whey protein

1. MONGOLIAN STATE UNIVERSITY OF AGRICULTURE
SCHOOL OF ANIMAL SCIENCE AND BIOTECHNOLOGY
Borkhuu Nomin-Erdene
THE RESEARCH OF WHEY PROTEIN
Supervisor’s: /N.Pagmadulam (Ph.D)/
/Du Ming( )/
Opponent’s /B.Tuyaagerel(Ph.D)/
Ulaanbaatar
2020

2. Topic: The research of whey protein
Introduction: Separate proteins from whey and determine its chemical
composition. The amino acid is determined by thin- layer chromatography
in the separated protein and the protein structure is determined by infrared
spectrophotometer.
Keywords: whey, protein, thin-layer chromatography, infrared spectrum, analysis

3. Title
Introduction
Rationale for the research
Novelty of the research:
The purpose of the research
CHAPTER1. PRESS REVIEW
1.1 Status of research
1.2 Composition and technological properties of whey
1.3 Whey protein
1.4 Significance of whey
1.5 Whey protein concentrate
1.6. Infrared spectroscopy (IRS)
1.7 Thin layer chromatographic method
CHAPTER 2. RESEARCH MATERIALS AND METHODOLOGY
2.1 Materials used in the study
2.2 Methodology for chemical analysis of whey
2.2.1 Determination of moisture by weight
2.2.2 Determination of ash by weight
2.2.3 Determination of proteins by the Kjeldahl method
2.2.4 Soxlet separation and determination of oils
2.2.5 Determination of dry matter by refractometry
2.3 Methods for testing whey protein
2.3.1 Separate whey protein and purify protein by dialysis
2.3.2 Chemical analysis of whey protein
2.3.3 Determination of total hydrocarbons by phenol-sulfuric acid reaction
2.3.4 Determination by Infrared Spectroscopy
2.3.5 Determination of protein amino acids by the thin layer
chromatography method
2.3.6 Determination of protein by biuret method

4. CHAPTER 3. RESEARCH RESULTS
3.1 Results of chemical analysis of whey
3.2 The amount of protein extracted from whey
3.3 Results of chemical analysis of whey protein
3.4 Determination of protein structure by infrared spectrophotometer
3.5 Amino acid determination by the AH method
3.6 Determination of protein by biuret method
Judicial proceedings
Conclusion
Gratitude
Appendix
References

5. Table list
Table1. Ingredients of whey dry matter… ………………………………................... 9
Table2. Composition and basic parameters of whey ............................................. 10
Table3. Whey protein and cream are ingredients ...................................... .. .... ... 12
Table 4. Whey protein amino acids ...... .... ................................... ........................ 12
Table 5. Amount of milk components ........................................... ……………....... 16
Table 6 . Directions for using whey protein concentrate ......................................... 16
Table.7 Results of chemical analysis of whey ......................................................... 35
Table.8 Protein separation from whey .......................................... .......................... 35
Table.9 Whey protein amino acids ................... ....................... ........ …………....... 39

6. List of figure
Figure 1. Molecular absorption energy diagram of a molecule ............................... 18
Figure 2. Spectrum of the substance . .................................................................... 19
Figure 3. “IK-14A” spectrometer block diagram ...................................................... 21
Figure 4 . General scheme of research work .............................................. .......... 23
Figure 5 . Kjeldahl method and distillation equipment ..................... ...................... 27
Figure 6 . Spectrogram of isolated whey protein NTP ............................................ 37
Figure 7 . Chromatogram of whey protein amino acids m .................................... 38
Figure 8. Vacuum evaporator ............................................. ..... ..................... .. .... 43
Figure 9. A freeze-dried whey ...................... ...... . .......... .. . . ........................... ... 43
Figure 10. dialysis protein isolates ........................................ .... .................... . . ..43
Figure 11. Kjeldal protein determination ....................... ..................... ...... . ... . . . 43
Figure 12. Determination of protein by biuret method ........................................... ...44
Figure 13. Sox years ............................................. ........ ......................................... 44
Figure.14 Determination of moisture ............................................. ............................44
Figure 15. Protein is determined by Kjeldahl .......................................................... 44

7. Introduction
There is a lot of research being done around the world on whey processing
technology, equipment design, biotechnology and nanotechnology. Today, whey is used
to make concentrates, dried concentrates, whey protein, and milk sugar, as well as in
the production of alcohol, bread, sugar, and feed. In addition, a special section is
separated from whey for use in pharmaceuticals and sanatoriums.
The whey is rich in lactic acid, enzymes and good quality protein. It is also a low-
calorie food that contains vitamins B6, B12, B2, calcium, potassium, phosphorus and
other amino acids. Whey has long been used by humans to improve appetite
and digestion. Modern Scientific studies show that these drinks in addition to preventing
high blood pressure, stroke, heart attack due to cholesterol increase balance, muscle
mass, and studied may prevent osteoporosis and improve the human body immune
system, reduces stress and adds energy
The technology of gel filtration of whey protein was first introduced in Goethe,
Sweden. A large-molecule solution obtained by fermenting sugar and altering its
structure is used as a filter to separate whey protein. When whey is passed through
such a solution, small amounts of molecules, such as lacto globulin and lacto albumin,
are introduced, which can separate whey protein from milk sugar and minerals. The
technology of making whey cheese and its products is widely used in
the Scandinavian countries. The whey is pre-thickened and then mixed with milk and
curds to produce a product with a unique taste and brown color. It has a technology of
constant stirring and rapid cooling.

8. Basis of research work
Our country is a nomadic country and has an ancient tradition of producing the widest
variety of sour milk products from its milk. According to the Ministry of Food and
Agriculture, the demand for dairy products in Mongolia in 2018 was 902.4 million liters,
of which 134.9 million liters of industrial processing produced 857 kg of whey waste
[1].
The production of curd, cheese, curds, food and technical casein produces large
amounts of whey, which is a valuable raw material containing 5-7% of dry matter, such
as lactoglobulin, lactalbumin, milk sugar, minerals and water-soluble vitamins. . Whey
contains high quality proteins that activate the immune system and have antioxidant
properties. Whey protein contains immune-boosting substances β-globulin α-
lactoalbumin, glycomacropeptide and immunoglobulins. They activate the immune
system and improve the body's resistance.
The issue of full and proper use of whey has not been resolved in the world, and it is
also a problem for our country. In Mongolia, whey is not used for health purposes
except for a small amount of animal feed. However, it is unfortunate that proteins that
are of special importance to the human body are not used for food.
One of the issues to be addressed in the dairy sector of our country is to process whey
on a scientific basis to separate products, create technology to use them in addition to
other food products, and increase production efficiency.

