INTRODUCTION

VELP E-BOOK
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BARLEY AND FEED

N/Protein Determination and
Fiber Determination

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TABLE OF CONTENTS

INTRODUCTION ............................................................... 3
BARLEY AND FEED: A STRONG BOND ................................................... 4

MATERIALS, METHODS & RESULTS ............................ 7

PROTEIN DETERMINATION ..................................................................... 8
DUMAS METHOD .................................................................................................. 9
KJELDAHL METHOD ........................................................................................... 10
N/PROTEIN RESULTS ........................................................................................ 11
FIBER DETERMINATION ......................................................................... 12
FIBER DETERMINATION METHODS ................................................................. 13
FIBER RESULTS ................................................................................................. 15

CONCLUSIONS............................................................... 19
PROTEIN ANALYSIS ............................................................................... 20
FIBER ANALYSIS ..................................................................................... 21

VELP SOLUTIONS .......................................................... 22

VELP ELEMENTAL ANALYZERS ............................................................ 23
VELP KJELDAHL ANALYZERS................................................................ 24
VELP FIBER ANALYZERS ....................................................................... 25
INTRODUCTION

INTRODUCTION
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BARLEY AND FEED: A

STRONG BOND

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INTRODUCTION

BARLEY AND FEED: A STRONG BOND

Barley ranks fourth in importance among cereals, after wheat, corn and rice.
It represents two-thirds of the fodder grains demanded by the world and most
of it is used to feed livestock, with industrial consumption remaining
practically stable.

Barley is considered a medium-energy grain, low in starch and high in fiber.

Regarding protein levels, barley is similar to wheat and superior to corn, the
level of this nutrient can vary between 9% and 12-13%. It is an excellent
source of some B vitamins (thiamine, riboflavin, pyridoxine, pantothenic acid)
and niacin. Barley has a high fiber content, higher than that of corn and
wheat, which results in a lower nutritional value for species sensitive to fiber
content. Barley is one of the grains most commonly used in feed for dairy
cows and fattening cattle. Due to its high ruminal digestibility, barley has
high metabolizable energy values for ruminants.

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INTRODUCTION

BARLEY – OVERVIEW AND FIGURES

▪ High ruminal digestibility resulting in high

metabolizable energy values for ruminants.
▪ Most commonly used in feed for dairy cows and
fattening cattle.

Proteins
The level of proteins can vary between 9% and 12-13%.

Fiber
High fiber content, the level of Crude Fiber is around 4% and the level of
Neutral Detergent Fiber is around 16%.

Vitamins
Excellent source of B vitamins and niacin.

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INTRODUCTION

Feed represents 70% of the costs of a livestock operation, therefore,

knowledge of different nutritional alternatives (alternative cereals) is an
essential tool to maintain the profitability of the system. Feed is made up of
many components, which are fats, oils, carbohydrates, protein, vitamins,
minerals and product quality enhancement. In order to obtain a nutritionally
balanced diet it is important to know in which amount these elements should
be included in the diet, and in every single component of the feed, as for
example, barley.

Within this scenario, Proficiency Testing Programs organized by

institutions like BIPEA allow the comparison of several laboratories’
results, in order to evaluate their analytical performance on the same
homogeneous sample. By taking part in such programs on a regular basis,
the performance of VELP instruments is constantly evaluated. Samples
provided by BIPEA are analyzed in accordance with the official reference
ISO and AOAC and the obtained results are compared with the given
tolerance range.

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MATERIALS, METHODS & RESULTS

MATERIALS, METHODS & RESULTS

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PROTEIN AND FIBER

ANALYSIS: EVIDENCE
APPLYING DIFFERENT
METHODS

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MATERIALS, METHODS & RESULTS

PROTEIN DETERMINATION
To determine the quantity of proteins it is possible to analyze the sample
according to the Dumas method and the Kjeldahl method. Both are
primary methods working in accordance to international standards such as
AOAC, AACC, ASBC, ISO, IFFO, OIV.

The Dumas method for nitrogen determination, developed in 1831, is

older than the Kjeldahl, 1883, but more convenient in many aspects such
as speed, safety, cleanliness, productivity and cost per analysis.

The problem in the past was that it was not easy to reproduce the
conditions required by the Dumas method and for this reason, the Kjeldahl
technique took the lead and became considered as the classical method
for nitrogen/protein determination.

Nowadays, thanks to steps forward in technology, the Dumas nitrogen

determination is becoming more widespread.

Results obtained with the Dumas nitrogen determination are usually a

little bit higher than with Kjeldahl since nitrogen compounds like nitrates,
nitrites and heterocyclic compounds are detected.

