoilage from bacteria, molds, Fruit sauces and jellies, beverages, Ascorbic acid, citric acid, sodium benzoate, calcium
fungi, or yeast (antimicrobials); slow or prevent baked goods, cured meats, oils and propionate, sodium erythorbate, sodium nitrite, calcium
changes in color, flavor, or texture and delay margarines, cereals, dressings, sorbate, potassium sorbate, BHA, BHT, EDTA, tocopherols
rancidity (antioxidants); maintain freshness snack foods, fruits and vegetables (Vitamin E)
Sweeteners Add sweetness with or without the extra Beverages, baked goods, Sucrose (sugar), glucose, fructose, sorbitol, mannitol, corn
calories confections, table-top sugar, syrup, high fructose corn syrup, saccharin, aspartame,
substitutes, many processed foods sucralose, acesulfame potassium (acesulfame-K), neotame
Color additives Offset color loss due to exposure to light, air, Many processed foods, (candies, FD&C Blue Nos. 1 and 2, FD&C Green No. 3, FD&C Red
temperature extremes, moisture and storage snack foods margarine, cheese, soft Nos. 3 and 40, FD&C Yellow Nos. 5 and 6, Orange B, Citrus
conditions; correct natural variations in color; drinks, jams/jellies, gelatins, Red No. 2, annatto extract, beta-carotene, grape skin extract,
enhance colors that occur naturally; provide pudding and pie fillings) cochineal extract or carmine, paprika oleoresin, caramel color,
color to colorless and “fun” foods fruit and vegetable juices, saffron (Note: Exempt color additives
are not required to be declared by name on labels but may be
declared simply as colorings or color added)
Flavors and spices Add specific flavors (natural and synthetic) Pudding and pie fillings, gelatin Natural flavoring, artificial flavor, and spices
dessert mixes, cake mixes, salad
dressings, candies, soft drinks, ice
cream, BBQ sauce
Flavor enhancers Enhance flavors already present in foods Many processed foods Monosodium glutamate (MSG), hydrolyzed soy protein,
(without providing their own separate flavor) autolyzed yeast extract, disodium guanylate or inosinate
Fat replacers (and Provide expected texture and a creamy Baked goods, dressings, frozen Olestra, cellulose gel, carrageenan, polydextrose, modified food
components of “mouth-feel” in reduced-fat foods desserts, confections, cake and starch, microparticulated egg white protein, guar gum, xanthan
formulations used to dessert mixes, dairy products gum, whey protein concentrate
replace fats)
Nutrients Replace vitamins and minerals lost in Flour, breads, cereals, rice, Thiamine hydrochloride, riboflavin (Vitamin B2), niacin,
processing (enrichment), add nutrients that macaroni, margarine, salt, milk, niacinamide, folate or folic acid, beta carotene, potassium
may be lacking in the diet (fortification) fruit beverages, energy bars, instant iodide, iron or ferrous sulfate, alpha tocopherols, ascorbic acid,
breakfast drinks Vitamin D, amino acids (L-tryptophan, L-lysine, L-leucine,
L-methionine)
15  Additives and Contaminants
Emulsifiers Allow smooth mixing of ingredients, prevent Salad dressings, peanut butter, Soy lecithin, mono- and diglycerides, egg yolks, polysorbates,
separation chocolate, margarine, frozen sorbitan monostearate
Keep emulsified products stable, reduce desserts
stickiness, control crystallization, keep
ingredients dispersed, and to help products
Food Additives

dissolve more easily


Stabilizers and Produce uniform texture, improve Frozen desserts, dairy products, Gelatin, pectin, guar gum, carrageenan, xanthan gum, whey
thickeners, binders, “mouth-feel” cakes, pudding and gelatin mixes,
texturizers dressings, jams and jellies, sauces
pH control agents Control acidity and alkalinity, prevent Beverages, frozen desserts, Lactic acid, citric acid, ammonium hydroxide, sodium
and acidulants spoilage chocolate, low acid canned foods, carbonate
baking powder
Leavening agents Promote rising of baked goods Breads and other baked goods Baking soda, monocalcium phosphate, calcium carbonate
Anti-caking agents Keep powdered foods free-flowing, prevent Salt, baking powder, confectioner’s Calcium silicate, iron ammonium citrate, silicon dioxide
moisture absorption sugar
Humectants Retain moisture Shredded coconut, marshmallows, Glycerin, sorbitol
soft candies, confections
Yeast nutrients Promote growth of yeast Breads and other baked goods Calcium sulfate, ammonium phosphate
Dough strengtheners Produce more stable dough Breads and other baked goods Ammonium sulfate, azodicarbonamide, L-cysteine
and conditioners
Firming agents Maintain crispness and firmness Processed fruits and vegetables Calcium chloride, calcium lactate
Enzyme Modify proteins, polysaccharides and fats Cheese, dairy products, meat Enzymes, lactase, papain, rennet, chymosin
preparations
Gases Serve as propellant, aerate, or create Oil cooking spray, whipped cream, Carbon dioxide, nitrous oxide
carbonation carbonated beverages
Adapted from: http://www.fda.gov/Food/IngredientsPackagingLabeling/FoodAdditivesIngredients/ucm094211.htm#types accessed 8/4/2016
535
536 15  Additives and Contaminants

yeasts than against bacteria, and more active Table 15.3 Applications of sorbates as antimicrobial
agents
against gram-positive than gram negative bacte-
ria. They are used in fruitcakes, pastries, and fruit Products Levels (%)
fillings. Methyl and propyl parabens can be used Dairy products: aged cheeses, processed 0.05–0.30
cheeses, cottage cheese, cheese spreads,
in soft drinks. Combinations of several parabens
cheese dips, sour cream, yogurt
are often used in applications such as fish prod- 0.03–0.30
Bakery products: cakes, cake mixes, pies,
ucts, flavor extracts, and salad dressings. fillings, mixes, icings, fudges, toppings,
doughnuts
Vegetable products: fermented vegetables, 0.02–0.20
Sorbic Acid pickles, olives, relishes, fresh salads
Fruit products: dried fruit, jams, jellies, juices, 0.02–0.25
fruit salads, syrups, purees, concentrates
Sorbic acid is a straight-chain, trans-trans unsat-
Beverages: still wines, carbonated and 0.02–0.10
urated fatty acid, 2,4-hexadienoic acid. As an noncarbonated beverages, fruit drinks,
acid, it has low solubility (0.15 g/100 mL) in low-calorie drinks
water at room temperature. The salts, sodium, or Food emulsions: mayonnaise, margarine, 0.05–0.10
potassium are more soluble in water. Sorbates are salad dressings
stable in the dry form; they are unstable in aque- Meat and fish products: smoked and 0.05–0.30
ous solutions because they decompose through salted fish, dry sausages
Miscellaneous: dry sausage casings, 0.05–0.30
oxidation. The rate of oxidation is increased at
semimoist pet foods, confectionery
low pH, by increased temperature, and by light
Source: Reprinted with permission from J.N. Sofos and
exposure. F.F. Busta, Sorbic Acid and Sorbates, in Antimicrobials in
Sorbic acid and sorbates are effective against Foods, RM. Davidson and A.L. Branen, eds., p. 62, 1993,
yeasts and molds. Sorbates inhibit yeast growth by courtesy of Marcel Dekker, Inc.
in a variety of foods including wine, fruit juice,
dried fruit, cottage cheese, meat, and fish prod- Table 15.4  Sources of SO2 and their content of active
ucts. Sorbates are most effective in products of SO2
low pH including salad dressings, tomato prod- Content of
ucts, carbonated beverages, and a variety of other Chemical Formula active SO2 (%)
foods. Sulfur dioxide SO2 100.00
The effective level of sorbates in foods is in Sodium sulfite, anhydrous Na2SO3 50.82
the range of 0.5–0.30%. Some of the common Sodium sulfite, Na2SO3·7 25.41
heptahydrate H2O
applications are shown in Table 15.3. Sorbates
Sodium hydrogen sulfite NaHSO3 61.56
are generally used in sweetened wines or wines
Sodium metabisulfite Na2S2O5 67.39
that contain residual sugars to prevent refermen-
Potassium metabisulfite K2S2O5 57.63
tation. At the levels generally used, sorbates do Calcium sulfite CaSO3 64.00
not affect food flavor. However, when used at
higher levels, they may be detected by some peo-
ple as an unpleasant flavor. Sorbate can be Sulfites
degraded by certain microorganisms to produce
off-flavors. Molds can metabolize sorbate to pro- Sulfur dioxide and sulfites have long been used
duce 1,3 pentadiene, a volatile compound with an as preservatives, serving both as antimicrobial
odor like kerosene. High levels of microorgan- substance and as antioxidant. Their use as preser-
isms can result in the degradation of sorbate in vatives in wine dates back to Roman times. Sulfur
wine and result in the off-flavor known as gera- dioxide is a gas that can be used in compressed
nium off-odor (Edinger and Splittstoesser 1986). form in cylinders. It is liquid under pressure of
The compounds responsible for the flavor defect 3.4 atm and can be injected directly in liquids. It
are ethyl sorbate, 4-hexenoic acid, 1-ethoxyhexa-­ can also be used to prepare solutions in ice cold
2,4-diene, and 2-ethoxyhexa-3,5-diene. The water. It dissolves to form sulfurous acid. Instead
same problem may occur in fermented vegetables of sulfur dioxide solutions, a number of sulfites
treated with sorbate. can be used (Table 15.4) because, when dissolved
Food Additives 537

in water, they all yield active SO2. The most


widely used of these sulfites is potassium metabi-
sulfite. In practice, a value of 50% of active SO2
is used. When sulfur dioxide is dissolved in
water, the following ions are formed:
SO2 ( gas) → SO2 (aq )
SO2 (aq ) → H 2 O → H 2 SO3
+

(
H 2 SO3 → H + + HSO3 − K l = 1.7 × 10 −2 )
(
HSO3 − → H + + SO32 − K 2 = 5 × 10 −6 )
− 2−
2HSO3 → S2 O5 + H 2 O
All of these forms of sulfur are known as free
sulfur dioxide. The bisulfite ion (HSO3−) can
react with aldehydes, dextrins, pectic substances,
proteins, ketones, and certain sugars to form
addition compounds.
Fig. 15.5  The various forms of SO2 in wine and their
activity

Table 15.5  Effect of pH on the proportion of active anti-


septic SO2 of wine containing 100 mg/L free SO2
pH Active SO2 (mg/L)
The addition compounds are known as bound 2.2 37.0
sulfur dioxide. Sulfur dioxide is used extensively 2.8 8.0
in wine making, and in wine acetaldehyde reacts 3.0 5.0
preferentially with bisulfite. Excess bisulfite 3.3 3.0
reacts with sugars. It is possible to classify bound 3.5 1.8
SO2 into three forms: aldehyde sulfurous acid, 3.7 1.2
glucose sulfurous acid, and rest sulfurous acid. 4.0 0.8
The latter holds the SO2 in a less tightly bound
form. Sulfites in wines serve a dual purpose: (1) The antiseptic activity of SO2 is highly depen-
antiseptic or bacteriostatic and (2) antioxidant. dent on the pH, as indicated in Table 15.5. The
These activities are dependent on the form of SO2 lower the pH, the greater the antiseptic action of
present. The various forms of SO2 in wine are SO2. The effect of pH on the various forms of
represented schematically in Fig. 15.5. The free sulfur dioxide is shown in Fig. 15.6.
SO2 includes the water-soluble SO2 and the Sulfurous acid inhibits molds and bacteria and
undissociated H2SO3 and constitutes about 2.8% to a lesser extent yeasts. For this reason, SO2 can
of the total. The bisulfite form constitutes 96.3% be used to control undesirable bacteria and wild
and the sulfite form 0.9% (all at pH 3.3 and yeast in fermentations without affecting the SO2-­
20 °C). The bound SO2 is mostly (80%) present tolerant cultured yeasts. According to Chichester
as acetaldehyde SO2, 1% as glucose SO2, and and Tanner (1968), the undissociated acid is 1000
10–20% as rest SO2. The various forms of sulfite times more active than HSO3—for Escherichia
have different activities. The two free forms are coli, 100–500 times for Saccharomyces cerevi-
the only ones with antiseptic activity. The anti- siae, and 100 times for Aspergillus niger.
oxidant activity is limited to the SO32− ion The amount of SO2 added to foods is self-­
(Fig. 15.5). limiting because at levels from 200 to 500 ppm
538 15  Additives and Contaminants

Fig. 15.6  Effect of pH


on the ionization of
sulfurous acid in water

the product may develop an unpleasant off-flavor. Nitrates and Nitrites


The acceptable daily intake (ADI) is set at
1.5 mg/kg body weight. Because large intakes Curing salts, which produce the characteristic
can result from consumption of wine, there have color and flavor of products such as bacon and
been many studies on reducing the use of SO2 in ham, have been used throughout history. Curing
wine making. Although some other compounds salts have traditionally contained nitrate and
(such as sorbic acid and ascorbic acid) may par- nitrite; the discovery that nitrite was the active
tially replace SO2, there is no satisfactory replace- compound was made in about 1890. Currently,
ment for SO2 in wine making. nitrate is not considered to be an essential com-
The use of SO2 is not permitted in foods that ponent in curing mixtures; it is sometimes sug-
contain significant quantities of thiamine, gested that nitrate may be transformed into
because this vitamin is destroyed by SO2. In the nitrite, thus forming a reservoir for the produc-
United States, the maximum permitted level of tion of nitrite. Both nitrates and nitrites are
SO2 in wine is 350 ppm. Modern practices have thought to have antimicrobial action. Nitrate is
resulted in much lower levels of SO2. In some used in the production of Gouda cheese to pre-
countries SO2 is used in meat products; such use vent gas formation by butyric acid–forming bac-
is not permitted in North America on the grounds teria. The action of nitrite in meat curing is
that this would result in consumer deception. SO2 considered to involve inhibition of toxin forma-
is also widely used in dried fruits, where levels tion by Clostridium botulinum, an important fac-
may be up to 2000 ppm. Other applications are in tor in establishing safety of cured meat products.
dried vegetables and dried potato products. Major concern about the use of nitrite was gener-
Because SO2 is volatile and easily lost to the ated by the realization that secondary amines in
atmosphere, the residual levels may be much foods may react to form nitrosamines, as
lower than the amounts originally applied. follows:
Food Additives 539

Fassett (1977) estimated that the nitrate intake


from 100 g of processed meat might be 50 mg
and from 100 g of high-nitrate spinach, 200 mg.
Wagner and Tannenbaum (1985) reported that
nitrate in cured meats is insignificant compared
to nitrite produced endogenously. Nitrate is pro-
duced in the body and recirculated to the oral
cavity, where it is reduced to nitrite by bacterial
The nitrosamines are powerful carcinogens, action.
and they may be mutagenic and teratogenic as
well. It appears that very small amounts of nitro-
samines can be formed in certain cured meat Hydrogen Peroxide
products. These levels are in the ppm or the ppb
range and, because analytical procedures are dif- Hydrogen peroxide is a strong oxidizing agent
ficult, there is as yet no clear picture of the occur- and is also useful as a bleaching agent. It is used
rence of nitrosamines. The nitrosamines may be for the bleaching of crude soya lecithin. The anti-
either volatile or nonvolatile, and only the latter microbial action of hydrogen peroxide is used for
are usually included in analysis of foods. the preservation of cheese milk. Hydrogen perox-
Nitrosamines, especially dimethyl-nitrosamine, ide decomposes slowly into water and oxygen;
have been found in a number of cases when cured this process is accelerated by increased tempera-
meats were surveyed at concentrations of a few ture and the presence of catalysts such as cata-
μg/kg (ppb). Nitrosamines are usually present in lase, lacto-peroxidase and heavy metals. Its
foods as the result of processing methods that antimicrobial action increases with temperature.
promote their formation (Havery and Fazio When hydrogen peroxide is used for cheese mak-
1985). An example is the spray drying of milk. ing, the milk is treated with 0.02% hydrogen per-
Suitable modifications of these process condi- oxide followed by catalase to remove the
tions can drastically reduce the nitrosamine lev- hydrogen peroxide. Hydrogen peroxide can be
els. Considerable further research is necessary to used for sterilizing food processing equipment
establish why nitrosamines are present only in and for sterilizing packaging material used in
some samples and what the toxicological impor- aseptic food packaging systems.
tance of nitrosamines is at these levels. There
appears to be no suitable replacement for nitrite
in the production of cured meats such as ham and Sodium Chloride
bacon. The ADI of nitrite has been set at 60 mg
per person per day. It is estimated that the daily Sodium chloride has been used for centuries to
intake per person in Canada is about 10 mg. prevent spoilage of foods. Fish, meats, and vege-
Cassens (1997) has reported a dramatic tables have been preserved with salt. Today, salt
decline in the residual nitrite levels in cured meat is used mainly in combination with other pro-
products in the United States. The current resid- cessing methods. The antimicrobial activity of
ual nitrite content of cured meat products is about salt is related to its ability to reduce the water
10 ppm. In 1975 an average residual nitrite con- activity (aw,), thereby influencing microbial
tent in cured meats was reported as 52.5 ppm. growth. Salt has the following characteristics: it
This reduction of nitrite levels by about 80% has produces an osmotic effect, it limits oxygen solu-
been attributed to lower ingoing nitrite, increased bility, it changes pH, sodium and chloride ions
use of ascorbates, improved process control, and are toxic, and salt contributes to loss of magne-
altered formulations. sium ions (Banwart 1979). The use of sodium
The nitrate-nitrite intake from natural sources chloride is self-limiting because of its effect on
is much higher than that from processed foods. taste.
540 15  Additives and Contaminants

Bacteriocins carboxylic acids, propionic and sorbic acids, are


used for their antimicrobial properties. Propionic
Nisin is an antibacterial polypeptide produced by acid is mainly used for its antifungal properties.
some strains of Lactococcus lactis. Nisin-like Propionic acid applied as a 10% solution to the
substances are widely produced by lactic acid surface of cheese and butter retards the growth of
bacteria. These inhibitory substances are known molds. The fungistatic effect is higher at pH 4
as bacteriocins. Nisin has been called an antibi- than at pH 5. A 5% solution of calcium propionate
otic, but this term is avoided because nisin is not acidified with lactic acid to pH 5.5 is as effective
used for therapeutic purposes in humans or ani- as a 10% unacidified solution of propionic acid.
mals. Nisin-producing organisms occur naturally The sodium salts of propionic acid also have anti-
in milk. Nisin can be used as a processing aid microbial properties.
against gram-positive organisms. Because its
effectiveness decreases as the bacterial load
increases, it is unlikely to be used to cover up Antioxidants
unhygienic practices.
Nisin is a polypeptide with a molecular weight Food antioxidants in the broadest sense are all of
of 3500, which is present as a dimer of molecular the substances that have some effect on prevent-
weight 7000. It contains some unusual sulfur ing or retarding oxidative deterioration in foods.
amino acids, lanthionine and β-methyl lanthio- They can be classified into a number of groups
nine. It contains no aromatic amino acids and is (Kochhar and Rossell 1990).
stable to heat. Primary antioxidants terminate free radical
The use of nisin as a food preservative has chains and function as electron donors. They
been approved in many countries. It has been include the phenolic antioxidants, butylated
used effectively in preservation of processed hydroxyanisole (BHA), butylated hydroxytolu-
cheese. It is also used in the heat treatment of ene (BHT), tertiary butyl hydroquinone (TBHQ),
nonacid foods and in extending the shelf life of alkylgalates, usually propylgallate (PG), and nat-
sterilized milk. ural and synthetic tocopherols and tocotrienols.
A related antibacterial substance is natamycin, Oxygen scavengers can remove oxygen in a
identical to pimaricin. Natamycin is effective in closed system. The most widely used compounds
controlling the growth of fungi but has no effect are vitamin C and related substances, ascorbyl
on bacteria or viruses. In fermentation industries, palmitate, and erythorbic acid (the D-isomer of
natamycin can be used to control mold or yeast ascorbic acid).
growth. It has a low solubility and therefore can Chelating agents or sequestrante remove
be used as a surface treatment on foods. metallic ions, especially copper and iron, that are
Natamycin is used in the production of many powerful prooxidants. Citric acid is widely used
varieties of cheese. for this purpose. Amino acids and ethylene
diamine tetraacetic acid (EDTA) are other exam-
Acids ples of chelating agents.
Acids as food additives serve a dual purpose, as Enzymic antioxidants can remove dissolved
acidulante and as preservatives. Phosphoric acid or head space oxygen, such as glucose oxidase.
is used in cola soft drinks to reduce the pH. Acetic Superoxide dismutase can be used to remove
acid is used to provide tartness in mayonnaise and highly oxidative compounds from food systems.
salad dressings. A similar function in a variety of Natural antioxidants are present in many
other foods is served by organic acids such as cit- spices and herbs (Lacroix et al. 1997; Six 1994).
ric, tartaric, malic, lactic, succinic, adipic, and Rosemary and sage are the most potent antioxi-
fumarie acid. The properties of some of the com- dant spices (Schuler 1990). The active principles
mon food acids are listed in Table 15.6 (Peterson in rosemary are carnosic acid and camosol
and Johnson 1978). Members of the straight-chain (Fig.  15.7). Antioxidants from spices can be
Table 15.6  Properties of some common food acids
Property Acetic acid Adipic acid Citric acid Fumaric acid Glucono-Delta lactone lactic acid Malic acid Phos-phoric acid Tartaric acid

Empirical formula C2H4O2 C6H10O4 C6H8O7 C4H4O4 C6H10O6 C3H6O3 C4H6O5 H3PO4 C4H6O6
Physical form Oily Liquid Crystalline Crystalline Crystalline Crystalline 85% Water Solution Crystalline 85% Water Solution Crystalline
Molecular weight 60.05 146.14 192.12 116.07 178.14 90.08 134.09 82.00 150.09
Equivalent weight 60.05 73.07 64.04 58.04 178.14 90.08 67.05 27.33 75.05
Physical form Oily Liquid Crystal-­line Crystal-­line Crystal-line Crystal-line 85% Water Crystal-­line 85% Water Solution Crystal-­line
Molecular weight 60.05 146.14 192.12 116.07 178.14 90.08 134.09 82.00 150.09
Equivalentweight 60.05 73.05 64.04 58.04 178.14 90.08 67.05 27.03 75.05
Sol. in water (g/100 ml) ∞ 1.4 181.00 0.63 59.0 ∞ 144.0 ∞ 147.0
K1 8 × 10−5 3.7 × 10−5 8.2 × 10−4 1 × 10−3 2.5 × 10−4 (gluconic acid) 1.37 × 10−4 4 × 10−4 7.25 × 10−3 1.04 × 10−3
K2 2.4 × 10−6 1.77 × 10−5 3 × 10−5 9 × 10–6 6.23 × 10−8 5.55 × 10−3
K3 3.9 × 10−6 3 × 10−13
542 15  Additives and Contaminants

Fig. 15.7 Chemical
structure of the active
antioxidant principles in
rosemary

obtained as extracts or in powdered form by a Monoglycerides are produced by transesterifi-


process described by Bracco et al. (1981). cation of glycerol with triglycerides. The reaction
The level of phenolic antioxidants permitted proceeds at high temperature, under vacuum and
for use in foods is limited. U.S. regulations allow in the presence of an alkaline catalyst. The reac-
maximum levels of 0.02% based on the fat con- tion mixture, after removal of excess glycerol, is
tent of the food. known as commercial monoglyceride, a mixture
Sometimes the antioxidants are incorporated of about 40% monoglyceride and di- and triglyc-
in the packaging materials rather than in the food erides. The di- and triglycerides have no emulsi-
itself. In this case, a larger number of antioxi- fying properties. Molecular distillation can
dants is permitted, provided that no more than increase the monoglyceride content to well over
50 ppm of the antioxidants become a component 90%. The emulsifying properties, especially
of the food. HLB, are determined by the chain length and
unsaturation of the fatty acid chain.
Hydroxycarboxylic and fatty acid esters are
Emulsifiers produced by esterifying organic acids to mono-
glycerides. This increases their hydrophilic prop-
With the exception of lecithin, all emulsifiers erties. Organic acids used are acetic, citric,
used in foods are synthetic. They are character- fumarie, lactic, succinic, or tartaric acid.
ized as ionic or nonionic and by their hydrophile/ Succinylated monoglycerides are synthesized
lipophile balance (HLB). All of the synthetic from distilled monoglycerides and succinic anhy-
emulsifiers are derivatives of fatty acids. dride. They are used as dough conditioners and
Lecithin is the commercial name of a mixture crumb softeners (Krog 1981). Acetic acid esters
of phospholipids obtained as a byproduct of the can be produced from mono- and diglycerides by
refining of soybean oil. Phosphatidylcholine is reaction with acetic anhydride or by transesterifi-
also known as lecithin, but the commercial cation. They are used to improve aeration in
product of that name contains several phospho- foods high in fat content and to control fat crys-
lipids including phosphatidylcholine. Crude tallization. Other esters may be prepared: citric,
soybean lecithin is dark in color and can be diacetyl tartaric, and lactic acid. A product con-
bleached with hydrogen peroxide or benzoyl taining two molecules of lactic acid per emulsi-
peroxide. Lecithin can be hydroxylated by treat- fier molecule, known as stearoyl-2-lactylate, is
ment with hydrogen peroxide and lactic or ace- available as the sodium or calcium salt. It is used
tic acid. Hydroxylated lecithin is more in bakery products.
hydrophilic, and this makes for a better oil-in- Polyglycerol esters of fatty acids are produced
water emulsifier. The phospholipids contained by reacting polymerized glycerol with edible
in lecithin are insoluble in acetone. fats. The degree of polymerization of the glycerol
Food Additives 543

and the nature of the fat provide a wide range of Sweeteners


emulsifiers with different HLB values. Sweeteners can be divided into two groups, non-
Polyethylene or propylene glycol esters of nutritive and nutritive sweeteners. The nonnutri-
fatty acids are more hydrophilic than monoglyc- tive sweeteners include saccharin, cyclamate,
erides. They can be produced in a range of aspartame, acesulfame K, and sucralose. Plant
compositions. extracts from Stevia and Monk Fruit extract
Sorbitan fatty acid esters are produced by where the main sweetener is Mogroside V are
polymerization of ethylene oxide to sorbitan fatty plant extracts that have been approved for food
acid esters. The resulting polyoxyethylene sorbi- uses. The nutritive sweeteners are sucrose; glu-
tan esters are nonionic hydrophilic emulsifiers. cose; fructose; invert sugar; and a variety of poly-
They are used in bakery products as antistaling ols including sorbitol, mannitol, maltitol, lactitol,
agents. They are known as polysorbates with a xylitol, and hydrogenated glucose syrups are dis-
number as indication of the type of fatty acid cussed in Chap. 2.
used (e.g., lauric, stearic, or oleic acid). The chemical structure of the most important
Sucrose fatty acid esters can be produced by nonnutritive sweeteners is shown in Fig. 15.8.
esterification of fatty acids with sucrose, usually Saccharin is available as the sodium or calcium
in a solvent system. The HLB varies, depending salt of orthobenzosulfimide. The cyclamates are
on the number of fatty acids esterified to a sucrose the sodium or calcium salts of cyclohexane sulfa-
molecule. Monoesters have an HLB value greater mic acid or the acid itself. Cyclamate is 30–40
than 16, triesters less than 1. When the level of times sweeter than sucrose, and about 300 times
esterification increases to over five molecules of sweeter than saccharin. Organoleptic comparison
fatty acid, the emulsifying property is lost. At of sweetness indicates that the medium in which
high levels of esterification the material can be the sweetener is tasted may affect the results.
used as a fat replacer because it is not absorbed or There is also a concentration effect. At higher
digested and therefore yields no calories. concentrations, the sweetness intensity of the syn-
thetic sweeteners increases at a lower rate than
that which occurs with sugars. This has been
Bread Improvers ascribed to the bitterness and strong aftertaste that
appears at these relatively high concentrations.
To speed up the aging process of wheat flour, Cyclamates were first synthesized in 1939 and
bleaching and maturing agents are used. Benzoyl were approved for use in foods in the United
peroxide is a bleaching agent that is frequently States in 1950. Continued tests on the safety of
used; other compounds—including the oxides of these compounds resulted in the 1967 finding
nitrogen, chlorine dioxide, nitrosyl chloride, and that cyclamate can be converted by intestinal
chlorine—are both bleaching and improving (or flora into cyclohexylamine, which is a carcino-
maturing) agents. Improvers used to ensure that gen. Apparently, only certain individuals have the
dough will ferment uniformly and vigorously ability to convert cyclamate to cyclohexylamine
include oxidizing agents such as potassium bro- (Collings 1971). In a given population, a portion
mate, potassium iodate, and calcium peroxide. In are nonconverters, some convert only small
addition to these agents, there may be small amounts, and others convert large amounts.
amounts of other inorganic compounds in bread Aspartame is a dipeptide derivative,
improvers, including ammonium chloride, ammo- L-aspartyl-L-phenylalanine methyl ester, which
nium sulfate, calcium sulfate, and ammonium and was approved in the United States in 1981 for
calcium phosphates. Most of these bread improv- use as a tabletop sweetener, in dry beverage
ers can only be used in small quantities, because mixes, and in foods that are not heat processed.
excessive amounts reduce quality. Several com- This substance is metabolized in the body to
pounds used as bread improvers are actually phenylalanine, aspartic acid, and methanol. Only
emulsifiers and are covered under that heading. people with phenylketonuria cannot break down
544 15  Additives and Contaminants