9. Novelty of the research:
The novelty of the research is that the protein, which is rich in minerals, proteins and
vitamins, is extracted from the waste and whey.
Purpose of the research: Structural analysis of whey protein separation.
Objective:
1. Determine the composition of whey
2. Distinguish whey protein
3. Determining the chemical composition of proteins
4. Compare with the results of standards and other researchers

10. CHAPTER1. PRESS REVIEW
1.1 Status of research
Technological methods used in many parts of the world are mainly used to extract
whey protein to increase the nutritional value of other foods and to use the remaining
sugar for other purposes.
Whey protein is 15-22% of all milk proteins, and 0.7-1.3% of total milk protein is serum
albumin, 7-12% β-globulin, 2-5% α-lactalbumin, and 1.9-3.3%. immunoglobulin and 2-
6% of proteozo-peptone. Casein-precipitated whey contains glycomacropeptides due to
the action of renin [2].
Goat milk yield, composition, properties, goat color and milk relationship were
studied. The study also compared the price of black and white goat's milk with this [3].
Research on the use of whey protein in food production. A comparison was
made between Mongolian cheese, curd, lemon, whey protein from industrial whey, and
other ingredients [4].
A study of the decomposition of gonads by the action of casein and whey protein
enzymes. The possibility of extracting whey protein and enzymatic action in the
production of food supplements containing biologically active peptides has
been studied [5].

11. 1.2 The technological characteristics of the composition of whey
One of the by-products of the dairy industry is whey. Whey contains 5-7% of dry matter,
including 50% of milk powder or finely dispersed milk fat, protein, nitrogenous
compounds, minerals, lactose, and various vitamins, especially water-soluble vitamins
B1, B2, C, enzymes, and trace elements. , is a valuable raw material containing organic
acids [6].
Whey was discovered 3,000 years ago as a result of carrying milk in a bag made of
dried calf's stomach. Under the influence of heat and the effect of ecchymosis, a
digestive enzyme in calves, milk is fermented to form whey and casein precipitates.
Over the centuries, these biological and chemical principles have been studied and
refined into art and science. Although the main component of whey is a by-product of
cheese production whey. The issue of full and proper use of whey has not been
resolved in the world, and it is a problem for our country as well. Therefore, it is
necessary to master the features, characteristics, uses and technology of whey
products. After the separation of milk fat and protein, a real solution or yolk remains. In
whey, lactose and water-soluble vitamins are formed in the form of molecules, while salt
is formed by electrolyte dissociation and ionic hydrate. The dry matter content can be
calculated from the average content of whey.
Table1. The composition of whey dry matter is shown below .
g / 100 ml %
Lactose 4.66 71.7
Protein substances 0.91 14.0
Minerals 0.50 7.7
Fat 0.37 5.7
Others 0.06 0.9
The bulk of whey dry matter is lactose (approximately 70 %) . Other ingredients (non-
sugar ) account for 30 % . Whey contains an average of 134 mg / ml of nitrogenous
compounds, of which 65 % are protein nitrogen compounds and 35 % are non-protein
compounds. The amount of protein nitrogenous compounds in whey is 0.7-1.1 % by
calculation. This amount is conditional because all nitrogen is included in the

12. protein. The nitrogen content of the true protein is 0.5-0.8 %, depending on the method
of precipitation of milk protein used to produce the main product.
Table2. Whey composition and basic parameters
Specifications
Types of whey Nutrient
conversion
rate,%
Cheese Cream Caseins
Dry matter,% 4.5-7.2 4.2-7.4 4.5-7.5 51.2
Carbohydrate 3.9-4.9 3.2-5.1 3.5-5.2 96.0
Protein 0.9-1.6 0.5-1.4 0.8-1.4 25.0
Mineral 0.3-0.8 0.5-0.8 0.3-0.9 71.5
Fat 0.2-0.5 0.05-0.4 0.02-0.1 5.6
Vitamin - - - 70.0
Calorific value, kJ - 101.3 - 36.1
Sour, º
T 15-25 50-85 50-120 -
Density, kg / m 3
1018-1027 1019-2016 1020-1026 -
According to the study of physical and chemical properties of whey:
Whey contains 25.5% of milk protein, 17.7% of fat and 95.78% of sugar. Whey proteins
differ from casein proteins in their precipitation. Whey protein does not decompose
under the action of acids and enzymes and precipitates on heating.

13. 1.3 Whey protein
The composition of whey varies depending on: For example, it depends on the
type of cheese and casein, the method of making curd, and the fat content of the
primary product. Conversion rate and soy milk, milk main component of this molecular
weight colloid p is defined as the quantity felt.
Casein-derived milk contains about 0.6% protein. This is called whey protein.
The multi-fractional protein is 15-22% of all milk proteins, and 0.7-1.3% of total milk
protein is serum albumin, 7-12% β-lacto globulin, 2-5% α-lacto albumin, 1.9-3.3% -
immunoglobulin and protease-peptone 2-6%. Casein-precipitated whey contains
glycomacropeptides due to the action of renin. α and β-immunoglobulin are essential
amino acids and sulfur-containing amino acids that are more nutritious than casein
proteins. Whey protein is small in size, but more biologically important than casein.
Whey protein can be separated at different pH values using MgSO4 or (NH4) 2SO4, -
lacto globulin and α-lacto albumin precipitate when 3% acetic acid is used. Whey
protein is separated for research purposes. In practice, all fractions of whey protein are
denatured together at 90-95 °C. Sugar is also extracted from whey. Although not
enough whey protein is used for food, it is more nutritious than casein due to its higher
content of essential amino acids and sulfur. The value of whey protein in terms of 100 is
70-80 casein. The body breaks down whey protein quickly and without residues. In the
pharmaceutical industry, protein is used as the main raw material for the production of
drugs. Whey protein differs from curd in its properties and composition. Whey protein is
rich in amino acids that regenerate muscle and is rapidly absorbed, making it more
important for active adults. Whey protein, which makes up about 20 percent of total milk
protein, plays a key role in lowering blood cholesterol and boosting the immune system.
The content of essential amino acids in whey protein is higher than in meat protein.
Whey protein supplements slow down the cancer process. Protein concentrate is
produced from whey and used for medical treatment and fitness [7].

