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Using Vis-NIR
Spectroscopy
for Predicting
Quality
Compounds
in Foods
Edited by
Mercedes Del Río Celestino and Rafael Font Villa
Printed Edition of the Special Issue Published in Sensors

www.mdpi.com/journal/sensors
Using Vis-NIR Spectroscopy for
Predicting Quality Compounds in
Foods
Using Vis-NIR Spectroscopy for
Predicting Quality Compounds in
Foods

Editors
Mercedes Del Rı́o Celestino
Rafael Font Villa

MDPI • Basel • Beijing • Wuhan • Barcelona • Belgrade • Manchester • Tokyo • Cluj • Tianjin
Editors
Mercedes Del Rı́o Celestino Rafael Font Villa
Agri-Food Laboratory Agri-Food Laboratory
CAGPDS CAGPDS
Córdoba Córdoba
Spain Spain

Editorial Office
MDPI
St. Alban-Anlage 66
4052 Basel, Switzerland

This is a reprint of articles from the Special Issue published online in the open access journal Sensors
(ISSN 1424-8220) (available at: www.mdpi.com/journal/sensors/special issues/NIR-Foods).

For citation purposes, cite each article independently as indicated on the article page online and as
indicated below:

LastName, A.A.; LastName, B.B.; LastName, C.C. Article Title. Journal Name Year, Volume Number,
Page Range.

ISBN 978-3-0365-7501-8 (Hbk)

ISBN 978-3-0365-7500-1 (PDF)

Cover image courtesy of Mercedes Del Rı́o Celestino

© 2023 by the authors. Articles in this book are Open Access and distributed under the Creative
Commons Attribution (CC BY) license, which allows users to download, copy and build upon
published articles, as long as the author and publisher are properly credited, which ensures maximum
dissemination and a wider impact of our publications.
The book as a whole is distributed by MDPI under the terms and conditions of the Creative Commons
license CC BY-NC-ND.
Contents

About the Editors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii

Preface to ”Using Vis-NIR Spectroscopy for Predicting Quality Compounds in Foods” . . . . ix

Mercedes del Rı́o Celestino and Rafael Font

Using Vis-NIR Spectroscopy for Predicting Quality Compounds in Foods
Reprinted from: Sensors 2022, 22, 4845, doi:10.3390/s22134845 . . . . . . . . . . . . . . . . . . . . 1

Araz Soltani Nazarloo, Vali Rasooli Sharabiani, Yousef Abbaspour Gilandeh, Ebrahim
Taghinezhad and Mariusz Szymanek
Evaluation of Different Models for Non-Destructive Detection of Tomato Pesticide Residues
Based on Near-Infrared Spectroscopy
Reprinted from: Sensors 2021, 21, 3032, doi:10.3390/s21093032 . . . . . . . . . . . . . . . . . . . . 5

Marı́a del Carmen Garcı́a-Garcı́a, Emilio Martı́n-Expósito, Isabel Font, Bárbara del Carmen
Martı́nez-Garcı́a, Juan A. Fernández and Juan Luis Valenzuela et al.
Determination of Quality Parameters in Mangetout (Pisum sativum L. ssp. arvense) by Using
Vis/Near-Infrared Reflectance Spectroscopy
Reprinted from: Sensors 2022, 22, 4113, doi:10.3390/s22114113 . . . . . . . . . . . . . . . . . . . . 19

Lucas de Paula Corrêdo, Leonardo Felipe Maldaner, Helizani Couto Bazame and José Paulo
Molin
Evaluation of Minimum Preparation Sampling Strategies for Sugarcane Quality Prediction by
vis-NIR Spectroscopy
Reprinted from: Sensors 2021, 21, 2195, doi:10.3390/s21062195 . . . . . . . . . . . . . . . . . . . . 39

Ofélia Anjos, Ilda Caldeira, Tiago A. Fernandes, Soraia Inês Pedro, Cláudia Vitória and
Sheila Oliveira-Alves et al.
PLS-R Calibration Models for Wine Spirit Volatile Phenols Prediction by Near-Infrared
Spectroscopy
Reprinted from: Sensors 2021, 22, 286, doi:10.3390/s22010286 . . . . . . . . . . . . . . . . . . . . . 63

Isabel Revilla, Ana M. Vivar-Quintana, Marı́a Inmaculada González-Martı́n, Miriam

Hernández-Jiménez, Iván Martı́nez-Martı́n and Pedro Hernández-Ramos
NIR Spectroscopy for Discriminating and Predicting the Sensory Profile of Dry-Cured Beef
“Cecina”
Reprinted from: Sensors 2020, 20, 6892, doi:10.3390/s20236892 . . . . . . . . . . . . . . . . . . . . 79

Guillermo Ripoll, Sebastiana Failla, Begoña Panea, Jean-François Hocquette, Susana Dunner
and Jose Luis Olleta et al.
Near-Infrared Reflectance Spectroscopy for Predicting the Phospholipid Fraction and the Total
Fatty Acid Composition of Freeze-Dried Beef
Reprinted from: Sensors 2021, 21, 4230, doi:10.3390/s21124230 . . . . . . . . . . . . . . . . . . . . 95

Juan Francisco Garcı́a Martı́n

Potential of Near-Infrared Spectroscopy for the Determination of Olive Oil Quality
Reprinted from: Sensors 2022, 22, 2831, doi:10.3390/s22082831 . . . . . . . . . . . . . . . . . . . . 109

Ahyeong Lee, Saetbyeol Park, Jinyoung Yoo, Jungsook Kang, Jongguk Lim and Youngwook
Seo et al.
Detecting Bacterial Biofilms Using Fluorescence Hyperspectral Imaging and Various
Discriminant Analyses
Reprinted from: Sensors 2021, 21, 2213, doi:10.3390/s21062213 . . . . . . . . . . . . . . . . . . . . 135

v
Hongyan Zhu, Aoife Gowen, Hailin Feng, Keping Yu and Jun-Li Xu
Deep Spectral-Spatial Features of Near Infrared Hyperspectral Images for Pixel-Wise
Classification of Food Products
Reprinted from: Sensors 2020, 20, 5322, doi:10.3390/s20185322 . . . . . . . . . . . . . . . . . . . . 151

Ziran Yuan, Yin Ye, Lifei Wei, Xin Yang and Can Huang
Study on the Optimization of Hyperspectral Characteristic Bands Combined with Monitoring
and Visualization of Pepper Leaf SPAD Value
Reprinted from: Sensors 2021, 22, 183, doi:10.3390/s22010183 . . . . . . . . . . . . . . . . . . . . . 171

Eshetu Bobasa, Anh Dao T. Phan, Michael Netzel, Heather E. Smyth, Yasmina Sultanbawa
and Daniel Cozzolino
The Use of a Micro Near Infrared Portable Instrument to Predict Bioactive Compounds in a
Wild Harvested Fruit—Kakadu Plum (Terminalia ferdinandiana)
Reprinted from: Sensors 2021, 21, 1413, doi:10.3390/s21041413 . . . . . . . . . . . . . . . . . . . . 191

Olga Escuredo, Laura Meno, Marı́a Shantal Rodrı́guez-Flores and Maria Carmen Seijo
Rapid Estimation of Potato Quality Parameters by a Portable Near-Infrared Spectroscopy
Device
Reprinted from: Sensors 2021, 21, 8222, doi:10.3390/s21248222 . . . . . . . . . . . . . . . . . . . . 201

Candela Melendreras, Sergio Forcada, Marı́a Luisa Fernández-Sánchez, Belén

Fernández-Colomer, José M. Costa-Fernández and Alberto López et al.
Near-Infrared Sensors for Onsite and Noninvasive Quantification of Macronutrients in Breast
Milk
Reprinted from: Sensors 2022, 22, 1311, doi:10.3390/s22041311 . . . . . . . . . . . . . . . . . . . . 213

vi
About the Editors
Mercedes Del Rı́o Celestino
Mercedes Del Rı́o Celestino, Ph.D. in Biological Sciences (2000), has been working in different
institutes and research centers in Spain (IAS-CSIC, Cordoba) and Belgium (Université Libre de
Bruxelles), and has been awarded a contract from the “Ramon y Cajal” Spanish post-doc program.
She was a permanent researcher in the Department of Plant Breeding and Biotechnology at the
IFAPA (Almerı́a, Spain) from 2009 to 2018. She is currently a research scientist at the Agri-Food
Laboratory of Cordoba (Spain). She has published over 80 peer-reviewed scientific papers and
various book chapters in the area of Plant Breeding. For several years, Dr. Del Rı́o Celestino has been
studying the genetic control of the fatty acids of the Ethiopian mustard seed and how to increase the
added value of the fruit of zucchini (Cucurbita pepo subsp. pepo) through nutritional quality. Her
achievements include obtaining materials from Ethiopian mustard with different profiles of the fatty
acid composition of the seed adapted to the semi-arid conditions of Southern Spain and the obtaining
of the first TILLING platform in zucchini fruit. Later, after the toxic waste spill of the Aznalcóllar mine
in 1999, she became a part of the CSIC expert Group and a member of the Bioremediation Network
for monitoring and recovering the ecosystem altered by metalloids. She has been a pioneer in
genotoxicity and cytotoxicity studies in complex biological matrices (horticultural products) in order
to quickly and economically determine its toxicity. Her research also focuses on the developments of
chemometric and NIR spectroscopy for determining quality components in horticultural products.

Rafael Font Villa

Rafael Font Villa, Ph.D. in Biological Sciences (2003) has been working in Spain (IAS-CSIC,
Cordoba) and has obtained a pre-doctoral mobility fellowship for short stays in different research
centers in Reino Unido (SAK, Aberdeen), Italy (University of Genoa), and Belgium (Universite Libre
de Bruxelles). He was a permanent researcher in the Department of Food Science and Health, IFAPA
Center La Mojonera (Almerı́a, Spain) from 2009 to 2018. He is currently a research scientist at
the Agri-Food Laboratory of Cordoba (Spain). He has published over 75 peer-reviewed scientific
papers and various book chapters in the the research areas of Plant Breeding and Postharvest. After
the toxic waste spill of the Aznalcollar mine in 1999, he became a pioneer for using chemometric
and spectroscopic methods to determine metalloids in different matrices (soil, plant, animals) and
quality components in horticultural products. For several years, Dr. Font has also studied the
genetic control of quality components (glucosinolates, fiber, fatty acids) of the Brassica seed and
how to increase the added value of the fruit of zucchini (Cucurbita pepo subsp. pepo) through
nutritional quality. He has worked on genotoxicity and cytotoxicity studies in complex biological
matrices (horticultural products) for quickly and economically determining its toxicity. His research
also focuses on postharvest technologies such as controlled ripening, edible coating, temperature
management, and chemical treatment methods that are potential tools to reduce fruit and vegetable
postharvest losses.

vii
Preface to ”Using Vis-NIR Spectroscopy for
Predicting Quality Compounds in Foods”
The development of affordable and more reliable methods of controlling food quality and
managing crops is imperative in order to maximize productivity and profitability and to minimize
the environmental impacts of agriculture.
For several years, visible and near-infrared (VIS–NIR) spectroscopy has contributed to
improving the control of food quality by providing the possibility to probe the internal quality of
fresh fruits, vegetables, cereals, and other edibles.
We aimed to provide the readership with a comprehensive summary of present state-of-the-art
NIR spectroscopy, current development trends, and future possibilities. We also believe that by doing
so, we will be able to provide an acceptable chance for all contributors to make their results and
methodologies more visible, as well as to highlight their current achievements in their respective
fields which have been made possible by the use of NIR spectroscopy. These articles cover a wide
range of topics related to NIR spectroscopy in a broad sense.
We would like to thank all of the authors and co-authors for their contributions, as well as all
of the reviewers for their time and effort in carefully analyzing the submissions. Last but not least,
we would like to express our gratitude to the Sensor journal’s editorial office for their cooperation in
preparing this Special Issue.

Mercedes Del Rı́o Celestino and Rafael Font Villa

Editors

ix
 sensors
Editorial
Using Vis-NIR Spectroscopy for Predicting Quality Compounds
in Foods
Mercedes del Río Celestino * and Rafael Font

Agri-Food Laboratory, CAGPDS, Avd. Menéndez Pidal, s/n, 14080 Córdoba, Spain;
[email protected]
* Correspondence: [email protected]

Over the past four decades, near-infrared reflectance spectroscopy (NIRS) has become
one of the most attractive and used technique for analysis as it allows for fast and simulta-
neous qualitative and quantitative characterization of a wide variety of food samples [1].
NIR spectroscopy is also essential in various other fields, e.g., pharmaceuticals [2], petro-
chemicals [3], textiles [4], cosmetics [5], medical applications [6], and chemicals such as
polymers [7].
The high level of interest in NIR spectroscopy among scientific and professional
sectors demonstrates its relevance. We hope that this Special Issue’s scope facilitates the
interchange of ideas and thereby aids in expanding the frontiers of this field of knowledge.
Furthermore, we aim to provide readers with a comprehensive summary of present state-
of-the-art NIR spectroscopy, trends in development, and future possibilities. We believe
that by doing so, we will be able to provide a chance for all contributors to make their
results and methodologies more visible, as well as to highlight current achievements in
their respective fields made possible by the use of NIR spectroscopy.
This Special Issue has had a resoundingly enthusiastic response, with several submis-
sions from academics and professional spectroscopists, resulting in a collection of 13 papers,
including one exhaustive review paper [8–20]. The articles submitted represent the variety
of the discussed field well, covering a wide range of topics related to NIR spectroscopy.
Citation: del Río Celestino, M.; Font,
R. Using Vis-NIR Spectroscopy for
The majority of the papers concentrate on applied qualitative and quantitative analysis in a
Predicting Quality Compounds in
variety of fields.
Foods. Sensors 2022, 22, 4845. New progress has been made in improving food quality thanks to the first investigation.
https://doi.org/10.3390/s22134845 Accordingly, it was determined that the use of variable selection algorithms provided a
better performance in predicting the amount of organophosphorus pesticide residues in
Received: 16 June 2022
tomatoes using NIRS than the use of all spectral data [8].
Accepted: 22 June 2022
The feasibility of measuring physicochemical quality parameters of mangetout pods
Published: 27 June 2022
by means of VIS-NIRS has also been demonstrated. The results revealed that the models
Publisher’s Note: MDPI stays neutral allow for an accurate quantification of protein and total polyphenol content and a rough
with regard to jurisdictional claims in screening method of the samples for color parameters (c* and h*), firmness, ascorbic acid
published maps and institutional affil- content and pH [9].
iations. In addition, despite the advantages of NIR nondestructive measurement, there is a
lack of basic studies comparatively evaluating various forms of sampling with and without
minimal processing. The analyses conducted in this Special Issue have showed that Vis-
NIR spectroscopy could be used as a quick method to assess the abundance of chemical
Copyright: © 2022 by the authors.
compounds (soluble solids content, saccharose (Pol), fiber, Pol of cane, and total recoverable
Licensee MDPI, Basel, Switzerland.
sugars) of sugarcane. Moreover, the performance of the models on defibrated cane and raw
This article is an open access article
distributed under the terms and
juice samples were similar, but defibrated cane samples involve less preparation as they do
conditions of the Creative Commons
not require juice extraction [10].
Attribution (CC BY) license (https://
For the first time, this research shows the applicability of NIR spectroscopy to assess
creativecommons.org/licenses/by/ volatile phenol contents (guaiacol, 4-methyl-guaiacol, eugenol, syringol 4-methyl-syringol
4.0/).

1
Sensors 2022, 22, 4845

and 4-allyl-syringol) and confirms the ability of this technique to quantify compounds that
contribute to the sensory quality of aged wine spirits [11].
NIRS technology can be a powerful tool to ensure the quality of food products and
prevent fraud. From the results obtained, it can be concluded that NIRS together with
artificial neural networks allow for the accurate prediction of almost all sensory parameters
selected for an exhaustive characterization of dry-cured beef meat—cecina—quality. It
would be possible to substitute the sensory panel with a faster, reliable, nondestructive and
cheaper instrumental technique that may be implemented on site [12].
In addition, this Special Issue showed that NIRS is a feasible and useful tool for
screening purposes, and it has the potential to predict most of the fatty acids of freeze-dried
beef [13].
Moreover, a comprehensive review of the state of the art in research and the actual
potential of NIRS for the analysis of olive oil has been included. It can be concluded that
the four most common physicochemical parameters that define the quality of olive oils,
namely free acidity, peroxide value, K232, and K270, can be measured using NIRS with high
precision. In addition, NIRS is suitable for the nutritional labeling of olive oil because of its
great performance in predicting the total fat, total saturated fatty acid, monounsaturated
fatty acid, and polyunsaturated fatty acid contents in olive oils [14].
Likewise, the potential of hyperspectral imaging can be also recognized on the basis of
the articles collected in this Special Issue [15–17]. Hyperspectral imaging (his) emerges as a
non-destructive and rapid analytical tool for assessing food quality, safety, and authenticity.
This technology can not only identify the physical chemistry characteristics of a substance
through spectroscopic analysis, but also simultaneously obtains information about the
spatial distribution of certain components through image analysis [21]. In this Special Issue,
we present the possibility of rapidly inspecting and detecting Escherichia coli and Salmonella
typhimurium on the surface of food processing facilities, which is a major global public
health problem [22], via fluorescence hyperspectral imaging and various discriminant
analysis techniques [15].
This Special Issue aims to investigate the potential of combining the spectral and
spatial features of HSI data with the aid of deep-learning approaches for the pixel-wise
classification of food products (sweet products and salmon fillets). The results demonstrated
that spectral pre-processing techniques prior to convolutional neural network model’s
development can enhance the classification performance. This work will open the door for
more research in the area of practical applications in food industry [16].
Important information is generated for the agrifood industry thanks to the new data
provided in this Special Issue. Hyperspectral imaging technology has been used to develop
a method for diagnosing the soil plant analysis development (SPAD) value and mapping
the spatial distribution of chlorophyll in leaves located at different positions during the
growth season of pepper plants. The results show that hyperspectral imaging is a very
promising technology and has great potential for the intuitive monitoring of crop growth,
laying the foundation for the development of hyperspectral field dynamic monitoring
sensors [17].
The growing applicability and importance of portable NIR spectrometers is reflected
by several articles, opening a new window for the utilization of these types of instruments
in the analysis and monitoring of the composition of foods. In this context, the ability of a
micro-near-infrared portable instrument to predict vitamin C in both whole and pureed
Kakadu plum fruit samples was demonstrated [18].
In this regard, the use of MicroNIR as a tool for estimating dry matter and reducing
sugars of fresh potato in a warehouses by directly measuring the tubers without chemical
treatment and destruction of samples has been demonstrated. The efficiency of such
automation techniques optimizes the management of industrial processing, guaranteeing
the quality of the potato tubers during in-line processing [19].
In this work, we also focused on the development of a real-time and simple methodol-
ogy to quantify the macronutrients (fat, raw protein and carbohydrates) in breast milk using

2
Sensors 2022, 22, 4845

a portable NIRS instrument. Notably, the implementation of this procedure requires the
use of low-cost and handheld NIRS instruments where expert personnel are not required
for analyzing samples, facilitating the quality-control procedure in the feeding of newborns
in neonatology units [20].
It should be noted that these contributions accurately reflect the diversity and dy-
namism of current NIR spectroscopy development trends.
This Special Issue is accessible through the following link: https://www.mdpi.com/
journal/sensors/special_issues/NIR-Foods (accessed on 24 June 2022). We would like to
thank all of the authors and co-authors for their contributions, as well as all of the reviewers
for their time and effort in carefully analyzing the submissions. Last but not least, we
would like to express our gratitude to the editorial office of Sensors for their cooperation in
preparing this Special Issue.