Fig. 15.8  Chemical structure of approved intense sweeteners

phenylalanine. Another compound, diketopiper- of its main advantages is heat stability, so it can
azine, may also be formed. However, no harmful be used in baking.
effects from this compound have been demon- Table 15.7 Summarizes the regulatory status,
strated. The main limiting factor in the use of brand names and applications of currently avail-
aspartame is its lack of heat stability (Homler able intense sweeteners.
1984). Stevioside and mogroside are glycosides of
Acesulfame K is the potassium salt of phenolic groups as can be seen in Fig. 15.6.
6-methyl-1,2,3-oxathiozine-4(3H)-one-2, Blending of nonnutritive sweeteners may lead to
2-dioxide (Fig. 15.8). It is a crystalline powder improved taste, longer shelf life, lower produc-
that is about 200 times sweeter than sugar. The tion cost, and reduced consumer exposure to any
sweetening power depends to a certain degree on single sweetener (Verdi and Hood 1993).
the acidity of the food it is used in. Acesulfame K
is reportedly more stable than other sweeteners. Phosphates
The sweet taste is clean and does not linger. These compounds are widely used as food addi-
Sucralose is a trichloroderivative of the C-4 epi- tives, in the form of phosphoric acid as acidulant,
mer galactosucrose. It is about 600 times sweeter and as monophosphates and polyphosphates in a
than sucrose and has a similar taste profile. One large number of foods and for a variety of purposes.
Food Additives 545

Table 15.7  Regulatory stauus, brand names, relative sweetness and daily intake for currently approved intense
sweeteners
Sweetness
Examples of brand intensity
names containing compared
Sweetener Regulatory status sweetener to sucrose ADI
Acesulfame Approved as a sweetener and flavor enhancer in Sweet One® 200× 15
potassium (Ace-K) foods generally (except in meat and poultry) 21 Sunett®
CFR 172.800
Advantame Approved as a sweetener and flavor enhancer in 20,000× 32.8
foods generally (except in meat and poultry) 21
CFR 172.803
Aspartame Approved as a sweetener and flavor enhancer in Nutrasweet® 200× 50
foods generally 21 CFR 172.804 Equal®
Sugar Twin®
Neotame Approved as a sweetener and flavor enhancer in Newtame® 7000– 0.3
foods generally (except in meat and poultry) 21 13,000×
CFR 172.829
Saccharin Approved as a sweetener only in certain special Sweet and Low® 200– 15
dietary foods and as an additive used for certain Sweet Twin® 700×
technological purposes 21 CFR 180.37 Sweet’N Low®
Necta Sweet®
Mogroside Siraitia SFGE containing 25%, 45% or 55% Mogroside V Nectresse® 100– NS
grosvenorii Swingle is the subject of GRAS notices for specific Monk Fruit in the 250×
extracts (SGFE) conditions of use GRAS Notice Inventory Raw®
PureLo®
Certain high purity ≥95% pure glycosides Subject of GRAS notices for Truvia® 200– 4
steviol glycosides specific conditions of use GRAS Notice Inventory PureVia® 400×
purified from the Enliten®
leaves of Stevia
rebaudiana(Bertoni)
Bertoni
Sucralose Approved as a sweetener in foods generally 21 Splenda® 600× 5
CFR 172.831
ADI Acceptable daily Intake in milligrams per kilogram body weight per day (mg/kg bw/d)
Adapted from: http://www.fda.gov/Food/IngredientsPackagingLabeling/FoodAdditivesIngredients/ucm397725.
htm#Luo_Han_Guo_fruit_extracts

Phosphates serve as buffering agents in dairy, meat,


and fish products; anticaking agents in salts; firm-
ing agents in fruits and vegetables; yeast food in
bakery products and alcoholic beverages; and melt-
ing salts in cheese processing. Phosphorus oxy-
chloride is used as a starch-­modifying agent.
The largest group of phosphates and the most
important in the food industry is the orthophos-
phates (Fig. 15.9). The phosphate group has three
replaceable hydrogens, giving three possible
sodium orthophosphates—monosodium, diso-
dium, and trisodium phosphate. The phosphates
can be divided into orthophosphates, polyphos-
phates, and metaphosphates, the latter having little Fig. 15.9  Structure of ortho- and polyphosphate salts
546 15  Additives and Contaminants

practical importance. Polyphosphates have two (Noonan 1968). As an example, FD&C red dye
or more phosphorus atoms joined by an oxygen no. 2 is known as amaranth outside the United
bridge in a chain structure. The first members of States. Over the years the originally permitted
this series are the pyrophosphates, which have fat-soluble dyes have been removed from the list
one P-O-P linkage. The condensed phosphate of approved dyes, and only water-soluble colors
with two linkages is tripolyphosphate. Alkali remain on the approved list (Newsome 1990).
metal phosphates with chain lengths greater than Color additives listed in 21 CFR Parts 74 must
three are usually mixtures of polyphosphates be analyzed and batch certified by FDA before
with varied chain lengths. The best known is they can be used in any food in the United States.
sodium hexametaphosphate. The longer chain This requirement applies to products imported
length salts are glasses. Hexametaphosphate is into this country as well as those manufactured
not a real metaphosphate, since these are ring domestically. Straight colors required to be certi-
structures and hexametaphosphate is a straight-­ fied are listed in 21 CFR Part 74. Most lakes are
chain polyphosphate. Sodium hexametaphos- provisionally listed under 21 CFR 81.1 for use as
phate has an average chain length of 10–15 listed in 21 CFR 82.51 (food, drugs, and cosmet-
phosphate units. ics), All FD&C Red No. 40 lakes are permanently
Phosphates are important because they affect listed under 21 CFR 74.340 (food),. FD&C
the absorption of calcium and other elements. Table  15.8 summarizes the approved colors
The absorption of inorganic phosphorus depends which are exempt from batch certification. Colors
on the amount of calcium, iron, strontium, and requiring batch certification are included in
aluminum present in the diet. Chapman and Table 15.9.
Pugsley (1971) have suggested that a diet con- Lakes are insoluble forms of the dyes and are
taining more phosphorus than calcium is as detri- obtained by combining the color with aluminum
mental as a simple calcium deficiency. The ratio or calcium hydroxide. The dyes provide color in
of calcium to phosphorus in bone is 2 to 1. It has solution, and the lakes serve as insoluble
been recommended that in early infancy, the ratio pigments.
should be 1.5 to 1; in older infants, 1.2 to 1; and
for adults, 1 to 1. The estimated annual per capita Food Irradiation
intake in the United States is 1 g Ca and 2.9 g P, Food irradiation is the treatment of foods by ion-
thus giving a ratio of 0.35. The danger in raising izing radiation in the form of beta, gamma, or
phosphorus levels is that calcium may become X-rays. The purpose of food irradiation is to pre-
unavailable. serve food and to prolong shelf life, as other pro-
cessing techniques such as heating or drying have
Coloring Agents done. For regulatory purposes irradiation is con-
In the United States two classes of color additives sidered a process, but in many countries it is con-
are recognized: colorants exempt from certifica- sidered to be an additive. This inconsistency in
tion and colorants subject to certification. The the interpretation of food irradiation results in
former are obtained from vegetable, animal, or great obstacles to the use of this process and has
mineral sources or are synthetic forms of natu- slowed down its application considerably. Several
rally occurring compounds. The latter group of countries are now in the process of reconsidering
synthetic dyes and pigments is covered by the their legislation regarding irradiation. Depending
Color Additives Amendment of the U.S. Food, on the radiation dose, several applications can be
Drug and Cosmetic Act. In the United States distinguished. The unit of radiation is the Gray
these color compounds are not known by their (Gy), which is a measure of the energy absorbed
common names but as FD&C colors (Food, Drug by the food. It replaced the older unit rad
and Cosmetic colors) with a color and a number (1 Gy = 100 rad).
Table 15.8  Color additives approved for use in human food Part 73, Subpart A: Color additives exempt from batch
certification
21 CFR Year
section Straight color approved Uses and restrictions
§73.30 Annatto extract 1963 Foods generally
§73.40 Dehydrated beets 1967 Foods generally
(beet powder)
§73.75 Canthaxanthin(3) 1969 Foods generally, NTE(7) 30 mg/lb of solid or semisolid food or
per pint of liquid food; May also be used in broiler chicken feed
§73.85 Caramel 1963 Foods generally
§73.90 β-Apo-8′-carotenal 1963 Foods generally, NTE(7): 15 mg/lb solid, 15 mg/pt liquid
§73.95 β-Carotene 1964 Foods generally
§73.100 Cochineal extract 1969 Foods generally
2009 Food label must use common or usual name “cochineal extract”;
effective January 5, 2011
Carmine 1967 Foods generally
2009 Food label must use common or usual name “carmine”; effective
January 5, 2011
§73.125 Sodium copper 2002 Citrus-based dry beverage mixes NTE(7) 0.2% in dry mix;
chlorophyllin(3) extracted from alfalfa
§73.140 Toasted partially defatted 1964 Foods generally
cooked cottonseed flour
§73.160 Ferrous gluconate 1967 Ripe olives
§73.165 Ferrous lactate 1996 Ripe olives
§73.169 Grape color extract(3) 1981 Non-­beverage food
§73.170 Grape skin extract 1966 Still & carbonated drinks & ades; beverage bases; alcoholic
(enocianina) beverages (restrict. 27 CFR Parts 4 & 5)
§73.200 Synthetic iron oxide(3) 1994 Sausage casings NTE(7) 0.1% (by wt)
2015 Hard and soft candy, mints and chewing gum
2015 For allowed human food uses, reduce lead from ≤20 ppm to ≤5 ppm
§73.250 Fruit juice(3) 1966 Foods generally
1995 Dried color additive
§73.300 Carrot oil 1967 Food generally
§73.340 Paprika 1966 Food Generally
§73.345 Paprika oleoresin 1966 Food generally
§73.350 Mica-based pearlescent 2006 Cereals, confections and frostings, gelatin desserts, hard and soft
pigments candies (including lozenges), nutritional supplement tablets and
gelatin capsules, and chewing gum
2013 Distilled spirits containing not less than 18% and not more than
23% alcohol by volume but not including distilled spirits mixtures
containing more than 5% wine on a proof gallon basis
2015 Cordials, liqueurs, flavored alcoholic malt beverages, wine
coolers, cocktails, nonalcoholic cocktail mixers and mixes and in
egg decorating kits
§73.450 Riboflavin 1967 Foods generally
§73.500 Saffron 1966 Foods generally
§73.530 Spirulina extract 2013 Candy and chewing gum
2014 Coloring confections (including candy and chewing gum), frostings, ice
cream and frozen desserts, dessert coatings and toppings, beverage
mixes and powders, yogurts, custards, puddings, cottage cheese, gelatin,
breadcrumbs, and ready-to-eat cereals (excluding extruded cereals)
§73.575 Titanium dioxide 1966 Foods generally; Not to exceed 1% by wt
§73.585 Tomato lycopene extract; 2006 Foods generally
tomato lycopene
concentrate(3)
§73.600 Turmeric 1966 Foods generally
§73.615 Turmeric oleoresin 1966 Foods generally
http://www.fda.gov/ForIndustry/ColorAdditives/ColorAdditiveInventories/ucm115641.htm#table4A
548 15  Additives and Contaminants

Table 15.9  Color additives approved for use in human food Part 74, Subpart A: Color additives subject to batch
certification
21 CFR section Straight color Year approved Uses and restrictions
§74.101 FD&C Blue No. 1 1969 Foods generally
1993 Added Mn spec
§74.102 FD&C Blue No. 2 1987 Foods generally
§74.203 FD&C Green No. 3 1982 Foods generally
§74.250 Orange B(3) 1966 Casings or surfaces of frankfurters and sausages; NTE(7)
150 ppm (by wt)
§74.302 Citrus Red No. 2 1963 Skins of oranges not intended or used for processing;
NTE(7) 2.0 ppm (by wt)
§74.303 FD&C Red No. 3 1969 Foods generally
§74.340 FD&C Red No. 40(3) 1971 Foods generally
§74.705 FD&C Yellow No. 5 1969 Foods generally
§74.706 FD&C Yellow No. 6 1986 Foods genera
http://www.fda.gov/ForIndustry/ColorAdditives/ColorAdditiveInventories/ucm115641.htm#table4A

Radiation sterilization produces foods that are vent deficiency diseases. Most of the additives in
stable at room temperature and requires a dose of this category are vitamins or minerals. Enrichment
20–70 kGy. At lower doses, longer shelf life may of flour and related products is now a well-recog-
be obtained, especially with perishable foods nized practice. The U.S. Food and Drug
such as fruits, fish, and shellfish. The destruction Administration (FDA) has established definitions
of Salmonella in poultry is an application for and standards of identity for the enrichment of
radiation treatment. This requires doses of wheat flour, farina, com meal, com grits, maca-
1–10 kGy. Radiation disinfestation of spices and roni, pasta products, and rice. These standards
cereals may replace chemical fumigants, which define minimum and maximum levels of addition
have come under increasing scrutiny in recent of thiamin, riboflavin, niacin, and iron. In some
years. Dose levels of 8–30 kGy would be cases, optional addition of calcium and vitamin D
required. Other possible applications of irradia- is allowed. Margarine contains added vitamins A
tion processing are inhibition of sprouting in and D, and vitamin D is added to fluid and evapo-
potatoes and onions and delaying of the ripening rated milk. The addition of the fat-­soluble vita-
of tropical fruits. mins is strictly controlled, because of the possible
toxicity of overdoses of these vitamins. The vita-
Nutrition Supplements min D enrichment of foods has been an important
There are two fundamental reasons for the addi- measure in the elimination of rickets. Another
tion of nutrients to foods consumed by the public: example of the beneficial effect of enrichment
(1) to correct a recognized deficiency of one or programs is the addition of iodine to table salt.
more nutrients in the diets of a significant number This measure has virtually eliminated goiter.
of people when the deficit actually or potentially One of the main potential deficiencies in the
adversely affects health; and (2) to maintain the diet is calcium. Lack of calcium is associated
nutritional quality of the food supply at a level with osteoporosis and possibly several other
deemed by modern nutrition science to be appro- ­diseases. The recommended daily allowance for
priate to ensure good nutritional health, assuming adolescents/young adults and the elderly has
only that a reasonable variety of foods are con- increased from the previous recommendation of
sumed (Augustin and Scarbrough 1990). 800–1200 mg/day to 1500 mg/day. This level is
A variety of compounds are added to foods to difficult to achieve, and the use of calcium citrate
improve the nutritional value of a product, to in fortified foods has been recommended by
replace nutrients lost during processing, or to pre- Labin-Goldscher and Edelstein (1996). Sloan
Food Additives 549

and Stiedemann (1996) highlighted the relation- Incidental Additives or Contaminants


ship between consumer demand for fortified
products and complex regulatory issues. Pesticides

 igration from Packaging Materials


M Contamination of food with residues of pesticides
When food packaging materials were mostly may result from the application of these chemi-
glass or metal cans, the transfer of packaging cals in agricultural, industrial, or household use.
components to the food consisted predomi- Nearly 300 organic pesticides are in use, includ-
nantly of metal (iron, tin, and lead) uptake. With ing insecticides, miticides, nematocides, rodenti-
the advent of extensive use of plastics, new cides, fungicides, and herbicides. The most likely
problems of transfer of toxicants and flavor and compounds to appear as food contaminants are
odor substances became apparent. In addition to insecticides, of which there are two main
polymers, plastics may contain a variety of classes—chlorinated hydrocarbon insecticides
other chemicals, catalysts, antioxidants, plasti- and organophosphorous insecticides.
cizers, colorants, and light absorbers. Depending The chlorinated hydrocarbon insecticides can
on the nature of the food, especially its fat con- be divided into three classes—oxygenated com-
tent, any or all of these compounds may be pounds, benzenoid nonoxygenated compounds,
extracted to some degree into the food (Bieber and nonoxygenated nonben-zenoid compounds
et al. 1985). (Table 15.10) (Mitchell 1966). In addition to the
Awareness of the problem developed in the pesticide compounds, there may be residues of
mid-1970s when it was found that mineral waters their metabolites, which may be equally toxic.
sold in polyvinyl chloride (PVC) bottles con- Two important properties of the chlorinated
tained measurable amounts of vinyl chloride hydrocarbons are their stability, which leads to
monomer. Vinyl chloride is a known carcinogen. persistence in the environment, and their solubil-
The Codex Alimentarius Committee on Food ity in fat, which results in their deposition and
Additives and Contaminants has set a guideline accumulation in fatty tissues. The structure of
of 1 ppm for vinyl chloride monomer in PVC some of the chlorinated hydrocarbon insecticides
packaging and 0.01 ppm of the monomer in food is given in Fig. 15.10. Aldrin is a technical com-
(Institute of Food Technologists 1988). Another
additive found in some PVC plastics is octyl tin Table 15.10 Classes of chlorinated hydrocarbon
mercaptoacetate or octyl tin maleate. Specific insecticides
regulations for these chemicals exist in the Class I—Oxygenated compounds
Canadian Food and Drugs Act. Chlorobenzilate Methoxychlor
The use of plastic netting to hold and Dicofol Neotran
shape meat during curing resulted in the Dieldrin Ovex
finding of N-nitrosodiethylamine and Endosulfan Sulfenone
N-nitrosodibutylamine in hams up to levels of Endrin Tetradifon
19 ppb (parts per billion) (Sen et al. 1987). Later Kepone
research established that the levels of nitrosa- Class II—Benzenoid, nonoxygenated compounds
BHC Perthane
mines present were not close to violative levels
Chlorobenside TDE
(Marsden and Pesselman 1993).
DDT Zectran
Plasticizers, antioxidants, and colorants are all
Lindane
potential contaminants of foods that are con-
Class III—Nonoxygenated, nonbenzenoid compounds
tained in plastics made with these chemicals.
Aldrin Mirex
Control of potential migration of plastic compo- Chlordan Strobane
nents requires testing the containers with food Heptachlor Toxaphene
simulants selected to yield information relevant
Source: From L.E. Mitchell, Pesticides: Properties and
to the intended type of food to be packaged Prognosis, in Organic Pesticides in the Environment,
(DeKruyf et al. 1983; Bieber et al. 1984). R.F. Gould, ed., 1966, American Chemical Society
550 15  Additives and Contaminants

Fig. 15.10  Structure of


some chlorinated
hydrocarbon pesticides

pound containing about 95% of the compound may occur at levels of up to 70% of the original
1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexa-­ DDT. It is usual to combine DDT, DDE, and TDE
hydro-exo-1,4-endo-exo-5,8-dimethanonaph-­ in one figure as “total DDT equivalent.”
thalene. It has a molecular weight of 365, formula Heptachlor contains about 75% of
C12H8Cl6, and contains 58% chlorine. Residues 1,4,5,6,7,10,10-heptachloro-4,7,8,9-tetrahyro-­
of this compound in animal and plant tissues are 4,7-methyleneindene, formula C10H5Cl7. It has a
converted into dieldrin by epoxidation. The epox- molecular weight of 373.5 and contains 67%
ide is the stable form and, thus, it is usual to con- chlorine. In animal and plant tissues, it ­epoxidizes
sider these compounds together. to heptachlor epoxide, which is analogous in
Dieldrin contains about 85% of the compound structure to HEOD (dieldrin).
1, 2, 3, 4, 10, 10-hexachloro-6, 7-epoxy-1, 4, 4a, The organophosphorous insecticides are
5, 6, 7, 8, 8a-octahydro-exo-l, 4-endo-exo-5, inhibitors of Cholinesterase and, because of their
8-dimethano-naphthalene (HEOD). It has a water solubility and volatility, create less of a
molecular weight of 381, formula C12H8Cl6O, problem as food contaminants than the chlori-
and contains 56% chlorine. DDT is a technical nated hydrocarbons. A large number of organo-
compound that contains about 70% of the active phosphorous insecticides are in use; these can act
ingredient pp'-DDT. In addition, there are other by themselves or after oxidative conversions in
isomers, including op'-DDT, as well as related plants and animals (Table 15.11). The water solu-
compounds such as TDE or rhothane. The insec- bility of these compounds varies widely, as is
ticide pp'-DDT is 1,1,1-trichloro-2,2-di-(4-­ indicated by Table 15.12. The organophospho-
chlorophenyl) ethane, formula C14H9Cl5. It has a rous insecticides may be subject to oxidation,
molecular weight of 334.5 and contains 50% hydrolysis, and demethylation (Fig. 15.11).
chlorine. Residues of DDT in animal tissue are Thiophosphates may be changed to sulfoxides
slowly dehydrochlorinated to pp’-DDE, which and sulfones in animals and plants.
Incidental Additives or Contaminants 551

Table 15.11 Classification of organophosphorous canning may remove large proportions of pesti-


insecticides
cide residues (Liska and Stadelman 1969;
Aliphatic derivatives Farrow et al. 1969). A summary of how different
Butonate Mevinphos food processing approaches can influence pesti-
Demeton Mipefox cide residue in foods are found in Table 15.13. It
Dichlorvos Naled is important to recognize that levels of pesticides
Dimefox Phorate can be lowered during processing but complete
Dimethoate Phosphamidon
removal is unlikely.
Dithiodemeton Schradan
It has been reported (Farrow et al. 1969) that
Ethion Sulfotepp
48% of DDT residues on spinach and 91% on
Malathion Tepp
tomatoes are removed by washing. Elkins (1989)
Methyl demeton Trichlorofon
reported that washing and blanching reduced car-
Aromatic (Cyclic) derivatives
Azinphosmethyl EPN
baryl residues on spinach and broccoli by 97%
Carbophenothion Fenthion and 98%, respectively. Washing, blanching, and
Diazinon Methyl canning reduced carbaryl pesticides on tomatoes
parathion and spinach by 99%. Although this pattern of
Dicapthon Parathion removal generally holds true, Peterson et al.
Endothion Ronnel (1996) have pointed out that there are exceptions.
Source: From L.E. Mitchell, Pesticides: Properties and Pesticides may accumulate in one part of an agri-
Prognosis, in Organic Pesticides in the Environment, cultural product. Friar and Reynolds (1991)
R.F. Gould, ed., 1966, American Chemical Society
reported that baking does not result in a decline
in thiabendazole residues in potatoes, and Elkins
Table 15.12  Water solubilities of some organophospho- et al. (1972) found that thermal processing does
rus insecticides not result in a reduction of methoxychlor resi-
Insecticide (ppm) dues on apricots. Sometimes processing may
Carbophenothion 2 cause a chemical to degrade, producing a com-
Parathion 24 pound that is more toxic than the original one.
Azinphosmethyl 33 The dietary intake of pesticide chemicals from
Diazinon 40 foods is well below the acceptable daily intake
Methyl parathion 50 (ADI) levels set by the FAO/WHO (Table 15.14).
Phorate 85 In recent years, severe restrictions on the use of
Malathion 145 many chlorinated hydrocarbon pesticides have
Dichlorvos 1000 been instituted in many areas. It can be seen that
Dimethoate 7000 in most cases the allowable daily intake is at least
Mevinphos ∞ one order of magnitude than the No Observed
Source: From L.E. Mitchell, Pesticides: Properties and Effect Level, constituting a reasonable degree of
Prognosis, in Organic Pesticides in the Environment, safety.
R.F. Gould, ed., 1966, American Chemical Society

In animal products, chlorinated hydrocarbon Dioxin


residues are predominantly present in the lipid
portion, organophosphates in both lipid and The term dioxin is used to represent two related
aqueous parts. In plant materials, the residue of groups of chlorinated organic compounds, poly-
chlorinated hydrocarbons are mostly surface chlorinated dibenzo-p-dioxins (PCDD) and poly-
bound or absorbed by waxy materials, but some chlorinated dibenzofurans (PCDF) (Fig. 15.12).
can be translocated to inner parts. Extensive A total of eight carbon atoms in each molecule
research has demonstrated that processing meth- can carry chlorine substitution, which produces
ods such as washing, blanching, heating, and 75 possible isomers for PCDD and 135 for PCDF.
552 15  Additives and Contaminants

Fig. 15.11 Oxidation, hydrolysis, and demethylation in Organic Pesticides in the Environment, R.F. Gould, ed.,
reactions of organophosphorous insecticides. Adapted 1966, American Chemical Society
from L.E. Mitchell, Pesticides: Properties and Prognosis,

These compounds are lipophilic, have low vola- Dioxins can be generated from chlorine
tility, and are extremely stable. They are also very bleaching of wood pulp in the paper- and
toxic, although the toxicity of each isomer may cardboard-­ making process. This can not only
vary widely. These compounds may exhibit acute lead to environmental contamination but also to
toxicity, carcinogenicity, and teratogenicity (birth incorporation of the dioxins in the paper used for
defects). They are ubiquitous environmental con- making coffee filters, tea bags, milk cartons, and
taminants and are present in human tissues. so forth. Dioxins can migrate into milk from car-
The dioxins are produced as contaminants in tons, even if the cartons have a polyethylene plas-
the synthesis of certain herbicides and other chlo- tic coating. Unbleached coffee filters and
rinated compounds, as a result of combustion and cardboard containers have been produced to
incineration, in the chlorine bleaching of wood overcome this problem, and there have also been
pulp for paper making, and in some metallurgical improvements in the production of wood pulp
processes (Startin 1991). Dioxins first attracted using alternative bleaching agents. The FDA
attention as a contaminant of the herbicide guideline for dioxin in fish is 25 parts per trillion
2,4,5-trichloro-phenoxyacetic acid (2,4,5-T). The (Cordle 1981). Dioxin is considered a very potent
particular compound identified was 2,3,7,8-­ toxin, but information on harmful effects on
TCDD, which was for some time associated with humans is controversial.
the name dioxin. This compound was present in
substantial concentration in the defoliant “Agent
Orange” used by U.S. forces during the war in Polychlorinated Biphenyls (PCBs)
Vietnam.
The various isomers, also known as conge- The PCBs are environmental contaminants that
ners, vary in toxicity with the 2,3,7,8-substituted are widely distributed and have been found as
ones being the most toxic. Humans appear to be residues in foods. PCBs are prepared by chlorina-
less sensitive than other species. tion of biphenyl, which results in a mixture of
Table 15.13  Influence of various types of food processing on reduction of residual pesticide levels in foods (adapted from Kaushik et al. 2009)
Final
Commodity Pesticide Process Initial ppm ppm % Loss by Dissipation Reason References
Potato Profenofos Baking 11.48 0.22 Loss of pesticide due to physico- Sharma et al. (2005)
microwave oven 0.19 chemical processes, e.g., evaporation,
codistillation, and thermal degradation
Wheat flour Endosulfan Bread making 70 Bread makingprocess involves Sharma et al. (2005)
Deltamethrin 63 yeast-­mediated fermentation and
baking which contribute to
Malathion 60
degradation of pesticides
Propiaconazole 52
Chlorpyriphos 51
Hexaconazole 46
Milk Leptophos Cheese making 100 1.84 Heating and salting stages in cheese Abu-Elamayem et al.
Incidental Additives or Contaminants

Leptophos-­oxon 0.76 making caused the greatest (1979)


degradation of leptophos
Phenol derivative 32.26
Rice Ekalux 25 EC Parboiling 0.078 49 Inactivation or degradation of the Krishnamurthy and
Dursbsan 25 EC 3 to 0.042 51 pesticides during parboiling at high Sreeramulu (1982)
temperature
Lebaycid ECO all (0.05%) 6 68
Rough rice Malathion Parboiling 14 0.013 Cogburn et al. (1990)
Maize and Malathion 12 Months 7.73 64 Volatilization and possible settling of Lalah and Wandiga
beans storage open 7.52 47 pesticide dust formulation to the (2002)
basket bottom and on the sides of basket
during storage in the open and windy
tropical laboratory
Tomatoes HCB Wash with acetic 42.9 Reduction by dissolution of phosalone Mergnat et al. (1995)
acid in water residue
Lindane 46.1 Effectiveness of washing in removing Holland et al. (1994)
residues depends upon:
p,p-DDT 27.2 1. Location of residue
Dimethoate 90.8 2. Age of the residue
Profenofos 82.4 3. Water solubility of the pesticide
Pirimiphos-­Methyl 91.4 4. Temperature and type of wash
5. Effectiveness of washing may be
improved addition of a detergent
Tomatoes HCB Wash with tap 9.62
Lindane water 15.3
p,p-DDT 9.17
Dimethoate 18.8
553

Profenofos 22.7
Pirimiphos-­Methyl 16.2
554 15  Additives and Contaminants

Table 15.14  The use, allowable daily intake and NOAEL levels for some pesticides and herbicides
Compound Use ADI (mg/kg bw) NOAEL
Aldrin Insecticide 0.0001 0.025
Carbamyl Insecticide 0.01 0.06
Chlordane Insecticide 0.0005 0.05
Cycloxydim Herbicide 0.07 7
DDT Insecticide 0.02 0.25
Diazinon Herbicide 0.002 0.025
Dieldrin Insecticide 0.0001 0.025
Diquat 0.002 0.19
Glyphosate Herbicide 0.3 31
Heptachlor-heptachlor epoxide Insecticide 0.0001 0.025
Lindane Insecticide 0.008 0.75
Malathion Insecticide 0.02 0.2
Methoxychlor Insecticide 0.1 10
Paraquat Insecticide 0.004 1.6
Parathion Insecticide 0.005 0.05
Pyrethrins Insecticide 0.04 10
2,4,D Herbicide 0.3 31
2,4,5-T Herbicide 0.03 3
Adapted from Lu 1995