14. Table3. Whey protein and curd composition
№ Components Ingredients,%
Whey protein Curds
1 Water 75-78 77-80
2 Dry matter 22-25 20-23
3 Protein 18.0 15.0
4 Lactose 2.0 3.0
5 Ash 1.2 0.5
6 Lactic acid 0.8 1.0
Under the action of temperature, organic polyelectrolytes are added to the denatured
whey protein, which quickly forms a coarse-grained structure. Whey protein is yellowish,
while curd is white.
Table 4. Whey protein amino acids,% [ 8 ]
Amino acids Immunoglobulin lactoalbumin lactoglobulin
Lucien 19.5 11.5 15.3
Isoleucine 6.2 6.8 8.4
Lysine 13.5 11.5 11.4
Threonine 20.6 5.5 5.8
Valine 19.8 4.7 5.8
Arginine 8.4 1.2 2.9
Aspartic acid - 18.7 11.4
Glycol
Tyrosine
- 3.2 1.4
Glutamic acid - 12.9 19.5
Histidine 4.0 2.9 1.6
Methionine 2.1 1.0 3.2
Phenylanine 7.4 4.5 3.5
Prolin - 1.5 4.1
Serin - 4.8 5.0
Tryptophan 5.1 7.0 1.9
Tyrosine - 5.4 3.8
-Lactalbumin : 18% whey protein, does not contain phosphorus in the molecular
composition , is not digested by gastric enzymes. Albumin is a bubble protein that
dissolves in water and dilute saline. This protein coagulates with heating. The second
type of milk protein is lacto albumin. Albumin is a bubble protein consisting of 575 amino
acid residues with the highest negative charge at the same electrical charge point of 4.7
and pH 8.6.

15. -Lactoglobulin : The molecule has a single thiol group, which is a natural protein inside
the molecule and is shielded. When heated, it undergoes semi-denaturation and the
thiol group is exposed to disulfide bonds, which combine with other milk proteins,
especially casein and lactalbumin. -lactoglobulin is a third type of milk protein. It has
immune-boosting properties. The relative molecular mass is 1800 kDa and
the isoelectric point is 5.35-5.5 [ 8 ] .
High-protein whey
Whey protein is made up of 20 essential amino acids and is rich in branched-chain
amino acids, which are involved in the formation of new muscle tissue. Whey protein, a
key component of an athlete's diet, is used by athletes and fitness athletes for a long
time to ensure the growth of dry muscle mass and to restore the body after strength
training and active exercise. Elderly people can prevent whey protein by consuming
whey protein. "Consumers are more likely to consume whey protein as a water-soluble
beverage," said Carrie O'Neill. Nutritional whey is a study that has improved nutritional
levels in food-deficient areas. Recent research has shown that malnourished children in
Malawi and Mozambique are given whey protein concentrates and permeates with
specially formulated supplements to compensate for their malnutrition and have a
positive effect on children's growth.
Whey from the production of cheese, casein and yogurt is an important source of
essential protein today. The world produced about 180 million tons of whey in 2013, of
which 1.5 million tons was valuable protein and 8.6 million tons was lactose, an
important source of carbohydrates. According to a recent study, whey protein is the
most nutritious protein and has led to unprecedented levels of investment in milk
production, including in the markets for athletes, sanatoriums and infant
nutrition. Despite the fact that a large part of the world's population lacks protein-rich
foods, 50 percent of the total whey produced that year is still discarded. From "natural
sweetness", consisting of soy milk -lactoglobulin, mother's milk protein content
equal -lacto albumin and lactoferrin, immune globulin, probiotic microbial growth
promoters galactooligosugar all necessary nutrients the human body is the main source
of substances [ 9] .

16. 1.4 The importance of whey
Certain properties of whey protein are interesting. They affect the quality of the
product in different technological ways. On the other hand, changes in whey protein
fraction can be inferred from milk processing. An important feature of the technology is
that whey protein is easily denatured by temperature. This has a strong effect on
product quality. Whey protein polypeptide chains have a high content of amino acids
and a high content of sulfur. When heated, the hydrogen and coil connections are
broken. The formation of new hydrogen bonds and disulfide bridges between whey
proteins causes denaturation by temperature. The amount of denaturation depends on
the temperature value and the service life. The most sensitive are immunoglobulins,
which are denatured at 60 degrees, followed by serum albumin and beta lactoglobulin,
and the most resistant to temperature are alpha-lactoglobulin. The protease-peptone
fraction begins to denature at -110° C [ 10 ] .
Whey contains high-quality protein, which stimulates the immune system and has
antioxidant properties. Whey protein contains substances that activate the immune
system - globulin, lactoalbumin, glycomacropeptide and immunoglobulins. They
activate the immune system and improve the body's resistance. During the processing
of dairy products, whey protein is easily denatured by temperature, which affects the
quality of the product. Whey protein is good for lean muscle growth, fat loss,
cardiovascular health and healthy metabolism. It also supports the immune system.
During the processing of protein products, a large amount of whey is released. It
improves liver and bile function, stimulates intestinal peristalsis and has a positive
effect on hematopoiesis. Elderly people can prevent whey protein by consuming whey
protein. Acidification of milk precipitates some of the whey protein with casein, reducing
the amount of protein left in the whey. When the milk was heated to 95-98 for
2.5 minutes, the nitrogen content of the whey was reduced by 25-30%. Under the action
of temperature, the ratio between -lactoalbumin -lactoglobulin changes due to the
formation of an intermolecular disulfide bridge. Changes in whey protein affect the
quality of dairy products. Dry milk is classified as high or low temperature due to the fact
that the solubility of denatured whey protein decreases due to temperature, and
precipitates with casein when salt is added. To distinguish between renin and acid-
derived whey, saline acid is determined from renin-derived whey. The content of
whey can be determined in a mixture of powdered and dried milk powder with the
content of saline acid . I used to be interested in the fat content of milk, but now I'm

17. interested in the protein content. There is a growing interest in low-fat and high-protein
milk. To do this, we try to include whey protein in the product as much as
possible. Soluble protein products are obtained by the osmotic method
called ultrafiltration [ 11 ] .
Table 5. The amount of milk components
Components Diameter, nm Molecular weight
Fat bubbles 100..10000 1012
.. 10 14
Casein micelles 10..300 107
..10 9
Whey protein 3..6 10 4
..10 5
Lactose ~ 1 342
Mineral ~ 0.4..1 20..200
Water molecules ~ 0.3 18
1nm = 10 9
m = 10 3
μm
1.5 Whey protein concentrate
The “whey protein concentrate” is produced by concentrating and drying the whey
protein through an ultra-fine filter. Based on the unique technological features of whey
protein concentrate, it is used to stabilize, whiten and strengthen food types. [ 12 ] .
Table 6. Directions for using whey protein concentrate
Properties of whey protein
concentrate
Directions for use
Biological and nutritional
value
In the production of dairy products similar to breast
milk, medical and health food for athletes, cheese,
yogurt, dairy products,
Soluble state
In the manufacture of beverages, meat, sausages,
chocolate and mayonnaise
Ability to form jelly
In the manufacture of bakery, mayonnaise, sausage,
milk, dairy and medical food
Emulsion formation
Mayonnaise, sausages, meat substitutes, health
products
Foaming ability
In the manufacture of ice cream, mayonnaise,
beverages, confectionery and cocktails