Author Contributions: Conceptualization, M.d.R.C. and R.F.; writing—review and editing, M.d.R.C.
All authors have read and agreed to the published version of the manuscript.
Funding: This research received no external funding.
Conflicts of Interest: The authors declare no conflict of interest.

References
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for Wine Spirit Volatile Phenols Prediction by Near-Infrared Spectroscopy. Sensors 2022, 22, 286. [CrossRef]
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Pixel-Wise Classification of Food Products. Sensors 2020, 20, 5322. [CrossRef]
17. Yuan, Z.; Ye, Y.; Wei, L.; Yang, X.; Huang, C. Study on the Optimization of Hyperspectral Characteristic Bands Combined with
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 sensors
Article
Evaluation of Different Models for Non-Destructive Detection of
Tomato Pesticide Residues Based on Near-Infrared Spectroscopy
Araz Soltani Nazarloo 1 , Vali Rasooli Sharabiani 1, * , Yousef Abbaspour Gilandeh 1 , Ebrahim Taghinezhad 2
and Mariusz Szymanek 3

1 Department of Biosystem Engineering, Faculty of Agriculture and Natural Resources,

University of Mohaghegh Ardabili, Ardabil 56199-11367, Iran; [email protected] (A.S.N.);
[email protected] (Y.A.G.)
2 Department of Agricultural Engineering and Technology, Moghan College of Agriculture and Natural
Resources, University of Mohaghegh Ardabili, Ardabil 56199-11367, Iran; [email protected]
3 Department of Agricultural, Forest and Transport Machinery, University of Life Sciences in Lublin,
Street Gł˛eboka 28, 20-612 Lublin, Poland; [email protected]
* Correspondence: [email protected]; Tel.: +98-914-451-9089

Abstract: In this study, the possibility of non-destructive detection of tomato pesticide residues
was investigated using Vis/NIRS and prediction models such as PLSR and ANN. First, Vis/NIR
spectral data from 180 samples of non-pesticide tomatoes (used as a control treatment) and samples
impregnated with pesticide with a concentration of 2 L per 1000 L between 350–1100 nm were
recorded by a spectroradiometer. Then, they were divided into two parts: Calibration data (70%) and
prediction data (30%). Next, the prediction performance of PLSR and ANN models after processing
was compared with 10 spectral preprocessing methods. Spectral data obtained from spectroscopy
Citation: Nazarloo, A.S.; Sharabiani,
were used as input and pesticide values obtained by gas chromatography method were used as
V.R.; Gilandeh, Y.A.; Taghinezhad, E.;
output data. Data dimension reduction methods (principal component analysis (PCA), Random frog
Szymanek, M. Evaluation of Different
(RF), and Successive prediction algorithm (SPA)) were used to select the number of main variables.
Models for Non-Destructive
According to the values obtained for root-mean-square error (RMSE) and correlation coefficient (R)
Detection of Tomato Pesticide
Residues Based on Near-Infrared
of the calibration and prediction data, it was found that the combined model SPA-ANN has the
Spectroscopy. Sensors 2021, 21, 3032. best performance (RC = 0.988, RP = 0.982, RMSEC = 0.141, RMSEP = 0.166). The investigational
https://doi.org/10.3390/s21093032 consequences obtained can be a reference for the development of internal content of agricultural
products, based on NIR spectroscopy.
Academic Editors: Mercedes Del
Río Celestino and Federico Angelini Keywords: pesticide residues; spectroscopy; PLS; soft computing; algorithm

Received: 24 March 2021

Accepted: 23 April 2021
Published: 26 April 2021 1. Introduction
Tomato (Solanum lycopersicum) is one of the most widely used crops in the world,
Publisher’s Note: MDPI stays neutral
which is rich in antioxidants such as carotenoids, total phenols, vitamin E, and vitamin
with regard to jurisdictional claims in
C [1]. Related empirical studies have shown that vitamin C affects the human immune
published maps and institutional affil-
system and prevents diseases such as Alzheimer’s [2]. In addition, the prevention of
iations.
illnesses by fruits and vegetables also depends on antioxidants [3].
Tomatoes need intensive pest management due to their low resistance to pests and
diseases. The need to use pesticides can leave harmful residues in the product. Organophos-
phorus pesticides can be stable for a considerable time even after washing and cooking in
Copyright: © 2021 by the authors.
the product if used without observing its pre-harvest interval [4–6].
Licensee MDPI, Basel, Switzerland.
Today, many countries have restricted the use of pesticides, requiring the pesticide
This article is an open access article
maximum residue limit (MRL) in food [7], and the amount is specified for each crop [8–11].
distributed under the terms and
Currently, there are several methods for determining the concentration of pesticides, includ-
conditions of the Creative Commons
ing GC, HPLC, thin layer chromatography, and capillary electrophoresis [12]. However,
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
due to time constraints and high costs, it is not possible to use these methods to control all
4.0/).
products [13].

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Sensors 2021, 21, 3032

Many studies are currently underway to develop safe, rapid, reliable, and low-cost
methods for determining pesticide residues that can prevent the use of organic solvents and
reduce operator exposure to toxic substances. Spectroscopy-based methods are a potential
method that can solve the problems mentioned above.
NIRS is suitable non-destructive method for quantitative and qualitative analysis in
agriculture, chemistry, medicine, and other sciences [14–18]. This technique is faster and
cheaper than conventional methods and environmentally friendly and can usually be used
without the need to prepare samples [19–22]. This technology is based on the absorption of
radiation in the infrared region near the electromagnetic spectrum, which can be used to
control the quality of food products [23–25]. Furthermore, in some studies, this technology
has been used to detect pesticide residues in agricultural products [26–28].
Jun et al. [29] examined cadmium residue in tomato leaves using hyperspectral imag-
ing. In this method, WT and LSSVR were used to choose the best wavelength and create
a detection model. The best prediction performance for the detection of cadmium (Cd)
content in tomato leaves was obtained using the second derivative preprocessing method.
Chen et al. [30] used NIRS to determine organophosphate chemicals. PLSR was used
to create the prediction models. The best prediction result was obtained using PLSR with
MSC and the first derivative as the preprocessing method.
Fen et al. [31] used NIRS and ANN for non-destructive detection of a common pesti-
cide on the Longan surface. The results showed that the correct diagnosis ratio was 93%.
Jiang et al. [32] combined deep learning and machine vision to predict the pesticide.
The consequences showed that when the training epoch is 10, the precision of the test
set detection will be 90.09% and the average picture bandwidth detection precision will
be 95.35%.
Wei et al. [33] offered a technique for removing residues of pesticide in apple juice.
This technique can precisely identify and classify data about residues of pesticide in apples.
Soltani et al. [18] used NIRS technology with multivariate regression analysis to predict
pesticide residues in tomato. The best prediction results were obtained using the PLS model
based on the smoothing + moving average method (Rcv = 0.92, RMSECV = 4.25).
Xue et al. [34] used the PSO algorithm to predict dichlorvos residue on the orange
surface by Vis-NIR spectroscopy. The PSO-PLS model was able to predict the dichlorvos
residue with a correlation coefficient of 0.8732. They have stated that the selection of
wavelengths through a PSO algorithm increases the ability to predict when using the
PLS model.
According to previous studies, the NIRS can be used to predict pesticide residues
from other crops. To the best of our knowledge, there is no research to determine the
organophosphorus pesticides and their prediction methods in tomatoes.
Therefore, in this paper, we use NIRS and chemometric methods to create a prediction
model without destruction to detect the tomato pesticide residues. Spectral data obtained
with a spectroradiometer and reference data obtained by a gas chromatography equipment
were used as input and output of the models used in this study, respectively. PCA, SPA,
and RF algorithms were utilized to select the variable as input for artificial neural network
(ANN) and PLSR. First, all spectral data without dimension reduction and then spectral
data obtained from variable selection algorithms were used to predict the amount of
pesticide in tomatoes. Then 8 combined modes (PLS, ANN, PCA-ANN, RF-ANN, SPA-
ANN, PLS-PCA, PLS-RF, and PLS-SPA) were developed for pesticides residues prediction.
The use of several algorithms for variable selection to predict organophosphorus pesticide
in tomatoes has not been evaluated in previous research. New progress can be made in
improving food quality by this investigation.

2. Materials and Methods

2.1. Sample Preparation
180 samples of tomatoes (Queen) were randomly harvested from a greenhouse where
almost all their produce was uniform in size and stored until 5 ◦ C until use. Pest control

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Sensors 2021, 21, 3032

in tomatoes was non-chemical from the beginning of planting to the harvest stage. In
order to achieve different pesticide residual concentrations, the samples were infected with
Profenofos 40% (EC 40%) (C11H15BrClO3PS) with a Pre-Harvest Interval (PHI) of 14 days.
Therefore, the solution of Profenofos pesticide with a concentration of 2 per 1000 L of water
was prepared and sprayed on the samples. The samples were divided into 6 categories:
The first group (P0) was used without any spraying as control and non-pesticide samples;
second group two hours (P-2H); third group two days (P-2D); the fourth group is the same
as the third category, except that it was washed after spraying (P-2D-W); the fifth group for
one week (P-1W) and the sixth group for two weeks (P-2W) were subjected to VIS/NIR
spectroscopy after spraying with the prepared solution. All samples reached equilibrium
temperature in the laboratory before completing the measurements.

2.2. Vis/NIR Spectroscopy

Vis/NIR spectroscopy tests was performed using a PS-100 spectroradiometer (Apogee
Instruments, INC., Logan, UT, USA) with CCD detector, 2048 pixels, 1 nm resolution
and halogen-tungsten light source in the wavelength range of 350–1100 nm. Prior to
spectroscopy, black and white (reference) spectra were first defined and stored. In this
way, first by turning off the light source, the dark spectrum was taken, then in the light
source mode, a standard Teflon disk with the ability to reflect above 97 in the range of 300
to 1700 nm was used to achieve the reference spectrum. For each tomato sample from 4
different points of each sample with 8 scans, within the spectral range of the equipment
used, spectroscopy was performed with software Spectra-Wiz Spectrometer OS v5.33
(c) 2014 and the data were recorded after averaging. To find the spectral regions in the
pesticide solution a quartz cell and two single-stranded fiber optics P400-2-VIS-NIR was
used (Figure 1) [35]. Reference measurements were performed one day after spectroscopic
analysis [18].

Figure 1. Measurement of Vis/NIR spectra of tomato samples in reflection mode and pesticide in passing mode.

2.3. Reference Measurements

After Vis/NIR spectroscopy, all tomatoes were prepared frozen to measure profenofos
by gas chromatographic reference method (Agilent 5977A Series GC/MSD—Santa Clara,
CA 95051, USA). To determine the retention time of the peak of the diagram obtained for
Profenofos pesticide, the Profenofos standard material (95%) prepared from Agricultural
Exir Company was injected into the chromatograph. For this purpose, sample preparation
was performed according to the British standard BS EN 15662 [36,37]. First, 10 g of the
homogenized sample was poured into a 50 mL centrifuge falcon. Then 10 mL of ethyl
acetate, 1.9 mL of distilled water and 5 g of nitrogen sulfate were added and stirred for
1 min. It was then centrifuged at 5000 rpm for 5 min and 6 mL of the extract formed on
top of the falcon was transferred to another glass falcon. It was shaken for 1 min and
centrifuged at 5000 rpm for 5 min. Then 4 mL of the upper extract of glass was poured into
another falcon and 50 μL of ethyl acetate was added. After filtration, 1 μL of extract was
injected into the equipment. The run conditions of the gas chromatography equipment are
fully described in Table 1.

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Sensors 2021, 21, 3032

Table 1. GC run conditions.

Analytical Column HP-5 ms Ultra Inert 30 m × 250 μm, 0.25 μm (p/n 19091S-433UI)
Injection volume 1 μL
Injection mode Spitless
Inlet temperature 280 ◦ C
Liner UI, split less, single taper, glass wool (p/n 5190-2293)
Plated seal kit Gold Seal, Ultra Inert, with washer (p/n 5190-6144)
Carrier gas Helium, constant flow, 1 mL/min
60 ◦ C for 1 min
then 40 ◦ C/min to 170 ◦ C
Oven program
then 10 ◦ C/min to 310 ◦ C
then hold for 2 min
Transfer line temperature 280 ◦ C

2.4. Remove the Outlier Data

The Monte Carlo cross-validation method was used to remove outliers. This method
can simultaneously detect spectral outliers and reference data [38]. Initially, the data were
randomly divided into two categories: Calibration set (70%) and prediction set (30%). Then,
PLS models were got with full cross-validation. When the RMSECV is minimized, the
best number of PC of the model is achieved. Next, the statistical characteristic parameters
of each model and the cumulative value of the sum of squares of predicted residual
errors of each sample were determined [39,40]. In this paper, outlier data (20 samples)
have been deleted by the method mentioned and the amount of R of the model has been
improved from 0.8113 to 0.8609 after their removal. Table 2 shows the reference values
(mean, standard deviation, and range) for the profenofos content (mg kg−1 ) in the tomato
samples used in this study. As can be seen, the values ranged from n.d (Not detected) to
42.9 mg/kg.

Table 2. Reference values (mean, standard deviation (SD) and range) for profenofos content (mg/kg).

Profenofos (mg/kg)
Number Range Mean Standard Deviation
calibration 112 n.d. *–42.9 14.0 10.1
validation 48 n.d.–34.0 13.7 8.9
* Not detected.

2.5. Variable Selection Method

2.5.1. Random frog (RF) Algorithm
The RF algorithm is generally used in the set of meta-heuristic algorithms. This algo-
rithm is a useful wavelength selection method that calculates the probability of selection
for each variable [40]. In short, the random frog algorithm consists of three steps [41,42]: (1)
The random initialization of a subset of variable V0 containing the variables Q; (2) creating
a subset of the variable V * including the variable Q *; accepting V * as V 1 with a certain
probability and considering V 0 = V 1 ; the above procedure is repeated until the end of N
and (3) calculating the probability of selecting each variable that can be used as a measure
of the importance of the variable. The schematic of the algorithm is shown in Figure 2.

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Sensors 2021, 21, 3032

Figure 2. Flowchart of a random frog algorithm.

Figure 3 shows the appropriate wavelengths attained by the RF algorithm. In order to

have the large part of impressible data in the main spectrum, the selection threshold was
determined experimentally by 20% trial and error method and the wavelengths above this
selection threshold were selected as the number of characteristic wavelengths. Therefore,
28 wavelengths above the dotted line were used as the final wavelengths to predict pesticide
residues in tomatoes.

Figure 3. The result of extracting useful wavelengths using the RF algorithm.

2.5.2. SPA
SPA is a forward selection method that uses simple operations in a vector space to
minimize the linearity of variables. The useful variable can be selected in spectral data
analysis for multivariate calibration using this new method. This technique is widely
used in optimizing specific spectral wavelengths that evaluate variable subsets based
on RMSEC [43]. According to the change curve of RMSEC in relation to the number
of wavelengths, it was determined that by selecting 14 characteristic wavelengths, the
value of RMSE attained a lowest value of 0.141 (Figure 4). Thus, 14 effective wavelengths
were applied as input to the prediction model. The selected characteristic wavelength
distributions across the whole spectrum are shown in Figure 5. Wavelengths close to
650–700, 750–800 and 960–1000 were chosen to build the model. These wavelengths were
in some cases like the wavelengths of the RF algorithm.

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Sensors 2021, 21, 3032

Figure 4. Change in RMSEC and Number of effective wavelengths.

Figure 5. Selected bands by SPA.

2.5.3. PCA
PCA is one of the most widely used multivariate statistical methods in chemistry [44,45].
The corresponding mathematical model for PCA is based on the decomposition of matrix
X into score matrix n × A (T) and loading matrix N × A (P) as Equation (1):

A
X = TP + F = ∑ t a p a + F (1)
a =1

where X is the spectral data matrix, T is the score matrix for X, P is the loading matrix
for X, F is the residual or model error matrix, ta is the sample score vector on each PC for
X, and pa is the variable loading vector on each PC for X. In this study, the share of the
first principal component (PC1), the second principal component (PC2), the third principal
component (PC3) and the fourth component were 55%, 18%, 8%, and 6%, respectively.
In total, the cumulative share rate of these four components reached 87.00%. To avoid
under-fitting of the prediction model due to lack of components, and to prevent over-fitting
due to information of redundant components, finally 14 main components were selected as
input to the prediction model of the amount of pesticide residues in tomatoes.