Fig. 15.12  Chemical structure of Polychlorinated Dibenzo-p-dioxins (PCDD) and Polychlorinated Dibenzofurans
(PCDF)

isomers that have different chlorine contents. In ered in fish and wildlife in Sweden in 1966, and
North America, the industrial compounds are they can now be found in higher concentrations in
known as Aroclor; these are used industrially as fish than organochlorine pesticides (Zitco 1971).
dielectric fluids in transformers, as plasticizers, as PCBs decompose very slowly. It is estimated
heat transfer and hydraulic fluids, and so forth. that between 1929 and 1977, about 550 million
The widespread industrial use of these com- kg of PCBs were produced in the United States.
pounds results in contamination of the environ- Production was stopped voluntarily after a seri-
ment through leakages and spills and seepage ous poisoning occurred in Japan in 1968. Large
from garbage dumps. The PCBs may show up on amounts are still present in, for example, trans-
chromatograms at the same time as chlorinated formers and could enter the environment for
hydrocarbon pesticides. The numbering system many years. Federal regulations specify the fol-
used in PCBs and the prevalent substitution pat- lowing limits in foods: 1.5 ppm in milk fat,
tern are presented in Fig. 15.13. Table 15.15 pres- 1.5 ppm in fat portion of manufactured dairy
ents information on commercial Aroclor products, 3 ppm in poultry, and 0.3 ppm in eggs.
compounds. In the years prior to 1977 production The tolerance level for PCB in fish was reduced
of PCBs in North America amounted to about 50 from 5 to 2 ppm in 1984. Although there has been
million pounds per year. PCBs were first discov- a good deal of concern about the possible toxicity
Incidental Additives or Contaminants 555

Fig. 15.13 The
numbering system used
in PCBs and the
prevalent substitution
patterns of chlorine

Table 15.15  Information on aroclor preparations animals. The latter problem has created the great-
Average Average est concern. The use of antibiotics in therapy,
number of Cl molecular prophylaxis, and growth promotion of animals
Aroclor % Cl per molecule weight may result in residues in foods. These residue
Aroclor 1221 21 1.15 192 levels rarely exceed the range of 1–0.1 ppm
Aroclor 1232 32 2.04 221
(where toxicological interest ceases). However,
Aroclor 1242 42 3.10 261
levels well below those of toxicological interest
Aroclor 1248 48 3.90 288
may be important in food processing, for exam-
Aroclor 1254 54 4.96 327
ple, in cheese making, by preventing starter
Aroclor 1260 60 6.30 372
development. The low levels may also be impor-
Aroclor 1262 62 6.80 389
Aroclor 1268 68 8.70 453
tant in causing allergies and development of
resistant organisms. Highly sensitized persons
may experience allergic reactions from milk that
of PCBs, there is now evidence that PCBs are contains extremely low amounts of penicillin.
much less toxic than initially assumed (American The various antibiotics used in agriculture,
Council on Science and Health 1985). including some used in food processing, are
Zabik and Zabik (1996) have reviewed the listed in Table 15.16 The tetracyclines, CTC and
effect of processing on the removal of PCBs from OTC, are broad-spectrum antibiotics and act
several foods. In the processing of vegetable oil the against both gram-positive and gram-negative
PCB present in the crude oil was completely bacteria. The action is bacteriostatic and not bac-
removed; some was removed by the hydrogenation tericidal. The tetracyclines have been used to
catalyst, but most was lost by deodorization. The delay spoilage in poultry and fish. Their effec-
PCB was recovered in the deodorizer distillate. tiveness seems to decrease quite rapidly, because
the contaminating flora quickly become resistant.
Nisin, which is one of the few antibiotics not
Antibiotics used in human therapeutics, has been found to be
effective as an aid in heat sterilization of foods. It
Growth-retarding or antimicrobial substances is a polypeptide with a molecular weight of about
may be present in foods naturally, may be pro- 7000 and contains 18 amino acid residues. It is
duced in a food during processing, or may occur active against certain gram-positive organisms
incidentally through the treatment of diseased only, and all spores are sensitive to it.
556 15  Additives and Contaminants

Table 15.16  Antibiotics used in animal production


Drug Desired effect in animals References
Bambermycin Improves feed conversion ratio and weight gain in chickens, beef Reinhardt (2012) and
cattle, swine, and turkeys Allen and Stanton (2014)
Lasalocid Improves feed conversion ratio and weight gain in beef cattle Reinhardt (2012) and
Allen and Stanton (2014)
Monensin Increase feed conversion and weight gain in beef cattle and sheep Reinhardt (2012)
Improves milk production in dairy cows Allen and Stanton (2014)
Salinomycin Increase weight gain in chickens Reinhardt (2012)
Virginiamycin Improved feed conversion and weight gain in chickens, swine, Reinhardt (2012) and
turkeys, and beef cattle Allen and Stanton (2014)
Bacitracin Increase weight gain in chickens, turkeys, beef cattle, and swine; Reinhardt (2012) and
promotes egg production in chickens Allen and Stanton (2014)
Carbadox Increases feed conversion and weight gain in swine Allen and Stanton (2014)
Laidlomycin Increase feed conversion and weight gain in beef cattle Allen and Stanton 2014
Lincomycin Increase feed conversion ratio and weight gain in chickens and swine Allen and Stanton (2014)
Neomycin/ Increase weight gain and feed conversion ratio in chickens, turkeys, Allen and Stanton (2014)
oxytetracyclinee swine, and beef cattle
Penicillin Increase feed conversion ratio and weight gain in chickens, turkeys, Allen and Stanton (2014)
and swine
Roxarsone Increase feed conversion ratio and weight gain in chickens and Allen and Stanton (2014)
turkeys
Tylosin Increase feed conversion ratio and weight gain in chickens and swine Allen and Stanton (2014)

Trace Metals Natural sources of mercury in the environ-


ment include elemental mercury vapor from vol-
A variety of trace metals (such as mercury and canoes and forest fires and the release of inorganic
lead) may become components of foods through mercury from movement of water (Bose-O’Reilly
industrial contamination of the environment. et al. 2010). Environmental sources of Mercury
Some trace metals (such as tin and lead) may be include burning of coal and fossil fuels, mining
introduced into foods through pickup from equip- of mercury, precious metal refinement, electrical
ment and containers (especially tin cans). and automotive part manufacture, and chemical
processing, and release through waste incinera-
tion, landfills, and industrial contamination of
Mercury water systems. Methylmercury (MeHg) contami-
nation of fish is still a significant source of human
Large amounts of mercury are released into the exposure to mercury compounds in fish-eating
environment by several industries. Major mer- populations. As much as 90% of ingested MeHg
cury users are the chloralkali industry, where is absorbed through the intestine and forms thiol
mercury is used in electrolytic cells; traditional (-SH) complexes with proteins and amino acids
gold extraction and the pulp and paper industry. in the liver. Some MeHg enters the general circu-
Mercury (Hg) occurs naturally in the Earth’s lation, and is distributed throughout all tissues
crust and is present in the environment and atmo- including the brain. However, most of the
sphere at low levels. Concerns about Mercury are absorbed MeHg is incorporated into bile, secreted
related to its release through anthropogenic emis- into the intestine, and reabsorbed through the
sions. Mercury is detrimental to humans, animals enterohepatic circulation (Martinez-Finley and
and plants (Eisler 2004). Aschner 2014).
Incidental Additives or Contaminants 557

Fig. 15.14  The environmental conversion of inorganic mercury and some mercury-containing compounds to methyl
mercury. Source: https://www.ec.gc.ca/mercure-mercury/default.asp?lang=En&n=67E16201-1#mercurymethylation

The discoveries of biomethylation and bioac- total mercury present. The present interest in mer-
cumulation aroused intense interest in the environ- cury and its effect on humans and wildlife origi-
mental fate of mercury and its pathways to human nated with the discovery of mercury as the
exposure. Methyl mercury is found in most fish causative agent in the Minamata disease in Japan.
species and in fish consuming animals including Near the town of Minamata, a chemical industry
humans. The cyscle of mercury in the environment used mercury compounds as catalysts for the con-
ae illustrated in Fig. 15.14 where elemental mer- version of acetylene into acetaldehyde and vinyl
cury is biomethylated by microorganisms in sedi- chloride. Organic mercury compounds were
ments found in both fresh and ocean water. released into the waters of Minamata Bay and con-
Historically environmental mercury came from taminated fish and shellfish. Many cases of mer-
Chloralkali plants which discharged large amounts cury poisoning occurred, resulting in the death of
of inorganic mercury into rivers, lakes, and ocean close to 50 patients. This event triggered research
bays. Other sources included paper pulp factories. into mercury contamination in many areas of the
These practices have been eliminated, however world. As a result, there is now much improved
gold mining operations can result in large quanti- control of environmental release of mercury. It
ties of liquid mercury being deposited in river beds should be noted that some natural sources such as
(Pfeiffer and Lacerda 1988). volcanic eruptions still contribute to volatile mer-
Formation of the more volatile dimethyl mer- cury being released to the environment.
cury is favored at alkaline pH. The less volatile Fresh and saltwater seafood remain the largest
monomethyl form is favored at acid pH. Because dietary source of mercury. The higher the species
much of the mercury pollution ends up in rivers in the food chain the greater the bioaccumulation.
and lakes where it is converted into methyl mer- The US. FDA has summarized th mercury levels
cury, contamination of fish with mercury has been in commercial fish and shell fish over the period
a great concern. In many animal tissues, methyl of 1990–2012. Table 15.17 summarizes the FDA
mercury may comprise as much as 99% of the report.
558 15  Additives and Contaminants

Table 15.17  Mercury concentration found in fish. http://www.fda.gov/Food/FoodborneIllnessContaminants/Metals/


ucm115644.htm
Mean Minimum Maximum Num. of
Species (PPM) (PPM) (PPM) samples Data source
Anchovies 0.016 ND 0.049 15 FDA 2007–2009
Bass (Saltwater, Black, striped, rockfish) 0.167 ND 0.96 101 FDA 1991–2010
Bass chilean 0.354 ND 2.18 74 FDA 1994–2010
Bluefish 0.368 0.089 1.452 94 FDA 1991–2009
Buffalofish 0.137 0.032 0.43 17 FDA 1992–2008
Butterfish 0.058 ND 0.36 89 NMFS report 1978
Carp 0.110 ND 0.271 14 FDA 1992–2007
Catfish 0.024 ND 0.314 59 FDA 1991–2010
Clam 0.009 ND 0.028 15 FDA 1991–2010
Cod 0.111 ND 0.989 115 FDA 1991–2010
Crab 0.065 ND 0.61 93 FDA 1991–2009
Crawfish 0.033 ND 0.051 46 FDA 1991–2007
Croaker atlantic (Atlantic) 0.069 ND 0.193 90 FDA 2002–2011
Croaker white (Pacific) 0.287 0.18 0.41 15 FDA 1997
Flatfish 0.056 ND 0.218 71 FDA 1991–2009
Grouper (all species) 0.448 0.006 1.205 53 FDA 1991–2005
Haddock (Atlantic) 0.055 ND 0.197 50 FDA 1991–2009
Hake 0.079 ND 0.378 49 FDA 1994–2009
Halibut 0.241 ND 1.52 101 FDA 1992–2009
HERRING 0.078 ND 0.56 27 FDA 2005–2012
Jacksmelt 0.081 0.011 0.5 23 FDA 1997–2007
Lobster (Northern/American) 0.107 ND 0.23 9 FDA 2005–2007
Lobster (Species unknown) 0.166 ND 0.451 71 FDA 1991–2008
Lobster (Spiny) 0.093 ND 0.27 13 FDA 1991–2005
Mackerel atlantic (N.Atlantic) 0.05 0.02 0.16 80 NMFS report 1978
Mackerel chub (Pacific) 0.088 0.03 0.19 30 NMFS report 1978
Mackerel king 0.73 0.23 1.67 213 Gulf of Mexico report 2000
Mackerel spanish (Gulf of Mexico) 0.454 0.07 1.56 66 NMFS report 1978
Mackerel spanish (S. Atlantic) 0.182 0.05 0.73 43 NMFS report 1978
Mahi mahi 0.178 ND 0.45 29 FDA 1991–2005
Marlin 0.485 0.1 0.92 16 FDA 1992–1996
Monkfish 0.161 ND 0.289 11 FDA 1994–2007
Mullet 0.050 ND 0.27 20 FDA 1991–2008
Orange roughy 0.571 0.265 1.12 81 FDA 1991–2009
Oyster 0.012 ND 0.25 61 FDA 1991–2009
Perch (Freshwater) 0.150 ND 0.325 19 FDA 1991–2007
Perch ocean 0.121 ND 0.578 31 FDA 1991–2010
Pickerel 0.095 ND 0.31 16 FDA 1991–2007
Pollock 0.031 ND 0.78 95 FDA 1991–2008
Sablefish 0.361 0.09 1.052 26 FDA 2004–2009
Salmon (Canned) 0.014 ND 0.086 19 FDA 1993–2009
Salmon (Fresh/frozen) 0.022 ND 0.19 94 FDA 1991–2009
Sardine 0.013 ND 0.083 90 FDA 2002–2010
Scallop 0.003 ND 0.033 39 FDA 1991–2009
(continued)
Incidental Additives or Contaminants 559

Table 15.17 (continued)
Mean Minimum Maximum Num. of
Species (PPM) (PPM) (PPM) samples Data source
Scorpionfish 0.233 0.098 0.456 6 FDA 2006–2007
Shad 0.038 ND 0.186 15 FDA 2007–2011
Shark 0.979 ND 4.54 356 FDA 1991–2007
Sheepshead 0.090 ND 0.17 8 FDA 1992–2007
Shrimp 0.009 ND 0.05 40 FDA 1991–2009
Skate 0.137 0.04 0.36 56 NMFS REPORT 1978
Snapper 0.166 ND 1.366 67 FDA 1991–2007
Squid 0.024 ND 0.07 36 FDA 2005–2009
Swordfish 0.995 ND 3.22 636 FDA 1990–2010
Tilapia 0.013 ND 0.084 32 FDA 1991–2008
Tilefish (Atlantic) 0.144 0.042 0.533 32 FDA 1994–2004
Tilefish (Gulf of Mexico) 1.123 0.65 3.73 60 NMFS report 1978
Trout (Freshwater) 0.071 ND 0.678 35 FDA 1991–2008
Tuna (Canned, albacore) 0.350 ND 0.853 451 FDA 1991–2009
Tuna (Canned, light) 0.126 ND 0.889 545 FDA 1991–2010
Tuna (Fresh/frozen, albacore) 0.358 ND 0.82 43 FDA 1992–2008
Tuna (Fresh/frozen, All) 0.386 ND 1.816 420 FDA 1991–2010
Tuna (Fresh/frozen, bigeye) 0.689 0.128 1.816 21 FDA 1993–2005
Tuna (Fresh/frozen, skipjack) 0.144 0.022 0.26 3 FDA 1993–2007
Tuna (Fresh/frozen, species unknown) 0.410 ND 1.3 122 FDA 1991–2010
Tuna (Fresh/frozen, yellowfin) 0.354 ND 1.478 231 FDA 1993–2010
Weakfish (Sea trout) 0.235 ND 0.744 46 FDA 1991–2005
Whitefish 0.089 ND 0.317 37 FDA 1991–2008
Whiting 0.051 ND 0.096 13 FDA 1991–2008
Source of data: FDA 1990–2012, “National Marine Fisheries Service Survey of Trace Elements in the Fishery Resource”
Report 1978, “The Occurrence of Mercury in the Fishery Resources of the Gulf of Mexico” Report 2000
ND-mercury concentration below detection level (Level of Detection (LOD) = 0.01 ppm)
N/A-data not available

Lead and  Tin lead-­containing glazes. Both lead and tin may be
taken up by foods from the tin of cans and from
The presence of lead in foods may be the result the solder used in their manufacture. The
of environmental contamination, pickup of the amounts of lead and tin taken up depend on the
metal from equipment, or the solder of tin cans. type of tin plate and solder used and on the com-
It has been estimated that nearly 90% of the position and properties of the canned foods. In a
ingested lead is derived from food (Somers and study on the detinning of cans by spinach,
Smith 1971). However, only 5% of this is Lambeth et al. (1969) found that detinning was
absorbed. In the early 1970s, the average North significantly related to the oxalic acid content
American car was reported to emit 2.5 kg of lead and pH of the product. Detinning in excess of
per year (Somers and Smith 1971), and Zuber 60% was observed during 9 months’ storage of
et al. (1970) reported that crops grown near busy high-­oxalate spinach.
highways had a high lead content (in some cases, The present levels of lead that humans ingest
exceeding 100 ppm of lead in the dry matter). cause concern because ADI calculations range
The removal of lead from gasoline has elimi- from 0.1 to 0.8 mg lead per day. The average
nated this source of contamination. Lead can daily intake is in the vicinity of 0.4 mg lead per
also be picked up by acid foods such as fruit day. This means that lead is one of the few toxic
juices that are kept in glazed pottery made with food components for which the acceptable daily
560 15  Additives and Contaminants

intake is approached or exceeded by the general based products, however, the Joint FAO-WHO
population (Clarkson 1971). Codex Alimentarius Commission in July 2014
established a maximum level of 200 ppb for inor-
ganic arsenic in polished rice. Based on its test-
Cadmium ing, the FDA on April 1, 2016 proposed an action
level, or limit, of 100 parts per billion (ppb) for
As are lead and mercury, cadmium is a nones- inorganic arsenic in infant rice cereal. This level,
sential trace metal with high toxicity. Crustaceans which is based on the FDA’s assessment of a
have the ability to accumulate cadmium as well large body of scientific information, seeks to
as other trace metals, such as zinc. Cadmium lev- reduce infant exposure to inorganic arsenic.
els in oysters may reach 3–4 ppm, whereas in http://www.consumerreports.org/cro/maga-
other foods, levels are only one-tenth or one-­ zine/2012/11/arsenic-in-your-food/index.htm
hundredth of these (Underwood 1973). https://medicalxpress.com/news/2015-01-ar-
senic-food.html#jCp
https://medicalxpress.com/news/2015-01-ar-
Arsenic senic-food.html#jCp

Arsenic is found in the environment from both


natural and human sources. Environmental  olycyclic Aromatic Hydrocarbons
P
Aarsenic can come from erosion of arsenic-­ (PAHs)
containing rocks, volcanic eruptions, contamina-
tion from mining and smelting ores. Environmental These compounds form a large group of materials
arsenic can contaminate food products from that are now known to occur in the environment.
absorption through soil and water. Foods that are The structural formulas of the major members of
produced following prolonged contact with water, this group are presented in Fig. 15.15. Several of
such as seafood and rice frequently have higher these, especially benzo(a)pyrene (3,4-benzopy-
levels of arsenic contamination. rene), have been found to be carcinogenic. Usually,
Arsenic is used as a pesticide primarily to pre- the polycyclic hydrocarbons occur together in
serve wood from rot and decay. Historically, foods, especially in smoked foods, because the
arsenic was also used in rat poisons, ant poisons aromatic hydrocarbons are constituents of wood
and weed killers. Agricultural soils can contain smoke. Trace quantities of PAHs have been found
high levels of arsenic resulting from its former in a variety of foods, and this may be the result of
agricultural uses. Most forms of arsenic tend to environmental contamination.
stick to soil or sediment particles however, some The PAHs may be carcinogenic and muta-
arsenic can dissolve in water, leaching into lakes, genic. The level of carcinogenicity may vary
rivers, or ground water. widely between different members of this
Exposure to arsenic can come from many dif- group. Minor constituents of PAH mixtures may
ferent foods and beverages. Consumer awareness make large contributions to the carcinogenic
of dietary arsenic exposure in a publication by activity of the mixture. Certain methylchry-
Consumer Reports magazine analyzing arsenic senes, particularly the 5-isomer, which is one of
content of commercially available fruit juices the most ­carcinogenic compounds known, may
(January, 2012) and rice-based products dominate the carcinogenic activity of a mixture
(November, 2012). In July 2013, the FDA pro- (Bartle 1991).
posed an action level of 10 parts per billion (ppb), Rhee and Bratzler (1968) analyzed hydrocar-
which is similar to the U.S. Environmental bons in smoke, and the amounts found in smoke
Protection agency (EPA) drinking water stan- and in the vapor phase (smoke filtered to remove
dard, for inorganic arsenic in apple juice. The particles) are listed in Table 15.18. Small amounts
FDA has not yet set an arsenic standard for rice- of these smoke constituents may be transferred to
Incidental Additives or Contaminants 561

Fig. 15.15 Chemical
structure of some
polycyclic aromatic
hydrocarbons

Table 15.18  Aromatic polycyclic hydrocarbons in wood are listed in Table 15.19. These compounds have
smoke and in wood smoke vapor phase
also been found in unsmoked foods, as is shown
Amount, μg/45 kg sawdust in Table 15.20. Higher levels than those found in
Hydrocarbon whole smoke vapor phase
smoked food may occur as a result of barbecuing
Phenanthrene 51.5 28.4
or charcoal broiling. Roasting of coffee and nuts
Anthracene 3.8 1.9
results in formation of PAHs. The levels present
Pyrene 5.5 4.1
in roasted coffee increase with more intense
Fluoranthene 5.7 4.2
roasting; this is shown in Table 15.21, which is
1,2-Benzanthracene 7.0 4.3
Chrysene 2.6 0.3
based on results obtained by Fritz (1968). PAHs
3,4-Benzopyrene 1.2 0.4 occur in vegetables (Grimmer and Hildebrand
1,2-Benzopyrene 0.9 Trace 1965), and the levels are thought to be related to
Source: From K.S. Rhee and L.J. Bratzler, Polycyclic the leaf area and the relative level of atmospheric
Hydrocarbon Composition of Wood Smoke, J. Food Sci., pollution.
Vol. 33, pp. 626–632, 1968 Surprisingly, the largest proportion of the total
human intake of PAHs does not come from
foods during smoking. Howard and Fazio (1969) smoked or roasted foods, but from other common
reported the levels of aromatic polycyclic hydro- products. Bartle (1991) has stated that cereals are
carbons in foods, and results for smoked foods likely to be a greater hazard, especially in the
562 15  Additives and Contaminants

Table 15.19  Polycyclic aromatic hydrocarbons found in smoked food products (ppb)
Benzo Benzo Benzo Benzo 4-Methyl-
Food Product (a)-anthracene (a)-pyrene (e)-pyrene (g,h,i,)-perylene Fluoranthene Pyrene pyrene
Beef, chipped 0.4 0.6 0.5
Cheese, Gouda 2.8 2.6
Fish
Herring 3.0 2.2
Herring (dried) 1.7 1.0 1.2 1.0 1.8 1.8
Salmon 0.5 0.4 3.2 2.0
Sturgeon 0.8 2.4 4.4
White 4.6 4.0
Ham 2.8 3.2 1.2 1.4 14.0 11.2 2.0
Frankfurters 6.4 3.8
Pork roll 3.1 2.5
Source: From J.W. Howard and T. Fazio, A Review of Polycyclic Aromatic Hydrocarbons in Foods, Agr. Food Chem.,
Vol. 17, pp. 527–531, 1969, American Chemical Society

Table 15.20 Polycyclic aromatic hydrocarbons in


unsmoked food products
Food Product Fluoranthene (ppb) Pyrene (PPb)
Cheese, cheddar 0.8 0.7
Fish, haddock 1.6 0.8
Fish, herring (salted) 0.8 1.0
Fish, salmon (canned) 1.8 1.4
Source: From J.W. Howard and T. Fazio, A Review of
Polycyclic Aromatic Hydrocarbons in Foods, Agr. Food
Chem., Vol. 17, pp. 527–531, 1969, American Chemical
Society Fig. 15.16  The structure of caffeine 1,3,7- trimethylxanthine

Table 15.21 Polycyclic Aromatic Hydrocarbons in


Coffee (μg/kg)
Heavy Normal form of flour, than smoked or barbecued foods.
Compound roasting roasting Although cereal has a much lower PAH content
Anthracene 6.2 1.5 than smoked or roasted foods do, cereal is con-
Phenanthrene 74.0 28.0 sumed in much greater amounts.
Pyrene 28.0 3.5
Fluoranthene 34.0 3.9
1,2-Benzanthracene 14.2 1.5
Caffeine
Chrysene 14.8 –
3,4-Benzopyrene 5.8 0.3
Caffeine is a naturally occurring chemical,
1,2-Benzopyrene 7.0 0.7
1,3,7-trimethylxanthine (Fig. 15.16), which is
Perylene 0.6 –
found in the leaves, seeds, and fruits of more than
11, 12-Benzfluoranthene 1.8 –
63 species of plants growing all over the world. It
Anthanthrene 0.9 –
occurs as a constituent of coffee, tea, cocoa, and
1, 12-Benzperylene 2.2 –
chocolate, and is an additive in soft drinks and
3,4-Benzfluoranthene 1.2 –
Coronene 0.9 –
other foods. Because humans have used it for
Indenopyrene 0.7 –
thousands of years, caffeine has GRAS status in
the United States. Roberts and Barone (1983)
Source: From W. Fritz, Formation of Carcinogenic
Hydrocarbons During Thermal Treatment of Foods, have estimated that daily caffeine consumption in
Nahrung, Vol. 12, pp. 799–804, 1968 the United States is 206 mg per person. Caffeine
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Appendix A: Moisture Analysis

Moisture content is one of the most commonly accuracy, cost, speed, sensitivity, specificity, ease
measured properties of food materials. It is of operation, etc. The choice of an analytical pro-
important to food scientists for a number of dif- cedure for a particular application depends on the
ferent reasons: nature of the food being analyzed and the reason
the information is needed.
• Legal and Labeling Requirements. There are
legal limits to the maximum or minimum
amount of water that must be present in cer- Properties of Water in Foods
tain types of food.
• Economic. The cost of many foods depends on The moisture content of a food material is defined
the amount of water they contain - water is an through the following equation:
inexpensive ingredient, and manufacturers
often try to incorporate as much as possible in %Moisture = (mw/msample) × 100
a food, without exceeding some maximum
legal requirement. Where mw is the mass of the water and msample
• Microbial Stability. The propensity of micro- is the mass of the sample. The mass of water is
organisms to grow in foods depends on their related to the number of water molecules (nW)
water content. For this reason many foods are by the following expression: mw = nwMw/NA,
dried below some critical moisture content. where Mw is the molecular weight of water
• Food Quality. The texture, taste, appearance (18.0 g per mole) and NA is Avadagro’s number
and stability of foods depends on the amount (6.02 × 1023 molecules per mole). In principle,
of water they contain. the moisture content of a food can therefore be
• Food Processing Operations. A knowledge of the determined accurately by measuring the number
moisture content is often necessary to predict the or mass of water molecules present in a known
behavior of foods during processing, e.g. mixing, mass of sample. It is not possible to directly mea-
drying, flow through a pipe or packaging. sure the number of water molecules present in a
sample because of the huge number of molecules
It is therefore important for food scientists to involved. A number of analytical techniques
be able to reliably measure moisture contents. commonly used to determine the moisture content
A number of analytical techniques have been of foods are based on determinations of the mass
developed for this purpose, which vary in their of water present in a known mass of sample.