In the Kjeldahl method, such compounds are converted into the

ammonium ion incompletely or not at all. The opposite could also happen
(rarely) because in this kind of analysis there are lots of variables that
could influence the final result.

Indeed, there are many minor variants of the Kjeldahl method, involving
the use of different catalysts, heating times, volumes and distribution of
sulfuric acid and masses of test portion: this shows that the Kjeldahl

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MATERIALS, METHODS & RESULTS

procedure may be influenced by experimental errors. Recovery is the

same for both the methods (≥ 99.5%), while the detection limit is lower for
Dumas than for Kjeldahl (0.001 mg N absolute vs. ≥ 0.1 mg N absolute).

DUMAS METHOD
The Dumas method starts with a combustion furnace (CF) to burn the
sample, obtaining elemental
compounds. Water is removed by a
first physical trap (WT1) and a
second chemical one (WT2).
Between the two, the elemental
substances pass through a
reduction furnace (RF). The auto-
regenerative CO2 absorbers let only
the elemental nitrogen pass, which
is detected by Thermal Conductivity
Detector (TCD).

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MATERIALS, METHODS & RESULTS

KJELDAHL METHOD
The Kjeldahl method consists in a procedure of catalytically supported
mineralization of organic material in a boiling mixture of sulfuric acid and
sulfate salt at digestion temperatures higher than 400 °C. During the
process the organically bonded nitrogen is converted into ammonium
sulfate. Alkalizing the digested solution liberates ammonia which is
quantitatively steam distilled and determined by titration.

• Homogenization
• Weighing
SAMPLE • Sulphuric Acid and KjTabsTM Catalyst addition
PREPARATION

• Digestion system configuration with fumes neutralization system

• Method setting (time and temperature ramps)
• Digestion at 420 °C
DIGESTION • Cooling

• VreceiverTM boric acid preparation and addition to the receiving flask

• Tube positioning with the digested sample
• Distiller method setting
• Addition of H2O - NaOH - VreceiverTM (different automation level available)
DISTILLATION
• Removal of residues (diffierent automation level available)

• Manual or automatic
• Colorimetric titration according to AOAC included in atomatic distillers
• External potentiometric titration available for selected models (electrode
TITRATION requires continuose calibration)

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MATERIALS, METHODS & RESULTS

N/PROTEIN RESULTS
Materials:

▪ NDA 702 – N/Protein Elemental Analyzer

▪ DKL 20 – Digestion Unit
▪ JP Pump and SMS Scrubber for fumes neutralization
▪ UDK 159 – Distillation Unit

Table 1 – Kjeldahl and Dumas Nitrogen and Protein results

Sample quantity Nitrogen % Protein %

Technique (mg)
199.97 1.319 8.244

199.84 1.327 8.294

Dumas
199.89 1.315 8.219

Average ± SD% 1.320 ± 0.006 8.252 ± 0.038

1.0030 g 1.264% 7.900%

1.0011 g 1.287% 8.044%

Kjeldahl
1.0048 g 1.282% 8.012%

Average ± SD% 1.278 ± 0.010 7.985 ± 0.075

BIPEA Protein Assigned Value - Dumas: 8.2 ± 0.2 % P

BIPEA Protein Assigned Value - Kjeldahl: 8.1 ± 0.2 % P

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MATERIALS, METHODS & RESULTS

FIBER DETERMINATION
Fiber is a component of the vegetable cell walls and as a primary function
of plant support. From a nutritional perspective we can define it as the
hydrolytically indigestible partially fermentable components of feed. A
chemical perspective defines fiber as a variable mixture of predominantly
cellulose, hemicelluloses, lignin, and soluble dietary fibers (e.g., pectins).

The demand for fiber content determination in feed is constantly

growing because it is a fundamental parameter for the assessment of
Feed's Quality.

The fiber amount of feed has impact on livestock productivity and the cost
of the feed. For example, the calculation of the correct amount of fiber in
the feedstuff influences milk production, milk quality and fat content of
milk.

“Because there is no guarantee of direct correspondence between

chemical solubility and nutritional availability, in reality, fiber is defined by
the method used to isolate it. The actual definition of fiber becomes
method-dependent”. (AAFCO - Critical Factors in Determining Fiber in
Feeds and Forages)

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MATERIALS, METHODS & RESULTS

This passage of an AAFCO document suggests that is important to follow

official method exactly in order to obtain reproducible results and that any
modification leads to incomparability of tests.