2.6. Prediction Models

2.6.1. PLSR
PLSR is a method for relating two matrices X (predictor) and Y (response), by a linear
multivariate model, which also models the structure of X and Y [46]. It works well for
analyzing large, noisy, and collinear data. In this model, by increasing the number of
variables and related observations, the accuracy of the model parameters improves [47].

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Sensors 2021, 21, 3032

This method, the least squares solution, is applied to several orthogonal components that
are a linear combination of independent variables and are created alternately with the aim
of maximizing the covariance of the linear transformation of independent variables and
dependent variables. It is very important to select the main factor when using PLSR for
regression analysis. Wrong selection of the number of main factors causes the model to
under-fitting or over-fitting, thus reducing the model prediction accuracy [48]. In this study,
the mentioned method in the wavelength range of 300–1100 nm was used for modeling
and analysis of spectral data. The fully cross-validation method was used to enhance the
selection and the number of main factors RF, SPA, and PCA were 28, 14, and 14, respectively.

2.6.2. BP-ANN
BP-ANN, a multilayer feed-forward neural network trained by the post-propagation
error algorithm, is today the most widely used reductive neural network [40,49]. In this
paper, a BP feed-forward neural network with one and two hidden layers was modeled.
“tansig”, “logsig”, and “purlin” were used in the hidden and output layers as transfer
functions. The training function used in this model was “trainlm” and the maximum
number of repetitions was 3000. The optimal number of hidden layer neurons for RF-BP,
PCA-BP, and SPA-BP combined models was obtained by trial-and-error method, 8, 12, and
14, respectively.

2.7. Model Validation

Validation methods are important to assess calibration precision and avoid data over-
fitting. The predictive power of a calibration model can be evaluated by the R, RMSEP and
RMSEC between the predicted value and the measured value in the validation set [50]. In
this research, we used R and RMSEC-RMSEP values to evaluate the accuracy and overall
strength of the model, respectively. These indicators are defined as follows:


 ∑n (ŷi − yi )2
R =  n i =1 2
(2)
∑i=1 (ŷi − ymean )

nc
1
RMSEC =
nc ∑ (ŷi − yi )2 (3)
i =1


 1 np
RMSECV = RMSEP =  ∑ (ŷi − yi )2 (4)
np i =1

ŷi : Predicted value of ith observation.

yi : Measured value of ith observation.
ymean : Mean of the prediction or calibration set.
n, nc , n p : The number of observations in the data set, calibration and prediction set,
respectively.
In general, a good model should have higher correlation coefficients, lower RMSEC,
lower RMSEP [51,52].

3. Results and Discussion

Pre-Processing Spectra
Due to the presence of noise in the initial and final parts of the diagram of absorption
spectra of tomato samples with different concentrations of pesticides, the spectrum range
from 460–1050 nm was considered (Figure 6). The following 10 spectral preprocessing
methods were applied to stabilize the models: Moving average, gaussian filter, median
filter, S-Golay, Maximum normalize, derivative-S-Golay, SNV, MSC, (Gaussian filter) +
(median filter), Normalize + Gaussian.

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Sensors 2021, 21, 3032

Figure 6. Absorption spectra of tomato samples with different concentrations of pesticides.

The residual reference values of the pesticide obtained by the GC-MS destructive
test are between 42.9–“n.d” percent. Moreover, according to the prediction results of
the combined models, the values of pesticide residues in the calibration and prediction
data were between “n.d” up to 62.75%. The spectral diagram for tomatoes with different
concentrations of pesticides is shown in Figure 6. In the diagram, the peak points in the
visible and infrared region are closely visible. The peak points between 650–700, 750–800,
and 960–1000 are related to the absorption of red pigments, the second and third overtone
vibrations of OH and the first and second overtone vibrations of OH are related to water
absorption. The results of PLS models obtained with different preprocessing methods to
predict the Profenofos pesticide residues in tomato samples were shown in Table 3. Most
of the developed calibration models had an acceptable ability to predict pesticide residues
in samples with an RCV above 0.8. However, the best prediction results were obtained
using the PLS model based on the Smoothing + moving average method (Rcv = 0.92,
RMSECV = 4.25). Hence, this model was selected for further analysis. Shan et al. (2020),
Soltani et al. (2021), Yi et al. (2010) and Sharabiani et al. (2019) also used the method used in
this study to predict the amount of soil atrazine uptake, residual pesticides in strawberries,
the amount of nitrogen in orange leaves and the amount of wheat protein, respectively,
and achieved acceptable results [16,18,53,54].

Table 3. Results of different preprocessing methods for predicting Profenofos residues.

Pre-Processing RMSECV RCV LV

No preprocessing 5.7129 0.8609 15
Smoothing-moving average 4.2562 0.9254 13
Smoothing-gaussian filter 4.2680 0.9251 14
Smoothing-median filter 5.2481 0.8847 13
Smoothing 4.1379 0.9295 15
Maximum normalize 5.5788 0.8679 11
1derivative (S-Golay) 7.6328 0.7522 15
SNV 6.8656 0.7978 13
MSC 7.1441 0.7828 15
(Smoothing-Gaussian) + (smoothing median) 7.0276 0.7778 11
Normalize + Gaussian 5.9218 0.8490 10

Figure 7 shows the correlation diagrams of the predicted values versus the main
values of the models used.
The use of NIRS technology in the detection of pesticide residues in fruits and veg-
etables, as well as their qualitative prediction, provides the researcher with a myriad of
spectral data for analysis. Large amounts of spectral data complicate analysis, prediction
errors, as well as over-fitting and under-fitting correlation curves. As a result, we need
to reduce the data dimension. In this paper, it was found that the combined models used
to predict the amount of Profenofos pesticide residues in tomato based on RF, SPA, and

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Sensors 2021, 21, 3032

PCA can achieve the same performance using only a few characteristic spectra, and in
some cases achieve better performance than the mode based on all spectral data (Figure 7).
Some variables selected using the frog (28 wavelength) and SPA (14 wavelengths) al-
gorithms were similar and the rest of the characteristic wavelengths were close to each
other. Table 4 shows the results of model evaluation indicators. According to the results
obtained in ANN-based combined models, using SPA algorithm with values of Rc = 0.989,
Rp = 0.982, RMSEC = 0.141 and RMSEP = 0.166 and using total spectral data with values
of Rc = 0.86, Rp = 0.81, RMSEC = 0.521 and RMSEP = 0.561, respectively, had the best
and worst performance in predicting Profenofos pesticide in tomatoes. Also, in PLS-based
combined models, the modes of using SPA, RF, PCA, and total spectral data had the best
and worst performance in predicting, respectively. In general, according to the results
obtained in terms of validation parameters, the best model proposed in this paper is the
SPA-ANN model.

Figure 7. Correlation diagrams of the predicted values versus the main values of the models used.

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Sensors 2021, 21, 3032

Table 4. Results of validation parameters of combined models.

Validation Parameters
Combined Models
Rc Rp RMSEC RMSEP
PLS 0.79 0.85 0.66 0.62
PCA-PLS 0.88 0.85 0.53 0.55
SPA-PLS 0.89 0.80 0.46 0.59
RF-PLS 0.91 0.91 0.40 0.36
ANN 0.86 0.81 0.52 0.56
PCA-ANN 0.93 0.89 0.36 0.40
SPA-ANN 0.98 0.98 0.14 0.16
RF-ANN 0.91 0.89 0.40 0.54

In a similar study the feasibility of using NIRS to detect the soluble solids content
(SSC) of Malus micro malus Makino were studied using SPA, SVR, PLSR, and BP-ANN.
The comparison studies confirmed that the optimal fusion model of SPA-SVR had the best
performance (RC = 0.9629, RP = 0.9029, RMSEC = 0.199, RMSEP = 0.271) [42]. In other
study, a new method of variable interval selection based on random frog (RF), known
as Interval Selection based on Random Frog (ISRF), is developed. The results show that
the proposed method is very efficient to find the best interval variables and improve the
model’s prediction performance and interpretation [55]. The results of various studies show
that the use of soft computing has been an effective method in the qualitative diagnosis of
products. This is evidence of the confirmation of the results obtained from our study.

4. Conclusions
In this paper, a rapid and non-destructive near-infrared method was used to predict the
profenofos pesticide residues in tomatoes. Spectral data obtained with a spectroradiometer
and reference data obtained by a gas chromatography equipment were used as input and
output of the models used in this research, respectively. PCA, SPA, and RF algorithms
were used to select the variable as input for artificial neural network (ANN) and PLSR.
First, all spectral data without dimension reduction and then spectral data obtained from
variable selection algorithms were used to predict the amount of pesticide in tomatoes.
Afterwards, 8 combined modes (pls, ANN, PCA-ANN, RF-ANN, SPA-ANN, PLS-PCA,
PLS-RF, and PLS-SPA) were developed for prediction. Finally, the prediction accuracy of
different combined models was compared and the best case was introduced. Based on what
was said in the previous sections of the paper, it was found that it is possible to predict the
amount of pesticide residues in tomatoes using the spectrum in the range of 460–1050 nm.
Accordingly, it was determined that the use of variable selection methods had a better
performance in predicting the amount of pesticide residues than the use of all spectral data.
Finally, according to the results of the validation parameters of the combined models used,
the SPA-ANN combined model with values of Rc = 0.989, Rp = 0.982, RMSEC = 0.141 and
RMSEP = 0.166 had the best performance in predicting Profenofos pesticide in tomatoes.
At the end of the article, it can be mentioned that NIRS technology, in addition to
advantages such as a non-destructive method, low cost measurement, high speed and
online uses in the processes of quality determination. However, this method has some
disadvantages, which are: NIRS requires chemometric techniques to provide the reference
data for calibration and validation of experimental results, and the large number of samples
with large variations to extract accurate information.

Author Contributions: Methodology, V.R.S., A.S.N. and Y.A.G.; formal analysis, V.R.S., E.T. and M.S.;
investigation, V.R.S. and Y.A.G.; experiment conduction, V.R.S. and Y.A.G.; statistical analysis, V.R.S.
and A.S.N.; Writing—original draft preparation, V.R.S., A.S.N., E.T., Y.A.G. and M.S. writing—review
and editing, A.S.N., V.R.S., Y.A.G., E.T. and M.S. All authors have read and agreed to the published
version of the manuscript.
Funding: This research received no external funding.

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Sensors 2021, 21, 3032

Institutional Review Board Statement: Not applicable.

Informed Consent Statement: Not applicable.
Data Availability Statement: Data is contained within the article.
Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations

MRL Maximum Residue Limit

EU European Union
VIS/NIRS Visible/Near InfraRed Spectroscopy
PCA Principal Component Analysis
PLSR Partial Least Squares Regression
ANN Artificial Neural Network
R Regression Coefficient
RMSE Root Mean Square Error
GC Gas Chromatography
HPLC High-Performance Liquid Chromatography
WT Wavelet Transform
LSSVR Least-Square Support Vector Machine Regression
PSO Particle Swarm Optimization
RPD Residual Prediction Deviation
PLS-DA Partial Least Squares-Discriminant Analysis
PHI Pre-Harvest Interval
LOD Limit of detection

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 sensors
Article
Determination of Quality Parameters in Mangetout (Pisum
sativum L. ssp. arvense) by Using Vis/Near-Infrared
Reflectance Spectroscopy
María del Carmen García-García 1, *, Emilio Martín-Expósito 1 , Isabel Font 2 , Bárbara del Carmen Martínez-García 3 ,
Juan A. Fernández 4 , Juan Luis Valenzuela 5 , Pedro Gómez 6 and Mercedes del Río-Celestino 7, *

1 Department of Agro-Food Engineering and Technology, IFAPA Centro La Mojonera, CAGPDS,

04745 Almería, Spain; [email protected]
2 ETSIIT, Campus Aynadamar, University of Granada, 18071 Granada, Spain; [email protected]
3 ESI, Higher Engineering School, University of Almería, 04120 Almería, Spain; [email protected]
4 Department of Agronomical Engineering, Technical University of Cartagena, 30203 Murcia, Spain;
[email protected]
5 Department of Biology and Geology, Higher Engineering School, University of Almería,
04120 Almería, Spain; [email protected]
6 Department of Plant Breeding and Biotechnology, IFAPA Centro La Mojonera, CAGPDS,
04745 Almería, Spain; [email protected]
7 Agri-Food Laboratory, CAGPDS, Avda, Menéndez Pidal, s/n, 14080 Córdoba, Spain
* Correspondence: [email protected] (M.d.C.G.-G.);
[email protected] (M.d.R.-C.)

Abstract: Pisum sativum L. ssp. arvense, is colloquially called tirabeque or mangetout because it is
eaten whole; its pods are recognized as a delicatessen in cooking due to its crunch on the palate
Citation: García-García, M.d.C.; and high sweetness. Furthermore, this legume is an important source of protein and antioxidant
Martín-Expósito, E.; Font, I.; compounds. Quality control in this species requires the analysis of a large number of samples using
Martínez-García, B.d.C.; Fernández,
costly and laborious conventional methods. For this reason, a non-chemical and rapid technique
J.A.; Valenzuela, J.L.; Gómez, P.;
as near-infrared reflectance spectroscopy (NIRS) was explored to determine its physicochemical
Río-Celestino, M.d. Determination of
quality (color, firmness, total soluble solids, pH, total polyphenols, ascorbic acid and protein content).
Quality Parameters in Mangetout
Pod samples from different cultivars and grown under different fertigation treatments were added
(Pisum sativum L. ssp. arvense) by
Using Vis/Near-Infrared Reflectance
to the NIRS analysis to increase spectral and chemical variability in the calibration set. Modified
Spectroscopy. Sensors 2022, 22, 4113. partial least squares regression was used for obtaining the calibration models of these parameters.
https://doi.org/10.3390/s22114113 The coefficients of determination in the external validation ranged from 0.50 to 0.88. The RPD
(standard deviation to standard error of prediction ratio) and RER (standard deviation to range) were
Academic Editor: Luca Fiorani
variable for quality parameters and showed values that were characteristic of equations suitable for
Received: 7 April 2022 quantitative prediction and screening purposes, except for the total soluble solid calibration model.
Accepted: 26 May 2022
Published: 28 May 2022 Keywords: mangetout; pea pod; near-infrared reflectance spectroscopy; quality parameters
Publisher’s Note: MDPI stays neutral
with regard to jurisdictional claims in
published maps and institutional affil-
iations. 1. Introduction
Vegetable proteins are appearing as a sustainable source for human consumption [1].
Demand for protein is likely to increase significantly over the next few decades to keep pace
with a growing population, which is projected to reach nearly ten billion by 2050 [2]. The
Copyright: © 2022 by the authors.
trend of animal protein consumption is increasing in recent decades [3,4], with production
Licensee MDPI, Basel, Switzerland.
of animal source foods responsible for a significant proportion of global greenhouse gas
This article is an open access article
(GHG) emissions, water consumption and land use [5]. However, the proportion of protein
distributed under the terms and
consumption that the World Health Organization recommends is 75% vegetable and 25%
conditions of the Creative Commons
animal [6]. In this context, legumes, including soybeans, peanuts, beans, peas, fava beans
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
and lentils, among others, have a higher protein content than most plant foods and about
4.0/).
twice the protein content of cereals [7]. The high protein content of legumes may be related

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Sensors 2022, 22, 4113

to their association with nitrogen-fixing bacteria in their roots, which converts the unusable
nitrogen into ammonium that is used for protein synthesis [8].
At present, the consumer demands new products on the supermarket shelves and
is also attracted by local markets and products. A segment of the population considers
itself a green consumer [9], in its different variants, and values healthy and quality foods.
Legumes, for all the above exposed, satisfy the current market trends [10].
Several species have been the subject of research for the diversification of vegetables
in the agricultural system of the province of Almería (Southeast Spain), with more than
32,000 hectares of greenhouses [11], more than 60% of cultivated vegetables belonging to
the Solanaceae family. The species tested to diversify these horticultural crops are sweet
cucumber, berries, pitahaya, passion fruit, fig tree and a wide range of legumes, among
some of them Pisum sativum L. ssp. arvense, colloquially called tirabeque or mangetout [12].
This species is recognized as a delicatessen in cooking due to its crocanti on the palate
and high sweetness. Whole mangetout pods are cooked and eaten, this being possible by
the absence of “parchment” in the pod walls, hence its pod is indehiscent. The external
appearance of pods, particularly their color, is also of great importance when considering
the fruits destined for fresh products.
Previous studies have also revealed the nutritional potential of mangetout, not only for
its protein content but also for its content of total soluble solids and antioxidant compounds
such as polyphenols, ascorbic acid, fiber, phytoprostanes and phytofurans [12–14].
Overall, the methodology used for the determination of phenolic compounds and
ascorbic acid content is based on spectrophotometric and chromatographic techniques;
however, these techniques require expensive equipment and usually use hazardous and
pollutant reagents [15,16]. Another relevant method includes colorimetric and titration
measurements, since it represents a relatively simple method for measuring total phenolic
compounds and ascorbic acid content, respectively.
The need to carry out screening in breeding programs, quality controls, traceability
studies and/or obtaining rapid information for labelling in a large number of samples using
conventional methods, leads to high costs, labour input and delays in the rapid decision
making. For this reason a non-chemical (producing no chemical waste) and rapid technique,
near-infrared reflectance spectroscopy (NIRS), which has been successfully applied in vari-
ous fields from life sciences to environmental issues, is explored here to screen quality in
mangetout pods [17]. Near-infrared spectroscopy is a technique that uses the radiation
absorbed by a set of samples in the region from 780 to 2500 nm (near-infrared region-
NIR spectroscopy in combination with chemometric analyses can be used for analysis of
numerous components (protein, carbohydrates, carotenoid, minerals, glucosinolates, phe-
nolics) and parameters of the sample (firmness, Brix, acidity, color) to be analyzed [18–23].
NIRS depends on the number and type of C-H, N-H and O-H bonds in the material being
analyzed, then spectral features are combined with reliable compositional or functional
analyses of the material in a predictive statistical model. This model is then used to predict
the composition of new or unknown samples [24].
Recently, the use of NIRS models for predicting the quality of vegetables has been
reported, several of which have addressed zucchini [19,20], pepper, rocket leaves, blackber-
ries [16,21,22] and Ethiopian mustard leaves [23], among others. The seed quality of various
legume species has also been analyzed using NIRS such as lentils [25], chickpeas [26] and
pea accessions from different germplasm collections [27,28]. Other studies have focused on
predicting the sensory quality and maturity of peas [29,30] using NIRS. To the best of our
knowledge, there is no research that predicted the quality in mangetout pods.
NIRS calibration models have been developed using a variety of linear regression
approaches, including modified partial least squares regression (MPLS). The modified
partial least squares (MPLS) is an improved version of traditional PLS that was developed
by Shenk and Westerhaus [31]. The MPLS procedure copes more effectively with non-
analyte interference in multicomponent determinations. This regression approach is a
soft-modeling method for generating predictive models when the factors are many and