© Springer International Publishing AG 2018 567


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8
568 Appendix A: Moisture Analysis

Nevertheless, as we will see later, there are a surrounded only by other water molecules.
number of practical problems associated with It therefore has physicochemical properties
these techniques that make highly accurate deter- that are the same as those of pure water, e.g.,
minations of moisture content difficult or that melting point, boiling point, density, com-
limit their use for certain applications. For these pressibility, heat of vaporization, electromag-
reasons, a number of other analytical methods netic absorption spectra.
have been developed to measure the moisture • Capillary or trapped water. Capillary water is
content of foods that do not rely on direct mea- held in narrow channels between certain food
surement of the mass of water in a food. Instead, components because of capillary forces.
these techniques are based on the fact that the Trapped water is held within spaces within a
water in a food can be distinguished from the food that are surrounded by a physical barrier
other components in some measurable way. that prevents the water molecules from easily
An appreciation of the principles, advantages escaping, e.g., an emulsion droplet or a bio-
and limitations of the various analytical techniques logical cell. The majority of this type of water
developed to determine the moisture content of is involved in normal water-water bonding
foods depends on an understanding of the molec- and so it has physicochemical properties simi-
ular characteristics of water. A water molecule lar to that of bulk water.
consists of an oxygen atom covalently bound to • Physically bound water. A significant fraction
two hydrogen atoms (H2O). Each of the hydrogen of the water molecules in many foods are not
atoms has a small positive charge (δ+), while the completely surrounded by other water mole-
oxygen atom has two lone pairs of electrons that cules, but are in molecular contact with other
each has a small negative charge (δ−). food constituents, e.g. proteins, carbohydrates
Consequently, water molecules are capable of or minerals. The bonds between water mole-
forming relatively strong hydrogen bonds cules and these constituents are often signifi-
(O-Hδ+  ∙  δ−O) with four neighboring water mole- cantly different from normal water-water
cules. The strength and directionality of these bonds and so this type of water has different
hydrogen bonds are the origin of many of the unique physicochemical properties than bulk water
physicochemical properties of water. The develop- e.g., melting point, boiling point, density,
ment of analytical techniques to determine the compressibility, heat of vaporization, electro-
moisture content of foods depends on being able to magnetic absorption spectra.
distinguish water (the “analyte”) from the other • Chemically bound water. Some of the water
components in the food (the “matrix”). The charac- molecules present in a food may be chemically
teristics of water that are most commonly used to bonded to other molecules as water of crystalli-
achieve this are: its relatively low boiling point; its zation or as hydrates, e.g. NaSO4 ∙ 10H20. These
high polarity; its ability to undergo unique chemical bonds are much stronger than the normal water-
reactions with certain reagents; its unique electro- water bond and therefore chemically bound
magnetic absorption spectra; and, its characteristic water has very different physicochemical prop-
physical properties (density, compressibility, elec- erties to bulk water, e.g., lower melting point,
trical conductivity and refractive index). higher boiling point, higher density, lower com-
Despite having the same chemical formula pressibility, higher heat of vaporization, different
(H2O) the water molecules in a food may be electromagnetic absorption spectra.
present in a variety of different molecular envi-
ronments depending on their interaction with the Foods are heterogeneous materials that contain
surrounding molecules. The water molecules in different proportions of chemically bound, physi-
these different environments normally have dif- cally bound, capillary, trapped or bulk water. In
ferent physiochemical properties: addition, foods may contain water that is present in
different physical states: gas, liquid or solid. The
• Bulk water. Bulk water is free from any other fact that water molecules can exist in a number of
constituents, so that each water molecule is different molecular environments, with different
Appendix A: Moisture Analysis 569

physicochemical properties, can be problematic for food before and after the water is removed by
the food analyst trying to a­ ccurately determine the evaporation:
moisture content of foods. Many analytical proce-
M INITIAL − M DRIED
dures developed to measure moisture content are %Moisture = ×100
more sensitive to water in certain types of molecu- M INITIAL
lar environment than to water in other types of Here, MINITIAL and MDRIED are the mass of the
molecular environment. This means that the mea- sample before and after drying, respectively. The
sured value of the moisture content of a particular basic principle of this technique is that water has a
food may depend on the experimental technique lower boiling point than the other major compo-
used to carry out the measurement. Sometimes nents within foods, e.g., lipids, proteins, carbohy-
food analysts are interested in determining the drates and minerals. Sometimes a related parameter,
amounts of water in specific molecular environ- known as the total solids, is reported as a measure
ments (e.g., physically bound water), rather than of the moisture content. The total solids content is
the total water content. For example, the rate of a measure of the amount of material remaining
microbial growth in a food depends on the after all the water has been evaporated:
amount of bulk water present in a food, and not
M DRIED
necessarily on the total amount of water present. %Total Solids = ×100
There are analytical techniques available that can M INITIAL
provide some information about the relative frac- Thus, %Total solids = (100 − %Moisture). To
tions of water in different molecular environ- obtain an accurate measurement of the moisture
ments (e.g., DSC, NMR, vapor pressure). content or total solids of a food using evaporation
methods it is necessary to remove all of the water
molecules that were originally present in the
Sample Preparation food, without changing the mass of the food
matrix. This is often extremely difficult to achieve
Selection of a representative sample, and preven- in practice because the high temperatures or long
tion of changes in the properties of the sample times required to remove all of the water mole-
prior to analysis, are two major potential sources cules would lead to changes in the mass of the
of error in any food analysis procedure. When food matrix, e.g., due to volatilization or chemi-
determining the moisture content of a food it is cal changes of some components. For this reason,
important to prevent any loss or gain of water. the drying conditions used in evaporation meth-
For this reason, exposure of a sample to the atmo- ods are usually standardized in terms of tempera-
sphere, and excessive temperature fluctuations, ture and time so as to obtain results that are as
should be minimized. When samples are stored accurate and reproducible as possible given the
in containers it is common practice to fill the con- practical constraints. Using a standard method of
tainer to the top to prevent a large headspace, sample preparation and analysis helps to mini-
because this reduces changes in the sample due to mize sample-to-sample variations within and
equilibration with its environment. The following between laboratories.
section provides an overview on the topic with Evaporation Devices The thermal energy used
some key methods. Please note these are some to evaporate the water from a food sample can be
key methods but not all the methods available. provided directly (e.g., transfer of heat from an
oven to a food) or indirectly (e.g., conversion of
electromagnetic radiation incident upon a food
Evaporation Methods into heat due to absorption of energy by the water
molecules).
Principles
Convection and Forced Draft Ovens
These methods rely on measuring the mass of Weighed samples are placed in an oven for a
water in a known mass of sample. The moisture specified time and temperature (e.g. 3 h at 100 °C)
content is determined by measuring the mass of a and their dried mass is determined, or they are
570 Appendix A: Moisture Analysis

dried until they reach constant mass. The thermal of microwave methods over other drying methods
energy used to evaporate the water is applied is that they are simple to use and rapid to carry out.
directly to the sample via the shelf and air that Nevertheless, care must be taken to standardize the
surround it. There are often considerable temper- drying procedure and ensure that the microwave
ature variations within convection ovens, and so energy is applied evenly across the sample. A
precise measurements are carried out using number of microwave oven drying methods are
forced draft ovens that circulate the air so as to officially recognized.
achieve a more uniform temperature distribution
within the oven. Samples that contain significant Infrared Lamp Drying
quantities of carbohydrates that might undergo The sample to be analyzed is placed under an
chemical changes or volatile materials other than infrared lamp and its mass is recorded as a func-
water should not be dried in a convection or tion of time. The water molecules in the food
forced draft oven. Many official methods of anal- evaporate because they absorb infrared energy,
ysis are based on forced draft ovens. which causes them to become thermally excited.
One of the major advantages of infrared drying
Vacuum Oven methods is that moisture contents can be deter-
Weighed samples are placed under reduced pres- mined rapidly using inexpensive equipment, e.g.,
sure (typically 25–100 mmHg) in a vacuum oven 10–25 min. This is because the IR energy pene-
for a specified time and temperature and their dried trates into the sample, rather than having to be
mass is determined. The thermal energy used to conducted and convected inwards from the sur-
evaporate the water is applied directly to the sample face of the sample. To obtain reproducible mea-
via the metallic shelf that it sits upon. There is an air surements it is important to control the distance
inlet and outlet to carry the moisture lost from the between the sample and the IR lamp and the
sample out of the vacuum oven, which prevents the dimensions of the sample. IR drying methods are
accumulation of moisture within the oven. The boil- not officially recognized for moisture content
ing point of water is reduced when it is placed under determinations because it is difficult to standard-
vacuum. Drying foods in a vacuum oven therefore ize the procedure. Even so, it is widely used in
has a number of advantages over conventional oven industry because of its speed and ease of use.
drying techniques. If the sample is heated at the
same temperature, drying can be carried out much Advantages and Disadvantages
quicker. Alternatively, lower temperatures can be • Advantages: Precise; Relatively cheap; Easy
used to remove the moisture (e.g. 70 °C instead of to use; Officially sanctioned for many appli-
100 °C), and so problems associated with degrada- cations; Many samples can be analyzed
tion of heat labile substances can be reduced. A simultaneously
number of vacuum oven methods are officially • Disadvantages: Destructive; Unsuitable for
recognized. some types of food; Time consuming

Microwave Oven
Weighed samples are placed in a microwave oven Distillation Methods
for a specified time and power-level and their dried
mass is weighed. Alternatively, weighed samples Principles
may be dried until they reach a constant final
mass - analytical microwave ovens containing bal- Distillation methods are based on direct measure-
ances to continuously monitor the weight of a food ment of the amount of water removed from a food
during drying are commercially available. The sample by evaporation: %Moisture = 100
water molecules in the food evaporate because (MWATER/MINITIAL). In contrast, evaporation ­methods
they absorb microwave energy, which causes them are based on indirect measurement of the amount of
to become thermally excited. The major advantage water removed from a food sample by evaporation:
Appendix A: Moisture Analysis 571

%Moisture = 100 (MINITIAL −  MDRIED)/MINITIAL. • Disadvantages: Destructive; Relatively time-­


Basically, distillation methods involve heating a consuming; Involves the use of flammable sol-
weighed food sample (MINITIAL) in the presence of vents; Not applicable to some types of foods.
an organic solvent that is immiscible with water.
The water in the sample evaporates and is col-  hemical Reaction Methods
C
lected in a graduated glass tube where its mass is Reactions between water and certain chemical
determined (MWATER). reagents can be used as a basis for determining
the concentration of moisture in foods. In these
methods a chemical reagent is added to the food
Dean and Stark Method that reacts specifically with water to produce a
measurable change in the properties of the sys-
Distillation methods are best illustrated by exam- tem, e.g., mass, volume, pressure, pH, color, con-
ining a specific example: the Dean and Stark ductivity. Measurable changes in the system are
method. A known weight of food is placed in a correlated to the moisture content using calibra-
flask with an organic solvent such as xylene or tion curves. To make accurate measurements it is
toluene. The organic solvent must be insoluble important that the chemical reagent reacts with
with water; have a higher boiling point than all of the water molecules present, but not with
water; be less dense than water; and be safe to any of the other components in the food matrix.
use. The flask containing the sample and the A method that are commonly used in the food
organic solvent is attached to a condenser by a industry are the Karl-Fisher titration
side arm and the mixture is heated. The water in
the sample evaporates and moves up into the con- Karl-Fisher Method
denser where it is cooled and converted back into The Karl-Fisher titration is often used for deter-
liquid water, which then trickles into the gradu- mining the moisture content of foods that have low
ated tube. When no more water is collected in the water contents (e.g. dried fruits and vegetables,
graduated tube, distillation is stopped and the confectionary, coffee, oils and fats). It is based on
volume of water is read from the tube. the following reaction:

Practical Considerations 2H2O + SO2 + I2 = H2SO4 + 2HI


There are a number of practical factors that can
lead to erroneous results: (1) emulsions can This reaction was originally used because HI
sometimes form between the water and the sol- is colorless, whereas I2 is a dark reddish brown
vent which are difficult to separate; (2) water color, hence there is a measurable change in color
droplets can adhere to the inside of the glassware, when water reacts with the added chemical
(3) decomposition of thermally labile samples reagents. Sulfur dioxide and iodine are gaseous
can occur at the elevated temperatures used. and would normally be lost from solution. For
this reason, the above reaction has been modified
Advantages and Disadvantages by adding solvents (e.g., C5H5N) that keep the
• Advantages: Suitable for application to foods S2O and I2 in solution, although the basic princi-
with low moisture contents; Suitable for appli- ples of the method are the same. The food to be
cation to foods containing volatile oils, such analyzed is placed in a beaker containing solvent
as herbs or spices, since the oils remain dis- and is then titrated with Karl Fisher reagent (a
solved in the organic solvent, and therefore do solution that contains iodine). While any water
not interfere with the measurement of the remains in the sample the iodine reacts with it
water; Equipment is relatively cheap, easy to and the solution remains colorless (HI), but once
setup and operate; Distillation methods have all the water has been used up any additional
been officially sanctioned for a number of iodine is observed as a dark red brown color (I2).
food applications. The volume of iodine solution required to titrate
572 Appendix A: Moisture Analysis

the water is measured and can be related to the It is an alternative to typical NMR, commonly
moisture content using a pre-prepared calibration used in structural analysis. In TD-NMR, it is pos-
curve. The precision of the technique can be sible to use permanent magnets as it works at low
improved by using electrical methods to follow magnetic fields. Permanent magnets are simple,
the end-point of the reaction, rather than observ- inexpensive and do not need extensive cooling by
ing a color change. Relatively inexpensive com- liquid gases. However, the low magnetic field
mercial instruments have been developed which results in a low resolution. This is inadequate for
are based on the Karl-Fisher titration, and some obtaining Fourier-transformation frequency
of these are fully automated to make them less spectra. Hence TD-NMR receives the data from
labor intensive. the analysis of dependence of signal intensity on
time. The signal time-dependency is either fitted
directly with appropriate equations or relaxation
Physical Methods time constants are obtained from it. Figure X
illustrates the various regions of the electromag-
A number of analytical methods have been devel- netic spectra. Two regions are used for the deter-
oped to determine the moisture content of foods mination of moisture: the Near Infrared, described
that are based on the fact that water has apprecia- as NIR and the mid-Infrared region noted as
bly different bulk physical characteristics than the IR. The NIR region encompasses the region from
food matrix, e.g. density, electrical conductivity or 2500 to 700 nm. NIR measurements are made
refractive index. These methods are usually only with either a filter based instrument on an FT-NIR
suitable for analysis of foods in which the compo- instrument. In NIR the first second and third
sition of the food matrix does not change signifi- overtones of the OH stretch are seen at 1450, 970
cantly, but the ratio of water-to-food matrix and 760 nm respectively. The position of these
changes. For example, the water content of oil-­in-­ bands are temperature and hydrogen bonding
water emulsions can be determined by measuring Furthermore there are combination bands at 1940
their density or electrical conductivity because the and 1190 nm from OH stretching and bending.
density and electrical conductivity of water are Generally, the band at 1940 nm is used for low
significantly higher than those of oil. If the compo- moisture foods while a band 1450 for high mois-
sition of the food matrix changes as well as the ture foods. In the case of FTIR which can be
water content, then it may not be possible to accu- defined as the region from about 600–4000 wave-
rately determine the moisture content of the food numbers peaks at 3300 wavenumbers (broad OH)
because more than one food composition may give and 11,650 wavenumbers (HOH bend) are used.
the same value for the physical property being
measured. In these cases, it may be possible to use References
a combination of two or more physical methods to
determine the composition of the food, e.g., den- Determination of moisture and total solids; peo-
sity measurements in combination with electrical ple.umass.edu/~mcclemen/581Moisture.html.
conductivity measurements. Kasaturi, P., & Vedula. (1998). Near infrared
spectroscopy development for temperature
Spectroscopic Methods control in the baking process in seminars.
Spectroscopic methods utilize the interaction of Food Analysis, 136–172. www.bruker.com.
electromagnetic radiation with materials to Amir, R. M., Anjum, F. M., Khan, M. I., Khan,
obtain information about their composition. M. R., Pasha, I., & Nadeem, M. (2013).
Three of the more established methods are Application of Fourier transform infrared
TD-NMR,NIR and mid-IR. (FTIR) spectroscopy for the identification of
TD-NMR (Time Domain) is also called Pulsed wheat varieties. Journal of Food Science and
NMR TD-NMR is a highly powerful technique, Technology, 50, 1018–1023.
 ppendix B: Units and Conversion
A
Factors

The International System (SI) of the Units rests appendix is given for convenience in converting
upon seven base units and two supplementary these units, Table B.4.
units as shown in Table B.1. From the base units,
derived units can be obtained to express various Temperature
quantities such as area, power, force, etc. Some
of these have special names as listed in Table B.2. 0 °C = 273 K
Multiples and submultiples are obtained by using Celsius was formerly called Centigrade
prefixes as shown in Table B.3. 100 °C = (100 × 1.8) + 32 °F = 212 °F
Older units in the metric system and the avoir- 0 °C = 32 °F
dupois system are still widely used in the litera- °F = (°C × 1.8) + 32
ture, and the information supplied in this °C = (°F – 32) ÷ 1.8

Table B.1  Base units and supplementary units Table B.2  Derived units with special names
Quantity Unit Symbol Quality Unit Symbol Formula
Base units Force newton N kg.m/s2
 Length meter m Energy joule J N.m
 Mass kilogram kg Power watt W J/s
 Time second s Pressure pascal Pa N/m2
 Electric current ampere A Electrical potential volt V W/A
 Temperature kelvin K Electrical resistance ohm Ω V/A
 Luminous intensity candela cd Electrica conductance siemens S 1/Ω
 Amount of substance mole mol Electrical charge coulomb C A∙s
Supplementary units Electrical capacitance farad F C/V
 Plane angle radian rad Magnetic flux weber Wb V∙s
 Solid angle steradian sr Magnetic flux density tesla T Wb/m2
Inductance henry H Wb/A
Frequency hertz Hz 2π/s
Illumination lux lx cd∙sr/m2
Luminous flux lumen lm cd∙sr

© Springer International Publishing AG 2018 573


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8
574 Appendix B: Units and Conversion Factors

Table B.3  Multiples and submultiples


Multiplier Exponent form Prefix SI symbol
1,000,000,000,000 1012 tera T
1,000,000,000 109 giga G
1,000,000 106 mega M
1000 103 kilo k
100 102 hecto h
10 10−1 deca da
0.1 10−1 deci d
0.01 10−2 centi c
0.001 10−3 milli m
0.000001 10−6 micro μ
0.000000001 10−9 nano n
0.000000000001 10−12 pico p

Table B.4  Conversion factors


To Convert → Into Multiply By
0.30480 meters (m) feet (ft) (= 12 in) 3.28084
0.09290 m2 ft2 10.76391
0.02832 m3 cu ft. (ft3) 35.31467
28.31685 dm3—liters (L) ft3 0.03531
3.78541 liter (= 1000 cc) US gal (= 128 US fl. oz) 0.26417
4.54609 liter (= 1000 mL) Imp gal (= 160 I fl. oz) 0.21997
35.2383 liter (L) US bushel 0.02838
0.06 m3/h L/min 16.66667
1.69901 m3/h cu ft./min 0.58858
0.22712 m3/h USGPm 4.40287
0.27277 m3/h IGPM 3.66615
0.10197 kg (= 1000 g) Newton (N) 9.80665
0.45359 kg lb (av) (= 16 oz) 2.20462
0.90718 Metric ton (MT) Short ton (= 2000 lbs) 1.10231
1.01605 M ton (= 1000 kg) Long ton (= 2240 lbs) 0.96421
0.01602 kg/dm3 lb/ft3 62.42789
0.06895 bar (= 10 N/cm2) psi 14.50377
0.001 bar mbar (= 100 Pascals) 1.0 103
0.09807 bar mH2O 10.19716
1.33331 mbar mmHg (torr) 0.75001
33.77125 mbar inHg (60 °F) 0.02961
4.1868 kJ (kiloJoule) kcal 0.23885
1.05504 kJ BTU 0.94783
3.6 103 kJ kWh 0.27778 10−3
0.85985 103 kcal kWh 1.16300 10−3
1.16300 10−3 kW kcal/h 0.85985 103
0.29307 10−3 kW (kJ/s) BTU/h 3.41219 103
0.25199 kcal/h BTU/h 3.96838
0.746 kW HP (electr.) 1.34048
0.73550 kW Metric hp 1.35962
0.0935 foot-candle (ft-c) lux 10.76
1 centipoise (cp) mPa.s 1
1 centisokes (cSt) mm2/s 1
Multiply By Into ← To Convert
Appendix C: Greek Alphabet

Greek character Greek name Roman equivalent


Aα alpha Aa
Bβ beta Bb
Γγ gamma Gg
Δδ delta Dd
E ε, ∈ epsilon Ĕĕ
Ζζ zeta Zz
Ηη eta Ēē
Θ φ, θ theta Th th
Ιι iota Ii
K ℵ, κ kappa Kk
Λλ lambda Ll
Μμ mu Mm
Nν nu Nn
Ξξ xi Xx
Οο omicron Ŏö
Π π, ϖ pi Pp
Ρρ rho Rr
Σ σ, ς sigma Ss
Ττ tau Tt
ϒυ upsilon Yy
Φ φ,ϕ phi Ph ph
Χχ chi Ch ch
Ψψ psi Ps ps
Ωω omega Ōō

© Springer International Publishing AG 2018 575


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8
Index

A monoacyl, 39, 90, 97, 408, 409, 411


A bands, 149 triacyl (see Triglycerides)
Acceptable daily intake (ADI), 538, 539, 545, 551, Additives
554, 559 and contaminants, 527–563
Acesulfame-K, 178, 534, 543–545 incidental, 531, 549–563
Acetaldehyde, 317, 319, 320, 322, 324, 461, 485, 503, intentional, 531–533
537, 557 Additives, incidental contaminants
Acetals, 172, 324 antibiotics, 555–556
Acetic acid, 43, 142, 207, 295, 317, 322, 324, 425, 461, bacterial and fungal toxins, 528
467, 498, 500, 503, 533, 540–542 dioxin, 551–552
Acetophenoses, 310 from industry, 556, 557
Acetylated sugars, 168 natural toxins, 528, 529
Acetylformoin, 184 pesticides, 549–554, 560
Acetyl glucosamine, 178 polychlorinated biphenyls (PCB), 552–555
Achromatic colors, 258 polycyclic aromatic hydrocarbons (PAHs),
Acid detergent fiber (ADF), 223, 225 560–562
Acids trace metals, 556, 560
as acidulants, 540, 544 Additives, intentional, classes
as preservatives, 540 anticaking, 545
properties, 43, 50, 99, 139, 140, 295, 540, 541, 543 firming, 535, 545
Acidulants, acids glazes, 559
acetic, 43, 142, 184, 204, 207, 295, 311, 317, 322, humectants, 535
324, 455, 461, 467, 498, 503, 533, 540–542 polishes, 560
adipic, 295, 540, 541 propellants, 518, 535
citric, 85, 88, 112, 142, 177, 215, 239, 289, 295, 301, Additives, intentional, classification, see Food additives
303, 379, 403, 439, 454, 498, 534, 535, Additives, intentional, major classes
540–542 acidulants, 535, 540, 546
fumaric, 295, 462, 541 antimicrobial agents, 533, 536
lactic, 295, 317, 454–455, 500, 535, 540–542 antioxidants, 532–534, 536, 537, 540–542, 549
malic, 247, 295, 380, 439, 497, 498, 500, 540, 541 bleaching agents, 539, 543, 552
phosphoric, 60, 61, 88, 117, 206, 232, 238, 239, 243, bread improvers, 543
294, 295, 385, 403, 540, 544 coloring agents, 546
succinic, 319, 498, 540, 542 emulsifiers, 532, 533, 535, 542–543
tartaric, 295, 439, 495, 497, 498, 501, 540–542 flavor enhancers, 534, 545
Actin, 125, 148, 149, 152 flavors, 527, 532, 534, 536, 538, 545, 549
Active methylene group, 83, 84 irradiation, 546–548
Active oxygen method (AOM), 73, 81, 85, 92, 93 nutritional supplements, 547
Actomyosin, 148, 149, 152, 241 phosphates, 533, 535, 544–546
Acute toxicity, 552 preservatives, 532–534, 536, 539, 540
Acyclic aldehyde, 168 spices, 527, 532–534, 540, 548
Acylglycerols sweeteners, 532, 534, 543–545
diacyl, 39, 40, 90, 97, 409 Adenosine triphosphate (ATP), 148, 149, 161, 238,
glycerides, 39–43, 48, 53–63, 67–72, 83, 84, 90, 321, 485
95–97, 103–105, 322, 352, 407, 408, 411 Adhesiveness, 111, 334, 348

© Springer International Publishing AG 2018 577


J.M. deMan et al., Principles of Food Chemistry, Food Science Text Series,
https://doi.org/10.1007/978-3-319-63607-8
578 Index

ADI, see Acceptable daily intake (ADI) sorghum, 122


Adipic acid, 295, 541 wheat, 118
Adipic anhydride, 204 Amino acids, general
Adsorption, heat of, 21–23 acidic, 117, 145, 151
Adsorption isotherm, 20, 24 amides, 305
Aeration, 103, 140, 542 aromatic, 439, 466, 540
Aftertaste, 292, 303, 315, 543 basic, 117, 145, 300, 304
Agar, 217, 219, 359 cyclic, 439
Agaropectin, 220 essential, 68, 117, 118, 120, 122, 128, 139, 143,
Agarose, 219, 220, 521 156–158
Agent Orange, 552 hydroxyl, 542
Aggregated gels, 142 polar, 124
Aggregation, protein, 127 structure, 117, 125, 127
Aglycone, 172, 300, 440, 477 sulfur, 117, 127, 137, 138
Agriculture and Agri-food Canada, 477 Amino acids, individual
Agrobacterium, 517, 518 alanine, 120, 121, 128, 143, 301, 320
AH,B theory, 293 arginine, 118, 120, 121, 128, 143
Albumins, 117, 120, 125–127, 137, 143, 146, 147, asparagine, 120, 143, 292
154–158, 160, 304 aspartic, 120, 121, 128, 143, 148, 157, 301
Alcoholic beverages, 320, 324–325, 483, 484, 545, 547 cysteine, 118, 127, 249
Alcohol, sugars, see Sugar alcohols cystine, 121, 127, 128, 138, 151, 249
Aldehyde oxidase, 246 glutamic acid, 120, 121, 128, 143, 145, 292, 301, 304
Aldehydes, 79, 81, 83–85, 90, 93, 95, 132, 134 glutamine, 117, 118, 120, 143
Aldehyde sulfurous acid, 537 glycine, 118, 120, 121, 128, 143
Aldehydo-glycerides, 84 histidine, 118, 120, 121, 128, 143, 292, 301
Aldoseamine, 129 hydroxy proline, 128
Aldoses, 128–130, 135, 166–168 isoleucine, 118, 120, 121, 128, 143, 292, 301, 320, 504
Aldosylamine, 132 leucine, 118, 120, 121, 128, 143, 301, 320, 491, 504
Aleurone grains, 158 lysine, 118, 120–122, 128, 137, 143, 155
Alginate, 197, 218, 220, 236 methionine, 118, 120, 121, 128, 143, 249, 292, 320
Aliphatic acids, 319, 439, 467 ornithine, 135
Aliphatic alcohol, 63, 64, 466 phenylalanine, 118, 120, 121, 128, 143, 292, 301, 320
Alkaline taste, 302 proline, 118, 120–122, 128, 143, 157, 301, 404
Alkaloids, 280, 290, 294, 299, 461, 463, 474, 529 serine, 120, 121, 128, 143, 292, 301, 320
Alkanones, 321 threonine, 118, 120–122, 128, 143, 301, 320
Allele pairs, 511 tryptophan, 118, 120–122, 143, 156, 291, 292, 301
Allergen, 161, 162, 421 valine, 118, 120, 121, 128, 143, 292, 301, 491, 504
Allergenic properties, 419 Amino acids, sulfur
Allergic reactions, 555 cysteine, 118, 120, 127, 138, 249
Alliin, 463 cystine, 121, 127, 128, 138, 249
Allium, 458 methionine, 118, 120, 121, 128, 137, 138, 143, 155,
Allspice, 458, 460, 461, 496 158, 249, 292, 301, 320
Alpha amylase, 199–202, 282, 405, 406, 411–413 Amino acrylic acid, 138
Alpha helix, 120, 123, 124, 146, 148, 157, 159 Amino-nitro-propoxybenzene, 291
Aluminum, 247, 263, 443, 546 Amino-propanol, 388
phosphate, 298 Amino sugars, 129, 136, 168, 171–172, 191
Alzheimer’s disease, 247, 478 Ammonium chloride, 543
Amadori rearrangement, 129, 131 Ammonium phosphate, 242, 535
Amaranth, 546 Ammonium sulfate, 123, 143, 535, 543
Amino acid, composition Amorphous foods, 29
beef, 49, 50 Amphiphilic, 61, 140, 206
casein, 128, 145, 404 Amygdalin, 177
corn, 122 Amylase(s), 31, 32, 143, 177, 193, 201, 397, 398, 411,
egg, 53, 54, 127 413, 484, 486, 489
gelatin, 128, 152 Amyl esters, 322
milk, 123, 143 Amyloglucosidase, 202, 282, 406
millet, 122 Amylopectin, 192–197, 200–202, 354, 411, 412
peas, 118 Amylose, 192–203, 354, 411
rice, 118 Amylose complexing, 109
serum albumin, 127, 128 Angle of tilt, 98, 101, 102
Index 579