FIBER DETERMINATION METHODS

Crude Fiber, CF
The method is based on the solubilization (digestion) of non-cellulosic
compounds by sulfuric acid and potassium hydroxide solutions. Crude
fiber is the loss on ignition of the dried residue remaining after digestion
of the sample and is determined by weight difference.
This method is applicable to grains, meals, flours, feeds, and fiber-bearing
material from which fat can be extracted to leave workable residue.

Neutral Detergent Fiber, NDF

The sample is digested in the Neutral Detergent Solution NDS with heat-
stable α-amylase-treated enzyme to separate the neutral detergent
soluble fraction (sugars, starches and pectin soluble, filtered) from the
neutral detergent insoluble fraction (cell walls substances, hemicellulose,
cellulose and lignin, residues). The cell contents are highly digestible
(about 98 %) and include various sugars, starches, pectins and other
soluble carbohydrates, proteins, non-protein nitrogenous compounds,
lipids, water-soluble minerals and vitamins.
The remaining dry matter is estimated and the proportion gives the neutral
detergent fiber (NDF).

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MATERIALS, METHODS & RESULTS

Acid Detergent Fiber, ADF

The Acid detergent solution (ADS) solubilizes the hemicellulose while
lignin and cellulose remain insoluble. The residue is weighed for the
determination of ADF. It includes cellulose and lignin.

Acid Detergent Lignin, ADL

The remaining residue from the ADF analysis, is solubilized by 72%
sulfuric acid, leaving the lignin (ADL) which is determined gravimetrically.

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MATERIALS, METHODS & RESULTS

FIBER RESULTS
Materials:
▪ FIWE Advance – Fully Automatic Fiber Analyzer
▪ COEX – Cold Extractor

Crude Fiber calculation

To calculate this parameter, it is necessary to subtract the weight of the

crucible after the dry process minus the weight of the crucible after the
incineration in the muffle.

% Crude Fiber = (Mdry - Mash) *100/Msample

Mdry = sample weight after drying

Mash = sample weight after ashing

Msample = sample weight

Table 2 – Crude Fiber results

Msample (g) Mdry (g) Mash (g) CF %
1.0349 30.4385 30.394 4.1067

1.0511 31.221 31.1732 4.3573

1.0043 30.5511 30.5073 4.1621

Average ± SD% 4.21 ± 0.13

BIPEA Crude Fiber Assigned Value: 4.2 ± 0.4 %

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MATERIALS, METHODS & RESULTS

Neutral Detergent Fiber calculation

To calculate this parameter, it is necessary to subtract the weight of the

crucible after the time in the oven minus the weight of the crucible after the
incineration process in the muffle.

aNDF % = (Mdry - Mash - (Bdry - Bash)) *100/Msample

Mdry = sample weight after drying

Mash = sample weight after ashing

Msample = sample weight

Bdry = blank weight after drying

Bash = blank weight after ashing

Table 3 – Neutral Detergent Fiber results

Msample (g) Mdry (g) Mash (g) aNDF %
0.5009 30.101 30.0205 16.15%

0.5026 30.7516 30.669 16.43%

0.5068 31.0118 30.9294 16.26%

Average ± SD% 16.28 ± 0.39

BIPEA NDF Assigned Value: 16.7% ± 0.8%

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MATERIALS, METHODS & RESULTS

Acid Detergent Fiber calculation

To calculate this parameter, it is necessary to subtract the weight of the

crucible before the analysis minus the weight of the crucible after the
digestion end the dry process.

ADF % = (Mdry - Mtare - (Bdry - Btare)) *100/Msample

Mdry = sample weight after drying

Mtare = tare of the sample

Msample = sample weight

Bdry = blank weight after drying

Btare = tare of the blank

Table 4 – Acid Detergent Fiber results

Mtare (g) Msample (g) Mdry (g) ADF %
30.4512 1.0342 30.459 4.844%

30.992 0.9972 30.9987 4.8335

31.1974 1.0085 31.2067 4.9380

Average ± SD% 4.87 ± 0.06

BIPEA ADF Assigned Value: 5.0% ± 0.8%

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MATERIALS, METHODS & RESULTS

Acid Detergent Lignin calculation

To calculate this parameter, it is necessary to subtract the weight of the

crucible after the dry process minus the weight of the crucible after the
incineration process in the muffle.