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Sensors 2022, 22, 4113

very collinear. It allows us to develop a model that is then evaluated on external samples to
estimate the predictive ability of the model. The mathematical procedure’s end goal is to
decrease the large amount of spectral data points (1050 data points from 400 to 2500 nm
wavelength range, every 2 nm) and remove the correlation presented by neighboring
wavelengths. As a result, the model developed only takes into account the most significant
factors, with the “noise” encapsulated in the less important factors, hence the accuracy of
NIRS analysis is improved.
At present, the purpose of the producers and the Andalusian Administrations involved
in the cultivation of mangetout is to apply for a “Protected Geographical Indication” (PGI)
for the Dalías Valley (Almería, Southeast Spain). This European Indication distinguishes
the quality attributes of the products grown in a certain region, and the NIRS technique
is a suitable tool that could contribute quickly and accurately to verify the quality of
the productions.
The objective of this paper was to investigate the feasibility for measuring physico-
chemical quality parameters (color, firmness, total soluble solids, pH, total polyphenols,
ascorbic acid and protein content) of mangetout pods by means of VIS-NIRS. For this
purpose, different cultivars of mangetout grown under organic cultivation and two fertiga-
tion regimes were tested to generate the highest variability for the development of NIRS
prediction models.

2. Material and Methods

2.1. Plant Material
The vegetal material consisted of a local landrace (germplasm maintained by local
growers in Almería Province, Southeast Spain) and 7 commercial cultivars of mangetout
(Figure 1, Table 1).

Figure 1. Pea pods of the different cultivars of mangetout analyzed. From left to right: Local
landrace (a), AR-24007 (b), Capuchino (c), Tirabeque IS (d), Tirabí (e), Pea Zuccola (f), Pea Delikata (g)
and Bamby (h).

Table 1. Cultivars, companies and growth habit of mangetout used in this study.

Cultivars Companies Growth Habit

Local landrace Growers production Indeterminate climbing
AR-24007 Ramiro Arnedo Indeterminate climbing
Capuchino Batlle Indeterminate climbing
Tirabeque IS Intersemillas Indeterminate climbing
Tirabí Fitó Indeterminate climbing
Pea Zuccola Tozer Determinate climbing
Pea Delikata Tozer Determinate climbing
Bamby Gautier Deterninate postrate

Edible pods of Pisum sativum L. spp. arvense (tirabeque or mangetout) were grown
in an organic greenhouse of 800 m2 , at Instituto de Investigacion y Formacion Agraria y
Pesquera (IFAPA) Center “La Mojonera” (36◦ 48 N, 2◦ 41 W; altitude 142 m). The crop
(Figure 2) was carried out according to European ecological regulations [32]. The crop cycle
took place between October 2020 and March 2021. Two treatments, T100 (100% fertigation
treatment) and T50 (50% of water and fertilizers applied), were arranged in a randomized
complete block design with 3 replicates, for each cultivar and fertigation treatment, being
the planting density of 4 plants per m2 [13]. T100 consisted of water and fertilizer provided

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Sensors 2022, 22, 4113

according to fertigation management. The fertigation treatments allowed us to have a

larger number of samples with physicochemical variability (different qualities) to develop
NIRS predictive models.

Figure 2. Detail of flower, leaves (left up) and pod (left down) of mangetout. Panoramic view of
field trial (right).

A random monitoring of disease and pest symptoms was conducted weekly. In T100,
the consumption of irrigation water was 100 L m−2 , applying ecological fertilizers so that
the average nutrient solution reached 2.3 mS cm−1 . Pods were harvested when reached
standard commercial sizes.

2.2. Physicochemical Parameters

The parameters considered to assess the physical quality in mangetout fruit were
firmness and skin color, whereas the parameters of chemical quality were total soluble
solids content, pH, total vitamin C, total polyphenol content and protein content. All these
characters were determined on the fruit of fresh mangetout except the protein content. For
each cultivar (8), treatment (2) and replicate (3), three samples were used. Each sample was
composed of 5 pods from 3 plants selected at random, which were then averaged (n = 144).

2.2.1. Firmness
Texturometer XTPlus (Texture Analyzer, Surrey, UK) was used to obtain pod firmness
(Figure 3). Shear force was measured by the Warner-Bratzler test. The pod was cut
perpendicular with a Warner-Bratzler blade at 1 mm s−1 during 5 s. The result was
expressed in Newton (N).

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Sensors 2022, 22, 4113

Figure 3. Detail of Texturometer XTPlus Texture Analyzer (a); Smart-1 digital refractometer (b);
Automatic Metrohm 862 Titrosampler (c); Freeze-drying equipment (d); Mill (e); NIRS sample
holder (f); Spectrometer Model 6500 Foss-NIRSystems (g).

2.2.2. Color
CM-700d Konica Minolta portable colorimeter was used. Chroma and Hue angle were
measured externally, in two different pod locations, in the central plane.

2.2.3. Total Soluble Solids and pH

The soluble solid content (TSS) of the pods was obtained through measurement with
a Smart-1 digital refractometer (Atago, Japan) (Figure 3), and the previous sample was
homogenized for 30 s at 700 Braun CombiMax. The result was expressed in Brix. The pH
was obtained by automatic Metrohm 862 Titrosampler (Metrohm, Riverview, Florida, USA)
(Figure 3)

2.2.4. Total Polyphenol Content

In total, 10 g of the pods was homogenized with 10 mL of ethanol in PT3100 Polytron
(Littau, Switzerland) and then centrifuged for 10 min at 4 ◦ C in J2-21M/E Beckman (Fuller-
ton, CA, USA). The pellet was resuspended in 10 mL 70% methanol in water (v/v) and
centrifuged again. Finally, the supernatant was diluted with 25 mL of 70% methanol. This
extract was used to determine the TPC according to the Folin–Ciocalteu procedure [33].
In total, 200 μL of the extract, 1 mL of Folin–Ciocalteu solution (diluted 1:10 in water)
and 800 μL of Na2 CO3 (7.5%) were mixed vigorously, then the mixture was incubated
in the dark at room temperature. After 1 h, absorbance at 765 nm was determined on
ThermoSpectronic (Thermo Fisher Scientific, Waltham, MA, USA. The quantification of
TPC was expressed in Gallic acid equivalents (mg GAE kg−1 Fw).

2.2.5. Vitamin C
The reference values for ascorbic acid content (AAC) were obtained using the iodine
titration method by means of an automatic Metrohm 862 Titrosampler [34]. In total, 5 g of
sample juice was mixed with distilled deionized water until reaching 50 g of final weight,
mixing with 2 mL of glyoxal solution (40%). We proceeded to a brief stirring briefly and
5 min of rest. Once 5 mL of sulfuric acid (25%) was added, it was titrated with iodine
(0.01 mol L−1 ) to the end point (EP1). Pure ascorbic acid (AA) was used as an external
standard to determine the linearity of the method. For each standard solution, valuations
were performed in triplicate. The values of the regression equation and the regression
coefficient (r2 = 0.9998) were obtained. The ascorbic acid content was expressed as mg
100 g−1 fresh weight (fw).

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2.2.6. Protein Content

The nitrogen (N) content of the dried and ground pod samples was determined by the
Kjeldahl method using a distillation apparatus (k314, Büchi Labortechnik GmbH, Essen,
Germany) and then converted to protein content by multiplying it by 6.25. The protein
content was expressed as g 100 g−1 dry weight (dw).

2.3. Statistical Analysis

Analysis of variance (ANOVA) was used to compare differences among treatments for
total marketable yield. Previously, normality and homoscedasticity were tested using the
Shapiro–Wilk and Levenne tests, respectively. For these analyses, Fisher’s least significant
difference (LSD) test was used to compare the treatments, using the 5% level of significance.
Data were analyzed using the Statistical Package for the Social Sciences (SPSS) 24.0 software
package (LEAD Technologies, Inc., Chicago, IL, USA).

2.4. VIS-NIRS Analysis

Six replicate spectra were recorded for each sample (n = 144) and the average of the
spectra was calculated. The samples were lyophilized using freeze-drying equipment
(Telstar LyoQuest, Terrassa, Spain), then ground in a mill (Janke & Kunkel, model A10,
IKA® -Labortechnik) for about 20 s to pass through a 0.5 mm screen and stored at −80 ◦ C
until analysis. The samples were freeze-dried to eliminate the strong absorbance of water in
the infrared spectral region, which overlaps with important bands of nutritional compounds
that are present in low concentration. Samples were placed in the NIRS sample holder
(3 cm diameter) until it was 34 full (weight ∼
= 3.50 g) and were scanned (Spectrometer Model
6500 Foss-NIRSystems, Inc., Silver Spring, MD, USA). Their NIR spectra were acquired
over a wavelength range from 400 to 2500 nm (VIS + NIR regions) at 2 nm intervals.
Principal component analysis (PCA) was used to detect and remove possible spectral
outliers (spectra with a standardized Mahalanobis distance (H) from the mean spectrum of
the population greater than 3) [35].
Then, laboratory values were added to the spectra files. The reference values were
plotted as the dependent variable and the predicted NIRS values plotted as the indepen-
dent variable. The raw optical data (as log 1/R, being R = reflectance) or first or second
derivatives of the log 1/R data, with several combinations of derivative (gap) sizes and
segment (smoothing) were used to develop calibration equations [36,37]. Modified partial
least squares was used as regression method to correlate the spectral information (raw
optical and the different spectral treatments) of the samples and the quality components.
The applied pre-treatments to correct baseline offset due to spectral dispersion effects
(differences in particle size between samples) were standard normal variate and detrending
(SNV-DT) transformations.

2.5. Cross-Validation
Cross-validation is an internal validation method [38] and is useful because all samples
can be used to perform the calibration equation without the need to maintain separate
calibration sets and validation [39]. The method involves dividing the calibration set into M
segments (six) and calibrating M times, each time assessing a different part of the set of cal-
ibration (1/M) [40]. This number was proposed by WinISI software (Infrasoft International,
Port Matilda, PA, USA), five groups being used as the calibration set and then tested on the
remaining samples, performing a validation. This process continued until each group of
the six was used as a validation group. WinISI software uses principal component analysis
as a tool for selecting samples (spectra) to establish the calibration and validation groups.
Thus, both groups comprised samples representative of the whole spectral variability of
the population with similar mean and standard deviations for each trait.
Thus, cross-validation was conducted on the calibration set to establish the optimum
number of terms to be used in building the calibration equations and to identify spectral (H)
or chemical (T) outliers. “T” outliers are samples with high residuals when predicted by the

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Sensors 2022, 22, 4113

model build in the cross-validation. T values of greater than 2.5 are considered significant
and those NIR analyses which have large T values may possibly be outliers. The H outlier
identifies a sample that is spectrally different from other samples in the population and has
a standardized H value of greater than 3.0. The outlier elimination pass was set to allow
the software to remove outliers twice before completing the final calibration [41].
The performances of the different calibration equations obtained were determined
from cross-validation. Thus, the prediction ability of the equations obtained for each
quality component was determined on the basis of two mathematical relationships, which
are the standard error of cross-validation (SECV) [42] to standard deviation (SD) ratio
(RPD = relative percent difference).

2.6. External Validation

To evaluate the precision and accuracy of the equations obtained in the calibration
models, an external validation procedure in 30 independent samples was completed. Thus,
having ordered the sample set by spectral distance using the CENTER algorithm (Winisi),
the 30 samples forming the validation set were selected by taking approximately 1 of every
5 samples in the final 144 sample set. The calibration set thus comprised the remaining
114 samples.
The statistical methods applied in this study included the coefficient of determination
calculated in cross-validation (R2 CV) and external validation (R2 V), the root mean square
error of calibration (RMSEC), the root mean square error of cross-validation (RMSECV)
and the root mean square error of prediction (RMSEP). Moreover, the ratio of prediction
to deviation (RPD), which indicated the correlations between the SD of the standard wet
chemical analyzed data and prediction data by NIRS model (RMSECV or RMSEP) [42],
was applied to estimate the prediction ability of the model.
NIR models can be classified depending on the R2 value from the external valida-
tion [43] as: models (0.26 < R2 v < 0.49) with a low correlation; models (0.50 < R2 v
< 0.64) that can be used to discriminate between low and high values of the samples;
models (0.65 < R2 v < 0.81) that can be used for rough predictions of samples; models
(0.82 < R2 v < 0.90) with good correlations; and models (R2 v > 0.90) with excellent precision.
The RPD statistic demonstrates how well the calibration model predicts data. The RPD
value >3 is desirable for excellent calibration equations, while equations with an RPD <1.5
are unsuitable, according to the guideline used for defining performance calibrations [43].
With regard to the range error ratio (RER), values in the 4 to 8 range indicate the ability
to discriminate between high and low values, and RER values from 8 to 12 establish the
ability to predict quantitative data [44,45].
The mathematical expressions of these statistics are as follows:
 1/2 −1
n
−1
RPD = SD ∑ (yi − ŷi ) 2
( N − K − 1)
i =1

where yi = lab reference value for the ith sample; ŷ = NIR measured value; N = number of
samples; K = number of wavelengths used in an equation; and SD = standard deviation.
The coefficient of determination in the cross-validation (R2 ):
  −1
n n
R =
2
∑ (ŷ − y) 2
∑ ( yi − y ) 2

i =1 i =1

where ŷ = NIR measured value; y= mean “y” value for all samples; yi = lab reference value
for the ith sample.
 1/2 −1
n
−1
RER = range ∑ (yi − ŷi ) 2
( N − K − 1)
i =1

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Sensors 2022, 22, 4113

where yi = lab reference value for the ith sample; ŷ = NIR measured value; N = number of
samples; and K = number of wavelengths used in an equation.

3. Results and Discussion

3.1. Marketable Yield
Figure 4 shows the total marketable yield of the diverse varieties in response to different
fertigation treatments which ranged from 0.54 to 2.49 kg m−2 . Significant differences were
found between the different cultivars of mangetout, the most productive being the varietal
types of the indeterminate climbing growth plant, corresponding to the local Landrace (T50
2.49 kg m−2 , T100 2.44 kg m−2 ), AR-24009 (T50 2.22 kg m−2 , T100 2.05 kg m−2 ) and Tirabeque
IS (T50 1.76 kg m−2 , T100 2.05 kg m−2 ), followed by the varieties Tirabí (T50 1.57 kg m−2 , T100
1.58 kg m−2 ), Pea Zuccola (T50 1.42 kg m−2 , T100 1.34 kg m−2 ), Capuchino (T50 1.37 kg m−2 ,
T100 1.60 kg m−2 ), Pea Delikata (T50 1.37 kg m−2 , T100 1.34 kg m−2 ) and lastly the Bamby
variety which shows the prostrate growth (T50 0.57 kg m−2 , T100 0.54 kg m−2 ).

Figure 4. Total marketable yield (kg m−2 ) of the different varieties of mangetout under different
fertigation treatments (T50 and T100). Bars with different lowercase letters were significantly different
at p < 0.05 (Tukey’s multiple range test).

The production data obtained in the field trial for most of cultivars were higher than
those described previously in mangetout by García-García [46] (0.55–0.65 kg m−2 ), and
similar to those indicated by Estrada and Ibáñez [47] (1.5–2 kg m−2 ) in Mediterranean
greenhouse conditions.
Increasingly, the use of organic production regulations [48] as well as appropriate
fertigation management play an important role in enhancing crop quality and economizing
water [13] according to the Sustainable Development Goals (SDGs) by 2030. In this regard,
previous studies have showed that yield and quality of snap pods can be significantly
affected by different compositions of fertilization [49,50] and by different doses of water
in the fertigation solution [13,51]. In order to obtain the highest possible physicochemical
variability to develop NIRS predictive models, two fertigation treatments and different
mangetout cultivars were used.

3.2. Physicochemical Profiles

The samples analyzed varied in all variables as shown by the range and coefficient of
variation (CV) of the calibration set (Table 2). The highest values for the CV were observed
for C* chromatic value, firmness, ascorbic acid content and total polyphenol content (>20%),
possibly due to the different fertigation treatments and varieties used.

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Table 2. Mean, range and standard deviation (n = 144) for quality parameters of the mangetout
samples used in this study.