Anhydride formation, 181 Aroma


Anhydro base, 279 character, 457, 496
Anhydro sugars, 168, 183, 184 wheel for beer, 504
Animal depot fat, 50, 55 wheel for wine, 315–317, 506
Animal proteins, 118, 142–147, 154, 387 Aromatic acids, 279, 466
Anisaldehyde oxime, 293, 294 Ascorbic acid, 237, 245, 278, 366, 368, 379–382, 385,
Aniseed, 458, 460 392–394, 399, 422, 436, 444, 449, 453, 454,
Anisidine value, 81, 84 461, 462, 533, 534, 538, 540
Anisotropic particles, 357 oxidase, 379, 399
Annatto, 265, 266, 276, 534, 547 Ascorbic L, acid, see Vitamin C
Anomers, 167, 168, 170, 172, 177, 184, 214, 291 Ascorbyl palmitate, 88, 376, 392, 393, 540
Anthocyanidins as synergist, 376
antioxidants, 301 Ashing, 236
cyanidin, 277, 278, 442 Asparagus, 179, 249, 275, 316, 368, 437, 438, 444, 445,
delphinidin, 277, 278, 442 450–452, 454, 502
destruction, 278 Aspartame, 178, 287, 534, 543–545
lakes, 279 Aspergillus. spp
malvidin, 277, 278, 442 A. flavus, 528–530
oxidation, 240, 398 A. niger, 178, 406, 411, 415, 417
pelargonidin, 277, 278, 442 A. oryzae, 406, 413, 417, 432, 484
peonidin, 277, 278 Astaxanthin, 265, 266, 276
petunidin, 277, 278 Astringency, 280, 301, 302, 316, 317, 441,
spectrum UV, 277 494, 501
Anthocyanins, 262, 265, 266, 276–279, 317, 380, 441, Asymmetric carbon, 282, 375, 391
444, 451, 497, 498, 501 Atmospheric condition, 6, 448
Antibiotics, 153, 517, 540, 555–556 Autoxidation, 73, 78, 80, 81, 84, 87, 89, 91, 426
Antibiotics as contaminants Avidin, 124, 153, 154, 391
causing allergies, 555 Avogadro’s number, 109
causing resistant organisms, 555
used in agriculture, 555
Anticaking agents, 545 B
Antifirming effect, 109 Baccillus subtilis (B. subtilis), 413
Antimicrobial agents, see Preservatives Bacillus coagulans, 432
Antioxidants Bacillus licheniformis, 419
ascorbyl palmitate, 88 Back extrusion, 347
BHA, 88, 540 Bacterial growth, aw, 30, 31
BHT, 88 Bacteriocins
chelating agents, 88, 540 in milk, 540
enzymic, 540 natamycin, 540
natural, 85, 88, 540 nisin, 540
oxygen scavengers, 540 pimaricin, 540
PG, 540 as preservative, 540
phenolic, 80, 87, 88, 404, 540, 542 Barbecuing, 561
spices and herbs, 540 Barium, 14
TBHQ, 88, 540 Basic tastes
tocopherols, 85, 88, 375, 540 acid, 285, 304
tocotrienol, 540 salt, 285, 290
Antiparallel hydrogen bonding, 293 sour, 290
Antiparallel sheets, 146 umami, 286, 290
AN/TN ratio, 419 Basil, 458
Apo carotenal, 266 Bay tree fruits and leaves, 458
Apoenzyme, 400 Beans, 2, 35, 41, 62, 95, 111, 118, 179, 192, 195, 202,
Apolar molecules, 16 216–219, 234, 245, 248, 249, 270, 275, 316,
Apparent viscosity, 332, 341, 344, 345 317, 323, 324, 347, 353, 368, 377, 386, 387,
Applesauce, 249 392, 398, 399, 421, 427, 438, 439, 442–445,
Apricot, 179, 216, 274–276, 295, 316, 382, 398, 450, 451, 453, 454, 465–467, 528, 529, 553
437–439, 442, 444, 445, 447, 454, 496, 551 Beef fat, see Fats and oils, individual
Arabinoxylans, 211 Beer haze, 301, 418, 487
Arachidonic acid, 46, 53, 83 Bell peppers, 313, 316, 458, 498, 502
Aroclor, 554, 555 Benzidine value, 79–81
580 Index

Benzoic acid, 461, 533 Brittleness, 333, 334, 348, 351


as preservative, 533 Brookfield viscometer, 345
Benzopyrone nucleus, 279 Brothy taste, 322
Benzoyl peroxide, 542, 543 Brownian movement, 355–357
Benzopyrene, 560–564 Browning, nonenzymatic, see Nonenzymatic browning
Benzyl alcohol, 293, 323 Buccal proteins, 317
Beryllium salt, 290, 295 Buffering capacity, 295
Betacyanins, 266, 281 Bulking agent, 111, 112, 165, 214, 217
β-amino alanine, 139 Bulk sweeteners, 174, 178
β-elimination, 129, 138 Burgers model, 339
β-fructofuranosidase, 178 Butter, 1, 2, 39, 42, 44, 49, 51, 55–59, 66, 70, 71, 93,
β-furanoside, 168 95–97, 102, 103, 106, 111, 234, 309, 310, 316,
Beta-ionone, 276 317, 319, 339, 340, 343, 347, 352, 370, 372,
Betalains, 281–282 374, 377, 408–410, 467, 489, 496, 500, 528,
Betanin, 281, 282 535, 540
Betaxanthins, 266, 281 flavor, 310
BET isotherm, 21, 23 thixotropic hardness, 343
BHA, see Butylated hydroxy anisole (BHA) Butylated hydroxy anisole (BHA), 85, 87, 88, 534, 540
BHT, see Butylated hydroxy toluene (BHT) Butylated hydroxy toluene (BHT), 85, 87, 88,
Bifidus factor, 178 534, 540
Bile acids, 64 Butyric acid, 42–45, 310, 311, 317, 467, 503, 538
Bimolecular lipid layers, 109
Binding energy, 6
Bingham body, 340, 341 C
Biopolymers, 28, 359 Cabernet Sauvignon grapes, 278
Biotechnology, 397, 405, 515, 516, 520 Cabernet Sauvignon wine, 497, 501, 502, 504
Biotin Cadmium, 243, 560
avidin, 153 Caffeine, 289, 291, 299, 317, 562–563
sources, 391 Calcium
stability, 391 caseinate phosphate complex, 144
structure, 391, 392 channels, 288
Birefringence, 196, 197 sensitive casein, 145
Bisulfite, 135, 279, 425, 537 sulfate, 535, 543
Bitter peptides, 298, 300, 321, 322, 417 Calcium-chelate, 241
Bitter receptor, 304 Callus, 517, 518
Bitter taste, 184, 200, 286, 288–291, 298–301, 321, 486, Campesterol, 64, 67, 462
491, 492 Camphoraceous, 313
Bixa orellana, 272 Canadian Food and Drugs Act, 551
Bixin, 270, 272, 273, 427 Cancer, 247, 528, 529
Black body, 255 Candida antarctica, 409
Black pepper, 302, 316, 458, 460, 463–464, 502 Candida cylindracae, 411
Blanching, 276, 370, 380, 388, 390, 391, 397, 399, 403, Canned foods, metal uptake, 247–249
424, 425, 427, 449, 451–454, 465, 551 Canning, 275, 320, 321, 383, 386, 387, 448, 452,
Bleaching, 67, 427, 451, 539, 543, 552 454, 551
Blood pressure, 237, 238, 295, 296, 469, 529 Canola oil, see Fats and oils, individual
Blood serum albumin, 147 Canthaxanthin, 266, 276, 546
Bloom, 102, 103 Caper, 458
Bond angles, 5, 7, 120, 167 Capillary condensation, 24
Bonito, 305 Capillary viscometers, 345
Botulinum toxins, 538 Capillary water, 23, 24, 30, 31, 610
Branched chain fatty acids, 43 Capsaicin, 302, 462, 473
Branched dextrins, 202 Capsicum, 302, 458, 469
Brassica species, 458, 516, 520 Caramel
Brassicasterol, 64, 67 color production, 282
Bread flavor, 319, 320 flavor, 182, 184
Bread improvers Caramelization, 131, 135, 182–184, 253, 313, 320,
bleaching agents, 543 321, 449
inorganic compounds, 543 Caraway, 291, 458–460, 462
Bread, specific heat, 319 Carbanion, 139
Brevibacterium ammoniagenes, 432 Carbinol base, 279
Index 581

Carbohydrates, 1, 10, 27–29, 110, 111, 121, 123, 128, in oranges, 275, 276
131, 153, 159, 160, 165–225, 233, 238, 282, in palm oil, 275
286, 287, 313, 324, 329, 353, 355, 359, 365, in peaches, 274
382, 398, 400, 416, 418, 435–438, 446, 449, in salmon, 273
453, 483, 486, 489, 610–612 stability, 276
Carbohydrates, classification synthetic, 276
cellulose, 165, 166, 170, 171, 177, 192, 437 in tomatoes, 274, 275
cyclodextrins, 212–215 Zeaxanthin, 273, 275, 276
dextrins, 199, 201, 202, 206, 207 Carrageenan, 111, 152, 197, 218, 220–221, 534, 535
disaccharides, 165, 166, 175–178, 182–184 Carrots, 2, 28, 41, 166, 179, 209, 216, 234, 247, 249,
fiber, 198, 199, 212, 215, 222–225 262, 266, 270, 275, 276, 279, 334, 353, 368,
glycogen, 165, 166, 177, 208–209 370, 377, 379, 381, 414, 437–439, 442, 444,
gums, 165, 173, 217–219, 226 445, 449, 452–454, 502, 547
hemicelluloses, 165, 211–212, 223–226 Carry-through properties, 85
hexoses, 166–168 Carvacrol, 460–462
hydrocolloids, 197 Carvone, 291, 459–461
lignin, 209, 211–212, 223–225 Casein, 123, 125, 127, 128, 142–145, 147, 240, 300,
monosaccharides, 165–171, 174, 175, 178, 192 304, 418
oligosaccharides, 165, 175–191, 201, 202 See also Proteins, casein
pectins, 192–196, 200–202, 215–219, 222–225, 437 Casein, α, α3, αS2, κ, 143–145
pentosans, 211–212, 225 Caseinates
polyols, 172, 177, 178 agglomeration, 221
polysaccharides, 165, 166, 168, 175, 177, 178, particles, stability tests, 144, 221, 240, 241
191–225, 437, 452 Casein, genetic variants, 144, 145
starch, 165, 166, 170, 174, 175, 177–179, 192–209, Casein hydrolysates, 419, 420
211, 217, 220, 222, 225, 436, 437 Casein micelle, 144, 145, 240, 416
Carbohydrates, content in foods, 222 Casson model, 332
Carbohydrates, physical properties Catalase heat stability, 424
gum gels, 217–220, 226 Catalysts
lactose, 176–178, 188–191 hydrogenation (see Hydrogenation, fats)
pectin gels, 215–216, 223, 225 interesterification, 71
starch crystallinity, 194 Catechin, 280, 422, 442, 462, 501
starch gels, 196, 197 Catecholase activity, 421, 422
starch paste, 196, 197 Celery, 173, 174, 311, 312, 334, 437–439
sucrose, 166, 168, 170, 173–179, 182, 184–186, 188, Cellobiose, 176, 177, 436
190, 200, 202 Cellular component, 225, 237, 446–447
Carbonates, 14, 236, 239, 535 Cellular structure, 2, 9, 353
Carbonic anhydrase, 245 Cellulose
Carbonium ion, 279 crystallinity modified, 209
Carbon number, glycerides, 43 molecular weight, 209, 219
Carbonyls, 70, 79, 81, 123, 129, 136, 137, 166, 175, 238, molecule, 219
303, 319–323, 450, 451, 466 Cephalin, 60
Cardamom, 458, 468–469 Cereal germ oils, 375
Carmelen, 183 Cereal proteins, 118, 156, 158
Carnosic acid, 461, 462, 540 Certified colorants (synthetic), 263
Carnosol, 462 Chain elongation, 42
Carotene, 88, 270, 272, 273, 275, 367, 370, 372, 374, Chain fission, 83, 84
427, 444 Chain reaction, 78, 80, 83, 90, 161, 392, 461
Carotenoids Chain shortening, fatty acids, 42
annatto, 265, 266 Chalcones, 279, 294, 441
astaxanthin, 265, 266 Charcoal broiling, 561
canthaxanthin, 266 Charge frequency, 140–142
capsanthin, 462 Cheese flavor, 319
carotene, 272 Chelates, 85, 231, 233, 235–237, 241, 245, 247
in corn, 275 Chemisorption, 72, 74
cryptoxanthin, 276 Chewiness, 140, 334, 348
in eggs, 266, 273, 372 Chicken fat, 58
lutein, 275, 276 Chili (Tobasco), 458
lycopene, 273–275 Chilies, 332, 462, 496
in milk fat, 88 Chiral, 50, 54
582 Index

Chiral center, 166, 375 Coconut, see Fats and oils, individual
Chives, 458 Coconut oil, 70, 104, 375
Chlorine Codex Alimentarius, 549, 560
bleaching, 552 Codmium contamination in crustaceans, 560
dioxide, 552 Coenzyme, 365, 382, 384, 386, 389
Chlorogenic acids, 278, 422, 440, 441, 461, 462 Coenzyme A
Chlorophyll, 33, 88, 123, 232, 246, 266, 268–270, 274, sources, 391
425, 427, 444, 446, 448–451, 497, 547 stability, 391
Chlorophyllase, 269, 449–451 structure, 391
Chlorophyllin, 268, 547 Cofactor, 400, 418, 485, 505
Chloroplastids, 268 Coffee, 33, 111, 140, 170, 245, 306, 309, 312, 316,
Chocolate 323–325, 398, 399, 421, 494, 552, 561,
aroma, 312, 502 562, 613
microstructure, 356 flavor, 323, 324, 494
milk, 106, 222 Cognac, 324, 325
Cholecalciferol, 372 Cohesion, 333, 452
Cholesterol Cohesiveness, 140, 141, 203, 334, 348, 354, 447
acetate, 66 Collagen, 119, 122, 124, 125, 149–152
esters, 64, 66, 405 Colloidal dispersions, 196, 355
Chondrus crispus, 220 Colloidal particles, 239, 356, 357
Chroma, 258, 260, 261 Colloids
Chroman ring, 373, 374 anisotropic particles, 357
Chromaticity coordinates, 257, 258 definition, 355
Chromatogram, 50–52, 75, 77, 323, 324, 554 dimensions, 356
Chromen sulfonic acid, 279 particle size, 355, 356
Chromium, 243, 245 Color and food colorant
intake, 246–247 anthocyanins, 262, 266, 276–283
Chromoproteins, 123 betalains, 266, 281–282
Chromosome, 512, 517, 518, 524, 525 betanidin, 281, 282
CIE system, 253–259, 261, 262 caramels, 253, 258, 265, 266, 282–283
Cinnamaldehyde, 457, 459–461, 473, 474, 476 Colorants
Cinnamon, 457–461, 473–477 not requiring certification, 263–268
Circumvallate papillae, 286 synthetic, 263
Cis-cis-1,4 diene system, 93 Color difference, 261
Citral, 322, 459–461, 470 Coloring agents, 546
Citrates, 239 Coloring agents, additives, 546
Citric acid as synergist, 376 FD&C colors
Citronellyl acetate, 322, 460, 470 lakes, 263, 279, 546
Citrus flavor, 322 nature-identical, 267
Citrus fruit, 273, 279, 300, 322, 367, 379, 390, 413, 438, permitted, 546
439, 441, 459 synthetic dyes, 546
Clathrates, 16 Color measurements
Clausius Clapeyron law, 3, 22, 23 chromaticity coordinates, 258
Clostridium botulinum, 538 CIE system, 253–258, 260
Cloud separation, 413 complementary, 258
Cloves, 316, 457–461 gloss, 262
Clupein, 122 Hunter, 253, 256, 260–261
Coating fats, 60 Lovibond, 253, 261–262
Cobalt, 243, 245, 388, 389 Munsell, 253, 258–261
Cocarboxylase, 382 primary color, 255
Cocoa, 57, 132, 245, 312, 316, 356, 398, spectrophotometry, 253
464, 562 tristimulus values, 255, 256, 258
Cocoa butter wavelength, 253–258, 270, 271, 275, 282
equivalents, 106, 409 Color, pigments
fatty acids, 49, 57, 70 anthocyanins, 262, 266, 276–280
improvers, 382 benzopyran, 266, 276–280
isosolids blends, 70 betalains, 281–282
polymorphism, 100 caramel, 282–283
solid fat profiles, 96 carotenoids, 270–276
substitutes, 70 chlorophylls, 268–270
Index 583

cholemyoglobin, 268 Conjugation, 319, 427


dyes, 263, 264 Connective tissue, 148, 149, 151, 152, 349
flavonoids from artifacts, 266, 276–280 Consistency, 44, 104, 197, 332, 333, 346, 349, 350,
hemoglobin, 265, 266 352, 532
isoprenoid, 266, 270–276 Contaminants, 243, 527–563
myoglobin, 266–269 See also Additives, incidental
nitrosohemochrome, 268 Continuous phase, 1, 108, 197, 355–358
nitrosomyoglobin, 268 Contractile elements, 152, 349
not requiring certification, 263–268 Contraction, 95, 148, 149, 238, 492
oxymyoglobin, 267, 269 Contributory flavor compounds, 319
pheophytins, 269, 270 Cooked flavor, 146
sulfmyoglobin, 268 Cooling effect, 17, 302
synthetic, 263 Coolness, 97, 285, 302
tannins, 280–281 Coordination bonds, 266
tetrapyrrole, 266–268 Copolymerization, 429, 430
titanium dioxide, 265, 266 Copper, 78, 85, 87, 234, 235, 243, 245, 247, 248, 262,
xanthophylls, 270, 273–275 269, 303, 370, 379, 421, 422, 443, 451, 494,
Color purity, 261 540, 547
Color rendition, 255 Coprecipitates, 147
Colors Coriander, 458–460, 462
corn, 265, 266, 282 Corn
crude palm oil, 275 starch, 193, 197–199, 202, 282, 354–356, 412,
eggs, 266, 273 413, 486
fruits, 262, 265, 266, 268, 269, 276, 278–280 sweeteners, 200, 202–203
honey, 258, 259 syrup, 173, 175, 177, 199, 202, 203, 412, 534
maple syrup, 258, 259 Corn oil, see Fats and oils, individual
meat, 266–269, 281 Corrosion of tin cans, 249
milk fat, 88, 276 Cottonseed, 49, 56, 66, 73, 93, 104, 118, 266, 304, 332,
oranges, 263–265, 275, 276, 278 372, 377, 528, 547
paprika, 265, 266, 276 Cottonseed oil, see Fats and oils, individual
peaches, 274, 275, 278 Cottonseed protein, 118
raspberries, 278 Coumaryl alcohol, 211
red fish, 276 Covalent bonds, 4, 5, 14–16, 212, 293, 351, 357, 437
salmon, 266, 273 Creep curve, 337, 338, 340, 341
tomato, 274, 275 Cresolase activity, 421, 422
vegetables, 261, 262, 264, 266, 268, 273, 275, 276, Critical nucleation temperature, 9
278, 279 Crocetin, 270, 273
Color triangle, 256 Cross bonding of starch, 205
Complementary colors, 258 Crosshead, texture measurement, 343
Complex ions, 240 Cross linking, 145, 151, 156, 205, 207, 211, 317, 359,
Complex light types, 255 429, 430
Component fatty acids, 42, 43, 50–52, 54, 56, 75 Cross striation, 148
Composite foods, aw, 33 Cruciferae, 292
Compression cell, 347 Crucifera oils, 292
Compression force, 140 Crustacea, 171, 172, 191, 266, 273, 276, 560
Conalbumin, 153 Cryoprotection, 28
Condensed tannin, 280, 441 Cryptoxanthin, 273, 275, 276, 367
Confectionery fats, 100, 105 Crystal growth water, 8, 9
Configurations Crystallization, fats
aldoses, 166–168 crystal formation, 98
Fischer formula, 167 crystal polymorph, 97–99
Haworth formula, 167 crystal size, 97, 98
tautomeric forms, 168, 169 latent heat, 7–8, 95
Congeners, 552 nucleation, 97, 98
Conglycinin, 159 super cooling, 7, 25
Coniferaldehyde, 325 Crystallization, sugars
Coniferyl alcohol, 211 lactose, 176–177
Conjugase, 390 sorbitol, 174, 178
Conjugated dienes, 83 sucrose, 177–179
Conjugated double bonds, 74, 270, 275 Crystallization velocity, water, 9
584 Index

Crystal size of fats, 9, 352, 359 Deoxyribose-2, 525


Crystal structure, fats Deoxy sugars, 168
cocoa butter, 96, 97, 102, 103 Derived lipids, 83
double chain, 100 Desaturation, fatty acids, 42
hexagonal(α), 99 Desorption, hydrogenation fats, 72–74
hydrogenated palm oil, 92, 104, 105 Desorption isotherm, 19–21
hydrogenated sunflower oil, 92, 93, 103–105 Deterioration, 29, 33, 34, 72, 78, 79, 81, 88, 91, 100, 289,
long spacings, 100 322, 381, 392, 435, 445–448, 452, 461, 540
melting points, polymorphs, 99 Detinning, 247, 249, 559
orthorombic (β), 99, 101 Dextrinization, 196, 204, 206
palm oil triglycerides, 100, 103, 104 Dextrose equivalent, 199, 200
PEP, 104 DHA, see Docosahexaenoic acid (DHA)
polymorphism, 98, 100 Diabetic foods, 174
PSP, 104, 105 Diacetyl, 134, 184, 314, 324, 500, 503, 542
short spacings, 98, 101, 102 Diamino sugar, 129
transition, 102, 103 Diels-Alder reaction, 93
triclinic (β), 99 Dienal-2,4, 319
triple chain, 98–100 Dietary fiber
Crystal structure, water, 7 definition, 222
Cubic phase, 109 methods, 223
Cubic structure, water, 7 Differential scanning calorimetry, 8, 101, 198, 330
Cucumbers, 95, 310, 314, 317, 319, 427, 428, 432, 435, Diffused reflection, 262
436, 438, 447, 450, 454, 516 Dihydroperoxides, 83
Cuminaldehyde, 461 Dihydropyrazines, 312
Cutin, 223–225, 409 Diisopropylphosphorofluoridate, 418
Cyanidin, 277, 278, 442, 462 Diketo gulonic acid, 478–479
Cyanocobalamine, 388–389 Dilatancy, 332, 337
See also Vitamin B12 Dilatometry, 96
Cyclic diketones, 324 Dill, 458, 460, 462
Cyclic monomeric glycerides, 90 Dimethyl mercury, 557
Cyclic monomers, 89, 93 Dimethyl nitrosamine, 539
Cyclodextrin, 212–215 Dimethyl sulfide, 17, 321
Cyclohexylamine, 543 Dioxin, 551–552
Cyclopentenyl ring, 93 Dioxin, contaminants
Cysteine sulfoxides, 137 from agent orange, 552
Cytochrome C reductase, 426 from combustion, 552
Cytochrome oxidase, 379, 380 from herbicides, 552
from packaging, 549
from pulp and paper, 552
D Dioxygenase, 367
Dashpot in texture, 352 Diphenols, 422, 463
Decadienal-2,4, 84, 460 Directed interesterification, 68, 70
Deep frying fats, 91, 92 Disaccharides
See also Fat, deep frying cellobiose, 177
Deformation in texture, 331 lactose, 176–177
Dehydrated foods, 18, 33, 34, 169, 316 maltose, 177
Dehydration, 99, 100, 172, 176, 221–224, 419, 421, 436, melibose, 178
476, 488, 489, 492, 493 sucrose, 175–176
Dehydro-ascorbic acid, 380 sugar alcohols, 172–175
Dehydrocholesterol, 412 Disperse phase, 108, 356, 357
Dehydro retinol, 366 Disperse systems, 329, 342, 355–357
Delphinidin, 277, 278, 442, 462 Distarch adipate, 205
Denaturation, 111, 120, 125–127, 142, 146, 147, 153, Distarch phosphate, 199, 205
154, 403, 449, 450, 452 Distilled monoglyceride, 72, 110, 542
Dental health, 246 Distribution coefficients, 256
Denture tenderometer, 348 Disulfide bonds, 120, 124, 127, 141, 142, 155–157, 350,
Deodorization, oils, 90 351, 427
Deoxynivalenol, 530, 531 DNA, 161, 162, 397, 512, 514, 515, 517, 518, 520–525
Deoxyosulose, 130 Docosahexaenoic acid (DHA), 46, 52, 54, 411
Deoxyribonucleic acid, 526 Dominant wavelength, color, 257, 258
Index 585

Double bond migration in fatty acids, 75 238, 253–256, 258, 285, 290, 314, 334, 336,
Double chain length, X-ray diffraction, 99 341, 342, 350, 397, 436, 437, 445, 477, 485,
Double helix, 148, 193, 195, 201 534, 546
in starch, 195 Enolization, 181, 182
Dough, 32, 140–142, 156, 211, 212, 349–351, 392, 399, Entanglement, 352
427, 535, 542, 543 Entrainment, 105, 106
Drip loss, meat, 10 Entropy, 16, 25, 233–235
Dulcin, 291 Environmental Protection Agency (EPA), 46, 52–54, 560
Dynamic viscosity, 331, 332 Enzymatic activity, aw, 32
Enzymatic antioxidant superoxide dismutase, 540
Enzymatic browning, flavonoids, 478
E Enzyme
EDTA, see Ethylenediamine tetraacetic acid (EDTA) activity, 19, 30–32, 78, 119, 125, 148, 400, 403, 417,
Effective dose, in toxicity, 528 418, 449, 451
Effect of processing, 555 classification, 405
Egg, 2, 3, 24, 35, 39, 53–55, 63, 111, 118, 120, 121, 123, deactivation, 397
125–127, 142, 147, 153–154, 171, 191, 234, in food processing, 397, 398, 403, 430, 432
245, 247, 266, 273, 276, 304, 367, 369, 372, immobilization methods, 430
374, 377, 382, 383, 385, 386, 389–392, 399, sources, 406
423, 511, 534, 535, 547, 554, 556 specificity, 215, 313, 397, 404, 405, 407–409, 411,
protein, 111, 118, 142, 153–154 415, 417–419, 427
yolk, 24, 53–55, 63, 123, 142, 153, 154, 276, 367, Enzyme kinetics
374, 377, 386, 389, 391, 535 effect of concentration, 400–402
Egg yolk fat, see Fats and oils, individual effect of inhibitor, 403–404
Eicosapentaenoic acid (EPA), 46, 52, 76 effect of pH, 403
Einstein equation, 356 effect of temperature, 403
Elaidic acid, 42, 47, 52, 104 enzyme positional specificity, 405, 407
Elastic body, 337–340 Lineweaver-Burk plot, 402
Elasticity, texture, 334 Michaelis-Menten equation, 402
Elastin, 122 specificity of enzyme, 404
Electron microscopy, 158, 330 substrate concentration, 400–402
Electrophoresis, 153, 154, 158, 159, 425, 521 Enzyme reactor, 430
Electroplating, 244 Enzymes
Ellagic acid, 280, 441, 461 acid protease, 416–417
Ellagitannins, 280 aldehyde oxidase, 246
Emulsification, 108, 110, 112, 140–142, 209, 210, 239 alkaline phosphatase, 429
Emulsifiers alliinase, 461
fatty acid esters, 542, 543 amino peptidase, 419
hydrophile/lipophile balance (HLB), 542, 543 amylase, 32, 143, 159, 177, 193, 397, 483, 484
hydroxylated lecithin, 542 amylase α (endo), 202
monoglycerides and esters, 542, 543 amylases, α-, β-, gluco-, 170
polyglycerol esters, 542 amyloglucosidase, 202, 282, 406
Emulsifiers, nonionic ascorbic acid oxidase, 379, 399
atmul, 109 carbonic anhydrase, 245
myrj, 109 carboxypeptidase, 415, 416, 418, 419
span, 109 catalase, 399, 406, 424
tween, 109 cellulase, 397, 398, 406
Emulsion(s), 1, 10, 28, 106–110, 112, 140–142, 195, chlorophyllase, 449–451
204, 207, 332, 356–358, 367, 405, 536 chymosin, 416, 417, 535
destabilization, 10 chymotrypsin, 416–418
flocculation, 10 classification, 405
stability, 141, 195, 204 cocarboxylase, 382
Enantiomer, 48, 54, 57, 166 coenzyme A, 391
Ena1-2-trans, 319 coenzyme FAD, 365, 382, 389
Endo-α-amylase, 411 coenzyme FMN, 386
Endothia parasitica, 417 cytochrome C reductase, 384
Enediol, 182, 378 cytochrome oxidase, 379, 380
Energy, 1, 2, 6, 7, 9, 13–15, 25, 40, 47, 62, 74, 78, 79, dextran, sucrase, 398
88, 98, 100, 101, 120, 121, 124, 142, 148, 165, elastase, 417, 418
176–178, 185, 187, 208, 209, 232, 233, 237, esterase, 405–409
586 Index