18. The use of whey for food:
The crystallization of lactose from whey and its use in the pharmaceutical industry or
directly in animal feed has led to inefficient use of important nutrients in its composition.
Therefore, the issue of finding the best way to use the basic ingredients of whey for food
and nutrition is introduced in almost all countries of the world. One of the achievements
of scientists and experts in this field is film filtration technology. In the milk industry,
whey filters are used to filter whey and extract valuable compounds such as protein-
soluble preparations and lactose. Industrial experiments have shown that thickening
food through a film filter is 30% cheaper than using a vacuum evaporator, and that
equipment maintenance costs are 20-25% lower. The biologically active compounds
and substances released by this technology, such as proteins, amino acids and
vitamins, are not exposed to chemical, physical and biological factors during the
technological process, so they fully retain their biological activity and natural state. It has
already been established in production practice that the film filters can completely
separate the compounds and substances that make up the raw materials, so the
production efficiency is high and there is no harm to the environment [13]. There is a lot
of research being done around the world on the use of whey processing technology,
equipment design, biotechnology and nanotechnology. Today, whey is used to make
concentrates, dried concentrates, whey protein, and milk sugar, as well as in the
production of alcohol, bread, sugar, and feed. In addition, a special section is separated
from whey for use in the pharmaceutical industry and for medical purposes [14]. More
than 40 different substances are now used around the world instead of sugar, with corn
starch, whey and fruit flavors predominating. However, US scientists have recently
found that such substances can cause obesity. In the United States, 61% of the sugar
needed by the cannery and 55% of the beverage industry's needs are substituted.
Especially in the 1970s, when the world was once again experiencing a shortage of
sugar, the production of sugary sweets intensified. The main methods of using whey
components can be adapted to the specific situation of our country, but firstly, there are
insufficient raw material resources, and secondly, it is expensive because we import the
equipment needed for production, and thirdly, milk production in Mongolia is seasonal.

19. There is little reason to believe that the current situation will provide reliable economic
results. Based on this, in the Mongolian context, it may be possible to separate whey
protein from sugar and use it to improve the taste and nutritional value of other foods as
a whole before using sugar drying. Experience in other countries has shown that skim
milk, buttermilk, and whey are used extensively to improve the taste of bread [15].
1.6. Infrared spectral method
To serve in the light of atoms, molecules studied changes in the energy transferred
into the can. This change is expressed in the energy diagram in the absorption
spectroscopic analysis.
Figure 1. Molecular light absorption energy diagram
As can be seen from the figure, the oscillations of the atomic molecules of matter
in the wavelength = 700-1200 nm, the transitions corresponding to the changes in the
oscillation-rotational energy (electron transitions may occur slightly), make the resulting
spectrum invisible to the naked eye. Therefore, this spectrum is called the Purple
Radiation Spectrum. Divide by the wavelength of the violet (near = 700-1.5 • 10 nm, the
middle = 1.5 • 10 -7.5 • 10 nm, the far = 7.5 • 10 -10 nm). Since the (molecular)
spectrum of violet radiation is an absorption spectrum, it is governed by the constitution
of light absorption and is expressed in their units. This can be illustrated by the following
graph.
100
50
0
4000 3000 2000 1600 1200 800 600 300
Figure2. Infrared spectrum of matter

20. As can be seen from the figure, the analytical signal is given by the functional
groups in the compound. For example -ON, = NH-groups, v= 300-3700 cm, -SH groups
v = 2400-2600cm, etc. These are taken from the directory.
Volatility of polyatomic molecules is divided into valence and deformation. They are
divided into symmetrical and asymmetric. When the length of a molecular bond
changes, the valence fluctuates, and when the angle of the bond changes, the
deformation fluctuates. Let's look at the -CH3 radical group.
1
Ѵs = 2872cm -1
x symmetrical valence fluctuations.
2
= 2962 cm -1
Ѵas=2962cm-1
corresponding asymmetric valence
fluctuations
3
δs= 1830cm -1
x symmetrical deformation fluctuations
4
δ as= 1460cm -1
x symmetrical deformation variation
Organic solvents are widely used in the spectral analysis of infrared light. These
solvents should not overlap with the absorption of the test substance. Therefore, this
principle (from the reference) should be taken into account when selecting solvents. In
addition, the selected solvent should be clear (colorless) in the given range and weakly
react with the solute molecules. It depends on the polarity of the solvent.
Infrared spectral instruments:
The infrared spectrum of a substance is recorded using an instrument called an
infrared spectrophotometer, and the absorption of the corresponding peaks is compared
with the data in an atlas or table to analyze the quality of the substance [16].

21. The NFL infrared spectrophotometer is usually a two-channel spectrophotometer.
The difference between UV and Visual Light Spectrophotometers is that the sample is
hard-pressed with potassium bromide in a special cuvette, mixed with Vaseline oil, or
dissolved in appropriate solvents (to reduce light scattering) and placed in front of the
monochromator. It should be noted that glass and quartz cuvettes should not be used in
the analysis of violet spectra. This is because they have the disadvantage of absorbing
purple light. Also, do not use aqueous solutions. Therefore, optical components such as
the prism, lens, and cuvette of the Nile Spectrometer are made of pure optical salts of
alkali metals. Different brands of spectrometers can be used for NV spectrum analysis,
but the general principle of operation is the same as the optical circuit, so the Russian
IX 14A spectrometer block diagram in Figure 3 is usually a two-channel
spectrophotometer. The difference between UV and Visual Light Spectrophotometers is
that the sample is hard-pressed with potassium bromide in a special cuvette, mixed with
Vaseline oil, or dissolved in appropriate solvents (to reduce light scattering) and placed
in front of the monochromator. It should be noted that glass and quartz cuvettes should
not be used in the analysis of violet spectra. This is because they have the
disadvantage of absorbing purple light. Also, do not use aqueous solutions. Therefore,
optical components such as the prism, lens, and cuvette of the Nile Spectrometer are
made of pure optical salts of alkali metals. Different brands of spectrometers can be
used for NV spectrum analysis, but the general principle of operation is the same as the
optical scheme, so the Russian “IX 14A” spectrometer block diagram is shown in Fig 3.
Figure 3. Block diagram of “IK-14A” spectrometer
1 Light source ( Globar SiC t °= 1300°C )
2,2 ' - mirrors,
3 - modulators
4,5 - cuvettes
6- monochromatic ( prism made of NaCl, NaBr, NaF salts )
7- Detector (bolometer)
8- Indicator recording device.