Parameters Mean Range SD

C* 27.87 15.20–35.58 6.15
h* 109.46 105.13–112.91 1.49
Firmness (N) 43.62 20.59–67.52 12.74
TSS (Brix) 7.53 6.08–8.85 0.65
pH 6.80 5.99–7.28 0.27
Protein (g 100 g−1 dw) 23.48 11.50–29.75 3.02
AAC (mg 100 g−1 fw) 43.82 19.75–68.86 10.82
TPC (mg GAE kg−1 fw) 389.09 202.30–685.05 111.52

Based on the results of this study, the chromatic parameters (C* and h*) varied from
15.20 to 35.58 and 105.13 to 112.91, respectively. The h* values correspond to the color green.
Green color of fresh pods is one the key factors for deciding the commercial acceptance of
snap bean as a fresh vegetable. Similar results have been previously found in snap pods
with values ranged from 107 to 111 for h* parameter, but a narrow variation range (27 to
33) for C* chromatic parameter [13,52,53].
Texture is a quality attribute in mangetout fruits very important for consumers since its
singular quality of edible crunchy pod is highly appreciated. From our study, the firmness
values in mangetout pods ranged from 20.59 to 67.52 N. Although information is lacking
for the comparison of firmness with other mangetout cultivars from the literature, our
previous research results showed that mangetout “Tirabí” showed values included in the
range mentioned above [13].
Vitamin C is essential in both plants and animals. The main suppliers of this vitamin
in the diet are fruits and vegetables [54]. Legumes are considered an important source of
vitamins, especially rich in ascorbic acid content in the pods [55]. Considerable variation
was found for AAC which ranged from 19.75 to 68.86 mg AA 100 g−1 fw in mangetout
pods. Previous studies revealed AA content within the range of AAC showed in this work
for three pea varieties (26 to 31 mg AA 100 g−1 ) [56]. Our findings are also in agreement
with those of Rickman et al. [57] and Avilés and Cruz [58], who described AA values of
40 and 27 mg 100 g−1 fw in peas and pea pods, respectively. Mangetout pods can be
considered a rich source of vitamin C, since orange and lemon contain 30–50 mg of ascorbic
acid 100 g−1 fw [54].
The pH of foods is an important parameter related to the taste perceived by consumers.
In our study, the pH ranged from 5.99 to 8.85. The values obtained agree with previous
studies on legumes [59–61], but lower than those obtained in French bean pods (5.84–5.96)
by Segura et al. [62].
TSS is another taste quality determinant [63], and cultivars with higher TSS have
higher taste quality. Mangetout pods are rich in TSS content (6.29–8.83 Brix) in comparison
with other legume pods; thus, cowpea accessions from different Mediterranean countries
showed lower sweetness (range 5.07–7.57 Brix) [55] in relation to our results.
On the other hand, the fresh mangetout pod TSS content in our study was lower
compared to those previously reported in the scientific literature [12]. This previous work
revealed that the TSS of fresh pods ranged from 9.1 to 11.3 under specific fertigation
treatments demonstrating that the environmental factors such as available water had a
highly significant effect on this quality parameter. According to the Brix reference values of
the main greenhouse vegetables, the mangetout pods analyzed showed a higher sweetness
than California green pepper fruits (4.03–6.31 Brix) and similar to red California pepper
fruits (7.37–8.85 Brix) [64].
The presence of polyphenols in plants is very varied, depending on the plant species,
variety, part of the plant, growing conditions, etc. More than 8000 phenolic compounds
with a very varied structure have been identified from simple molecules, such as phenolic
acids and complex polymers of high molecular mass such as tannins [65]. In our study,

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mangetout exhibited higher total polyphenol content (202.30 to 685.05 mg GAE kg−1 fw)
that those reported for other snap pods, such as the French bean with 300 mg GAE kg−1
fw [46]. Our results agree with those of Devi et al. [66] who found a wide variation range
(126.3–1286.3 mg GAE kg−1 fw) in pea pods from 22 different genotypes. On the other
hand, the consumer increasingly appreciates fruits with antioxidant properties due to
the health benefits. A source of phenolic compounds is identified as a chemopreventive
agent since it eliminates free radicals and has a preventive effect on degenerative diseases,
among others [67]. Mangetout pods have a high potential to be used in the development of
functional foods or nutraceutical products and unlike pea pods they would not require any
processing as the whole pod is edible.
Our results showed a wide variability for protein content (11.50–29.75 g 100 g−1 dw)
and our results agree with those of Hood-Niefer et al. [68] (24.4 to 27.5 g 100 g−1 dw), but
are higher than the results obtained by Mateos-Aparicio et al. [69] (10.8 ± 0.3 g 100 g−1 dw)
in pea pods. Overall, in the pea, both the seeds (20.5–22.6%) and pods (13.37%) are a rich
source of protein [70]. A diet rich in vegetable protein is increasingly important nowadays
due to its health benefits and thus it is recommended that people reduce their consumption
of animal protein. In addition, pea pods have protein-denaturing properties that show
anti-inflammatory effects and anti-cholinesterase activity because of the strong antidiabetic
properties of peas [71].

3.3. VIS-NIRS Analysis

3.3.1. Raw Spectra on Mangetout
Raw spectra of the calibration set samples are shown in Figure 5. A remarkable
variability in the VIS region (400–850 nm) absorbance spectra was observed because of pig-
ments. The peak around 640–700 nm illustrated the color transition of pea pods correlated
with the chlorophyll content that absorbs radiation in this region [72].

Figure 5. Raw spectra for dried mangetout samples.

Absorption bands in the region from 1300 to 2000 nm have been assigned to the third
overtones of C-N (amines); C=O (ketones, amino acids); and C-O (long-chain fatty acids,
phenols). From 2200 to 2400 nm, absorptions of C-N (primary amines) and C-O (alcohols)
have been assigned to the third overtones of these compounds, while in the same region,
C-H (asymmetrical deformation) and C-O (symmetrical vibrations) have been assigned to
the second overtones of these molecules. Finally, the second overtones of C-H deformation
and C-N (amides) have been reported in the 2400–2500 nm region [73].

3.3.2. Second Derivative Spectra of Mangetout

The second derivative and SNT-DT (standard normal variate and de-trending) algo-
rithms to the raw spectra led to a substantial correction (Figure 6) of the baseline shift

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produced by differences in path length and particle size. The increase in the complexity of
the derivative spectra resulted in a clear separation between peaks which overlap in the
raw spectra.

Figure 6. Second derivative spectra (2, 5, 5, 2; SNV + DT) of the raw optical data for mangetout
samples in the range of 400 to 2500 nm, together with the most relevant absorption bands.

Absorption maxima bands (λmax) were observed between 400 and 700 nm (at 444,
546 nm and 670 nm) in the spectra attributed to pod pigments that absorb in visible
region (Figure 6). From all pigments that can be found in plants, chlorophylls are used for
photosynthesis (“a” and “b”), which absorb preferentially violet-blue light (400–500 nm)
and red light (600–700 nm), respectively [74].
Pigment–protein complex molecules could be responsible for some of the traits that
determine the VIS region at longer wavelengths. Thus, binding proteins in chlorophyll a/b
absorb in the 498–568 nm range [74] and red absorbing pigments, particularly chlorophyll,
give the fruit its green color [75,76].
In the region NIRS of the spectra, peaks at 1208 nm (attributed to a CH second
overtone), 1726 nm and 1762 nm (assigned to CH first overtone), 2308 nm and 2348 nm
(attributed to CH stretch and deformation in a CH2 group) were detected which are
related to lipids [77,78]. Other peaks located at 1210 nm corresponded to absorption by
OH groups in carbohydrates [79,80]. Other peaks at 1512, 2056 and 2174 nm related to
protein, specifically to NH stretch, NH stretch and amide II, and amide I and amide III,
respectively [78]. The last significant peaks were observed at 1436 and 2270 nm, these
wavelengths corresponding to the deformation of the OH + CO cellulose groups [79].

3.3.3. Calibration Development

Tables 3 and 4 show the summary of the statistics obtained from calibration, cross-
validation and external validation models in mangetout samples, respectively. The full
available visible region and near-infrared region (400–2500 nm) were used.
The coefficients of determination (R2 ) achieved in calibration were higher than those
found in external validation models for mangetout, as expected. The coefficient of deter-
mination for cross-validation (R2 CV), oscillated between 0.55 for pH to 0.92 for protein
(Table 3), whereas RPDcv values ranges from 1.50 for pH to 3.45 for protein.
Based on the R2 values of the external validation, the models were as follows [39]:
models that can be used to discriminate between low and high values of the samples
(0.50 m< R2 < 0.64), in our work the models developed for AAC and TSS; models that can be
used for rough predictions of samples (0.65 < R2 < 0.81), in our case the calibrations achieved
for C* and h* color parameters, firmness and pH; and models with good correlations
(0.83 < R2 < 0.90), these values corresponding with models obtained for total polyphenol
content and protein.

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The SEP values of the validation were lower than their respective SD, which indicates
that NIRS is able to determine these traits in mangetout.

Table 3. Calibration and cross-validation statistics of quality compounds for mangetout.

Parameters Range 1 SD 2 R2 3 SEC 4 R2 CV 5 SECV 6 RPDcv 7 Treatment 8 Cv

C* 15.20–35.58 6.35 0.87 2.24 0.81 2.78 2.28 2,5,5,2 0.22
h* 106.41–112.10 1.41 0.80 0.62 0.71 0.75 1.88 1,4,4,1 0.01
Firmness (N) 21.75–67.52 10.09 0.71 5.46 0.71 5.93 1.70 1,4,4,1 0.21
9 TSS (Brix) 6.29–8.83 0.65 0.93 0.18 0.68 0.39 1.66 1,4,4,1 0.08
pH 6.01–7.28 0.27 0.60 0.17 0.55 0.18 1.50 1,4,4,1 0.04
Protein
15.69–29.75 2.80 0.97 0.48 0.92 0.81 3.45 2,5,5,2 0.13
(g 100 g−1 dw)
10 AAC
19.75–64.40 10.89 0.79 5.02 0.56 7.16 1.52 1,4,4,1 0.24
(mg 100 g−1 fw)
11 TPC
239.28–670.30 101.91 0.93 27.01 0.86 39.08 2.61 1,4,4,1 0.28
(mg GAE kg−1 fw)
1 SD: Standard deviation; 2 R2 : Coefficient of determination in calibration; 3 SEC: Standard error in calibration;
4 R2 : Coefficient of determination in cross-validation; 5 SECV: Standard error of cross-validation; 6 RPDcv: Ratio
of the standard deviation to standard error of cross-validation; 7 Mathematical treatment; 8 Coefficient of variation;
9 TSS: Total soluble solids; 10 AAC: Ascorbic acid content; 11 TPC: Total polyphenol content.

Table 4. Reference values and external validation statistics of the NIRS calibrations for quality
compounds in mangetout.

Reference Values (n = 30) External Validation

Parameters Range Mean 1 SD 2 Rv2 3 SEP 4 RPDp 5 RER
C* 15.20–34.89 25.50 7.33 0.78 3.34 2.19 5.89
H* 107.40–111.71 109.71 1.24 0.68 0.56 2.00 6.95
Firmness (N) 24.45–67.20 40.48 12.51 0.65 7.34 1.70 5.96
6 TSS (Brix) 6.29–8.76 7.54 0.69 0.52 0.51 1.35 4.84
pH 6.22–7.20 6.83 0.22 0.50 0.14 1.57 7.00
Protein (g 100 g−1 dw) 17.22–29.5 24.95 2.18 0.88 0.68 3.20 14.89
7 AAC (mg 100 g−1 fw) 22.71–63.47 45.69 8.82 0.50 8.82 1.50 7.03
8 TPC (mg GAE kg−1 fw) 250.89–570.21 360.89 80.37 0.84 29.46 2.72 10.84
TSS: Total soluble solids; AAC: Ascorbic acid content; TPC: Total polyphenol content; 1 SD: Standard deviation;
2 Rv2 : Coefficient of determination in external validation; 3 SEP: Standard error of prediction corrected for bias;
4 RPDp: Ratio of the standard deviation to standard error of prediction (performance); 5 RER: Ratio of the range

to standard error of prediction (performance); 6 TSS: Total soluble solids; 7 AAC: Ascorbic acid content; 8 TPC:
Total polyphenol content.

According to the guideline used for defining performance calibrations [43] when this
ratio is greater than 3, the calibration equation is very significant, and this was reached
in our study for protein content; if RPD values range between 2.5 < RPD < 3, predictive
models are considered very good, in our case corresponding to the TPC model; while RPD
range between 1.5 < RPD < 2.5 predictive models are appropriate for screening purposes,
which was achieved for AAC, pH, firmness and C* and h* color parameter models.
Figure 7 shows the relationship between the predicted reflectance spectroscopy in the
near infrared (NIRS) and reference values for all parameters (color parameters (chroma *
and hue angle), firmness, total soluble solids, pH, protein content, ascorbic acid and total
polyphenol content) in the mangetout validation set samples.
In reference to RER (ratio of the range to standard error of prediction) coefficients, this
dimensionless parameter is also used to evaluate the predictive ability of NIRS equations,
in this work ranged from 4.84–14.89.
Prediction models for C*, h*, firmness, TSS, pH, AAC showed RER values within the
range from 4 to 8, which suggest the possibility of discriminating between high and low
values; while RER values in the range of 8 to 12 represent the possibility of predicting
quantitative data [44,45] which was achieved for protein and TPC predictive models.

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Figure 7. Predicted versus reference values for calibration and external validation for all parameters:
c* (a); h* (b); firmness (c); total soluble solids (d); pH (e); protein content (f); ascorbic acid (g); total
polyphenol content in the mangetout (h).

Previous works have demonstrated the validity of the NIRS technique in evaluating
the accuracy of pea single seed protein with R2 = 0.94 and RPD = 3.7 in external valida-
tion [28], and also for predicting soybean single seed protein content with Rval2 = 0.84 and
RPDval = 2.28 values [81].
The estimation of protein and total polyphenol content in common beans (Phaseolus
vulgaris L.) by NIRS has also been previously assayed by several authors reaching signifi-
cantly good results in general. Thus, the high R2 obtained ranged from 0.91–0.94 and RPD
values above 3.5 [82–85]. Other authors supported the validity of the NIRS technique in
similar approaches, with R2 and RPD values for firmness of 0.61 and 1.7, respectively, in
soybean single seed [82]. Wang [86] used NIRS to predict the total polyphenol content in
ground faba bean (Vicia faba L.), with an R2 of 0.79, RMSECV of 0.40 and RPD of 2.20, and
also for the determination of protein in ground faba bean seed powder with an R2 of 0.94.
It should be noted that the prediction accuracies in all of the above-mentioned studies
were comparable to those reported for mangetout in this study. To our knowledge, this is
the first article dealing with the use of NIRS to predict pod quality traits in mangetout.
Modified PLS regression was employed to reduce the spectral information of the
mangetout samples by creating a much smaller number of new orthogonal variables
(factors) which retain the essential information needed to predict the composition of
the samples.

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3.3.4. Modified Partial Least Squares Loadings for Quality Equations

The scores of the best models for all quality parameters were plotted by their first
MPLS loadings (Figure 8) to identify those areas within the spectral range where variance
had influenced the model fitting, to a lesser or greater degree, as well as the direction
(negative or positive).

Figure 8. Modified partial least squares (MPLS) loading of the optimal calibrations for physicochemi-
cal compounds measured by NIRS.

The region of the spectrum which most influenced the fitting of the model was the
visible segment between 480 to 700 nm. Thus, the contribution of chlorophyll (672 nm)
showed the highest weight on first MPLS loading [75] (Figure 8). Other chromophores
absorbing at 496 and 512 nm also participated in the equations. With respect to NIR region,
previous studies have shown the contribution of this region to predict color parameters for
species such as fresh Ginkgo biloba leaves [87], green-leafy species [88] or Sassafras tzumu [89].
Some plant chemical compounds (e.g., phenolics and flavonoids) respond to the stress and
environmental changes and correlate in a secondary way with the color parameters. The
characteristic bands of phenolics and flavonoids can be detected in wavelength regions
from 1415 nm to 1512 nm, 1650 to 1750 nm and from 1955 to 2035 nm in the MPLS loadings
for the color parameters (Figure 8) [90]. Furthermore, the color is caused by the reflection
of helicoidally stacked cellulose microfibrils that form multilayers in the cell walls of the
epicarp [91]. Thus, the wavelengths at 1932 nm (O-H stretching plus O-H deformation)
could be related to the cellulose of the pod tissues which can be observed in the MPLS

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loadings of the optimal calibrations for the color parameters (Figure 8). Others main NIR
contributions were those at 2284 nm (C-H stretching plus C-H deformation), 2300 nm
(stretching–bending of CH—CH2 bonds and C—O bonds) and 2348 nm (C-H combination
of methylene groups) [77]. In addition, absorption bands in the NIR region that influenced
the fitting of the models were found at 1212, 1388, 1412 and 1990 nm (associated with
glucides and water absorptions), and the region around 2072 nm (N—H bonds) associated
with protein.

4. Conclusions
This work has showed that genetic variability exists for the quality parameters ana-
lyzed in mangetout cultivars. Many of the traits analyzed are of economic interest (color,
firmness, protein content and antioxidant compounds). These new understandings could
be useful in selecting parents for breeding programs aimed at enhancing physicochemical
parameters that respond to the new trends market.
Moreover, the result of the present investigation explores the potential of NIRS to
simultaneously determine eight quality traits in mangetout, as an alternative to reference
methods. The measurements with the reference methods of most of these parameters are
expensive, have laborious protocols and require a long analysis time. Utilizing NIRS, every
2 min, we can analyze all the quality parameters of a sample. The results reveal that the
models allow an accurate quantification of protein and TPC and a rough screening method
of the samples for color parameters (c* and h*), firmness, AAC and pH.
The inclusion future of mangetout cultivars from different geographical origins and
segregant populations in the calibration models will allow us to increase the robustness of
the equations for these parameters.
The performance of the calibration model for TSS was lower than that obtained for
the other quality parameters in this work. The low variability among mangetout cultivars
used in this work (6.29–8.76 Brix) could be based on the lower accuracy of the calibration
model for TSS. An increase in both the number of samples and trait variation can be crucial
factors for improving the accuracy of this calibration model.
It is interesting to focus attention on firmness (shear force). Pod firmness is an excellent
indicator of pod quality, but its quantification is time consuming and not easily measured.
Pods must be harvested before they become tough and develop poor culinary acceptance,
even if it means sacrificing maximum yield. From this point of view, the use of NIRS
instead of a texturometer could be clearly advantageous.
Spectral ranges associated with the absorbance of chromophores, carbohydrates, water
and protein were used by MPLS regression for the model fitting of quality equations
in mangetout.