Enzymes (cont.) lipoxygenases, 31, 32, 88, 426–428, 432, 450–453


ficin, 418 oxidoreductases, 421–432
fumarase, 432 pectic, 438, 453–455, 489
galactosidase, 32 proteases, 32, 399, 415–419
glucoamylase, 177, 201, 412–413 EPA, see Eicosapentaenoic acid (EPA); Environmental
glucose isomerase, 202, 406, 430, 432 Protection Agency (EPA)
glucose oxidase, 32, 399, 406, 423, 424, 540 Epicatechin, 280, 301, 442, 476
glycosyl transferase, 175 Epoxide, 275, 374, 376, 490, 501, 550, 554
hydrolases, 405–420 Equal point, 257
immobilized enzymes, 429–432 Equatorial hydroxyl, 167
invertase, 31, 124, 398, 403 Equilibrium relative humidity (ERH), 20, 33, 34
lactase, 176, 177, 398, 406, 413, 430, 432 Equivalency, 3, 18, 70, 106, 118, 199, 200, 209, 258,
lactoperoxidase, 424–426 266, 275, 290, 367, 372, 375, 384, 387, 409,
lipase, 31, 32, 70, 72, 321, 322, 399, 405–409, 411 498, 522, 541, 550
lipoxidase, 78, 85, 270, 426 Ergocalciferol, 372
lipoxygenase, 31, 32, 124, 159, 399, 426–428, Ergosterol, 372
450–453 Erucic acid, 75
lysozyme, 124, 147, 153 Erythorbic acid, 237, 245, 378, 540
mold catalase, 406 Escherichia coli, 432, 537
naringinase, 398 Essential amino acids, 117, 118, 120, 122, 128, 139, 143,
nuclease, 399 155–158
oxidoreductase, 421–429 Essential oils, 322, 457, 459, 460, 462, 463, 478
pancreatic lipase, 407–409 Ester interchange, interesterification, 72
papain, 140, 153, 397, 405, 406, 418, 431, 535 Ester phosphate, 145
pectate lyase, 413, 415 Ethereal, 313
pectinases, 216 Ethoxy hexadiene, 496
pectin esterase, 413 Ethyl decadienoate, 322
pectin methyl esterase, 413 Ethylene (C2H4), 233, 313, 425, 447, 448, 540, 543
pentosanase, 211, 212, 398 Ethylenediamine tetraacetic acid (EDTA), 235, 534, 540
pepsin, 415–417 Ethyl sorbate, 536
peroxidase, 31, 143, 379, 380, 399, 403, 424, 425, Eugenol, 457, 459–462, 466, 469, 473, 474
429, 451, 452, 539 Euglobulin, 147
phenolase, 379, 380, 421–423 European Union
phosphatase, 143, 397, 399, 429 recommendations, 174
polygalacturonase, 413, 414 regulations, 522
polymethyl galacturonase, 414 Evaporated milk, 9, 177, 381, 382, 548
polyphenolase, 245, 382, 421, 422 Even distribution, glycerides, 55
polyphenol oxidase, 399, 422 Excitation purity, 257, 258
pregastric esterase, 407, 408 Exempt from certification (Natural colorants),
proline specific peptidase, 404 263–266, 546
prorennin, 416 Explosive puff-dried, 453
protease, 32, 399, 415–419 Extensigraph, 350
proteolytic, 123, 144, 145, 321, 415, 416, 418, 419 Extractives, natural, 218
pullulanase, 193, 200, 201
rennin, 145, 239, 397, 406, 416, 417
serine protease, 416–419 F
subtilisin, 417 Fabricated foods, 39
sulfhydryl protease, 416, 418 F-actin, 148–150, 152
tannase, 398 Farinograph, 349, 350
thiaminase, 399 Fat content, 39, 41, 54, 70, 95–97, 105, 334, 335, 352,
thrombin, 417 383, 532, 542, 549
trans-eliminase, 415 Fat crystallization, 97, 101, 542
trypsin, 32, 136, 145, 153, 159, 406, 415, 417, Fat crystal network, 339, 353, 355, 358, 359
418, 449 Fat, deep frying chemical reactions, 613
xanthine dehydrogenase, 246 Fat, hydrogenation, 72–77
xanthine oxidase, 143, 246, 427–429 See also Hydrogenation, fats
Enzymes groups Fat microstructure, 94, 355–359
amylases, 143, 193, 199, 201, 398, 411, 413, 486 Fat, oxidation
esterases, 405, 407, 408 aldehyde formation, 83, 84
hydrolases, 32, 405–420 autoxidation, 78, 80, 81, 84
Index 587

free radical, 78, 80, 81, 83–85 Fats, texture, 350–353


during frying, 90–93 Fat texture, effect of
hydroperoxide formation, 78, 81 crystallization temperature, 105, 352
light induced, 88 temperature treatment, 321, 351, 452
lipoxygenase, 85, 88 Fatty acid composition
oxidized flavor, 83, 93 canola, 49, 103
peroxide formation, 80, 427 cocoa butter, 49
photooxidation, 88–89 coconut, 49, 104
products, 78–81, 83–85, 87, 90 cottonseed, 73
sensitizers, 88, 381 dog fish, 75
singlet oxygen, 88, 89 egg yolk, 54
Fat plasticity, 359 fish oils, blue whiting, 75
Fat replacers goat, 51
olestra, 112 milk fats, bovine, 50, 52
salatrim, 112 mustard, 308
Fats and oils, 39, 44, 48, 49, 52, 55, 63, 64, 66, 78, 87, olive, 32, 39, 49, 56, 66, 68
90, 93, 95, 112, 245, 375–377, 407, 411, 532 palm, 57, 60
content in foods, 2, 29, 33, 70, 381, 391 palm kernel, 39, 72, 93
Fats and oils, classes peanut, 73
fruit coat, 39 rapeseed, 64, 66, 67, 75
kernel, 39 rice bran, 66
mammal depot, 39 sheep, 50, 51
marine, 39 soybean, 39, 48, 56, 63, 64, 66, 73
milk fat, ruminant, 39 sunflower, 49, 67, 92, 93, 103
seed oils, 39 Fatty acids
Fats and oils, individual ante-iso, 50, 68
canola, 39 arachidonic, 46, 53, 83, 84
chicken, 2, 42, 50, 63 branch chain, 43, 44
cocoa, 49, 51, 55, 56 conjugated, 74
cocoa butter, 49, 51, 55–57 cyclic, 94
coconut, 49, 66 DHA, 52
corn, 2, 49 elaidic, 42, 52
cottonseed, 49, 66 erucic, 75
egg yolk, 24, 53, 55, 63 even carbon, 319
heated, 90, 93, 109 hydrogenated, 44, 48
herring, 53, 66 hydroxy, 43
human, 44, 50, 58, 66 iso, 68, 310, 492
lard (pig), 49, 50, 52, 55 isomers, 44, 47, 52
linseed, 64, 68 isomers-cis, 42, 44, 47, 52, 57, 75
mustard, 265, 308, 472 isomers-geometric, 52, 74
novel, 80 isomers-positional, 52, 75, 76, 291
olive, 39, 49, 56, 68, 70 isomers-trans, 52, 74, 75
palm kernel, 39, 72, 93, 97 keto, 83, 90, 95
palm oil, 56, 70, 71 linolenic, 42, 52, 78
palm olein, 56, 93 linolenic γ, 46
palm stearin, 72, 93 long chain, 43–45, 51, 56, 58, 66, 99, 409, 411
peanut, 2, 31, 49, 56, 63 medium chain, 44, 45
rapeseed, 64, 66, 67, 75, 95 methyl ester, 50, 53, 67, 70, 77
replacer, 110–112 nomenclature, 43, 46, 102
rice bran, 66 odd carbon, 44
safflower, 49, 56 oleic, 42, 53, 56, 61, 70
sesame, 56, 66 palmitic, 42, 45, 50, 56, 103, 469
sheep, 50, 51, 55 palmitoleic, 45
soybean, 39, 48, 49, 56, 63, 64, 66 petroselinic, 458
soybean, low linoleic, 32, 42, 45, 51, 53 polyunsaturated, 42, 44, 47, 49–52, 57, 61, 68, 73,
sunflower, 49, 67 75, 76, 80, 90, 378
tallow (beef), 48, 49, 64, 66 saturated, 43–45, 47, 52, 55, 61, 81, 99, 100, 104
teaseed, 64 short chain, 39, 43, 44, 51, 56, 102, 165, 199, 322,
turkey, 50, 63 409, 411, 439
vegetable, 39, 44, 56, 59, 64, 67 stearic, 45, 70, 73, 96, 103, 470, 543
588 Index

Fatty acids (cont.) odor and molecular structure, 310–314


unconjugated, 52 odor description, 308
unsaturated, 42–45, 47, 51–53, 55, 57, 72, 73, 80, 84, off-flavor, 48, 78, 93, 129, 139, 140, 275, 314,
88, 90–92, 137, 324, 409 317–319, 321, 322, 399, 405, 423, 425, 427,
vaccenic, 45 451, 452, 491, 536, 538
FD & C colors, 263–265, 282, 534, 546, 548, 552, 557 potentiation, 304
Federal Food Drug and Cosmetic Act (FFDCA), 263, profile method, 314, 335
531, 532 receptor mechanism, 288
Fehling solution, 175 release, 280, 286–288, 321
Fenugreek, 458, 460 reversion, 93–95
Fermentation, 178, 199, 295, 305, 319, 321, 324, 390, salty taste, 295–298
398, 399, 406, 450, 454–455, 483, 484, sour taste, 286–288, 294–295, 322
486–488, 493, 495, 498–500, 504, 536, 537, spices and herbs, 457, 458, 460
540, 553 sweet taste, 293–294
Ferrous sulfate, 245, 534 taste, 314, 315, 318, 319, 321, 322
Fiber tea, 322–323
analysis, 222 theories of olfaction, 314
bulking capacity, 226 threshold, 289, 290, 299, 302, 303, 305, 313, 317
components, 111, 225 umami, 304–305
crude fiber, 222, 223, 225 unsaturated aldehydes, 314, 317, 324
dietary, 222–226 vanilla, 457–468
sources, 225 vegetables, 451–452
total dietary, 223–226, 438 wine, 501–504
Fibrillar proteins, 147 Flavor, additives
Film formation, 140, 202, 203 artificial, 532
Filtration of beer, 488 natural, 532
Firming agents, 535, 545 nature identical, 540
Firmness, texture, 333 Flavor changes, 438, 450–452
Fischer formula, 167 Flavor compounds and description
Fish bread, 319–320
actin, 152 cheese, 321–322
actomyosin, 152 coffee, 323–324
flavor, 320–321 dairy products, 319
liver oil, 367, 372 Flavor enhancer, additives
myosin, 152 ibotenic acid, 306
protein, 152–153 monosodium glutamate (MSG), 533
protein concentrate, 152 nucleotides, 305, 320, 524, 525
tropomyosin, 152 tricholomic acid, 306
Flatulence, 178 Flavor enhancers
Flavin adenine dinucleotide (FAD), 384, 423, 424, 429 characteristics, 304
Flavin mononucleotide (FMN), 384 glutamic acid, 304
Flavonoids ibotenic acid, 306
hesperidin, 279 maltol, 306
quercetin, 279 MSG production, 533
structures, 280 nucleotides, 305, 306
Flavor synergistic effect, 305
analysis, 315 tricholomic acid, 306
astringency, 316–317 umami, 304–306
bitter taste, 298–301 Flavr Savr tomato, 515, 518, 520
chemical structure and taste, 301–303 Flavylium, 276
description, 95, 314, 317 Flocculation, 10, 126, 220, 332, 358
enhancements, 304–306 Floral, 313, 316, 457, 490, 494, 496, 501, 502
enhancers, 295, 304–306, 322, 399, 534, 545 Flour enrichment, 245, 548
of foods, 294, 295, 299, 304, 527 Flow, 111, 161, 186, 187, 198, 202, 209, 220, 222, 249,
fruit, 497 309, 330–334, 336, 338–342, 347, 351, 356,
interrelationships of taste, 301, 304 357, 398, 430, 431, 488, 498, 499, 521, 609
margarine, 247, 315, 317, 343, 352, 356 Fluidity, 43, 205
meat, 320 Fluorescence, 384, 385, 528
milk, 321 Fluorescent light, 89
odor, 306–310 Fluorine, 243, 246
Index 589

Foaming, 140–142 brandies, 324


Folacin, see Folic acid cherry, sweet, 244, 437, 438, 442, 445
Foliar penetration, 244 coat fats, 39
Foliate papillae, 286 freezing expansion, 9, 10
Folic acid (folacin) grape, 166, 216, 244, 266, 278, 280, 295, 300, 309,
sources, 390 310, 316, 436–439, 441, 442, 444, 445, 447,
stability, 390 483, 484, 490, 495–498, 500–502, 504, 547
structure, 389 mango, 436, 438, 447
Food additives, 43, 214, 238, 244, 245, 472, 527, melon, cantaloupe, 438, 444
531–563 mineral content, 243, 244, 286
Food additives, regulations orange, 438
definition, 531–532 peach, 437, 438
GRAS, 531–533 pear, 437, 438
prior approval, 263 pineapple, 438
purposes, 263 plum, 438
Food and Agricultural Organization (FAO), 118, 367, puree, 343, 345, 414, 422, 451
514, 520, 551 raspberry, 438
Food irradiation, 546–549 strawberry, 437
Food simulants, 549 watermelon, 438
Food supplements, 301 Frying, 48, 85, 90–93, 112, 322, 329, 377, 383
Formic acid, 43, 81, 85, 184, 311, 313 Fumaric acid, 462, 541
Fortification, 383, 384, 532, 534 Functional foods, 208, 210, 477
Fortification of cereal grain products, 384 Functionality, 142, 143, 193, 195, 203, 204, 206, 220,
FOSs, see Fructooligosaccharides (FOSs) 296, 298, 419, 450
Fractionation, fats Functional properties of proteins, 139–141
dry, 105 Fungal toxins, 528
milk fat, 105, 106 Furanones, 307, 313
multistage, 106 Furanose ring, 179, 180
palm kernel oil, 97 Furans, 324
palm oil, 105, 106 Furcellaran, 218
solvent, 105 Furda method, 224
tallow, 105 Furfural, 320, 324, 379, 449, 461
Fracture resistance, 356, 357 Fusarium, 530
Free fatty acid Fusel alcohols, 317, 324, 503, 504
oxidation, 81
polymerization, 91, 93, 543
stability of oils, 84 G
trans-formation, 80, 100, 101 GAB sorption isotherm, 22
Free radical, 78–81, 83–85, 88, 90, 93, 94, 137, 392, 393, G actin, 148–150
427, 461, 477, 540 Galactans, 215
Freeze drying, 3, 27, 453 Galacto-mannoglycan, 219
Freezing, 1–4, 6–12, 14, 25, 26, 28, 89, 103, 126, 154, Galactopyranose, 219
165, 197, 206, 300, 343, 448, 449, 451, 454, Galacturonic acid, 191, 215, 217, 413–415
458, 466, 488, 500 Gallic acid, 280, 441, 461, 462
Freezing, sucrose solution, 9 Gallotannins, 280, 441
Fructooligosaccharides (FOSs), 178, 180 Gamma linolenic acid, 46
Fructose, 133, 166, 168–170, 173, 175, 178, 179, Garlic flavor, 322
182–185, 202, 203, 294, 301, 321, 398, 431, Gas liquid chromatography, 50, 75, 312, 314, 323, 324
432, 436, 437, 485, 497, 501, 534, 543 GDL, see Glucono delta lactone; Glucono delta
Fruit lactone (GDL)
air-dried, 453 Gel(s)
apple, 2, 40, 166, 174, 179, 216, 217, 243, 244, 278, electrophoresis, 154, 158, 425, 521
295, 316, 329, 368, 379, 392, 421, 437–442, formation, 142, 151, 198, 200, 203, 206, 216, 218,
444, 445, 447, 448, 451, 454, 484, 520, 532 219, 221
apricot, 179, 216, 274–276, 295, 316, 382, 398, Gelatin gels, 142
437–439, 442, 444, 445, 447, 454, 496, 551 Gelatinization temperature, 197, 203, 204, 354
banana, 2, 173, 178, 179, 195, 199, 216, 244, 275, Gelatin types, commercial, 151, 152
316, 322, 421, 425, 436, 438, 439, 441–445, Gene insertion, 513, 514, 517, 518
447, 451, 492, 501 Generally recognized as safe (GRAS) list, 531–533,
blueberry, 278, 437, 438, 442, 447, 497 545, 562
590 Index

Genetically engineered plants cottonseed, 104


alfalfa, 516 interesterification, 68–72
apple, 516 milk fat, 57, 58
canola, 516 monoglycerides, 40, 48
maize, field, 516 natural fats, 48, 53, 55, 58, 72
maize, sweet, 516 olive, 104
papaya, 516 palm oil, 104, 105
plum, 516 palm olein, 105
potato, 516 peanut, 104
rice, 516 racemic mixture, 50, 54
soybean, 516 random, 55, 58
squash, 516 soybean, 104
sugar beet, 516 sunflower, 104
tomato, 516 tripalmitin, 42, 105
Genetically modified organism (GMO), 515, 518, Glycerol esters, 408
520–526 Glycinin, 122, 124, 159
Genetic engineering, 92, 417, 512, 517, 518, 524, 525 Glycogen, 165, 166, 177, 208–209, 411
Genetic variants, 144–146 granules, 148
Gentiobiose, 177, 184, 272 Glycomacropeptide, 416
Geometrical characteristics, 334, 335 Glycophore, 293
Geotrichum candidum, 409, 411 Glycoproteins, 123, 153, 156, 159, 175, 191, 304,
Geranium off-odor, 536 316, 391
Germination, 413, 486, 489 Glycosides, 168, 172, 177, 272, 273, 276, 279, 299, 300,
Gibbs–Donnan equilibrium, 242 411, 463, 465, 466, 478, 498, 502, 544, 545
Ginger, 302, 303, 458, 460, 462, 469–471 Glycosylamine, 128, 129, 131
Gingerol, 303, 469, 470 Glyphosate, 515, 516, 520, 525, 554
Glass transition, 27–30, 190 GMO, see Genetically modified organism (GMO)
temperature, 8, 27–30, 135, 190, 198, 200, 360, 361 GMP, see Good manufacturing practice (GMP)
Glassy lactose, 190 Goiter, 246, 548
Glazed pottery, 559 Goitrin, 292
Glazes, 559 Goitrogens, 292
Gliadin, 117, 122, 125, 154–158, 161 Good manufacturing practice (GMP), 34, 533
Globulins, 117, 122, 126, 143, 146, 147, 153–160 Gouda cheese, 538
Gloss, 97, 112, 207, 262 G-protein, 288, 289
Glucitol, 174 Graininess, 398
Glucono delta lactone (GDL), 160, 161, 541 Grapefruit juice, 249, 382
Glucopyranosides, 477 Green pepper, 311, 312, 368, 439, 444, 463
Glucosamine, 153, 159, 172, 191 Green Revolution, 513
Glucosan, 183, 184 Grit cells, 425
Glucose isomerase, 202, 406, 430, 432 Ground-state oxygen, 88
Glucose-6-phosphate, 321 Growth hormones, 448
Glucose sulfurous acid, 537 Guanylate, 305, 534
Glucose syrup, 177, 202, 203, 543 Guar gum, 111, 217–219, 534, 535
Glucose units, 192–195, 197, 201, 205, 208, 212, 214, Gum arabic, 111, 218–219
354, 355, 411, 412, 437 Gumminess, 334, 348
Glucosinolates, 75 Gums
Glutamates, 290, 304, 306, 504, 505 algin, 220
Glutamic acid taste, 292, 304 arabic, 111, 218–219
Glutelin, 122, 154, 156–158, 161 carrageenan, 197, 218, 220–222
subunits, 157 guar, 111, 217–219, 534, 535
Gluten, 28, 125, 141, 155, 156, 158, 161, 304, 350, locust bean, 111, 217–219
358, 427 modified celluloses, 209, 210
Gluten-sorption isotherm, 28 Gustducin, 288
Glyceraldehyde, 166 Gymnemagenin, 303
Glyceride(s), 39–43, 48, 54–59, 67–72, 83, 84, 90, 91,
95, 96, 105, 322, 352, 407, 408, 411
structure, 48, 90 H
Glyceride composition Half-life, 432, 492
canola, 104 Hardness, 97, 333, 334, 343, 346, 348, 352, 359
corn, 104 Hardstock, 72, 103, 104
Index 591

Haworth formula, 167 Hydrochloric acid, 172, 182, 202, 205, 206, 294, 295, 304
Hazard, 561 Hydrocolloids
Heat denaturation, 120, 125, 153, 154 agar, 219–220
Heated fats, 90–93 application, 217
Heat of sorption, 23 carboxymethyl cellulose, 210
Heavy metal salts, 290 function, 197
Helix formation in starch, 193 gums, 217–218
Hemagglutination, 153, 154 pectins, 215–217
Heme pigments, 244, 266–269 Hydrogenated starch, 174, 177
Hemicelluloses, 165, 211–212, 223–226, 437, 446 Hydrogenation, 44, 47, 48, 51, 68, 72–77, 90, 104, 172,
Hemochrome, 268 174, 178, 411, 444, 450, 555
Hemoglobin, 119, 123, 124, 232, 235, 266 Hydrogenation, fats
Henderson–Hasselbach equation, 239 canola oil, 75–77, 104
Herbicides, 513, 514, 516, 520, 524, 525, 549, 552, 554 catalysts, 72, 74, 75
Herbs, 457–478, 488, 527, 540, 613 CBS, 555
Hershel-Bulkley model, 332, 333 cottonseed oil, 73, 372
Hesperidin, 279, 300, 462 fish oils, 75
Heterocyclic compounds, 132, 321 frying fats, 90–93
Hexadienoic acid, 536 liquid oil polymorphism, 72, 104
Hexagonal crystal structure, 101 nonselective, 72, 73, 76
Hexahydroxydiphenic acid, 280 olefinic compounds, 75
Hexenal, 84, 95, 317, 502 palm kernel oil, 72, 104
Hexenoic acid, 536 palm oil, 104, 352
Hexenol, 310, 427 peanut oil, 73
Hexitols, 172 pressure, 73
Hexoses, 135, 166–168, 211, 220, 224, 225, 412, 437, rapeseed oil, 75
497, 504, 505 selective, 72, 73
Heyns rearrangement, 129, 132 soybean oil, 73, 77
High erucic rapeseed oil, 75 temperature, 72, 351
High fructose corn syrup (HFCS), 173, 175, 202–203, texture, 352
431, 534 trans-formation, 72
High lysine corn, 156 Hydrogen bonds, 4–8, 10, 13–16, 24, 25, 120, 123, 124,
High melting glycerides (HMGs), 97, 103 127, 128, 141, 142, 151, 155, 159, 185, 193,
High oleic sunflower oil, 92 194, 196, 199, 209, 212, 214, 293, 316, 351,
High-temperature–short-time (HTST) pasteurization, 435, 437, 610, 614
413, 425 Hydrogen peroxide, 204, 380, 424–426, 539, 542
Hilum, 195 Hydrogen sulfide, 138, 313, 317, 320–322,
Histidine, 118, 120, 121, 128, 137, 143, 146, 153, 157, 324, 451
292, 301, 321, 418, 462 Hydrolytic reactions, 30, 409
Histones, 122 Hydrolyzates, 200, 202–203
HLB, see Hydrophile-lipophile balance (HLB) Hydrophile-lipophile balance (HLB), 108, 109,
Holoenzyme, 400 542, 543
Honey, color, 258, 259 Hydrophobic bonds, 17, 124, 141, 159, 216
Hooke’s law, 337 Hydrophobic interaction, 14, 16–17, 123, 124, 142,
Hordein, 122 213, 357
Hormones, 64, 66, 120, 448, 518 Hydrophobicity, 125, 140–142, 159, 300
Horseradish, 424, 458 Hydrophobicity values, 125, 300, 301
Hot break method, 414 Hydrostatic compression, 336
Hotness, 285, 302 Hydroxybenzoic acid, 462, 466, 533
Hudson equation, 181 Hydroxy fatty acids, 311, 438
Hue, 1, 256, 258–261, 263, 270 Hydroxylated lecithin, 542
Human fat, see Fats and oils, individual Hydroxymethyl-furfural, 129, 130, 320, 449
Humectants, 535 Hydroxyproline, 117, 127, 128, 151
Humin, 183, 184 Hydroxypropyl starch ether, 205
Hunter system of color, 260–261 Hydroysable tannin, 280, 281
Hydrides, 6 Hygroscopicity, 20
Hydrocarbons, 15, 39, 43, 48, 58, 60, 63, 64, 68, 90, 93, Hypobromite, 151
98, 101, 109, 210, 270, 271, 322, 438, 444, Hypochlorite, 204, 206
445, 450, 461, 463, 469, 489, 549–551, 554, Hypoxanthine, 321, 427–429
560, 561 Hysteresis, 19, 22, 23, 219
592 Index

I Iron bioavailability, 237, 245


Ibotenic acid, 306, 529 Irradiated yeast, 372
Ice Irradiation, 372, 387, 546–550
coefficient of thermal expansion, 4 See also Food unit of radiation
cream, 97, 111, 112, 174, 184, 190, 217, 264, 265, Irreversible deformation, 336
306, 381, 398, 464, 472, 534, 547 Isobetanin, 281, 282
crystal size, 9 Isobutanol, 323, 352, 503
density, 4 Isoelectric focussing, 154
heat capacity, 8, 14 Isoelectric point, 143, 144, 153, 158, 161, 241, 415,
heat of fusion, 4, 7, 8 416, 427
heat of sublimation, 4 meat, 241
lattice structure, 6 Isomaltol, 307, 313
physical properties, 1–4 Isomaltose, 177, 203, 306
specific heat, 4 Isomenthol, 302
structure, 6–7 Isomeric fatty acids, 44, 75
vapor pressure, 4 Isomerization, 74, 82, 90, 181, 202, 275, 427, 450,
Ice-like cluster, 7 453, 492
Ideal solubility, 106 Isoprebetanin, 281
Immobilized enzymes, 429–432 Isoprene, 63, 64, 444
Immobilized lipase, 411 Isoprenoid side chain, 373
Immune globulins, 143, 147 Isosacchrosan, 184
Indole acetic acid, 425 Isothiocyanates, 75, 291, 292
Induced dipole effect, 193 Isovalerate, 322, 503
Induction period, fat oxidation, 79, 81, 85, 88
Inherent stability, 92, 93
Inheritance, 511, 524 J
Inhibitors, 16, 128, 135, 136, 153, 159, 219, 400, 403, Juniper berries, 458, 460
404, 416, 418, 449, 452, 453, 469, 516, Just noticeable difference (JND), 289
540, 550
Initiation, 78–88, 90, 196, 197, 289, 293, 392
Ink bottle theory, 22, 23 K
Inosinate, 305 Kaempferol, 442, 461, 462, 478
Inosine monophosphate, 321 Keratin, 122, 154
Inositol, 61, 243, 244 Kestose, 178
Inositol hexaphosphoric acid, 243 Ketones, 83, 90, 93, 95, 165, 166, 310, 320, 322, 324,
Insoluble dietary fiber, 223, 224 438, 450, 451, 459, 490, 537
Interesterification Ketoseamine, 129, 130
catalyst, 68, 70–72 Ketoses, 128, 130, 135, 166–168
directed, 68, 70 Kinematic viscosity, 332
enzymatic, 72
glycerol, 70–72
palm kernel oil, 72 L
palm oil, 70, 71 Lachrymatory factor, 322
palm stearin, 72 Lactalbumin, 120, 125, 143, 145–147
randomization, 68, 70 Lactase, 176, 177, 398, 406, 413, 430, 432, 535
Interfacial loading, 142 Lactic acid, 295, 317, 454–455, 484, 500, 535, 540–542
Interfacial tension, 108 Lactitol, 174, 177, 178, 543
Intermediate moisture foods, 18, 29, 31, 32, 34 Lactobacillus casei, 390, 500
International Units (IU), 367 Lactococcus lactis, 540
Invert sugar, 412, 543 Lactoglobulin, 122, 124–127, 143, 145, 146, 321
Invisible fat, 39 Lactones, 160, 280, 310–312, 319, 322, 323, 378,
Iodine 466, 541
reaction, 193, 613 Lactose
value, 50–52, 73, 76, 93, 105, 106 α-hydrate, 188–190
Ionic equilibria, 241 β-anhydride, 189
Ionone ring, 367 physical properties, 189
Iron, 78, 85, 153, 237, 238, 243–245, 247, 249, 262, Lakes, 263, 279, 546, 557, 560
266–268, 279, 303, 306, 365, 370, 379, 384, Lamellar units, 109
424–426, 429, 440, 443, 534, 535, 540, 546, Laminaria japonica, 304
547, 549 Langmuir equation, 21
Index 593

Lanthionine, 540 Loss modulus, 342


Lard, 39, 48–50, 52, 55, 57, 58, 64, 66, 71, 78, 80, Lovibond system of color, 253, 261–262
85–87, 93, 105, 351, 374, 376, 377 Low acid foods, 34, 35
Lard stearin, 105 Low density lipoprotein, 139
Latent heat of crystallization, 95 Low linolenic canola oil, 92
Lauric oils, 104 Low linolenic soybean oil, 92
LD50, 296 Low methoxyl pectin, 413
Lead, 29, 74, 83, 123, 126, 129, 130, 136, 137, 139, 156, Lumichrome, 381, 385
161, 196, 197, 238, 243, 246–248, 262, 290, Lumiflavin, 385
295, 303, 379, 380, 390, 423, 466, 478, 544, Luminosity curves, 253, 256
547, 549, 552, 556, 559–560, 611, 613 Lutein, 265, 275, 276, 462, 501, 502
Lead acetate, 290, 296 Luteolin, 461, 462
Lead and tin contamination Lycopene, 265, 270–275, 444, 547
acid food, 559 Lysine, 32, 118, 120–122, 128, 135–139, 143, 146,
detinning of cans, 559 153–158, 301, 418, 516
equipment, 559 Lysine loss, 129, 137
gasoline, 559 Lysine loss in proteins of
tin cans, 559 cottonseed, 118
Leaf proteins, 154 fish, 136, 143
Leathery state, 360 maillard reaction, 33, 135, 137
Lecithin, 60, 61, 63, 67, 108, 526, 535, 539, 542 milk, 143, 146
Legumelin, 122, 126 peanut, 118, 154
Leucosin, 122 plasma albumin, 137
Leukotrienes, 53 wheat, 154–156
Levoglucosan, 182–184 Lysinoalanine, 138, 139, 450
Levulosan, 184 Lysosomes, 148
Lewis acid and base, 232, 233 Lysozyme, 124, 147, 153
Ligand, 233, 234, 247, 290, 299
Light exposure, 78, 536
Lightness, 258–261 M
Light sensitive receptors, 260 Mackerel, 52, 122, 321, 377, 558
Light sources, 253–255, 258, 261, 389 Macrocyclic ketones, 310
Lignin, 209, 211–212, 223–225, 325, 427, 437, 446, 515 Macrocystis pyrifera, 220
Limit dextrin, 201, 411 Magness-Taylor pressure tester, 345
Limonene, 322, 459–461, 463, 469, 474 Maillard reaction, 32, 127, 129, 131, 133, 135–137, 282,
Lineweaver-Burke equation, 402 320, 321, 449, 452
Linoleic acid oxidation, 73, 82, 427 See also Nonezymatic browning
Linseed oil, 64, 68 Maize, proteins
Lipase, 31, 32, 70, 72, 321, 322, 398, 399, 405–411, glutenin, 156, 157
426, 451 zein, 156, 157
Lipid oxidation, 18, 30, 78–79, 81, 85, 90, 137, 370 Malic acid, 439, 497, 498, 500, 541
Lipid oxidation, aw, 19 Malolactic fermentation, 295, 499, 500
Lipid peroxides, 137, 370 Malting of barley, 413, 418, 486, 487
Lipids Maltodextrins, 30, 111, 199, 200, 202
definition, 39 Maltol, 306, 307, 313, 324
interrelationship, 39, 40 Maltose, 29, 170, 175, 177, 184, 200–202, 294, 398, 411,
milk, 51, 52, 408 412, 436, 437, 486
nonpolar, 17 Malvidin, 277, 278
phospholipids, 39, 40, 43, 60–63, 68 Mammal depot fat, 39, 50
polar, 48, 60, 61 Manganese, 137, 243, 245–246, 443
Lipoproteins, 61, 66, 123, 139, 154, 156, 269 Mannans, 219
Lipovitellenin, 154 Mannitol, 173, 174, 294, 534, 543
Lipovitellin, 154 Maple syrup, color, 258, 259
Lipoxygenase, 31, 32, 85, 88, 124, 159, 399, 426–428, Margarine
450–453 crystal structure, 100
Liver necrosis, 246 no trans, 106
Locus, 257, 258 texture, 9, 103, 104, 347, 352
Locust bean gum, 111, 217–220 Marine oils, 39, 52, 53
Long spacing, X-ray diffraction, 100 Marjorum, 458
Loosely bound water, 24, 30 Mass spectrometry, 312, 314
594 Index