22. 1.7 Thin-layer chromatographic method
This method is based on the phenomenon of separation of the components of a
substance into two phases: animated and motionless. The animated phase is usually a
solid or liquid. The motionless phase is mounted on a solid. The term chromatography
and the term chromatography were first coined in 1903 by the Russian scientist Tsvet.
The word chromatography is derived from the Greek words chromos-color and graph-
writing. When a plant extract was passed through a column filled with calcium carbonate
powder, its components were distributed in several colored strips inside the column. In
this case, the plant extract entering the column is the animated phase, and the calcium
carbonate that fills the column is the motionless phase. The animated phase (usually
the solvent) added to the column is called the eluent. The moving phase leaving the
column is called the eluate. The process of separating the components of a substance
by chromatography is based on the fact that the components of the sample move
through the column at different speeds and travel different paths over a period of time.
Collect and analyze the fractions of each component. A detector-recorder may be used
to determine the composition of the substance. The registrar's indications are
automatically recorded and a chromatogram is established.
Modern chromatographic methods can quickly and completely separate 20-30
components in a single sample. This method is ideal for separating organic compounds
with similar structures and inorganic compounds with similar chemical properties.
When the animated phase (m) containing component A passes through the
motionless phase (s), substance A is distributed over the two phases according to the
distribution coefficient KD [2].
K D =
[ ]
[ ]
- distribution coefficient
[A]s- concentration of component A in the motionless phase
[A]m- concentration of component A in the animated phase
The thin-layer chromatographic method is based on the separation of impurities on a
very thin layer of sorbent. Place the stationary phase on a thin plate in a very thin layer.
Thin-layer chromatography is now widely used in the chemical study of plant
substances. This method was first developed by NA Izailova and MS Shrybora, who
effectively used a thin layer of aluminum peroxide to separate alkaloids from medicinal
plants. Sometime later, Meinger and Hall isolated ions of inorganic matter in a thin layer
of alumina prepared with starch.

23. CHAPTER 2. RESEARCH MATERIALS AND METHODOLOGY
2.1 Materials used in the study
5 liters of cheese whey from the milk farm of the School of Animal Science and
Biotechnology in February 2020. Curd whey was prepared in February 2020 from 5
liters of Bayangol soum of Uvurkhangai province in Mongolia. The experiments were
performed in the laboratory of the Department of Pasture and Nutrition Chemistry.
Figure 4. General scheme of research work
Whey
Vacuum evaporator
Freezer dryer
Chemical analysis
of whey
Separate of whey protein
Purify the protein by dialysis
Determination
of
carbohydrates
Chemical
analysis
of
whey
protein
The
determination
of
amino
acid
in
Thin
–
layer
Chromatography.
Infrared
spectral
Biuret

24. 2 .2 Methods of chemical analysis of whey
2.2 .1 Determination of moisture by weight method
Rationale for the method: The method for determining the moisture content of plants is
based on the method of calculating the difference in weight by dehydrating the sample
by drying it in a drying oven at 105 ° C.
Procedure: Take 2 g of sample, grind it well, put it in an airtight container, cover it and
store it without changing the quality and composition. Stabilize the weight, weigh 2 g of
the sample to a precision of 0.0002 g in a pre-prepared package, place at 120 ° C for
120 min, place in a desiccator, cool and weigh.
Calculate the results of the survey :
X=
a is the weight of the pre-drying sample packet, g
b is the weight of the packet with the sample after drying,
c is the sample weight, g
2.2.2 Determination of ash by weight method
Ash determination is a method of determining the amount of minerals in a food
sample.
Calculate the difference in weight of the sample before and after incineration. The
crucible is thoroughly washed with distilled water, dried and numbered. Dry at 450 °
C
for 1 h, cool in a desiccator for 25 to 30 min and bake. Repeat this process until the
difference in weight does not exceed 0.001 g. Take 2 g of each sample of the
plant under study in 4 repetitions, bake in a muffle furnace at 450 °
C for 1 h, cool in a
desiccator for 1.5 to 2 h and weigh. Repeat this process until the weight difference does
not exceed 0.001 g. Calculate by weight difference.
Calculate the results of the survey:
X=
x is the amount of ash,%
a is the weight of the crucible before incineration, g
b is the weight of the crucible with the sample after incineration,
c is the weight of the sample, g

25. 2.2.3 Determination of proteins by the Kjeldahl method
The essence of the Kjeldahl method is that the oxidation of the product under analysis
with high concentrations of sulfuric acid produces nitrogen ammonium sulphate, which
is neutralized with alkali to release nitrogen in the form of ammonia, which determines
the nitrogen content. This is a method of determining the total amount of nitrogen.
Nitrogen is the method by which all nitrogen compounds except protein nitrogen are
determined together. When the sample is heated with concentrated sulfuric acid, the
organic matter decomposes into carbon dioxide, water, and ammonia. Ammonia reacts
with sulfuric acid to form ammonium sulfate. To accelerate the decomposition of organic
matter, a mixture of CuSO4 and K2SO4 is often added as a catalyst, as well as H2O2, a
strong oxidizer.
(NH4)2SO4 Free NH3 is decomposed by concentrated alkali in a special distillation
device called the Kjeldah apparatus. NH3 is expelled by water vapor in the receiving
flask and the titer combines with a specific sulfuric acid. Titrate the sulfuric acid that has
not reacted with NH3 with the same normal alkali. To determine the amount of ammonia
in the water, air, and reagents, the same amount of water as the sample is taken as a
control and burned, and all subsequent operations are performed in the same way. The
amount of ammonia-bound acid contained in the test and control samples is determined
by the difference, the total nitrogen content is calculated, and the protein content is
calculated.
Reaction equation:
a. Burning:
Nitrogen-containing organic matter (protein) CO2, H2O, NH3
2NH3 + H2SO4 (NH4)2 SO4
b. Distillation:
(NH4)2 SO4 + NaOH Na2SO4 + 2NH3 + H2O
c. In the receiveng flask:
NH3 + H2SO4 (NH4)2SO4 + Excess H2SO4 (in test and control samples) Excess
H2SO4 + 2NaOH Na2SO4 + H2O (specific titer 0.01n NaOH)
Identification process:
1. Incineration of the sample: Take 0.5 g of the sample in the Kjeldahl
flask and add 1g of catalyst (0.25g of CuSO 4 + 0.75g K2 SO4 ) to 2 ml of

26. concentrated sulfuric acid. As a control, take the same amount of water instead of
the sample and burn it. Combustion is carried out until a clear green color is
obtained. First, the flask is burned without a lid to completely evaporate the water,
and then with a special glass lid.
Distillation apparatus to prepare: steam flask 1 / 2 - 2 / 3 filled distilled water. It has two
tubes. One tube, 80-90 cm long, almost at the bottom of the flask, regulates the water
vapor pressure inside the flask, while the other end of tube 2, which forms an angle
of 135 ° at the moment with the flask, enters the steam distributor. Place a few pieces of
porcelain on the bottom of the steamer to bring them to a boil. The steam distributor has
three tubes. One tube came out of the steam generator and almost reached the
bottom. The secondary tube, which is inserted into the limb, is designed to expel water
and steam and has a rubber tube with a clamp at the end. The third tube is located in
the middle of the steam distributor and is connected to the nasadka of the distillation
flask. There is a U-shaped connection between the steam distributor and the nozzle 2. It
is connected to the steam distributor by a clamped rubber tube and is directly welded to
the nozzle. This connection is connected to a concentrated alkaline burette by a
clamped rubber tube. The two-ball nozzle is connected to the distillation flask by a
grinder, and the tube reaches the bottom of the flask. The top ball is attached vertically
to the tripod and the tip is connected to the refrigerator by dipping it into the receiving
flask . Ammonia from the air is expelled before the device is assembled and used. To
do this, clamp the tube out of the steam distributor, open the tube connected to the
distillation flask, and boil the water in the flask for 15 to 20 minutes. Then open the outer
tube and close the tube into the nozzle, pouring water from the distillation and receiving
flasks.
Figure 5 . Kjeldahl method and distillation tools