Author Contributions: M.d.R.-C. and J.A.F. conceived and designed the experiments; B.d.C.M.-G.
and E.M.-E. performed the field trials and laboratory analysis; J.L.V. and P.G. performed supervision;
I.F.: MPLS regression analysis; M.d.C.G.-G.: NIRS analysis and wrote the paper. All authors have
read and agreed to the published version of the manuscript.
Funding: This study was carried out with the support of the Projects ‘Innovación en cultivos prote-
gidos: biodiversidad y bioeconomía aplicada’ (PP.TRA.TRA2019.003) and ‘Nuevas alternativas de
cultivos a la producción hortícola protegida’ (PP.AVA.AVA2019.023). FEDER cofund 80% ‘Andalucía
se mueve con Europa’.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Data sharing not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

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90. Verardo, V.; Cevoli, C.; Pasini, F.; Gómez-Caravaca, A.M.; Marconi, E.; Fabbri, A.; Caboni, M.F. Analysis of oligomer proantho-
cyanidins in different barley genotypes using high-performance liquid chromatography-fluorescence detection-mass spectrometry
and near-infrared methodologies. J. Agric. Food Chem. 2015, 63, 4130–4137. [CrossRef]
91. Vignolini, S.; Rudall, P.J.; Rowland, A.V.; Reed, A.; Moyroud, E.; Faden, R.B.; Baumberg, J.J.; Glover, B.J.; Steiner, U. Pointillist
structural color in Pollia fruit. Proc. Natl. Acad. Sci. USA 2012, 109, 15712–15715. [CrossRef]

37
 sensors
Article
Evaluation of Minimum Preparation Sampling Strategies for
Sugarcane Quality Prediction by vis-NIR Spectroscopy
Lucas de Paula Corrêdo * , Leonardo Felipe Maldaner , Helizani Couto Bazame and José Paulo Molin

Precision Agriculture Laboratory, Biosystems Engineering Department, ‘Luiz de Queiroz’ College of Agriculture,
University of São Paulo, Av. Pádua Dias 11, 13418-900 Piracicaba, Brazil; [email protected] (L.F.M.);
[email protected] (H.C.B.); [email protected] (J.P.M.)
* Correspondence: [email protected]

Abstract: Proximal sensing for assessing sugarcane quality information during harvest can be affected
by various factors, including the type of sample preparation. The objective of this study was to
determine the best sugarcane sample type and analyze the spectral response for the prediction of
quality parameters of sugarcane from visible and near-infrared (vis-NIR) spectroscopy. The sampling
and spectral data acquisition were performed during the analysis of samples by conventional methods
in a sugar mill laboratory. Samples of billets were collected and four modes of scanning and sample
preparation were evaluated: outer-surface (‘skin’) (SS), cross-sectional scanning (CSS), defibrated
cane (DF), and raw juice (RJ) to analyze the parameters soluble solids content (Brix), saccharose (Pol),
fibre, pol of cane and total recoverable sugars (TRS). Predictive models based on Partial Least Square
Regression (PLSR) were built with the vis-NIR spectral measurements. There was no significant
difference (p-value > 0.05) between the accuracy SS and CSS samples compared to DF and RJ samples
for all prediction models. However, DF samples presented the best predictive performance values for
the main sugarcane quality parameters, and required only minimal sample preparation. The results
Citation: Corrêdo, L.d.P.; Maldaner, contribute to advancing the development of on-board quality monitoring in sugarcane, indicating
L.F.; Bazame, H.C.; Molin, J.P. better sampling strategies.
Evaluation of Minimum Preparation
Sampling Strategies for Sugarcane Keywords: chemometrics; proximal sensing; precision agriculture
Quality Prediction by vis-NIR
Spectroscopy. Sensors 2021, 21, 2195.
https://doi.org/10.3390/s21062195

1. Introduction
Academic Editor: Lammert Kooistra
Near-Infrared (NIR) spectroscopy is a well-established technique to monitor the qual-
ity of raw sugarcane received by sugar mills [1], and consequently, for pricing and trading
Received: 23 January 2021
Accepted: 19 March 2021
with producers and growers [2]. Crop quality is estimated based on physicochemical
Published: 21 March 2021
parameters related to physiological composition, such as soluble solids content (Brix),
water-insoluble solids (Fibre), and the apparent sucrose in the juice (Pol). Furthermore, all
Publisher’s Note: MDPI stays neutral
other parameters (purity, Pol of cane, reducing sugars, and total recoverable sugars) are cal-
with regard to jurisdictional claims in
culated based on the former parameters [3], from which total recoverable sugars (TRS) are
published maps and institutional affil- used for the pricing and trading of the raw material. Sugarcane quality parameters are de-
iations. termined by analytical methods and empirical equations described by the National Council
of Sugarcane Producers (CONSECANA), which are based on the International Commission
for Uniform Methods of Sugar Analysis (ICUMSA). Using calibration methods, it is now
possible to obtain some crop quality parameters by NIR spectroscopy [4].
Copyright: © 2021 by the authors.
Different wavelength regions of the electromagnetic spectrum can be used in spec-
Licensee MDPI, Basel, Switzerland.
troscopy, such as visible (400 to 750 nm), near-infrared (NIR, 750 to 2500 nm), shortwave
This article is an open access article
near-infrared (SWNIR, 750 to 1100 nm), and visible and near-infrared (vis-NIR, 400 to
distributed under the terms and 2500 nm) [5–8]. However, there is no consensus on the limits between these regions.
conditions of the Creative Commons Interaction between electromagnetic radiation and matter causes molecular vibrations
Attribution (CC BY) license (https:// involving heavy atom (C, N, O, and S) attached to a hydrogen atom [8]. This basic principle
creativecommons.org/licenses/by/ has allowed substantial scientific advances to predict organic compounds of agricultural
4.0/). products associated with its quality using vis-NIR spectroscopy as a nondestructive and

39
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spirit with which we are best acquainted? We observe certain
phænomena. We cannot explain them into material causes. We
therefore infer that there exists something which is not material. But
of this something we have no idea. We can define it only in
negatives. We can reason about it only by symbols. We use the
word: but we have no more of the things; and the business of
poetry is with images, and not with words. The poet uses words
indeed; but they are merely the instruments of his art, not its
objects. They are the materials which he is to dispose in such a
manner as to present a picture to the mental eye. And if they are
not so disposed, they are no more entitled to be called poetry than a
bale of canvas and a box of colours to be called a painting.
Logicians may reason about abstractions. But the great mass of
men must have images. The strong tendency of the multitude in all
a^es and nations to idolatry can be explained on no other principle.
The first inhabitants of Greece, there is reason to believe,
worshipped one invisible Deity. But the necessity of having
something more definite to adore produced, in a few centuries, the
innumerable crowd of Gods and Goddesses. In like manner the
ancient Persians thought it impious to exhibit the Creator under a
human form. Yet even these transferred to the Sun the worship
which, in speculation, they considered due only to the Supreme
Mind. The History of the Jews is the record of a continued struggle
between pure Theism, supported by the most terrible sanctions, and
the strangely fascinating desire of having some visible and tangible
object of adoration. Perhaps none of the secondary causes which
Gibbon has assigned for the rapidity with which Christianity spread
over the world, while Judaism scarcely ever acquired a proselyte,
operated more powerfully than this feeling. God, the uncreated, the
incomprehensible, the invisible, attracted few worshippers. A
philosopher might admire so noble a conception: but the crowd
turned away in disgust from words which presented no image to
their minds. It was before Deity embodied in a human form, walking
among men, partaking of their infirmities, leaning on their bosoms,
weeping over their graves, slumbering in the manger, bleeding on
the cross, that the prejudices of the Synagogue, and the doubts of
the Academy, and the pride of the portico, and the fasces of the
Lictor, and the swords of thirty legions, were humbled in the dust.
Soon after Christianity had achieved its triumph, the principle which
had assisted it began to corrupt it. It became a new Paganism.
Patron saints assumed the offices of household gods. St. George
took the place of Mars. St. Elmo consoled the mariner for the loss of
Castor and Pollux. The Virgin Mother and Cecilia succeeded to Venus
and the Muses. The fascination of sex and loveliness was again
joined to that of celestial dignity; and the homage of chivalry was
blended with that of religion. Reformers have often made a stand
against these feelings; but never with more than apparent and
partial success. The men who demolished the images in Cathedrals
have not always been able to demolish those which were enshrined
in their minds. It would not be difficult to show that in politics the
same rule holds good. Doctrines, we are afraid, must generally be
embodied before they can excite a strong public feeling. The
multitude is more easily interested for the most unmeaning badge,
or the most insignificant name, than for the most important
principle. From these considerations, we infer that no poet, who
should affect that metaphysical accuracy for the want of which
Milton has been blamed, would escape a disgraceful failure. Still,
however, there was another extreme, which, though far less
dangerous, was also to be avoided. The imaginations of men are in a
groat measure under the control of their opinions. The most
exquisite art of poetical colouring can produce no illusion, when it is
employed to represent that which is at once perceived to be
incongruous and absurd. Milton wrote in an age of philosophers and
theologians. It was necessary, therefore, for him to abstain from
giving such a shock to their understandings as might break the
charm which it was his object to throw over their imaginations. This
is the real explanation of the indistinctness and inconsistency with
which he has often been reproached. Dr. Johnson acknowledges that
it was absolutely necessary that the spirit should be clothed with
material forms. “But,” says he, “the poet should have secured the
consistency of his system by keeping immateriality out of sight, and
seducing the reader to drop it from his thoughts.” This is easily said;
but what if Milton could not seduce his readers to drop immateriality
from their thoughts? What if the contrary opinion had taken so full a
possession of the minds of men as to leave no room even for the
half belief which poetry requires? Such we suspect to have been the
case. It was impossible for the poet to adopt altogether the material
or the immaterial system. He therefore took his stand on the
debatable ground. He left the whole in ambiguity. He has, doubtless,
by so doing, laid himself open to the charge of inconsistency. But,
though philosophically in the wrong, we cannot but believe that he
was poetically in the right. This task, which almost any other writer
would have found impracticable, was easy to him. The peculiar art
which he possessed of communicating his meaning circuitously
through a long succession of associated ideas, and of intimating
more than he expressed, enabled him to disguise those incongruities
which he could not avoid.
Poetry which relates to the beings of another world ought to be at
once mysterious and picturesque. That of Milton is so. That of Dante
is picturesque indeed beyond any that ever was written. Its effect
approaches to that produced by the pencil or the chisel. But it is
picturesque to the exclusion of all mystery. This is a fault on the
right side, a fault inseparable from the plan of Dante’s poem, which,
as we have already observed, rendered the utmost accuracy of
description necessary. Still it is a fault. The supernatural agents
excite an interest; but it is not the interest which is proper to
supernatural agents. We feel that we could talk to the ghosts and
dæmons without any emotion of unearthly awe. We could, like Don
Juan, ask them to supper, and eat heartily in their company. Dante’s
angels are good men with wings. His devils are spiteful ugly
executioners. His dead men are merely living men in strange
situations. The scene which passes between the poet and Farinata is
justly celebrated. Still, Farinata in the burning tomb is exactly what
Farinata would have been at an auto da fe. Nothing can be more
touching than the first interview of Dante and Beatrice. Yet what is
it, but a lovely woman chiding, with sweet austere composure, the
lover for whose affection she is grateful, but whose vices she
reprobates? The feelings which give the passage its charm would
suit the streets of Florence as well as the summit of the Mount of
Purgatory.
The spirits of Milton are unlike those of almost all other writers.
His fiends, in particular, are wonderful creations. They are not
metaphysical abstractions. They are not wicked men. They are not
ugly beasts. They have no horns, no tails, none of the fee-faw-fum
of Tasso and Klopstock. They have just enough in common with
human nature to be intelligible to human beings. Their characters
are, like their forms, marked by a certain dim resemblance to those
of men, but exaggerated to gigantic dimensions, and veiled in
mysterious gloom.
Perhaps the gods and dæmons of Æschylus may best bear a
comparison with the angels and devils of Milton. The style of the
Athenian had, as we have remarked, something of the Oriental
character; and the same peculiarity may be traced in his mythology.
It has nothing of the amenity and elegance which we generally find
in the superstitions of Greece. All is rugged, barbaric, and colossal.
The legends of Æschylus seem to harmonize less with the fragrant
groves and graceful porticoes in which his countrymen paid their
vows to the God of Light and Goddess of Desire, than with those
huge and grotesque labyrinths of eternal granite in which Egypt
enshrined her mystic Osiris, or in which Hindostan still bows down to
her seven-headed idols. His favourite gods are those of the elder
generation, the sons of heaven and earth, compared with whom
Jupiter himself was a stripling and an upstart, the gigantic Titans,
and the inexorable Furies. Foremost among his creations of this class
stands Prometheus, half fiend, half redeemer, the friend of man, the
sullen and implacable enemy of heaven. Prometheus bears
undoubtedly a considerable resemblance, to the Satan of Milton. In
both we find the same impatience of control, the same ferocity, the
same unconquerable pride. In both characters also are mingled,
though in very different proportions, some kind and generous
feelings. Prometheus, however, is hardly superhuman enough. He
talks too much of his chains and his uneasy posture: he is rather too
much depressed and agitated. His resolution seems to depend on
the knowledge which he possesses that he holds the fate of his
torturer in his hands, and that the hour of his release will surely
come. But Satan is a creature of another sphere. The might of his
intellectual nature is victorious over the extremity of pain. Amidst
agonies which cannot be conceived without horror, he deliberates,
resolves, and even exults. Against the sword of Michael, against the
thunder of Jehovah, against the flaming lake, and the marl burning
with solid fire, against the prospect of an eternity of unintermitted
misery, his spirit bears up unbroken, resting on its own innate
energies, requiring no support from any thing external, nor even
from hope itself.
To return for a moment to the parallel which we have been
attempting to draw between Milton and Dante, we would add that
the poetry of these great men has in a considerable degree taken its
character from their moral qualities. They are not egotists. They
rarely obtrude their idiosyncrasies on their readers. They have,
nothing in common with those modern beggars for fame, who extort
a pittance from the compassion of the inexperienced by exposing the
nakedness and sores of their minds. Yet it would be difficult to name
two writers whose works have been more completely, though
undesignedly, coloured by their personal feelings. The character of
Milton was peculiarly distinguished by loftiness of spirit; that of
Dante by intensity of feeling. In every line of the Divine Comedy we
discern the asperity which is produced by pride struggling with
misery. There is perhaps no work in the world so deeply and
uniformly sorrowful. The melancholy of Dante was no fantastic
caprice. It was not, as far as at this distance of time can be judged,
the effect of external circumstances. It was from within. Neither love
nor glory, neither the conflicts of earth nor the hope of heaven could
dispel it. It turned every consolation and every pleasure into its own
nature. It resembled that noxious Sardinian soil of which the intense
bitterness is said to have been perceptible even in its honey. His
mind was, in the noble language of the Hebrew poet, “a land of
darkness, as darkness itself, and where the light was as darkness.”
The gloom of his character discolours all the passions of men, and
all the face of nature, and tinges with its own livid hue the flowers of
Paradise and the glories of the eternal throne. All the portraits of him
are singularly characteristic. No person can look on the features,
noble even to ruggedness, the dark furrows of the cheek, the
haggard and woful stare of the eye, the sullen and contemptuous
curve of the lip, and doubt that they belong to a man too proud and
too sensitive to be happy.
Milton was, like Dante, a statesman and a lover; and, like Dante,
he had been unfortunate in ambition and in love. He had survived
his health and his sight, the comforts of his home, and the
prosperity of his party. Of the great men by whom he had been
distinguished at his entrance into life, some had been taken away
from the evil to come; some had carried into foreign climates their
unconquerable hatred of oppression; some were pining in dungeons;
and some had poured forth their blood on scaffolds. Venal and
licentious scribblers, with just sufficient talent to clothe the thoughts
of a pandar in the style of a bellman, were now the favourite writers
of the Sovereign and of the public. It was a loathsome herd, which
could be compared to nothing so fitly as to the rabble of Cornus,
grotesque monsters, half bestial half human, dropping with wine,
bloated with gluttony, and reeling in obscene dances. Amidst these
that fair Muse was placed, like the chaste lady of the Masque, lofty,
spotless, and serene, to be chattered at, and pointed at, and grinned
at, by the whole rout of Satyrs and Goblins. If ever despondency and
asperity could be excused in any man, they might have been
excused in Milton. But the strength of his mind overcame every
calamity. Neither blindness, nor gout, nor age, nor penury, nor
domestic afflictions, nor political disappointments, nor abuse, nor
proscription, nor neglect, had power to disturb his sedate and
majestic patience. His spirits do not seem to have been high, but
they were singularly equable. His temper was serious, perhaps
stern; but it was a temper which no sufferings could render sullen or
fretful. Such as it was when, on the eve of great events, he returned
from his travels, in the prime of health and manly beauty, loaded
with literary distinctions, and glowing with patriotic hopes, such it
continued to be when, after having experienced every calamity
which is incident to our nature, old, poor, sightless and disgraced, he
retired to his hovel to die.
Hence it was that, though he wrote the Paradise Lost at a time of
life when images of beauty and tenderness are in general beginning
to fade, even from those minds in which they have not been effaced
by anxiety and disappointment, he adorned it with all that is most
lovely and delightful in the physical and in the moral world. Neither
Theocritus nor Ariosto had a finer or a more healthful sense of the
pleasantness of external objects, or loved better to luxuriate amidst
sunbeams and flowers, the songs of nightingales, the juice of
summer fruits, and the coolness of shady fountains. His conception
of love unites all the voluptuousness of the Oriental harem, and all
the gallantry of the chivalric tournament, with all the pure and quiet
affection of an English fireside. His poetry reminds us of the miracles
of Alpine scenery. Nooks and dells, beautiful as fairy land, are
embosomed in its most rugged and gigantic elevations. The roses
and myrtles bloom unchilled on the verge of the avalanche.
Traces, indeed, of the peculiar character of Milton may be found in
all his works; but it is most strongly displayed in the Sonnets. Those
remarkable poems have been undervalued by critics who have not
understood their nature. They have no epigrammatic point. There is
none of the ingenuity of Filicaja in the thought, none of the hard and
brilliant enamel of Petrarch in the style. They are simple but majestic
records of the feelings of the poet; as little tricked out for the public
eye as his diary would have been. A victory, an expected attack upon
the city, a momentary fit of depression or exultation, a jest thrown
out against one of his books, a dream which for a short time
restored to him that beautiful face over which the grave had closed
for ever, led him to musings, which, without effort, shaped
themselves into verse. The unity of sentiment and severity of style
which characterise these little pieces remind us of the Greek
Anthology, or perhaps still more of the Collects of the English Liturgy.
The noble poem on the Massacres of Piedmont is strictly a Collect in
verse.
The Sonnets are more or less striking, according as the occasions
which gave birth to them are more or less interesting. But they are,
almost without exception, dignified by a sobriety and greatness of
mind to which we know not where to look for a parallel. It would,
indeed, be scarcely safe to draw any decided inferences as to the
character of a writer from passages directly egotistical. But the
qualities which we have ascribed to Milton, though perhaps most
strongly marked in those parts of his works which treat of his
personal feelings, are distinguishable in every page, and impart to all
his writings, prose and poetry, English, Latin, and Italian, a strong
family likeness.
His public conduct was such as was to be expected from a man of
a spirit so high and of an intellect so powerful. He lived at one of the
most memorable eras in the history of mankind, at the very crisis of
the great conflict between Oromasdes and Arimanes, liberty and
despotism, reason and prejudice. That great battle was fought for no
single generation, for no single land. The destinies of the human
race were staked on the same cast with the freedom of the English
people. Then were first proclaimed those mighty principles which
have since worked their way into the depths of the American forests,
which have roused Greece from the slavery and degradation of two
thousand years, and which, from one end of Europe to the other,
have kindled an unquenchable fire in the hearts of the oppressed,
and loosed the knees of the oppressors with an unwonted fear.
Of those principles, then struggling for their infant existence,
Milton was the most devoted and eloquent literary champion. We
need not say how much we admire his public conduct. But we
cannot disguise from ourselves that a large portion of his
countrymen still think it unjustifiable. The civil war, indeed, has been
more discussed, and is less understood, than any event in English
history. The friends of liberty laboured under the disadvantage of
which the lion in the fable complained so bitterly. Though they were
the conquerors, their enemies were the painters. As a body, the
Roundheads had done their utmost to decry and ruin literature; and
literature was even with them, as, in the long run, it always is with
its enemies. The best book on their side of the question is the
charming narrative of Mrs. Hutchinson. May’s History of the
Parliament is good; but it breaks off at the most interesting crisis of
the struggle. The performance of Ludlow is foolish and violent; and
most of the later writers who have espoused the same cause,
Oldmixon for instance, and Catherine Macaulay, have, to say the
least, been more distinguished by zeal than either by candour or by
skill. On the other side are the most authoritative and the most
popular historical works in our language, that of Clarendon, and that
of Hume. The former is not only ably written and full of valuable
information, but has also an air of dignity and sincerity which makes
even the prejudices and errors with which it abounds respectable.
Hume, from whose fascinating narrative the great mass of the
reading public are still contented to take their opinions, hated
religion so much that he hated liberty for having been allied with
religion, and has pleaded the cause of tyranny with the dexterity of
an advocate while affecting the impartiality of a judge.
The public conduct of Milton must be approved or condemned
according as the resistance of the people to Charles the First shall
appear to be justifiable or criminal. We shall therefore make no
apology for dedicating a few pages to the discussion of that
interesting and most important question. We shall not argue it on
general grounds. We shall not recur to those primary principles from
which the claim of any government to the obedience of its subjects
is to be deduced. We are entitled to that vantage ground; but we
will relinquish it. We are, on this point, so confident of superiority,
that we are not unwilling to imitate the ostentatious generosity of
those ancient knights, who vowed to joust without helmet or shield
against all enemies, and to give their antagonists the advantage of
sun and wind. We will take the naked constitutional question. We
confidently affirm, that every reason which can be urged in favour of
the Revolution of 1688 may be urged with at least equal force in
favour of what is called the Great Rebellion.
In one respect, only, we think, can the warmest admirers of
Charles venture to say that he was a better sovereign than his son.
He was not, in name and profession, a Papist; we say in name and
profession, because both Charles himself and his creature Laud,
while they abjured the innocent badges of Popery, retained all its
worst vices, a complete subjection of reason to authority, a weak
preference of form to substance, a childish passion for mummeries,
an idolatrous veneration for the priestly character, and, above all, a
merciless intolerance. This, however, we waive. We will concede that
Charles was a good Protestant; but we say that his Protestantism
does not make the slightest distinction between his case and that of
James.
The principles of the Revolution have often been grossly
misrepresented, and never more than in the course of the present
year. There is a certain class of men, who, while they profess to hold
in reverence the great names and great actions of former times
never look at them for any other purpose than in order to find in
them some excuse for existing abuses. In every venerable precedent
they pass by what is essential, and take only what is accidental: they
keep out of sight what is beneficial, and hold up to public imitation
all that is defective. If, in any part of any great example, there be
any thing unsound, these flesh-flies detect it with an unerring
instinct, and dart upon it with a ravenous delight. If some good end
has been attained in spite of them, they feel, with their prototype,
that