Matrix protein, 157 2–methoxy–3–isobutylpyrazine, 311, 313, 460, 500


Matte, 262 Methoxyl content of pectin, 413
Maxwell body, 338–340 Methoxypyrazines, 501, 502
Mayonnaise, 1, 42, 112, 142, 332, 356, 410, 472, 536, 540 Methyl alphaethyl-n-caproate, 322
Meat, drip loss, 10, 241 Methyl cellulose, 210
Meat flavor, 320 Methyl ketones, 134, 322
Meat minerals, 241–242 Methyl lanthionine, 540
Meat proteins Methyl mercaptan, 310, 320, 322, 324
actin, 147–149 Methyl mercury, 557
actomyosin, 148, 149 Methyl pyrazines, 312
collagen, 149–152 Methyl tetrahydrofolate, 389
myoalbumin, 148 Metmyoglobin, 267–269
myosin, 148, 149 Michaelis constant, 430
stroma, 147, 148 Michaelis-Menten equation, 402
tropomyosin, 147 Microencapsulation, 429, 430
Meat shear cell, 347 Microorganisms, growth, 30, 34, 200, 436, 448, 453,
Meat tenderizing, 418 454, 488
Meat texture, 348–349 Microsomes, 429
Meat-water binding, 195, 241, 245 Microstructure
Meaty taste, 303, 320 chocolate, 97, 356
Mechanical body, 338–340 colloidal dispersions, 355
Mechanical harvesting, 330 dispersed phases, 355, 356
Mechanical properties, 359 emulsions, 356–358
Medium chain triglycerides, 44 fat, 356–359
Melanoidins, 128, 130, 132, 134, 136, 137, 266, 282–283 soybean curd, 358
Melibose, 178 Migration from packaging, 549
Meltdown properties, 103 Milk
Melting points, fats Ca, P content, 234
cocoa butter, 49, 55, 56, 59, 70, 97, 102, 103 clotting, 416
high melting glycerides, 97, 103 fat, 17, 39, 44, 50–52, 57, 58, 64, 66, 78, 88, 97, 101,
milk fat, 44, 52, 57, 64 105, 106, 112, 359, 405, 410, 554
palm oil, 70, 97, 103–105, 352 flavor, 317–318
palm olein, 105 minerals, 239–241
palm stearin, 103, 105 powder, 1, 32, 125, 399
sheep depot fat, 55 protein, 111, 123, 128, 142–147, 386, 398
shortening, 52, 68, 103, 104, 352 serum proteins, 143, 425
triglycerides, 44, 48, 49, 55, 68, 70, 95, 97, 104, 105 Minamata disease, 557
vegetable fats, 55, 66 Minerals
Melting range, 48, 55, 95 abundance of trace, 244
Membrane effect, 8 determination, 236
Menadione, 366, 378, 379 enzyme reactions, 246
Menthol, 285, 302, 308, 310, 317, 459, 497 essential, 232, 247
Mercaptans, 317, 504 in fruit, 243, 244
Mercury interactions, 236–237
mercury contamination in foods, 557 in meat, 241–242
methylmercury, 556 metal chelates, 235, 236
Minamata disease, 557 in milk, 239–241
pulp and paper, 556 nonnutritive, nontoxic, 232
structure Hg, compounds, 556 nonnutritive, toxic, 232
Meromyosin, 148 in plant products, 242–243
Mesomorphism, 109 in soybeans, 243
Metabolism, 63, 64, 238, 321, 365, 375, 382, 385, 422, trace, 244–247
424, 448, 475, 485, 490, 500, 501, 504, 505 uptake, canned food, 247–249
Metal deactivator, 85 in wheat grain, 242
Metallic taste, 303 Minor components, 39, 51, 66, 86, 97, 146, 193, 322,
Metal pickup, 556 365, 435, 473
Metal uptake, 247–249, 303 Mint flavor complex, 302
Metameric, 258 Miracle fruit, 303
Methanethiol, 321, 504 Mitochondria, 66, 148, 446
Methional, 134, 139, 320, 322 Mixed crystals, 97, 105
Index 595

Mixed gels, 219, 222 Neomenthol, 302


Mobility, 341 Nerve activity, 288, 290
Modified cellulose, 209, 210 Neural response, 288
Modified starch, 199, 203–208, 354 Neuraminic acid, 178
Molasses, 186, 316, 332, 484 Neuron, 286–287, 307, 308
Mold and yeast, growth, 30, 31 Neutral detergent fiber (NDF), 224, 225
Molecular cross-section area, 347 Neutralization, 67, 147
Molecular distillation, 72, 108, 542 Neutron diffraction, 185
Molecular packing, 185 Newtonian fluids, 330, 332, 337, 338, 343, 344
Mollusks, 171, 191 Newtonian materials, 329
Molybdenum, 243, 245–248, 429, 443 Niacin
Monochromatic light, 255 pellagra factor, 386–387
Monoglycerides, 39, 40, 48, 50, 54, 72, 108–110, 407, sources, 387
408, 542, 543 stability, 366, 387
Monomolecular water, 21 structures, 387
Monophenols, 421, 422 from tryptophan, 387, 461
Monosaccharides Niacytin, 387
amino sugars, 171–172 Nickel
Fischer formula, 167 catalyst, 72, 74, 75
fructose, 168, 169 content in foods, 48, 243, 306
galactose, 168 Nicotinic acid, 366, 386–388
glycosides, 172 Nicotinic amide, 366, 386–388
sugar alcohols, 172–175 Nisin, 540, 555
Monosodium glutamate (MSG), 304–306, 533, 534 Nitrates
Mucins, 154 ADI, 539
Mucor applications, 538
M. javanicus, 411 curing salts, 538
M. miehei, 409, 417 natural sources, 539
M. pusillus, 417 nitrite cured meat, 237, 245
Multilayer adsorption, 31 nitrosamines, 538, 539, 549
Multiple sclerosis, 247 Nitrocyclohexanes, 310
Multi-stage fractionation, 107 Nitrogen closure, 249
Multivariate analysis, 314 Nitrogenous compounds, 132, 183, 282, 438
Munsell notations, 261 Nitrogen oxides, 543
Munsell system of color, 253, 258–260 Nitrosamines, 538, 539, 549
Muscle fiber, 148, 149 Nitrosated heme, 237, 245
Muscle protein, 148, 152, 242 Nitrosodiethylamine, 549
Musks, 310 Nitrosohemochrome, 268
Musky, 310, 313, 496 Nitrosyl chloride, 543
Mustard oil, 308 Nitrotoluidine, 291
Mustard white, 458 No effect dose, 527, 548
Mutarotation, 170, 175, 177, 179–182 Nonadienal, 310
Mycotoxins, 528–530 Nonalactone, 310
Myofibrils, 148, 152, 245 Noncariogenicity, 174–175, 178
Myoglobin, 88, 123, 235, 266–269 Nonenal, 84
Myosin, 122, 124–126, 147–149, 152 Nonenzymatic browning
Myrcene, 310, 322, 460–463, 469, 474, 489, 490 aw, 29, 30, 34
Myricetin, 462 Amadori rearrangement, 129, 131
browning reaction, 32, 127, 128, 131–133, 135,
137, 183
N caramelization, 131, 135, 182–184, 449
Naringin, 300, 398 enolization, 181, 182
Natamycin, 540 at glass transition temperature, 29, 30, 135
National Academy of Sciences, 296, 367, 383, 532 glycosylamines, 128, 129
Natural colorants, 263 Heyns rearrangement, 129, 132
Natural toxicants, 563 hydroxymethylfurfural, 129, 130, 320
Nature identical colors, 24, 333, 496 inhibitors, 128, 135, 136
Neat phase, 109, 110 lysine loss, 129, 137
Neohesporidose, 300 Maillard reaction, 32, 127, 129, 131, 133, 135–137,
Neoisomenthol, 302, 310 282, 320, 321, 449, 450
596 Index

Nonenzymatic browning (cont.) Olefins, 72, 74


melanoidins, 128, 130, 132, 136, 137 Oleic acid oxidation, 81, 93, 427
pH, 135 Olein, 56, 93, 105, 106
products, 31, 34, 183, 321, 453 Oleoresins, 276, 463, 469, 472–474, 478, 534, 547
Schiff base, 130 Olestra, 112, 534
Strecker degradation, 131, 132, 320 Olfaction, 306, 309, 312–314
velocity, 184 Olfactormetry, 308
Nonfermentable sweeteners, 175 Olfactory organ, 309
Nonionic emulsifiers, 108, 109 Olifactory, 307
Non-Newtonian fluids Oligosaccharides
dilatant, 332, 337 cellobiose, 177, 183, 436
plastic, 332, 337 in cow’s milk, 177, 178
pseudoplastic, 332, 337 fructo, 178, 180, 329
Nonpolar oligomeric glycerides, 90, 93 gentiobiose, 177, 183, 184
Nonprotein nitrogen, 143, 160, 438 lactose, 165, 176–178, 184, 188–191
Nonselective hydrogenation, 73, 76 in legumes, 178
Nonspectral colors, 257 maltose, 177, 200–202, 294, 411, 412
Nootkatone, 310, 311 manninotriose, 174, 294
Norepinephrine, 288 raffinose, 175, 178, 436
No-trans margarine, 106 sophorose, 183, 184
Novel food regulation, 522 in soy milk, 178
Novel oils and fats, 80 stachyose, 175, 178, 436
Nuclear magnetic resonance, 24, 96 sucrose, 175–179, 183–186, 188, 294, 412, 436
Nucleation, 4, 8, 9, 27, 97, 98 trehalose, 175, 177, 183
Nucleation, heterogeneous, 8 verbascose, 175
Nucleoproteins, 123, 156 Olive oil, 32, 39, 56, 66, 70, 88, 106, 332
Nucleotides, 299, 305, 306, 320, 388, 399, 515, 524, 525 Omithinoalanine, 139
Nucleotides as flavor enhancers, 305, 306, 399 Onion flavor, 178, 308, 322, 438, 442, 451
Nutmeg, 458, 460, 462 O-outside form, 167
Nutraceuticals, 262 Opponent colors theory, 260
Nutrient loss, 452–454 Optical rotation, 168, 179
Nutrition labeling, 532, 547 Orange, 179, 216, 234, 244, 258, 263, 265, 275, 276,
Nutrition supplement, 548–549 278, 295, 300, 309, 369, 379, 390, 392, 439,
Nystose, 178 441, 444, 445, 447, 496, 548
Oregano, 458–462, 478
Organic acids, 70, 93, 177, 233, 247, 276, 294–295, 319,
O 382, 438–441, 447, 495, 503, 540, 542
Oak casks, 316 Ornithine, 135, 139
Octyl tin maleate, 549 Orthorhombic crystal structure, 102
Octyl tin mercaptoacetate, 549 Oryzenin, 122
Odd carbon number fatty acids, 44, 322 Osazones, 175
Odor Osmotic pressure measurement, 220
analyses, 308 Ovomucin, 123, 153, 154, 171, 191
aroma description of beer, 309 Ovomucoid, 153
aroma description of wine, 309 Oxalic acid, 249, 439, 559
aroma wheel for beer, 317, 506 Oxazoles, 324
aroma wheel for wine, 316, 317 Oxidation rate, 30
description, 310 Oxidized glycerides, 48, 67, 83, 84, 90, 93
intensity factor, 310 Oxidized sterols, 93
just noticeable difference, 289 Oxonium salts, 276, 279
membrane puncturing theory, 288 Oxygen permeability, 267
olfactory organ, 309 Oxygen scavengers, 540
potency, 310, 312 Oxymyoglobin, 267, 269
primary odors, 313
threshold concentration, 309 (see also Flavor)
Off-flavors, 48, 78, 93, 95, 129, 139, 140, 275, 314, P
317–319, 321, 322, 324, 381, 399, 405, 423, Packaging, 33–34, 78, 244, 247, 263, 267, 381, 385, 453,
427, 451, 452, 491, 532, 536, 538 486, 501, 531, 539, 542, 549, 609
Oil absorption, 91 Packaging materials, migration to foods, 244, 263, 549
O-inside form, 167 PAHs, see Polycyclic aromatic hydrocarbons (PAHs)
Index 597

Palatinose, 178 Pepperminty, 313


Palm kernel oil, 72, 97 Peptides, 117–120, 123–125, 127, 128, 136, 137, 143,
Palm oil 145, 151, 156, 162, 298–301, 320–322, 389,
mid fraction, 104, 106 397, 404, 405, 415, 416, 418, 419, 461
palm stearin, 72, 93, 104–106 Peptones, 123, 145, 419
super olein, 106 Peroxidase, 31, 143, 280, 379, 380, 399, 403, 424–426,
Panose, 306 429, 451, 452
Panthenol, 391, 392 Peroxidase test, 425
Pantothenic acid, 371, 391, 392 Peroxides, 78–82, 84, 92, 93, 424, 427
Papain immobilized, 406, 431 Peroxide value, 79–81, 92, 93
Papillae, 286, 287 Persicaxanthin, 275
Paprika (Bell Pepper), 265, 266, 451, 458, 478, 547 Pesticides
Paprika oleoresin, 276, 534, 547 ADI, 551, 554
Parabens as preservative, 533, 536 aldrin, 549–550, 554
Paracasein, 144 chlorinated, 549–551
Paraffins, 317 classes and names, 549
Parallel sheet structure, 124 DDT, 549–551
Parsley, 458–460 dieldrin, 549, 550, 554
Partial glycerides, 91, 322, 407, 408 heptachlor, 549, 550, 554
Particle size analysis, 330 hydrocarbon, 549–551, 554
Pastes, 196–198, 203–205 removal, 551
Pattern recognition, 314 stability, 549
Patulin structures, 549, 550
aroclor, 554, 555 Pesticides, classes
in fish, 530 ADI, 551, 554
hydraulic fluids, 554 chlorinated hydrocarbon insecticides, 549
transformers, 554 classes and names, 549
PCBs, see Polychlorinated biphenyls (PCBs) organophosphorous, 550
PCDF, see Polychlorinated dibenzofurans (PCDF) organophosphorous insecticides, 551
Peach, 173, 274, 275, 278, 311, 322, 372, 414, 421, structure, 549, 550
437–440, 442, 444, 445, 447, 454, 496 water solubility, 550, 551
Peak time, in texture, 349, 350 Petunidin, 277, 278
Peak viscosity, 354 Phase angle, 341, 342
Peanut oil, see Fats and oils, individual Phase diagram, 26–27, 105, 106
Peanut protein, 118, 154 Phenolase, 245, 379, 380, 421, 422, 451
Peanuts, 31, 49, 56, 63, 66, 68, 73, 93, 97, 104, 106, 132, Phenolic
166, 179, 313, 377, 387, 392, 426, 528 anethole, 459
Pectic substances cavacrol, 459
acid, 215, 414, 449 cinnamaldehyde, 457, 459
content, plants, 413, 446, 449, 452 estragole, 459
degree of methylation, 215, 216 eugenol, 457, 459, 461, 462, 466
gel strength, 216 thymol, 459
hairy regions, 122 vanillin, 457, 459, 467
LMHM pectin, 216 Phenolic acid, 139, 280, 439–441
smooth regions, 535 Phenolic antioxidants, 80, 86–88, 404, 540, 542
structure, 398, 413, 446 Phenolic compounds, 85, 280, 318, 380, 392, 398, 421,
Pectin, 33, 111, 191, 215–218, 223–226, 398, 413–415, 422, 440–443, 457, 463, 488, 497, 498
437, 449, 515, 519, 535 Phenoloxidase, 31, 32, 280
Pelargonidin, 277, 278 Phenols, 86, 124, 282, 324, 380, 424, 457, 459–460, 466,
Pellagra preventive factor, 386–387 467, 553
Penetrometers, 345, 346, 350 Phenylketonuria, 543–544
Penicillium Phenylthiourea, 291, 292
P. camembertii, 407, 409 Pheophorbides, 269, 270, 450, 451
P. roqueforti, 322, 407, 409 Pheophytin, 33, 269, 270, 450, 451
Pentadiene group, 427 Phosphates
Pentene radical, 95 additives, 533, 543–546
Pentitols, 172, 174 firming agent, 535, 545
Pentosans, 211–212, 225, 474 Na-hexa, 550
Peonidin, 277, 278 in nutrition, 246
Pepper, 179, 302, 312, 438, 447, 463, 469 ortho, 204, 206, 545
598 Index

Phosphates (cont.) Plastic viscosity, 341


poly, 239, 241, 380, 544–546 Polarized light microscopy, 192
pyro, 298, 546 Polishes, 60, 157, 158
structure, 545, 546 Polyacrylamide gel, 432
Phosphatidylcholine, 61, 62, 542 Polychlorinated biphenyls (PCBs), 552–555
Phosphatidyl ethanolamine (PE), 61–63 Polychlorinated dibenzofurans (PCDF), 551, 552, 554
Phosphatidyl serine (PS), 61–63 Polychlorinated dibenzo-p-dioxins (PCDD),
Phospholipids, 39, 40, 43, 60–63, 68, 86, 97, 108, 233, 551, 554
238, 405, 438, 542 Polycyclic aromatic hydrocarbons (PAHs)
Phosphopeptone, 145 barbecuing, 561
Phosphoproteins, 123, 142–143, 153, 231, 238, 245 roasted coffee, 561
Phosphoric acid, 60, 61, 88, 117, 206, 232, 238, 243, in smoke, foods, 560–562
294, 403, 540, 541, 544 structures, 560, 561
Phosphorus oxychloride (POCI), 505, 545 unsmoked food, 561, 562
Phosphorylation, 145, 149, 205, 305, 485 Polydextrose, 112, 214, 215, 534
Photooxidation, 88–89 Polyglutamate, 390
Phthalides, 311, 312 Polyglycerol esters, 542
Physical properties Polyhedron, 16
fats, 44, 48, 57, 66, 68, 95–104 Polymerization, 91, 93, 94, 160, 181, 183, 212, 215, 280,
hydrides, 6 542–543
ice, 1–4 sulfhydryl, 127
volume change, water, 9 Polymorphism, fat, 100
water, 1–4 Polynuclear chelates, 247
Physical properties, fats Polyols, 172, 177, 178, 543
hardness, 97 Polypeptides, 16, 120, 123, 124, 126, 140, 142, 145,
margarines, 97, 98, 100, 103, 104 155, 156, 159, 237, 245, 350, 351, 416, 418,
shortenings, 97, 98, 100, 103–104 540, 555
texture, 103, 104 (see also Plastic fats) Polyphenolase, 245, 382, 421, 422
Physical refining, 67, 90, 375 Polyphenols, 139, 301, 316, 318, 441, 491
Physiology, 208, 309, 323, 438, 446–447 Polyphosphates, 239, 241, 380, 544–546
Phytate, 160, 237, 243 Polysaccharides
Phytic acid, 225, 243, 244 cellulose, 192, 209–210, 215, 219, 223
Phytosterols, 64, 66–68 chitin, 191
Picric acid, 290 complex, 193, 213
Picrocrocin, 273 glycogen, 208–209
Pigments, 39, 67, 88, 129, 133, 137, 183, 184, 244, 260, gums, 197, 210, 217–218
263, 266–283, 367, 379, 392, 427, 444–445, hydrocolloids, 197
450, 451, 546, 547 molecular structure, 212, 217
Pimaricin, 540 sources, 192, 195, 199, 208, 212
Pinking, 279 starch, 192–198
Piperine, 302, 303, 462–464 unit length, 193–195, 197, 201, 211, 212, 214, 215
Plane of symmetry, 48, 53, 57 (see also Oligosaccharides)
Plant breeding, 92, 118, 511–513, 518 Polysorbates, 535, 543
Plant products, 28, 35, 117, 139, 242–243, 367, 372, 379, Polyunsaturated fatty acids, 42–44, 47, 50–52, 68, 73,
386, 452 75, 76, 79, 80, 90, 378
Plant proteins, 117, 118, 154 Polyvinyl chloride (PVC), 33, 549
Plastic, 33, 71, 73, 92, 104, 317, 334, 337, 339–341, 358, Popcorn, 359, 360
381, 455, 549 crispness, 359, 360
Plastic fats Porosity, milk powder, 9
cocoa butter, 96, 97 Post-hardening, 71
margarine, 72, 75, 97, 99, 103, 104, 352 Postharvest deterioration, 445–446
melting, 95, 352 Potassium bromide, 290
shortening, 72, 97, 98, 103, 104, 351, 352 Potassium iodide, 290, 534
solid fat content, 95, 352 Potassium metabisulfite, 500, 536, 537
solid fat index, 96 Potato-dehydrated-BET plot, 22
solidification, 95 Potato protein, 2, 22, 24, 111, 112, 118
tallow, 96, 351 Potato starch, 205, 413
Plasticizers, 1, 29, 198, 549, 554 Powdered milk, 381
Plastic netting, 549 Power law, 332
Plastic range, 71, 104 Prebetanin, 281
Index 599

Preservatives heating, 26, 159


acids, 34, 533, 540 lipid oxidation, 18, 19, 137
bacteriocins, 540 Maillard reaction, 128, 133, 135, 136, 321, 449, 450
benzoic acid, 533 polyphenol reaction, 139, 301, 316, 441, 491
hydrogen peroxide, 539 processing and storage, 140
nitrites, 538–539 sunlight, 139, 466, 514
parabens, 533–536 UHT sterilization, 137
sodium chloride, 539 Proteins, classes
sorbic acid, 536 conjugated, 123
sulfites, 536–538 contractile, 148
Primary colors, 255 derived, 123
Primary odors, 313 fibrillar, 147
Primary oxidation products, 78, 81, 90 lipo, 61, 66, 123, 154, 156, 269
Proanthocyanidins, 280, 301, 451, 491 phospho, 123, 142–143, 153, 231, 238, 245
Processing requirements, 34 serum, 126, 143, 425
Procyanidins, 280 simple, 120–122
Profile functional group concept, 324, 325 soluble, 148, 151, 152
Prolamins, 122, 157, 161 stroma, 147, 148
Proline, 117, 118, 120–122, 124, 127, 128, 143, 145, Proteins, conjugated, 120, 123
148, 151, 157, 301, 316, 320, 404 Proteins, derived, 120, 123
Proline-rich proteins (PRPs), 316–318 Protein, separation
Promoter, 515, 518, 521, 523, 525 milk, 143, 144
Propagation, 8, 9, 78–81, 83, 86, 90, 511 skim milk, 143
Propellants, 518, 535 wheat, 154, 155
Propionic acid as preservative, 43–44, 540 whey, 143, 144
Propylene glycol esters, 543 Proteins, functional properties
Propyl gallate (PG), 85, 87, 540 dispersibility, 140
Propyl methane-thiosulfonate, 322 dough making, 142
Propyl propane-thiosulfonate, 322 emulsification, 140–142
Prostanoids, 53 foaming, 140–142
Prosthetic groups, 123, 379, 424, 443 gel formation, 142
Protamines, 122 solubility, 140–142
Protein biological value, 118, 138, 153 surface activity, 140–142
Protein bodies, 147, 157, 158 wettability, 140
Protein bonds Proteins of foods
covalent, 16, 212, 358 albumin, 120, 122, 127
disulfide, 120, 124, 127, 141, 142, 155–157, casein, 126, 127, 143, 419
350, 427 cereal, 118, 156, 158
electrostatic, 124 egg, 111, 118, 142, 153–154
energy, 124 fibrinogen, 126, 127
hydrogen, 14, 16, 124, 128 fish, 126, 136, 142, 147, 152–153
hydrophobic, 17, 124 globulin, 117, 122, 126, 143, 146, 147, 153–160
hydrophobicity, 125, 140–142, 159, 301 legume, 122, 154
noncovalent, 14, 416 maize, 156–157
sulfhydryl, 127 meat, 126, 147–149
Protein coagulants, 126 milk, 111, 123, 128, 142–147, 386, 398
Protein content of foods, 118, 119, 154, 156, 157, 209, 438 myosin, 122, 124–126, 148, 149, 152
Protein crosslinking, 450 ovalbumin, 125, 142, 143, 153
Protein efficiency ratio (PER), 118 rice, 157–158
Protein gels soybean, 140, 158–161
bean curd, 142, 160, 358 wheat, 117, 154–157
clear, 142, 204 whey, 111, 125, 126, 142–147, 160, 222, 329, 419, 534
gelatin, 142 Proteins simple, 120–122
isoelectric point, 143, 153, 158 Protein structure
mayonnaise, 142 alpha helix, 120, 159
yogurt, 142 antiparallel, 146, 159, 293
Protein hydrolysates, 304, 311, 399, 419–420, 450 coagulation, 126, 127, 142, 145
Proteins, changes during denaturation, 125–127, 449
alkali treatment, 137–138, 221, 387 double helix, 148
ammonia treatment, 139 oligomeric, 124, 280
600 Index

Protein structure (cont.) Raffinose, 175, 178, 436, 437


primary, 120, 123, 125 Rancidity, 78, 399, 405, 449, 461, 534
quaternary, 120, 124, 125 Random coil structure, 120, 124, 151, 199, 219
secondary, 120, 123, 127, 159 Random distribution, glycerides, 55, 58
stability, 123, 125 Randomization, 68, 70, 71, 90
tertiary, 117, 120, 124, 127, 159 See also Interesterification
Proteolysis, 144, 153, 300, 416, 418 Raoult’s law, 12, 20
Proteoses, 123, 419 Rapeseed oil, 64, 67, 75, 95
Protocrocin, 273 Rapid detinning, 247, 249
Protohemin, 424 Raspberries, 10, 173, 179, 216, 262, 278, 316, 438–440,
Protopectin, 33, 449 447, 496
Provitamin, 365, 367, 372 Reaction rate and aw, 30, 31
Provitamin A, 88, 273, 275, 367, 370, 373, 444, 453 Receptor mechanism, 288
Pseudobase, 276 Recommended daily allowance (RDA), 238, 244, 245,
Pseudoglobulin, 147 296, 367, 387, 389, 390, 548
Pseudoplastic, 167, 331, 332, 337, 345 Refining, 67–68, 84, 90, 185, 186, 246, 275, 375, 542
Pullulanase, 193, 200, 201 Reflectance, 253, 254, 256, 258
Pulsed nuclear magnetic resonance, 96 Refractive index, 4, 148, 610, 614
Punch dimension, texture, 346, 347 Regeneration enzymes, 425
Pungency, 302–303, 463, 469 Regiospecificity, enzymes, 408–409
Pungent, 44, 132, 134, 302, 303, 311, 313, 317, 322, 324, Relative humidity, 17, 18, 20, 21, 33, 34, 387, 448, 530
463, 464, 469, 472, 473 Relative sweetness, 170, 171, 173–175, 294, 545
Punnett square, 511, 512 Relaxation, muscle, 149
Purples, 193, 257–259, 267, 268, 511 Relaxation time, texture, 337, 614
Putrid, 313 Rennet action, 144
PVC bottles, migration of, 549 Rennet casein, 144, 147
Pyranose ring, 170, 185 Rennin sources, 422, 435
Pyranoside, α and β, 167, 168 Replacer fat, see Fats and oils, individual
Pyrazines, 311–313, 324 Resistant organisms, 555
Pyridoxal, 366, 385, 386 Resonance hybrids in oxidation, 81, 84
Pyridoxamine, 385, 386 Respiration, 386, 435, 438, 445–448, 485
Pyridoxine, 385–386 Restricted random distribution, glycerides, 55
Pyroconversion, 206 Rest sulfurous acid, 537
Pyrolysis of carbohydrates, 313 Retarded elasticity, texture, 338
Pyropheophytin, 451 Retinal, 272, 366, 367
Pyrroles, 266, 324 Retinin, 272
Pyruvic acid, 220, 484 Retinol, 275, 366–372
Retrogradation, 197, 199, 200, 204, 206
Reverse osmosis (RO), 147
Q Reversion disaccharides, 182–184
Quaternary protein structure, 120, 124, 125, 159 Rhamnoglycosides, 300
Quenching, 88 Rheological testing systems
Quercetin, 249, 279, 280, 442, 462, 478 cone penetrometer (AOCS), 350
Quercetin-3-rutinoside, 249 denture tenderometer, 348
Quinine, 287, 290, 291, 299, 300, 317 extensigraph, 350
Quinones, 85, 374, 380, 392 Farinograph, 349, 350
FIRA-NIRD extruder, 350
General Foods Texturometer, 348
R Instron Universal Testing Machine, 345, 350, 353
Racemic mixture, 50, 54, 304 Kramer shear press, 345, 347, 349, 350, 353
Racemization, 139, 450 Magness-Taylor fruit pressure tester, 345
Radiation disinfestation, 548 penetrometers, 345, 346, 350
Radiation pasteurization, 372 rotating knife tenderometer, 349
Radiation sterilization, 548 viscoamylo graph, 353–356
Radiation unit, 546 viscometers, 337, 343, 345
Radioactive isotopes Warner-Bratzler shear, 348
in food chain, 504 Rheology, 198, 335, 336, 342–348
half-life, 432, 492 Rhizopus
natural, 504 R. arrhizus, 409, 411
nuclear fall out, 514 R. oligosporus, 417
pathway, 557 Ribitol, 385
Index 601