27. 1. Steam flask
2. Water funnel
3. Re-suction fluid receiving and vapor transfer flask
4. Extracted waste liquid
5. Sampling flask
6. Steam pipe
7. Funnel to transfer the sample to the flask
8. Vapor conductive floor
9. Steam pipe
10.Condensing flask
11.Drip catcher
12.Atrium flask
13.Vacuum generator
2. Distillation: Take 50 ml of the diluted sample in the Kjeldahl flask, depending on
the amount of protein, 5 ml of 0.01 N sulfuric acid in the receiving flask, take 1-2
drops of the Tashir indicator, fill the refrigerator fork ( inner tube ) with acid and
transfer it to the distillation flask. Start distilling with 3 ml of concentrated
alkali. The released ammonia goes to the receiving flask together with the water
vapor. At this time, the nozzle is strongly heated. After heating for 5 minutes after
heating the top of the nozzle, distill the frosts for 3 minutes above the acid. To
avoid errors, titrate the tip of the fork with a small amount of distilled water into the
receiving flask. Distillation and titration of test and control samples shall be
repeated.
3. Titration and Calculation: Titrate the liquid in the receiving flask with 0.01N
alkali. The difference between the titrated acids in the control and test samples is
the amount of acid bound to the ammonia in the test sample. 1 ml of 0.01N
H2 SO4 solutions is equivalent to 0.14 mg of nitrogen. Knowing the titer of sulfuric
acid, calculate the amount of nitrogen in the distilled sample, and further the
amount of nitrogen in the incinerated sample. The amount of protein is equal to
the amount of nitrogen multiplied by 6.25. This is due to the fact that 16% of the
protein is nitrogen (100: 16 = 6.25).

28. 2.2 .4 Soxlet separations and determination of fat
Rationale for the method: The total amount of dried tissue is determined by the
continuous extraction of lipids with organic solvents and the difference in weight before
and after extraction.
Tools and reagents: Soxhlet apparatus, drying oven, box, bag, filter paper, analytical
balance, crucible holder, forceps, water bath with constant heat, thermometer up
to 200°
C
The Soxhlet apparatus consists of an evaporating flask, an extractor, and a condenser,
and is mounted on a fixed heat bath in a water bath. The Soxhlet apparatus extractor is
an important part of it and has two thick and narrow tubes. The ether vapor travels
through the thick tube to the refrigerator, where it drips and is returned to the extractor,
and the narrow tube returns the ether collected in the extractor to the evaporator flask.
Procedure: To determine the oil content, weigh 2-3 g of the sample in a filter paper bag
with a fixed weight bucks, place it in the bucks, dry it at 100-105o
C for 5 hours and
remove the moisture from the hydroscopic moisture sample. The continuation is easy to
determine. In order to determine the oil content, a sample of a moisture-proof, filtered
paper bag is placed in the extractor of the Soxhlet apparatus, ether up to 2/3 of the
flask, the apparatus is assembled, the water is cooled and the water bath is kept at
+80°C. After the distillation process lasts 6 to 12 hours and the ether becomes
colorless, the sample is removed from the extractor, the ether is evaporated at room
temperature, dried in an oven at 100-105 °
C for 5 h, cooled for 15 min and weighed for
15 min.
Calculation method: The weight of the sample, paper, and bucks after extraction from
the weight of the non-moistened box or paper sample is expressed in grams of oil. The
total fat content is calculated as a percentage using the following formula.
X=
Including: X-total fat,%
m - bucks, paper weight before sample extraction,
c is the weight of the sample taken for analysis, g
m1 - weight of bucks and paper samples after extraction, g
100 - The conversion factor for

29. 2.2 .5 Determination of dry matter by refract meter
Principle of the method: The higher the concentration of dry matter in the solution, the
greater the refraction. The accuracy of the description is 0.2%. This method is usually
used to determine the dry matter content of fruits, vegetables and aqueous extracts.
Procedure: To determine the dry matter, place a few drops of the sample on the prism
for 1 to 2 minutes to bring the sample to the same temperature conditions as the
test. Pre-check that the refract meter is working properly with distilled water. Turn the
refract meter screw to mark the instrument's dashed line in the event that the black-and-
white light intersects the checkpoint. Measure one sample 3-5 times and take averages.
2.3 Methods for testing whey protein
2.3.1 Separate whey protein and purify the protein by dialysis
Principle: Semi-permeable membrane-quality natural and synthetic materials
with ( kollodi, tsyellofan, pyergamyent ) using protein molecular weight
substances ( organic and inorganic ) cleans. Protein molecules cannot pass through
low-molecular-weight compounds through semiconductors. In humans and animals,
there are hereditary membranes that do not transmit proteins (such as the Bowman-
Shumlyansky capsule and the epithelium of the gastrointestinal mucosa) .
A dialysis device is called a dialyzer. The simple dializator with a glass immersed in
the protein solution tsyellofan peanut dipping easy way to depositod
Procedure: 5 g of protein was placed in a cellophane dialysis bag in a glass beaker with
water for 48 hours . Calculate the yield by drying the protein solution.
2.3.2 Chemical analysis of whey protein
- Determination of moisture by weight
- Determination of ash by weight
- Determination of proteins by the Kjeldahl method
- Soxlet sorting and determination of fats