“Their labour must be to pervert that end,

And out of good still to find means of evil.”
 To the blessings which England has derived from the Revolution
these people are utterly insensible. The expulsion of a tyrant, the
solemn recognition of popular rights, liberty, security, toleration, all
go for nothing with them. One sect there was, which, from
unfortunate temporary causes, it was thought necessary to keep
under close restraint. One part of the empire there was so unhappily
circumstanced, that at that time its misery was necessary to our
happiness, and its slavery to our freedom. These are the parts of the
Revolution which the politicians of whom we speak, love to
contemplate, and which seem to them not indeed to vindicate, but in
some degree to palliate, the good which it has produced. Talk to
them of Naples, of Spain, or of South America. They stand forth
zealots for the doctrine of Divine Right which has now come back to
us, like a thief from transportation, under the alias of Legitimacy. But
mention the miseries of Ireland. Then William is a hero. Then
Somers and Shrewsbury are great men. Then the Revolution is a
glorious era. The very same persons who, in this country, never omit
an opportunity of reviving every wretched Jacobite slander
respecting the Whigs of that period, have no sooner crossed St.
George’s Channel, than they begin to fill their bumpers to the
glorious and immortal memory. They may truly boast that they look
not at men, but at measures. So that evil be done, they care not
who does it; the arbitrary Charles, or the liberal William, Ferdinand
the Catholic, or Frederic the Protestant. On such occasions their
deadliest opponents may reckon upon their candid construction. The
bold assertions of these people have of late impressed a large
portion of the public with an opinion that James the Second was
expelled simply because he was a Catholic, and that the Revolution
was essentially a Protestant Revolution.
But this certainly was not the case; nor can any person who has
acquired more knowledge of the history of those times than is to be
found in Goldsmith’s Abridgment believe that, if James had held his
own religious opinions without wishing to make proselytes, or if,
wishing even to make proselytes, he had contented himself with
exerting only his constitutional influence for that purpose, the Prince
of Orange would ever have been invited over. Our ancestors, we
suppose, knew their own meaning; and, if we may believe them,
their hostility was primarily not to popery, but to tyranny. They did
not drive out a tyrant because he was a Catholic; but they excluded
Catholics from the crown, because they thought them likely to be
tyrants. The ground on which they, in their famous resolution,
declared the throne vacant, was this, “that James had broken the
fundamental laws of the kingdom.” Every man, therefore, who
approves of the Revolution of 1688 must hold that the breach of
fundamental laws on the part of the sovereign justifies resistance.
The question, then, is this; Had Charles the First broken the
fundamental laws of England?
No person can answer in the negative, unless he refuses credit,
not merely to all the accusations brought against Charles by his
opponents, but to the narratives of the warmest Royalists, and to
the confessions of the King himself. If there be any truth in any
historian of any party who has related the events of that reign, the
conduct of Charles, from his accession to the meeting of the Long
Parliament, had been a continued course of oppression and
treachery. Let those who applaud the Revolution, and condemn the
Rebellion, mention one act of James the Second to which a parallel
is not to be found in the history of his father. Let them lay their
fingers on a single article in the Declaration of Right, presented by
the two Houses to William and Mary, which Charles is not
acknowledged to have violated. He had, according to the testimony
of his own friends, usurped the functions of the legislature, raised
taxes without the consent of parliament, and quartered troops on
the people in the most illegal and vexatious manner. Not a single
session of parliament had passed without some unconstitutional
attack on the freedom of debate; the right of petition was grossly
violated; arbitrary judgments, exorbitant fines, and unwarranted
imprisonments, were grievances of daily occurrence. If these things
do not justify resistance, the Revolution was treason; if they do, the
Great Rebellion was laudable.
 But, it is said, why not adopt milder measures? Why, after the
King had consented to so many reforms, and renounced so many
oppressive prerogatives, did the parliament continue to rise in their
demands at the risk of provoking a civil war? The ship-money had
been given up. The Star Chamber had been abolished. Provision had
been made for the frequent, convocation and secure deliberation of
parliaments. Why not pursue an end confessedly good by peaceable
and regular means? We recur again to the analogy of the Revolution.
Why was James driven from the throne? Why was he not retained
upon conditions? He too had offered to call a free parliament and to
submit to its decision all the matters in dispute. Yet we are in the
habit of praising our forefathers, who preferred a revolution, a
disputed succession, a dynasty of strangers, twenty years of foreign
and intestine war, a standing army, and a national debt, to the rule,
however restricted, of a tried and proved tyrant. The Long
Parliament acted on the same principle, and is entitled to the same
praise. They could not trust the King. He had no doubt passed
salutary laws; but what assurance was there that he would not
break them? He had renounced oppressive prerogatives; but where
was the security that he would not resume them? The nation had to
deal with a man whom no tie could bind, a man who made and
broke promises with equal facility, a man whose honour had been a
hundred times pawned, and never redeemed.
Here, indeed, the Long Parliament stands on still stronger ground
than the Convention of 1688. No action of James can be compared
to the conduct of Charles with respect to the Petition of Right. The
Lords and Commons present him with a bill in which the
constitutional limits of his power are marked out. He hesitates; he
evades; at last he bargains to give his assent for five subsidies. The
bill receives his solemn assent; the subsidies are voted; but no
sooner is the tyrant relieved, than he returns at once to all the
arbitrary measures which he had bound himself to abandon, and
violates all the clauses of the very Act which he had been paid to
pass.
 For more than ten years the people had seen the rights which
were theirs by a double claim, by immemorial inheritance and by
recent purchase, infringed by the perfidious king who had
recognised them. At length circumstances compelled Charles to
summon another parliament: another chance was given to our
fathers: were they to throw it away as they had thrown away the
former? Were they again to be cozened by le Roi le vent? Were they
again to advance their money on pledges which had been forfeited
over and over again? Were they to lay a second Petition of Right at
the foot of the throne, to grant another lavish aid in exchange for
another unmeaning ceremony, and then to take their departure, till,
after ten years more of fraud and oppression, their prince should
again require a supply, and again repay it with a perjury? They were
compelled to choose whether they would trust a tyrant or conquer
him. We think that they chose wisely and nobly.
The advocates of Charles, like the advocates of other malefactors
against whom overwhelming evidence is produced, generally decline
all controversy about the facts, and content themselves with calling
testimony to character. He had so many private virtues! And had
James the Second no private virtues? Was Oliver Cromwell, his
bitterest enemies themselves being judges, destitute of private
virtues? And what, after all, are the virtues ascribed to Charles? A
religious zeal, not more sincere than that of his son, and fully as
weak and narrow-minded, and a few of the ordinary household
decencies which half the tombstones in England claim for those who
lie beneath them. A good father! A good husband! Ample apologies
indeed for fifteen years of persecution, tyranny and falsehood!
We charge him with having broken his coronation oath; and we
are told that he kept his marriage vow! We accuse him of having
given up his people to the merciless inflictions of the most hot-
headed and hard-hearted of prelates; and the defence is, that he
took his little son on his knee and kissed him! We censure him for
having violated the articles of the Petition of Right, after having, for
good and valuable consideration, promised to observe them; and we
are informed that he was accustomed to hear prayers at six o’clock
in the morning! It is to such considerations as these, together with
his Vandyke dress, his handsome face, and his peaked beard, that
he owes, we verily believe, most of his popularity with the present
generation.
For ourselves, we own that we do not understand the common
phrase, a good man, but a bad king. We can as easily conceive a
good man and an unnatural father, or a good man and a treacherous
friend. We cannot, in estimating the character of an individual, leave
out of our consideration his conduct in the most important of all
human relations; and if in that relation we find him to have been
selfish, cruel, and deceitful, we shall take the liberty to call him a
bad man, in spite of all his temperance at table, and all his regularity
at chapel.
We cannot refrain from adding a few words respecting a topic on
which the defenders of Charles are fond of dwelling. If, they say, he
governed his people ill, he at least governed them after the example
of his predecessors. If he violated their privileges, it was because
those privileges had not been accurately defined. No act of
oppression has ever been imputed to him which has not a parallel in
the annals of the Tudors. This point Hume has laboured, with an art
which is as discreditable in a historical work as it would be admirable
in a forensic address. The answer is short, clear, and decisive.
Charles had assented to the Petition of Right. He had renounced the
oppressive powers said to have been exercised by his predecessors,
and he had renounced them for money He was not entitled to set up
his antiquated claims against his own recent release.
These arguments are so obvious, that it may seem superfluous to
dwell upon them. But those who have observed how much the
events of that time are misrepresented and misunderstood will not
blame us for stating the case simply. It is a case of which the
simplest statement is the strongest.
 The enemies of the Parliament, indeed, rarely choose to take issue
on the great points of the question. They content themselves with
exposing some of the crimes and follies to which public commotions
necessarily give birth. They bewail the unmerited fate of Stafford.
They execrate the lawless violence of the army. They laugh at the
Scripitural names of the preachers. Major-generals fleecing their
districts; soldiers revelling on the spoils of a ruined peasantry;
upstarts, enriched by the public plunder, taking possession of the
hospitable firesides and hereditary trees of the old gentry; boys
smashing the beautiful windows of cathedrals; Quakers riding naked
through the market-place; Fifth-monarchy men shouting for King
Jesus; agitators lecturing from the tops of tubs on the fate of Agag;
all these, they tell us, were the offspring of the Great Rebellion.
Be it so. We are not careful to answer in this matter. These
charges, were they infinitely more important, would not alter our
opinion of an event which alone has made us to differ from the
slaves who crouch beneath despotic sceptres. Many evils, no doubt,
were produced by the civil war. They were the price of our liberty.
Has the acquisition been worth the sacrifice? It is the nature of the
Devil of tyranny to tear and rend the body which he leaves. Are the
miseries of continued possession less horrible than the struggles of
the tremendous exorcism?
If it were possible that a people brought up under an intolerant
and arbitrary system could subvert that system without acts of
cruelty and folly, half the objections to despotic power would be
removed. We should, in that case, be compelled to acknowledge that
it at least produces no pernicious effects on the intellectual and
moral character of a nation. We deplore the outrages which
accompany revolutions. But the more violent the outrages, the more
assured we feel that a revolution was necessary. The violence of
those outrages will always be proportioned to the ferocity and
ignorance of the people; and the ferocity and ignorance of the
people will be proportioned to the oppression and degradation under
which they have been accustomed to live. Thus it was in our civil
war. The heads of the church and state reaped only that which they
had sown. The government had prohibited free discussion: it had
done its best to keep the people unacquainted with their duties and
their rights. The retribution was just and natural. If our rulers
suffered from popular ignorance, it was because they had
themselves taken away the key of knowledge. If they were assailed
with blind fury, it was because they had exacted an equally blind
submission.
It is the character of such revolutions that we always see the
worst of them at first. Till men have been some time free, they know
not how to use their freedom. The natives of wine countries are
generally sober. In climates where wine is a rarity intemperance
abounds. A newly liberated people may be compared to a northern
army encamped on the Rhine or the Xeres. It is said that, when
soldiers in such a situation first find themselves able to indulge
without restraint in such a rare and expensive luxury, nothing is to
be seen but intoxication. Soon, however, plenty teaches discretion;
and, after wine has been for a few months their daily fare, they
become more temperate than they had ever been in their own
country. In the same manner, the final and permanent fruits of
liberty are wisdom, moderation, and mercy. Its immediate effects are
often atrocious crimes, conflicting errors, scepticism on points the
most clear, dogmatism on points the most mysterious. It is just at
this crisis that its enemies love to exhibit it. They pull down the
scaffolding from the half-finished edifice: they point to the flying
dust, the falling bricks, the comfortless rooms, the frightful
irregularity of the whole appearance; and then ask in scorn where
the promised splendour and comfort is to be found. If such
miserable sophisms were to prevail there would never be a good
house or a good government in the world. Ariosto tells a pretty story
of a fairy, who, by some mysterious law of her nature, was
condemned to appear at certain seasons in the form of a foul and
poisonous snake. Those who injured her during the period of her
disguise were for ever excluded from participation in the blessings
which she bestowed. But to those who, in spite of her loathsome
aspect, pitied and protected her, she afterwards revealed herself in
the beautiful and celestial form which was natural to her,
accompanied their steps, granted all their wishes, filled their houses
with wealth, made them happy in love and victorious in war. Such a
spirit is Liberty. At times she takes the form of a hateful reptile. She
grovels, she hisses, she stings. But woe to those who in disgust shall
venture to crush her! And happy are those who, having dared to
receive her in her degraded and frightful shape, shall at length be
rewarded by her in the time of her beauty and her glory!
There is only one cure for the evils which newly acquired freedom
produces; and that cure is freedom. When a prisoner first leaves his
cell he cannot bear the light of day: he is unable to discriminate
colours, or recognise faces. But the remedy is, not to remand him
into his dungeon, but to accustom him to the rays of the sun. The
blaze of truth and liberty may at first dazzle and bewilder nations
which have become half blind in the house of bondage. But let them
gaze on, and they will soon be able to bear it. In a few years men
learn to reason. The extreme violence of opinions subsides. Hostile
theories correct each other. The scattered elements of truth cease to
contend, and begin to coalesce. And at length a system of justice
and order is educed out of the chaos.
Many politicians of our time are in the habit of laying it down as a
self-evident proposition, that no people ought to be free till they are
fit to use their freedom? The maxim is worthy of the fool in the old
story who resolved not to go into the water till he had learnt to
swim. If men are to wait for liberty till they become wise and good
in slavery, they may indeed wait for ever.
Therefore it is that we decidedly approve of the conduct of Milton
and the other wise and good men who, in spite of much that was
ridiculous and hateful in the conduct of their associates, stood firmly
by the cause of Public Liberty. We are not aware that the poet has
been charged with personal participation in any of the blameable
excesses of that time. The favourite topic of his enemies is the line
of conduct which he pursued with regard to the execution of the
King. Of that celebrated proceeding we by no means approve. Still
we must say, in justice to the many eminent persons who concurred
in it, and in justice more particularly to the eminent person who
defended it, that nothing can be more absurd than the imputations
which, for the last hundred and sixty years, it has been the fashion
to cast upon the Regicides. We have, throughout, abstained from
appealing to first principles. We will not appeal to them now. We
recur again to the parallel case of the Revolution. What essential
distinction can be drawn between the execution of the father and
the deposition of the son? What constitutional maxim is there which
applies to the former and not to the latter? The King can do no
wrong. If so, James was as innocent as Charles could have been.
The minister only ought to be responsible for the acts of the
Sovereign. If so, why not impeach Jefferies and retain James? The
person of a King is sacred. Was the person of James considered
sacred at the Boyne? To discharge cannon against an army in which
a King is known to be posted is to approach pretty near to regicide.
Charles, too, it should always be remembered, was put to death by
men who had been exasperated by the hostilities of several years,
and who had never been bound to him by any other tie than that
which was common to them with all their fellow-citizens. Those who
drove James from his throne, who seduced his army, who alienated