Riboflavin, 88, 153, 262, 266, 366, 368, 371, 381, Sequence, 99, 118–120, 123, 126, 128, 144, 145, 147,
383–385, 534, 547, 548 151, 152, 182, 184, 274, 418, 422, 465, 485,
Ribonucleic acid (RNA), 305, 523, 525 512, 515, 518, 521, 524, 525
Ribosomes, 148 Sequestrante, 540
Rice bran, 158, 377 Serum albumin, 120, 125–128, 143, 146, 147
Rice polish, 158 Sesame oil, see Fats and oils, individual
Rice proteins, 157–158 Sewage sludge, 244
Rice starch, 483 Shear force, 197, 331, 332, 346, 353
Ripening, 123, 268, 273–275, 299, 301, 321, 329, 391, Shear press, 345, 347, 349, 350, 353
399, 417, 425, 437, 445, 447, 448, 497, 515, Sheep depot fat, 55
519, 548 Short chain fatty acids, 39, 43, 44, 51, 57, 102, 165, 199,
Risk, safety, 494 322, 409, 411, 439
Roasted peanuts, 312, 313 Shortening
Rosemary, 458–462, 540, 542 crystal structure, 100
Rotating knife tenderometer, 349 texture, 71, 103, 104, 352
Roundup, 525 trans isomers, 52, 75
Rubbery flow, 360 Shorthand description fatty acids, 40–43
Rubbery state, 27, 28, 198 Short spacing, X-ray diffraction, 99, 101, 102
Rum, 324, 483, 484 Shrink temperature, 151
Rutin, 249, 462 Silicon, 243, 247, 443
Rutinose, 279, 300 Sinapaldehyde, 325
Sinapic residue, 279
Sinapyl alcohol, 211
S Singlet oxygen, 88, 89
Saccharification, 174, 202 Sintering fat crystals, 352
Saccharin, 287, 290, 291, 294, 303, 534, 543, 545 Sinusoidal strain, texture, 341
Saccharinic acids, 182 Sitosterol-β, 64, 462, 474
Saccharomyces Slip melting point (SMP), 105
S. cerevisiae, 486, 493, 537 Slow-set pectin, 216
S. fragilis, 432 Smoke, 560, 561
S. lactis, 432 Snap, 18, 97, 102, 103, 179, 368, 387,
Safflower oil, see Fats and oils, individual 445, 451
Saffron, 262, 265, 266, 272, 273, 458, 460, 468, Sodium benzoate, see Benzoic acid
534, 547 Sodium chloride, 11–14, 144, 236, 237, 240–242, 290,
Saffronal, 273 295–298, 501, 539
Sage, 458–462, 540 Sodium methoxide, 70
Salatrim, 112 Sodium stearate, 109, 110
Salmonella, 548 Sodium trimetaphosphate, 205
Salty taste, 288, 295–298 Soft rot, 415
Sandiness, 190 Solder, 247, 559
Sarcolemma, 148 Solid fat content (SFC), 70, 95–97, 105, 352
Sarcomere length, 349 Solid fat index (SFI), 71, 96
Saturation, color, 258 Solid fat profile, 96
Savory, 458, 473 Solid solutions, 97, 102, 105, 106
Schiff base, 130 Solubility diagram, 107
Sclereids, 425 Solvent fractionation, 105
Scleroproteins, 122 Sophorose, 183, 184
Scombrin, 122 Sorbic acid (sorbates) as preservatives
Seafood toxins, 528 applications, 536
Secondary oxidation products, 79, 81, 84 off-flavor, 536
Sectilometer, 350–351 yeasts and molds, 536
Sedimentation methods, microstructure, 330 Sorbitol, 112, 173, 174, 178, 215, 294, 436, 534,
Seeding, 9, 185, 191 535, 543
Seed oils, 39, 43, 50, 76, 92, 375 Sorption, 17–24, 28, 31, 33, 34, 359, 360
Selectivity, 72, 73 Sorption isotherm, 20–24, 28, 31, 33, 34, 359, 360
Selenite, 246 Sorption phenomena, 17–23
Selenium, 243, 245, 246, 462 Sour taste, 286–288, 294–295, 322
Semipermeable membrane, 12, 242, 430 Southgate method, 224, 225
Senescence, 243, 268, 380, 425, 446–448 Soya lecithin, 108, 539
Sensitizer, 88, 381 Soybean curd, 142, 358
Sensory panel tests, 330 Soybean flour in wheat flour, 427
602 Index

Soybean oil, 48, 56, 67, 73, 77, 89, 92, 93, 95, 104, 105, properties, 207
112, 332, 372, 544 substitution, 203, 205
See also Fats and oils, individual Starch sodium octenyl succinate, 205
Soybean proteins, 140, 158–161 Starch viscosity of suspensions
Soybeans, mineral content, 243 corn, 193, 197–200, 202, 282, 354–356, 412, 413, 486
Soy milk, 178, 358, 427 tapioca, 111, 112, 192, 198, 202, 354, 355
Soy protein isolate, 160 waxy corn cross-bonded, 354–356, 413
Soy sauce, 304, 305, 316, 399, 416, 417, 419, 533 Steam flow closure, 249
Spatial representation, glyceride, 54 Stearin, 105, 106
Specialty fats, 105 Stearoyl–2–lactylate, 544
Specific rotation, 179, 183 Stereochemical site theory, odor, 313
Specific surface area, dispersions, 356 Stereoisomers in taste, 291
Spectral light types, 253, 255, 257 Stereospecific analysis, glycerides, 55, 59
Spectrophotometry, 77, 249, 253 Stereospecific numbering, glycerides, 54, 57
Specular reflection, 262 Sterilization flavor, 321
Spices, 207, 285, 302, 319, 457–459, 461, 463, 465, 468, Steroids, 63–66, 310
469, 471–473, 478, 497, 527, 532–534, 540, Sterols, 40, 63–66, 86, 93, 97, 372
548, 613 Stickiness, 18, 29, 197, 333, 336, 535
Spices and herbs, 457–478, 540 Stigmasterol, 64, 67, 462
Spinach, 245, 246, 249, 275, 368, 372, 377–380, 385, Stimulus concentration, taste, 288, 289
387, 390, 392, 437, 439, 443–445, 450–452, Stone cells, 425
454, 539, 551, 559 Storage modulus, texture, 342, 359
Spoilage, water activity, 30–33 Strain–texture, 336–337
Spring, 338, 339, 348, 383 Strecker degradation, 131, 132, 320, 379, 450
Springer, 249 Streptomyces, 305, 431
Squalene, 63, 64, 462 Stress, 2, 10, 26, 198, 222, 330–333, 336–343, 345, 359,
Stachyose, 175, 178, 436, 437 379, 441, 523
Stainless steel, 246 Stress decay, 337
Staling, 197 Stroma proteins, 147, 148
Staphylococcus aureus, 411 Strontium, 546
Starch Structural triacylglycerols, 411
acid conversion, 200, 205 Struvite, 242
amylase, 32, 193 St. Venant body, 340
amylopectin, 192–195, 197, 200 Subcell, 99, 101
birefringence, 197 Substrate, enzymes, 31, 32, 279, 400
branched, 192 Subunit, 120, 124, 145, 148, 157, 159, 160, 288, 424, 524
cereal (corn, wheat), 195, 198, 199, 205 Subunit association, 124
crystallinity, 193, 195, 197 Succinylated monoglycerides, 542
double helix, 193, 195, 201 Succulometer cell, 347
enzymatic conversion, 203 Sucrose, 9, 10, 14, 18, 32, 112, 166, 168, 170, 173–179,
gelatinization temp, 197, 203, 204, 354 182–186, 188, 190, 200, 202, 287, 290, 291,
gel formation, 198, 200, 203, 206 294, 299, 332, 398, 412, 436, 437, 452, 462,
glass transition, 29, 30, 198, 200 497, 534, 543–545
granules, 192–199, 353–355, 413, 449, 452 Sucrose fatty acid esters, 112, 543
hydrolyzates, 202 Sucrose polyester, 280
linear, 205 Sugar acids, 168, 301, 436, 497
modified, 199, 203–208, 354 Sugar alcohols
polymorphs, 98 arabinitol, 173
retrogradation, 197 caramelization, 131, 135, 182–184, 253, 313, 320,
root tuber (potato), 195 321, 449
sorption isotherm, 34 dehydration, 181
swelling, 195, 197 enolization, 181, 182
Starch, modified galactitol, 173
acid, 202, 203, 206 hexitols, 172
adipate, 205 isomerization, 181, 202
cross-bonding, 203, 354 lactitol, 174, 177, 178, 543
dextrinization, 196, 204, 206 maltitol, 174, 177, 178, 543
gel formation, 198, 200, 203, 206 mannitol, 173, 174, 294, 534, 543
oxidation, 203, 204, 206, 207 mutarotation, 170, 175, 177, 179–182
phosphate, 205, 207 pentitols, 172, 174
Index 603

polymerization, 181, 183, 212, 215 Sweeteners, intense


polyols, 172, 177, 178, 543 acesulfame-K, 178, 534, 543, 544
sorbitol (glucitol), 173, 174, 178, 294, 436, 534, aspartame, 178, 287, 534, 543–545
535, 543 cyclamate, 290, 291, 294, 543
xylitol, 173, 174, 178, 543 cyclohexylamine, 543
Sugar, reactions, 179–184 isomalt, 174, 177, 178
Sugars maltitol, 174, 177, 178, 543
arabinose, 211, 293, 304, 436 Mogroside V, 543, 545
fructose, 133, 166, 168–170, 173, 175, 178, 179, relative sweetness, 170, 171, 173–175, 294, 545
183–185, 202, 294, 301, 321, 398, 432, 436, saccharine, 299
437, 485, 497, 501, 534, 543 sorbitol, 112, 173, 174, 178, 215, 294, 436, 534,
galactose, 32, 135, 178, 211, 219, 220, 276, 293, 294, 535, 543
398, 413, 416, 432, 485 sucralose, 534, 543–545
glucose, 32, 135, 153, 165–168, 170–172, 174, sweeteners, nonnutritive, 543, 544
175, 177–179, 182–184, 192–195, 197, Sweetness of sweeteners, 174, 543, 545
199–203, 205, 206, 208, 209, 212–214, 222, Swelling temperature, 195
273, 276, 279, 293, 294, 300, 301, 320, 321, Swordfish, 52, 236, 559
354, 355, 384, 397–399, 411–413, 423, 424, Synaptic area, 288
432, 436, 437, 465, 475, 483–486, 497, 501, Synergistic effect in flavors, 294, 305
537, 543 Synergists in antioxidants, 376
mannose, 135, 153, 159, 166, 178, 211, 217, 219, Synsepalum dulcificum, 303
293, 436
ribose, 321
sorbose, 436 T
xylose, 174, 184, 211, 293, 304, 436, 485 Tackiness, 218, 334
Sugar sources Tallow, 39, 48, 49, 64, 66, 93, 96, 97, 105
honey, 18, 35, 60, 166, 177, 182, 258, 259, 484 Tannins
maple syrup, 35, 166, 258, 259 ellagic acid based, 280
sugar beet juice, 166 gallic acid based, 280
sugar cane juice, 166 hexahydroxy diphenic acid based, 280
Sulfide staining, 249 nonhydrolyzable, 280
Sulfite oxidase, 246 Tapioca starch, 112, 198, 202, 354, 355
Sulfites as preservatives Taraxanthin, 276
activity, 536–538 Tarragon, 458
ADI, 538, 539, 545, 551, 554, 559 Tartaric acid, 295, 439, 495, 497, 498, 501, 541, 542
antimicrobial, 536 Taste
antioxidant, 536, 537 ability to, 291, 303
applications, 538 alkaloids, 290, 299, 463
effect on pH, 537 bitter, 184, 200, 286–291, 298–301, 304, 321, 486,
thiamin and, 538 491, 492, 501
Sulfmyoglobin, 268 buds, 286–288, 293, 294
Sulfones, 138, 550 chemical structure, 290–292
Sulfoxides, 137, 138, 550 inhibition, 303–304
Sulfur dioxide, 128, 135, 136, 188, 279, 317, 382, 383, peptides, 118–120
404, 498, 500, 536, 537, 613 salts, 286–288, 290, 291, 295–298, 301, 304, 305,
Sulfur modified catalyst, 75 321, 501
Sulfur poison, 75 salty, 286–288, 290, 291, 295–299, 301, 304, 305,
Sunflower oil, 67, 92–94, 103–105 321, 501
Sunlight flavor, 139 sensations, 290, 295, 296, 299, 301, 302,
Supercooling, 8, 9, 25–27, 97 304, 305
Super olein, 106 sensitivity of the tongue, 175, 286
Supersaturation sucrose, 184 sour, 111, 286–290, 294–295, 298, 299, 301,
Surface active proteins, 108, 356 303–305, 317, 322, 442, 467, 497, 500,
Surface activity, 61, 140–142, 306 501, 536
Surface area, emulsions, 109, 110, 357, 405 stereoisomers, 64, 270, 271, 291, 310, 380, 391
Surface tension, 3, 4, 6, 10, 11, 24, 108, 217, 356 stimulas, 288
Surimi, 153 sweet, 78, 95, 170, 172, 175, 176, 178, 286–296,
Sweetened wine, 536 298, 299, 301, 303–305, 321, 322, 441,
Sweeteners, 175, 199, 202, 209, 215, 290, 294, 299, 303, 501, 544
532, 534, 543–545 thresholds, 290, 299, 318
604 Index

Taste interrelationships physical properties, 329


astringency, 280, 301, 302, 316–317, 441, 494, 501 rheological parameters, 335
coolness, 97, 285, 302 shortening, 343, 349, 351
enhancers, 295, 304–306, 322, 399, 534, 545 starch suspensions, 353–355
hotness, 285, 302 terminology, 329
meaty, 303, 305, 320, 321 types of bodies, 337–348
metallic, 285, 295, 297, 303 viscosity, 334, 335
pungency, 302, 463, 469, 501 water activity and, 359–361
suppressants, 303 Texture characteristics
sweet-sour, 301 geometrical, 334, 335
Tautomers, 170 measurements, 335–336
TCL, see Triple chain length (TCL) mechanical, 334
Tea, 64, 170, 245, 246, 301, 302, 306, 309, 316, others, 334
322–323, 399, 421, 477, 490, 496, 552, 562 terminology, 334
Technical monoglyceride, 72, 108 Texture measurements
Tempering, cocoa butter, 97 by compression, 346, 347, 351, 352
Tension force, texture, 336 creep compliance, 337
Teosinte, 513 deformation and strain, 336–337
Terminator sequence, 518 by extrusion, 350
Terpene by flow, 335, 336
anethole, 459 force and stress, 336
cavacrol, 459 by penetration with punches, 346
cinnamaldehyde, 459 relaxation time, 337
estragole, 459 by shearing, 336
eugenol, 459 shearing stress, rate of shear, 336
thymol, 459 sinusoidal strain, 341
vanillin, 459 strain time, 335, 337
Terpenic alcohols, 63, 64 stress decay, 337
Terpinyl acetate, 322 stress strain, 337, 338
Tert-butyl hydroquinone, 87 stress time, 335, 337, 338
Tertiary oxidation products, 78, 79, 81, 84 by stretching, 350
Tetracyclines, 555, 556 yield stress, 332, 333, 339–341
Tetrahedral structure, 5, 6 yield value, 332, 339, 341, 345, 346, 352, 353, 357
Tetrahydronaphthalenes, 310 Texture profile
Tetrapyrrole pigments, 266–268 analysis, 329
Tetraterpenoids, 270 geometrical characteristics, 334, 335
Textural instruments, see Rheological testing systems mechanical characteristics, 334
Textural integrity, 10 other characteristics, 334
Texture Texture, type of bodies
apples, 329, 347 Bingham, 332, 340, 341
applications, 331 Burgers, 339
beans, 347 elastic, 337–338, 340
beets, 347 Hookean, 337, 338, 340
bread, 329 Maxwell, 338–340
butter, 339, 340 plastic, 337, 339–341
definition, 329 plasto-elastic, 339, 340
dough, 349–350 plasto-viscoelastic, 339, 340
dynamic behavior, 341 retarded elastic, 338
fats, 350–353 St. Venant, 340
of foods, 329 thixotropic, 341
fruit purees, 343, 345 viscoelastic, 338–340
fruits and vegetables, 330, 353 viscous, 338
hydrogenated fats, 352 Voigt-Kelvin, 338, 339
interrelationships, 329, 330 Theobromine, 299
margarines, 343, 347, 352 Thiamin, 366, 368, 382–384, 445, 453, 548
meat, 329, 348–349 Thiazoles, 324
mechanical properties, 334 Thiazolidine–2, 138
microstructure, 329, 330, 355–359 Thin boiling starch, 203
objective measurements, 335–336 Thiol-disulfide exchange, 350
palm oil, 352 Thiophenes, 324
Index 605

Thiophosphates, 550 fruit juices, in, 245


Thio–propanal s–oxide, 322 vegetables, in, 245, 246
Thiouracil, 292 Transglucosidation, 206
Thiourea, 291, 292 Transglycosylation, 178, 202, 215
Thixotropy, 341, 343, 352, 355 Trans-isomers, 42, 44, 47, 51, 52, 74–76, 310, 427, 492
Thomson equation, 24 Transition metals, 235
Three dimensional folding, protein, 146 Triacylglycerols, 39, 48, 51, 57, 59, 60, 69, 90, 91,
Three dimensional network, fat, 352 97–99, 101, 104, 105, 408, 409, 411, 426, 450
1–threonic acid, 381 Tricholomic acid, 306
Threshold value, 289, 299, 303, 305, 315, 490 Trichromatic system, 255
Thyme, 457–462 Triclinic crystal structure, 102
Thymol, 459–462 Tricyclic compounds, 99, 101
Tin, 247, 249, 279, 303, 381, 451, 549, 556, 559–560 Trimethylamine, 321
Tin contamination, see Lead and tin contamination Tripalmitin, 42, 99, 103, 105–107
Tin-iron couple, 247 Triple chain length (TCL), 98–100
Tin plate, 247, 249, 559 Triple helix, 149, 151
Titanium dioxide, 265, 266, 547 Triplet oxygen, 88
Titratable acidity, 294, 301, 497, 498, 500 Trisaturates, 70
Tocopherolquinone, 374 Tristimulus values, 255, 256, 258
Tocopherols, 67, 85, 86, 88, 366, 372–378, 392, 462, Tropocollagen, 150, 151
534, 540 Tropomyosin, 147, 152
Found as Alpha–tocopheronic acid, 376 Trypsin digest, 145
α–tocopheronolactone, 376 Tuna, 2, 52, 54, 63, 234, 236, 321, 367, 559
Tocotrienols, 373–375, 377, 540 Turmeric, 265, 266, 458, 460, 462, 471–473, 478, 547
Tofu, 142, 160, 161, 245, 358, 399
Tomahawk crystal shape, 190
Tomato, 2, 35, 178, 179, 249, 274, 275, 309, 317, 329, U
370, 372, 377, 379, 382, 385, 386, 390, 392, UHT sterilization, 137
398, 413, 414, 437–439, 444, 445, 447, Ultracentrifuge, 120, 148, 158, 159
450–453, 496, 515, 516, 518–520, 536, 547, Ultrafiltration, 147, 239
551, 553 Ultraviolet light, 372, 385, 386, 528
Tomato paste, 249, 332 Umami, 286, 289, 290, 304–306, 321
Tongue, 175, 285–287, 289, 290, 297, 299, 303, 316, γ–Undecalactone, 310
334, 407, 463, 501 Uniaxial stress, 336, 337
Toughness of fish, 10 Unit cell, 99, 185, 190
Toxicants, natural Universal testing machine, 345, 350, 353
caffeine, 289, 291, 299, 317, 562–563 Unsaponifiables, 63, 64
glucosinolates, 75 Uric acid, 427, 428
isothiocyanates, 75, 291, 292, 308
seafood, 53, 54, 242, 245–247, 305, 557, 560
Toxic chemicals in food V
natural, 528 Value, color, 261, 262
Toxicity, 263, 290, 296, 365, 527, 528, 548, 552, 554, 560 Vander Wal’s forces, 55
Toxins, bacterial and fungal Vanilla, 316, 457–461, 464–468, 496, 497, 527
aflatoxins, 528–530 Vegetable
cadmium, 243, 560 asparagus, 179, 249, 275, 316, 368, 437, 438, 444,
C. botulinum, 34, 538 445, 450–452, 454, 502
lead and tin, 559–560 beans, lima, 438
mercury, 236, 243, 262, 267, 271, 303, 556–560 beet, 438
mycotoxins, 528–530 broccoli, 2, 179, 234, 275, 368, 370, 372, 380, 392,
patulin, 530, 531 436–439, 443–445, 454, 551
in peanuts and foods, 63, 68, 166, 387, 392 brussels sprout, 275, 379, 390, 438, 444, 445, 450,
from staphylococcus, 411 451, 454
trace elements, 231, 232, 234, 236, 243–248, 559 cabbage, 248, 275, 278, 292, 317, 372, 377–379,
trace metals, 85, 244, 443, 556–563 381, 385, 390, 391, 437–439, 442, 444, 445,
vomitoxin, 530, 531 451, 454
Trace minerals carrot, 2, 28, 41, 166, 179, 209, 216, 234, 248, 249,
absorption Fe, 232, 243, 244, 276, 444 262, 265, 266, 270, 275, 276, 279, 334, 353,
abundance of, 244 368, 370, 377, 379, 381, 414, 437–439, 442,
enzyme reactions, 246 444, 445, 449, 452–454, 502, 547
606 Index

Vegetable (cont.) sources, 382


cauliflower, 173, 378, 379, 438, 439, 443–445, 454 stability, 366, 383
celery, 173, 174, 311, 312, 334, 437–439 structures, 382, 383
cucumber, 95, 310, 314, 317, 319, 427, 428, 435, Vitamin B2 (riboflavin)
436, 438, 447, 450, 454, 516 fluorescence, 384, 385
grapes, American, 438 sources, 385
kale, 234, 438, 450 stability, 366, 384
lettuce, green leaf, 438 structure, 385
lettuce, romaine, 438 Vitamin B6 (pyridoxine)
onion, 2, 166, 178, 179, 234, 308, 322, 368, 377, forms, 385
436–439, 442, 445, 451, 490, 504, 548 sources, 386
peas, 118, 179, 195, 234, 245, 249, 264, 347, 353, 368, stability, 366, 386
377–379, 383, 385–387, 392, 398, 424, 426, structures, 385
427, 438, 443, 450, 451, 453, 454, 511, 512 Vitamin B12 (cyanocobalamine), 235, 245, 366, 388–389
pepper, sweet, 438, 462 Vitamin C (L-ascorbic acid)
potato, 2, 22, 24, 28, 32, 90, 109, 111, 112, 118, 166, as antioxidant, 382
179, 192, 193, 195, 197–199, 202, 205, 216, destruction by enzymes, 379, 381
225, 234, 245, 248, 249, 265, 297, 312, 329, 368, destruction by light, 379, 381
370, 374, 377, 379, 382, 383, 387, 390–392, losses in dairy products, 381
411, 421, 426, 436–440, 443, 444, 448, 454, protective compounds of, 378
484, 516, 520, 528, 538, 548, 551, 553 sources, 378, 379
sweet corn, 42, 166, 437–439, 452, 454 stability, 366, 381, 382
sweet potato, 166, 179, 192, 275, 370, 411, 436, 437, structure, 380
440, 444, 445, 528 technical uses, 382
turnip, 166, 292, 424, 439, 450 Vitamin D (D1, D2, D3)
Vegetable flavor, 322 fortification, 372
Vegetable oils, 39, 44, 56, 59, 64, 67, 77, 85, 86, 95, 97, IU, 372
104, 105, 112, 261, 375, 377, 392, 555 sources, 372
Vegetables, air-dried, 453 stability, 366, 372
Vegetables, texture, 353 structures, 374
Vibrational theory of olfaction, 313 Vitamin E (tocopherols)
Vinyl chloride monomer, 549 activity, 374
Violaxanthin, 275, 501 as antioxidants, 374
Viscoamylograph, 353–356 sources, 375
Viscoeleastic body, 338–340 stability, 366, 377
Viscometers structures, 375
Brookfield, 343, 345 tocotrienols, 373–375, 377
capillary, 337, 345 Vitamin fortification of flour, 383
falling ball, 337, 343 Vitamin, individual
rotational, 337, 343 A, 272, 366–374, 399, 427, 444, 445, 452, 453
Viscosity B12, cyanocobalamine, 235, 245, 366, 388–389
apparent, 28, 332, 341, 344, 345 biotin, 153, 154, 366, 391–392
coefficient, 330, 332 B6, pyidoxine, 366, 384–387, 445
dynamic, 331, 332 B2, riboflavin, 383–385, 534
kinematic, 332 B1, thiamin, 382–383
of some foods, 332 C, 365, 366, 371, 378–382, 392, 399, 445, 452, 453,
Viscous isotropic phase, 109, 110 455, 540
Visible fat, 39 D, 64, 365, 366, 372, 374, 534, 548
Vitamer, 365, 366 E, 366, 372–378, 383, 392, 474, 534
Vitamin A (retinol) folic acid, 384, 389–391, 534
coenzyme for, 391 K, 366, 378, 379
fortification, 383, 384 niacin, 366, 368, 384, 386–388, 445, 452, 461, 534, 548
IU, 275, 367, 368, 370–372 pantothenic acid, 391, 392
provitamin, 273, 275, 367, 370, 373, 444, 453 Vitamin K (menadione)
sources, 367 sources, 378
stability, 366, 370, 374 stability, 366, 378
structure, 367, 373 structure, 379
Vitamin B1 (thiamin) Vitamin loss in flour after milling, 383
loss of, 382, 383 Vitamins, 365–394
SO2 destruction, 382 food ingredients, antioxidants, 392
Index 607

Vitamins, classification crystallization, 8–10, 25, 29


fat soluble, 39, 48, 365–378, 548 density, 3, 4, 6, 14, 610, 614
water soluble, 365, 368, 371, 378–392, 452 dielectric constant, 4, 6
Vitamin stability, general, 366 glucose, 32
Vitellenin, 154 Haworth representation, 167
Vitellin, 154 heat of vaporation, 1, 3, 19, 20
Vodka, 324, 483, 484 hydration of, 19
Voigt Kelvin body, 338, 339 mannose, 135, 211, 219
Volatile compounds molecular structure, 4–6
coffee, 323 phase diagram, 26–27
spices, 457–478 physical properties, 1–4
Vomitoxin, 530, 531 refractive index, 4
Vulgaxanthins, 282 specific heat, 4, 14
structure, 4–6
surface tension, 3, 4, 6, 10–11
W tautomeric form, 168, 179
Water thermal conductivity, 4
activity (aw), 17–24, 28–35, 137, 237, 359–361, 451, types, 24–26
453, 530, 539 Wheat, 28, 31, 32, 40, 41, 63, 64, 66, 111, 112, 117,
bound loosely, 24, 30 118, 121, 122, 140, 154–158, 161, 179,
bound tightly, 13 192, 195, 198, 202, 205, 211, 223–225,
bound total, 24, 611 234, 242, 243, 245–247, 304, 319, 320,
cages, 16 349–351, 369, 376–379, 383, 384, 386,
caloric properties, 112, 173, 175, 177 387, 392, 398, 399, 407, 411–413, 426,
coefficient of thermal expansion, 4 427, 486, 489, 512–514, 525, 528, 530,
content, 1, 9, 18, 20, 22, 24, 26, 28–31, 33, 129, 135, 190, 543, 548, 553
359, 407, 409, 436, 453, 489, 609, 611, 613, 614 Wheat, dwarf, 513, 514

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