30. 2.3.3 Determination of total hydrocarbons by phenol-sulfuric acid reaction
Principle : In hot acidic environments, glucose dehydrates to form hydroxymethyl
furfural. This form forms a brownish-yellow compound with phenol and measures light
absorption at 490 nm.
Reagents used: 5% phenol, 96% sulfuric acid, 1 mg / ml glucose , UV 1100
spectrophotometer
Procedure : Standard glucose: Prepare working solutions with a concentration of 20, 40,
60, 80, 100 μg / ml from a solution with a concentration of 1 mg / ml. Prepare a solution
with a concentration of 2 mg / ml 20; Dilute to 100 μg / ml. To 1 ml of 5% phenol solution
add 1 ml of a solution containing 10 to 100 mg / ml of carbohydrates. Add 5 ml of
concentrated sulfuric acid solution, allow to stand at room temperature for 10 min and
leave at 20 ° C for 20 min. Measure the absorbance at 490 nm.
2 .3.4 Determination by Infrared Spectroscopy
Principle of the method: Numerous physical methods can be used to study the
molecular structure of organic matter, but spectroscopic methods can provide more
reliable information about the structure of the molecule.
Of the many types of optical spectroscopy methods, the most widely used by chemists
today is violet spectroscopy. An instrument that determines the absorption spectrum of
a compound is called an infrared spectrometer or spectrophotometer. In this way, it is
possible to directly determine the structural units and functional groups, regardless of
the state or size of the substance.
Process: A dedicated small spoon substance KBr than 3, and 1-g sample nukhaad
better atmosphere mortar pumped shakhagchid take part . Take a sample from
the compressor and insert it into the instrument to measure the spectrum. The
functional group of the resulting protein and pure albumin standard is determined by NU
T at 400-4000 nm [17].
2.3. Determination of protein amino acids by thin-layer chromatography
Principle of the method: It is based on the hydrolysis of a protein by breaking the
peptide bond into a solution of amino acids using a fixed phase distribution factor to
separate the acid. In addition to hydrochloric acid, sulfuric and other minerals, alkalis,

31. and enzymes are often used to break down proteins. By hydrolysis, 6 hydrochloric acid
can be obtained in a 10: 1 to 1000: 1 ratio with a protein, and the need to obtain a large
amount of acid to increase the amount of protein reduces the breakdown losses. The
results show that when the protein is boiled with a high concentration of acid, tryptophan
is completely decomposed and methionine, cystine and cysteine are significantly
reduced.
It is important to use a well-purified protein in the hydrolysis, as the carbohydrates in
the protein are broken down by acid to combine with the amino acids to form a colored
humic substance, which loses amino acids.
Reagents used: Adjustable thermostat, amino acids for protein hydrolysis, 6N
hydrochloric acid
Commonly used solvents for amino acid separation: n-butyl alcohol: acetic acid:
water (4:1: 1, 4: 1: 5 ) 16 amino acids using a solvent
Calibration standard amino acids: amino acids collection ( produced in the former Soviet
Union nabor amino acids ) in addition to the amino acids involved. Cystine, threonine,
proline and tyrosine are available in pure form for analysis. Depending on the amino
acid distribution coefficient, a concentration of 0.01 m in 2-3 mixtures and 0.1N
hydrochloric acid and 10% isopropyl alcohol was suitable for long-term storage.
Process: a thoroughly cleaned 50 mg protein ( 1-2mg ) of the test weight on the
measure took sealing tubes ( ampuli ) in 10 mL of 6 N hydrochloric acid in addition to
the welded. Samples were taken for analysis and the moisture and nitrogen content
were determined. The sealed sample vials pre-set temperature warm authorities (the
incubator ) 100-105o
C 24 hours in laid during Unpack hydrolyzed protein . At this time,
gently stir 2-3 times and rinse gently. Upon completion of ongoing hydrolysis cool to
break the weld amino acid solution porcelain dishes Decant 50 ampoules add a little
distilled water, rinse water bath besides the closet than using a maximum temperature
of fan evaporate hydrochloric acid.
Once the hydrochloric acid has evaporated and the amino acid has evaporated, add
15-20 ml of distilled water 3-4 times, evaporate and dissolve the dry residue in 4 ml of
10% trace opropyl alcohol. To dissolve, first rinse with 1-2 ml of alcohol and then wash
the porcelain cup several times with the remaining alcohol, add to the previous solution,
fill and store in the refrigerator. After 15-17 hours, the topical clear solution is used for
analysis, as the humic substance settles on the bottom of the fingers .

32. Chromatography: Cut the chromatographic plate according to the diameter and height
of the chromatographic chamber. Use a pencil at a distance of 1-2 cm from the edge of
the plate to mark the hands and the finish, and mark the dots at a distance of 2-2.5 cm
from the starting line. At these points, add 20 μl of test solution and 5 to 40 μl of
standard solutions containing 0.05-0.4 μl of amino acids. Place the plate in the solvent
system in the chromatographic chamber. The solvent was removed and dried at the
finish line. Sprayed with a solution of ninhydrin, heated on a wet surface, and the stain
was clarified [18].
Analyze the amino acids by calculating the Rf.
Rf =
2.3.6 Determination of protein by biuret method
Principle of the method: When an alkaline solution of copper salt is added to a protein
solution, a pink color is formed which is directly related to the concentration of the
protein. This is a reaction in which a protein ion binds to a peptide in the structure of a
protein molecule to form a complex of copper biuret. The biuret reaction is colored by
two or more peptide-bound compounds, the trepeptide and larger peptides.
The peptide (amide) group is characterized by lactam-lactim tachomerism. In an
alkaline environment, the predominant form of the polypeptide, lactim (enol), interacts
with copper to form a stable color complex:
Process : . Add 1 drop of 1% 1% protein solution (gelatin, egg white or plasma albumin),
1 ml of 10% alkaline solution (NaOH or KOH) and 1 drop of 1% copper sulfate. Blue-
pink or reddish-purple spots appear.

33. CHAPTER3. RESEARCH RESULTS
3.1 Results of chemical analysis of whey
1 liter of whey was evaporated in a vacuum evaporator to 400 ml for 3 days. The
concentrated soy milk, 100 ml then freeze khataagchind 24 hours. 3 g of dry sample
was obtained.
Table.7 Results of chemical analysis of whey
Specifications In the press review Experimentally
Cheese Curd Cheese Curd
Dry matter,% 4.5-7.2 4.2-7.4 6.8 5.7
Protein 0.9-1.6 0.5-1.4 1.2 0.6
Fat 0.2-0.5 0.05-0.4 0.23 0.0
Moisture 1-1.5 0.8-1.2 1.2 1.2
Ash 0.3-0.5 0.4-0.8 0.8 1
The results of the experiments were compared with those of the press review for dry
matter, protein, fat and moisture. O Spirit literature review [19 ] compared to show that
the rich nutrients of milk Mongolian livestock .
3.2 The amount of protein extracted from whey
Table.8 Protein separation from whey
Sample Whey, volume ml Separated protein, ml Output,%
Whey 1000ml 280ml 40%

34. 3.3 Chemical analysis of whey protein
Graph1. Chemical analysis of whey protein
The chemical composition of whey protein is high in protein, fat, ash and carbohydrates
compared to the press review, indicating that Mongolian animal milk is rich in nutrients.
The moisture content was lower than in the press review [20] due to the fact that the
whey was frozen and dried for further study.
3.4 Determination of protein structure by infrared spectrophometer
Physical methods are widely used to study the molecular mass and structure of high-
molecular compounds. Among the backs of the optical spectrum , the infrared spectrum
is the most widely used fast method. It directly determines the functional groups
contained in a substance, regardless of its state or size. Figure 3 shows the
spectrogram of the isolated whey protein.
0
10
20
30
40
50
60
70
Protein Moisture Fat Ash Carbohydrate
Standart Experimental analysis