ADL % = (Mdry - Mash - (Bdry - Bash)) *100/Msample

Mdry = sample weight after drying

Mash = sample weight after ashing

Msample = sample weight

Bdry = blank weight after drying

Bash = blank weight after ashing

Table 5 – Acid Detergent Lignin results

Msample (g) Mdry (g) Mash (g) ADL %
1.0342 30.459 30.4441 1.440

0.9972 30.9987 30.9836 1.5142

1.0085 31.2067 31.1924 1.4179

Average ± SD% 1.46 ± 0.05

BIPEA ADL Assigned Value: 1.2% ± 0.5%

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CONCLUSIONS

CONCLUSIONS
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RESULTS DISCUSSION

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CONCLUSIONS

The results obtained are reliable, reproducible in accordance with the

expected values of the proficiency testing program organized by
BIPEA.

It means that VELP instruments are able to determine accurate and

precise parameters for feed analysis in barley, in order to obtain a
complete diet for animals.

PROTEIN ANALYSIS
The obtained results for barley fell within the expected value range
indicated by BIPEA, demonstrating the high performances of VELP
Kjeldahl analytical instruments and VELP N/Protein Elemental
Analyzers, for protein determination. Both techniques are efficient and
capable of analyzing barley samples with high accuracy and
repeatability.

VELP N/Protein Elemental Analyzers with high productivity and non-stop

performances, are indeed ideal for high throughput, being fully automated
and requiring just 3-4 minutes per analysis.

VELP Kjeldahl system, is still a robust solution for protein determination

in food and feed field.

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CONCLUSIONS

FIBER ANALYSIS
The fiber results obtained are reliable and in accordance with the expected
values. The use of an extraction apparatus purposely devised for this
method as VELP Fiber Analyzer, makes the standardization of analytical
conditions very easy.

▪ Fast analysis (2 hours with VELP Fiber Analyzers vs. 6 hours manually)
▪ Ease of use with convenient filtration, with pump and air pressure
▪ High reproducibility of the results, ±1% relative or better
▪ Results in accordance with official procedures

It is a great advantage that a single company as VELP Scientifica, is able

to design, produce and support these three series of instruments to
determine the crucial feed parameters in barley.

Furthermore, VELP Scientifica internally produces the main

consumables to offer the most suitable turnkey solution for your
analyses guaranteeing a considerable advantage over the competition,
since all the instruments and consumables to optimize the performance of
the laboratory come from a single source.

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VELP DEDICATED SOLUTIONS

VELP SOLUTIONS
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ELEMENTAL ANALYZERS
KJELDAHL ANALYZERS
FIBER ANALYZERS

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VELP DEDICATED SOLUTIONS

VELP ELEMENTAL ANALYZERS

NDA 701 – Dumas Elemental Analyzer

▪ Nitrogen and Protein determination in

3-4 minutes
▪ Accurate and precise
▪ Versatile and cloud-enabled
▪ Helium as carrier gas

NDA 702 – Dumas Elemental Analyzer

▪ Nitrogen and Protein determination in

3-4 minutes
▪ Accurate and precise
▪ Unmatched LOD of 0.001 mgN (with
He as carrier gas)
▪ Versatile and cloud-enabled
▪ Helium and Argon as carrier gas

CN 802 – Elemental Analyzer

▪ Carbon, Nitrogen, C:N ratio, TOC and

TIC determination
▪ Fast analysis in 3-5 minutes
▪ Versatile and cloud-enabled
▪ Helium and Argon as carrier gas

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VELP DEDICATED SOLUTIONS

VELP KJELDAHL ANALYZERS

UDK Series – Distillation Units

▪ Full range of distillers with different

automation to match any lab requirement
▪ Exclusive patented titanium condenser
and steam generator to maximize
accuracy and efficiency
▪ Robust and chemical resistant

DKL Series – Automatic Kjeldahl Digesters

▪ Fully automatic digesters in 8, 12, 20 and

42 positions
▪ Stable and homogeneous temperature
(±0.5 °C)
▪ Fast, flexible and safe
▪ Robust and compact

DK Series – Kjeldahl Digesters

▪ Semi-automatic digester in 6, 8, 18, 20

and 42 positions
▪ Stable and homogeneous temperature
(±0.5 °C)
▪ Intuitive and easy-to-use

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VELP DEDICATED SOLUTIONS

VELP FIBER ANALYZERS

FIWE Advance – Automatic Fiber

Analyzer

▪ Crude and detergent fiber

determination to official standards
(ISO and AOAC)
▪ Fully automatic solution with reagent
loading and pre-set methods
▪ Safe, fast and cloud-enabled

FIWE – Fiber Analyzer

▪ Crude and detergent fiber

determination to official standards
(ISO and AOAC)
▪ Semi-automatic solution with timer
and acoustic signal
▪ High reproducibility

COEX – Cold Extractor

▪ Preliminary defatting of the sample

prior to fiber determination
▪ Reliable and easy-to-use

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VELP DEDICATED SOLUTIONS

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