his friends, who first imprisoned him in his palace, and then turned
him out of it, who broke in upon his very slumbers by imperious
messages, who pursued him with fire and sword from one part of
the empire to another, who hanged, drew, and quartered his
adherents, and attainted his innocent heir, were his nephew and his
two daughters. When we reflect on all these things, we are at a loss
to conceive how the same persons who, on the fifth of November,
thank God for wonderfully conducting his servant William, and for
making all opposition fall before him until he became our King and
Governor, can, on the thirtieth of January, contrive to be afraid that
the blood of the Royal Martyr may be visited on themselves and their
children.
 We disapprove, we repeat, of the execution of Charles; not
because the constitution exempts the King from responsibility, for we
know that all such maxims, however excellent, have their
exceptions; nor because we feel any peculiar interest in his
character, for we think that his sentence describes him with perfect
justice as “a tyrant, a traitor, a murderer, and a public enemy;” but
because we are convinced that the measure was most injurious to
the cause of freedom, He whom it removed was a captive and a
hostage: his heir, to whom the allegiance of every Royalist was
instantly transferred, was at large. The Presbyterians could never
have been perfectly reconciled to the father: they had no such
rooted enmity to the son. The great body of the people, also,
contemplated that proceeding with feelings which, however
unreasonable, no government could safely venture to outrage.
But though we think the conduct of the Regicides blameable, that
of Milton appears to us in a very different light. The deed was done.
It could not be undone. The evil was incurred; and the object was to
render it as small as possible. We censure the chiefs of the army for
not yielding to the popular opinion; but we cannot censure Milton for
wishing to change that opinion. The very feeling which would have
restrained us from committing the act would have led us, after it had
been committed, to defend it against the ravings of servility and
superstition. For the sake of public liberty, we wish that the thing
had not been done, while the people disapproved of it. But, for the
sake of public liberty, we should also have wished the people to
approve of it when it was done. If any thing more were wanting to
the justification of Milton, the book of Salmasius would furnish it.
That miserable performance is now with justice considered only as a
beacon to word-catchers, who wish to become statesmen. The
celebrity of the man who refuted it, the “Æneæ magni dextra,” gives
it all its fame with the present generation. In that age the state of
things was different. It was not then fully understood how vast an
interval separates the mere classical scholar from the political
philosopher. Nor can it be doubted that a treatise which, bearing the
name of so eminent a critic, attacked the fundamental principles of
all free governments, must, if suffered to remain unanswered, have
produced a most pernicious effect on the public mind.
We wish to add a few words relative to another subject, on which
the enemies of Milton delight to dwell, his conduct during the
administration of the Protector. That an enthusiastic votary of liberty
should accept office under a military usurper seems, no doubt, at
first sight, extraordinary. But all the circumstances in which the
country was then placed were extraordinary. The ambition of Oliver
was of no vulgar kind. He never seems to have coveted despotic
power. He at first fought sincerely and manfully for the Parliament,
and never deserted it, till it had deserted its duty. If he dissolved it
by force, it was not till he found that the few members who
remained after so many deaths, secessions, and expulsions, were
desirous to appropriate to themselves a power which they held only
in trust, and to inflict upon England the curse of a Venetian
oligarchy. But even when thus placed by violence at the head of
affairs, he did not assume unlimited power. He gave the country a
constitution far more perfect than any which had at that time been
known in the world. He reformed the representative system in a
manner which has extorted praise even from Lord Clarendon. For
himself he demanded indeed the first place in the commonwealth;
but with powers scarcely so great as those of a Dutch stadtholder, or
an American president. He gave the Parliament a voice in the
appointment of ministers, and left to it the whole legislative
authority, not even reserving; to himself a veto on its enactments;
and he did not require that the chief magistracy should be hereditary
in his family. Thus far, we think, if the circumstances of the time and
the opportunities which he had of aggrandizing himself he fairly
considered, he will not lose by comparison with Washington or
Bolivar. Had his moderation been met by corresponding moderation,
there is no reason to think that he would have overstepped the line
which he had traced for himself. But when he found that his
parliaments questioned the authority under which they met, and that
he was in danger of being deprived of the restricted power which
was absolutely necessary to his personal safety, then, it must be
acknowledged, he adopted a more arbitrary policy.
Yet, though we believe that the intentions of Cromwell were at
first honest, though we believe that he was driven from the noble
course which he had marked out for himself by the almost irresistible
force of circumstances, though we admire, in common with all men
of all parties, the ability and energy of his splendid administration,
we are not pleading for arbitrary and lawless power, even in his
hands. We know that a good constitution is infinitely better than the
best despot. But we suspect, that at the time of which we speak, the
violence of religious and political enmities rendered a stable and
happy settlement next to impossible. The choice lay, not between
Cromwell and liberty, but between Cromwell and the Stuarts. That
Milton chose well, no man can doubt who fairly compares the events
of the protectorate with those of the thirty years which succeeded it,
the darkest and most disgraceful in the English annals. Cromwell
was evidently laying, though in an irregular manner, the foundations
of an admirable system. Never before had religious liberty and the
freedom of discussion been enjoyed in a greater degree. Never had
the national honour been better upheld abroad, or the seat of justice
better filled at home. And it was rarely that any opposition which
stopped short of open rebellion provoked the resentment of the
liberal and magnanimous usurper. The institutions which he had
established, as set down in the Instrument of Government, and the
Humble Petition and Advice, were excellent. His practice, it is true,
too often departed from the theory of these institutions. But, had he
lived a few years longer, it is probable that his institutions would
have survived him, and that his arbitrary practice would have died
with him. His power had not been consecrated by ancient
prejudices. It was upheld only by his great personal qualities. Little,
therefore, was to be dreaded from a second protector, unless he
were also a second Oliver Cromwell. The events which followed his
decease are the most complete vindication of those who exerted
themselves to uphold his authority. His death dissolved the whole
frame of society. The army rose against the parliament, the different
corps of the army against each other. Sect raved against sect. Party
plotted against party. The Presbyterians, in their eagerness to be
revenged on the Independents, sacrificed their own liberty, and
deserted all their old principles. Without casting one glance on the
past, or requiring one stipulation for the future, they threw down
their freedom at the feet of the most frivolous and heartless of
tyrants.
Then came those days, never to be recalled without a blush, the
days of servitude without loyalty and sensuality without love, of
dwarfish talents and gigantic vices, the paradise of cold hearts and
narrow minds, the golden age of the coward, the bigot, and the
slave. The King cringed to his rival that he might trample on his
people, sank into a viceroy of France, and pocketed, with
complacent infamy, her degrading insults, and her more degrading
gold. The caresses of harlots, and the jests of buffoons, regulated
the policy of the state. The government had just ability enough to
deceive, and just religion enough to persecute. The principles of
liberty were the scoff of every grinning courtier, and the Anathema
Maranatha of every fawning dean. In every high place, worship was
paid to Charles and James, Belial and Moloch; and England
propitiated those obscene and cruel idols with the blood of her best
and bravest children. Crime succeeded to crime, and disgrace to
disgrace, till the race accursed of God and man was a second time
driven forth, to wander on the face of the earth, and to be a by-
word and a shaking of the head to the nations.
Most of the remarks which we have hitherto made on the public
character of Milton, apply to him only as one of a large body. We
shall proceed to notice some of the peculiarities which distinguished
him from his contemporaries. And, for that purpose, it is necessary
to take a short survey of the parties into which the political world
was at that time divided. We must premise, that our observations
are intended to apply only to those who adhered, from a sincere
preference, to one or to the other side. In days of public commotion,
every fiction, like an Oriental army, is attended by a crowd of camp-
followers, an useless and heartless rabble, who prowl round its line
of march in the hope of picking up something under its protection,
but desert it in the day of battle, and often join to exterminate it
after a defeat. England, at the time of which we are treating,
abounded with fickle and selfish politicians, who transferred their
support to every government as it rose, who kissed the hand of the
King in 1640, and spat in his face in 1649, who shouted with equal
glee when Cromwell was inaugurated in Westminster Hall, and when
he was dug up to be hanged at Tyburn, who dined on calves’ heads,
or stuck up oak-branches, as circumstances altered, without the
slightest shame or repugnance. These we leave out of the account.
We take our estimate of parties from those who really deserve to be
called partisans.
We would speak first of the Puritans, the most remarkable body of
men, perhaps, which the world has ever produced. The odious and
ridiculous parts of their character lie on the surface. He that runs
may read them; nor have there been wanting attentive and
malicious observers to point them out. For many years after the
Restoration, they were the theme of unmeasured invective and
derision. They were exposed to the utmost licentiousness of the
press and of the stage, at the time when the press and the stage
were most licentious. They were not men of letters; they were, as a
body, unpopular; they could not defend themselves; and the public
would not take them under its protection. They were therefore
abandoned, without reserve, to the tender mercies of the satirists
and dramatists. The ostentatious simplicity of their dress, their sour
aspect, their nasal twang, their stiff posture, their long graces, their
Hebrew names, the Scriptural phrases which they introduced on
every occasion, their contempt of human learning, their detestation
of polite amusements, were indeed fair game for the laughers. But it
is not from the laughers alone that the philosophy of history is to be
learnt. And he who approaches this subject should carefully guard
against the influence of that potent ridicule which has already misled
so many excellent writers.
 “Ecco il fonte del riso, ed ecco il rio
Che mortali perigli in se contiene:
Hor qui teuer a fren nostro desio,
Ed esser cauti niolto a noi conviene.”

Those who roused the people to resistance, who directed their

measures through a long series of eventful years, who formed, out
of the most unpromising materials, the finest army that Europe had
ever seen, who trampled down King, Church, and Aristocracy, who,
in the short intervals of domestic sedition and rebellion, made the
name of England terrible to every nation on the face of the earth,
were no vulgar fanatics. Most of their absurdities were mere external
badges, like the signs of freemasonry, or the dresses of friars. We
regret that these badges were not more attractive. We regret that a
body to whose courage and talents mankind has owed inestimable
obligations had not the lofty elegance which distinguished some of
the adherents of Charles the First, or the easy good-breeding for
which the court of Charles the Second was celebrated. But, if we
must make our choice, we shall, like Bassanio in the play, turn from
the specious caskets which contain only the Death’s head and the
Fool’s head, and fix on the plain leaden chest which conceals the
treasure.
The Puritans were men whose minds had derived a peculiar
character from the daily contemplation of superior beings and
eternal interests. Not content with acknowledging, in general terms,
an overruling Providence, they habitually ascribed every event to the
will of the Great Being, for whose power nothing was too vast, for
whose inspection nothing was too minute. To know him, to serve
him, to enjoy him, was with them the great end of existence. They
rejected with contempt the ceremonious homage which other sects
substituted for the pure worship of the soul. Instead of catching
occasional glimpses of the Deity through an obscuring veil, they
aspired to gaze full on his intolerable brightness, and to commune
with him face to face. Hence originated their contempt for terrestrial
distinctions. The difference between the greatest and the meanest of
mankind seemed to vanish, when compared with the boundless
interval which separated the whole race from him on whom their
own eyes were constantly fixed. They recognised no title to
superiority but his favour; and, confident of that favour, they
despised all the accomplishments and all the dignities of the world.
If they were unacquainted with the works of philosophers and poets,
they were deeply read in the oracles of God. If their names were not
found in the registers of heralds, they were recorded in the Book of
Life. If their steps were not accompanied by a splendid train of
menials, legions of ministering angels had charge over them. Their
palaces were houses not made with hands; their diadems crowns of
glory which should never fade away. On the rich and the eloquent,
on nobles and priests they looked down with contempt: for they
esteemed themselves rich in a more precious treasure, and eloquent
in a more sublime language, nobles by the right of an earlier
creation, and priests by the imposition of a mightier hand. The very
meanest of them was a being to whose fate a mysterious and
terrible importance belonged, on whose slightest action the spirits of
light and darkness looked with anxious interest, who had been
destined, before heaven and earth were created, to enjoy a felicity
which should continue when heaven and earth should have passed
away. Events which short-sighted politicians ascribed to earthly
causes, had been ordained on his account. For his sake empires had
risen, and flourished, and decayed. For his sake the Almighty had
proclaimed his will by the pen of the Evangelist, and the harp of the
prophet. He had been wrested by no common deliverer from the
grasp of no common foe. He had been ransomed by the sweat of no
vulgar agony, by the blood of no earthly sacrifice. It was for him that
the sun had been darkened, that the rocks had been rent, that the
dead had risen, that all nature had shuddered at the sufferings of
her expiring God.
Thus the Puritan was made up of two different men, the one all
self-abasement, penitence, gratitude, passion, the other proud,
calm, inflexible, sagacious. He prostrated himself in the dust before
his Maker: but he set his foot on the neck of his king. In his
devotional retirement, he prayed with convulsions, and groans, and
tears. He was half-maddened by glorious or terrible illusions. He
heard the lyres of angels or the tempting whispers of fiends. He
caught a gleam of the Beatific Vision, or woke screaming from
dreams of everlasting fire. Like Vane, he thought himself intrusted
with the sceptre of the millennial year. Like Fleetwood, he cried in
the bitterness of his soul that God had hid his face from him. But
when he took his seat in the council, or girt on his sword for war,
these impestuous workings of the soul had left no perceptible trace
behind them. People who saw nothing of the godly but their uncouth
visages, and heard nothing from them but their groans and their
whining hymns, yet laugh at them. But those had little reason to
laugh who encountered them in the hall of debate or in the field of
battle. These fanatics brought to civil and military affairs a coolness
of judgment and an immutability of purpose which some writers
have thought inconsistent with their religious zeal, but which were in
fact the necessary effects of it. The intensity of their feelings on one
subject made them tranquil on every other. One overpowering
sentiment had subjected to itself pity and hatred, ambition and fear.
Death had lost its terrors and pleasure its charms. They had their
smiles and their tears, their raptures and their sorrows, but not for
the things of this world. Enthusiasm had made them Stoics, had
cleared their minds from every vulgar passion and prejudice, and
raised them above the influence of danger and of corruption. It
sometimes might lead them to pursue unwise ends, but never to
choose unwise means. They went through the world, like Sir
Artegal’s iron man Talus with his flail, crushing and trampling down
oppressors, mingling with human beings, but having neither part nor
lot in human infirmities, insensible to fatigue, to pleasure, and to
pain, not to be pierced by any weapon, not to be withstood by any
barrier.
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knowledge seekers. We believe that every book holds a new world,
offering opportunities for learning, discovery, and personal growth.
That’s why we are dedicated to bringing you a diverse collection of
books, ranging from classic literature and specialized publications to
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More than just a book-buying platform, we strive to be a bridge

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Join us on a journey of knowledge exploration, passion nurturing, and

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ebookbell.com