35. Figure 6 . Infrared spectrogram of isolated whey protein
Figure 5 shows extracted from whey protein than (-NH-) amino group of the 3422 cm-
1
region has strong absorption and valence fluctuations, (C = O) carbonyl group of
the 1654 cm -1
mu jid weak absorption by valence words tethering with these amido
group of key functional groups of the protein. (CH (sp 3
)), a group of 2900 cm -1
to weak
absorption region, with valence is the prospect , and CO 2, a group of 2,400 cm -1
to
weak absorption range of RS on spectroscopic instrument togtvorjoogüigees
air CO 2 gas double has been detected.
3.5 Amino acid determination by thin-layer chromatography
According to the results of the analysis, tryptophan is broken down by high
concentrations of protein, methionine , cystine and cysteine are significantly reduced. It
is important to use a well-purified protein in the hydrolysis, as the carbohydrates in the
protein are broken down by acid to combine with the amino acids to form a colored
humic substance, which loses amino acids.

36. Figure 7 . Chromatogram of whey protein amino acids m
Figure 7 Whey protein decomposition by the NUH method 16 histamine, lysine,
arginine, serine, alanine, glutamic acid, cysteine valine, methionine, leucine, isoleucine,
phenylalanine, tryptophan, glycine, tyrosine, aspartic acid, etc. found in the sample. To
confirm this, the Rf is the same when calculating the Rf of standard amino acids and
samples in Table 9. This means that there are 16 amino acids.
Table .9 Whey protein amino acids
№
Essential
amino acids
Marked with
3 letters Rf №
Substitute
amino acids
Marked with
3 letters Rf
1 Isoleucine With 0.53 9 Alanine Ala 0.32
2 Leucine Leu 0.49 10 Serin Ser 0.26
3 Valin Val 0.47 11 Glutamic acid Glu 0.29
4 Methionine Met 0.47 12 Arginine Arg 0.12
5 Tryptophan Trp 0.59 13 Cysteine Cys 0.50
6 Lysine Lys 0.10 14 Tyrosine Tyr 0.25
7 Phenylalanine Phe 0.52 15 Glycine Gly 0.30
8 Histidine His 0.09 16 Aspartic Asp 0.10

37. 3. 6 Determination of protein by biuret method
When copper salt CuSO4 is added to the protein solution, a purple color is
formed. It recognizes the presence of a peptide bond. As a result of the biuret reaction,
the solution turned purple, indicating that the protein had been isolated experimentally.

38. JUDICIAL PROCEEDINGS
The chemical composition of whey and whey protein in this study is in line with the
results of this study “Casein and whey protein enzyme degradation into peptides (NUM
N.Uranchimeg, B.Tuyaagerel, B.Munkhjargal)”.
The determination of amino acids by thin-layer chromatography is equivalent to that
written by S.Tsendsuren and Ts.Erdenebileg in the 2015 book “Traditional and modern
production of technologies for the use of biological resources”.

39. Conclusion
In this study, proteins were isolated from whey, chemical composition was
determined, protein amino acids were determined by thin-layer chromatography, and
protein structure was determined by infrared spectrophometer. The chemical
composition of cheese and whey was also determined.
1. 280 ml of whey protein was extracted from whey with 40% yield.
2. The chemical composition of cheese and whey was determined. The cheese
whey contained 6.8% dry matter, 1.2% protein, 1.2% minerals, 0.23% fat, 1.2%
moisture, and 0.8% ash. Curd contains 5.7% dry matter, 0.6% protein, 1% mineral,
0.00% fat, 1.2% moisture, and 1% ash.
3. The chemical composition of whey protein was determined. Whey protein was
found to contain 4% moisture, 64.6% protein, 11.5% fat, 4.3% ash, and 2.2%
carbohydrates.
4. Thin-layer chromatograph protein analysis includes histidine, lysine, arginine,
serine, alanine, glutamic acid, cysteine valine, methionine, leucine, isoleucine,
phenylalanine, tryptophan, glycine, tyrosine, aspartic acid, etc. detected.
5. In the analysis of whey protein by infrared spectrophometer, the amino group
of protein (-NH-) was strongly absorbed in the range of 3422 cm-1, with valence
fluctuations, (C = O) carbonyl group was weakly absorbed in the range of 1654 cm-1,
valence fluctuates.
6. The purple color of the solution as a result of the biuret reaction indicates that
a protein has been isolated.

40. Thank you
We would like to express our sincere gratitude to Supervisior’s Du Ming,
N.Pagmadulam, J.Oyuntsetseg /Senior Research Fellow of the Institute of Chemistry
and Chemical Technology of the Academy of Sciences/ and B.Tuyaagerel /Professor of
the National University of Mongolia/ for their dedication, knowledge and valuable time. I
would like to express my deep gratitude to the staff of the Department of Pasture and
Nutrition Chemistry of the School of Animal Science and Biotechnology of the
Agricultural University for providing the necessary materials, equipment and facilities for
the research.

41. Appendix
Figure.8 Vacuum evaporator Figure . 9 Freeze -drying of whey
Figure .10 Protein separation by dialysis Figure.11 Protein determination by Kjeldahli

42. Figure12 Protein was determined by biuret method Figure. 13 Soxlet
Figure.14 Determination of moisture Figure.15 Protein determination by
Kjeldahli

43. Appendix 2. Supervisor's evaluation of the dissertation
The Supervisor’s: Dr. N.Pagmadulam ( Ph.D)
Student's name: Borkhuu Nomin-Erdene
Research work topic: RESEARCH OF WHEY PROTEIN
№ Work performed Percentage
in the
assessment
Teacher
evaluation /
percent /
Note
1 whether the research plan was
written in a logical sequence,
accurately, and in accordance
with the writing guidelines
10 10
The research was written in
accordance with the writing guidelines,
the plan was made in a logical and
realistic manner, and the experiment
was successfully completed at the
place where the practice was planned.
2 Follow the teacher's
instructions and write the
research methodologically 10 10
The dissertation was written according
to the methodology
3 Did you get acquainted with the
research on the topic according
to the teacher's instructions
20 20
Acquainted with the research on the
topic
4 Follow the instructions for
writing the research paper and
write the introduction, materials
and appendices correctly
20 20
The introduction, materials used, and
appendix are written correctly
5 Proper selection and use of
research methods and
methodologies within the scope
and objectives of the research
20 20
Within the framework of the research
goals and objectives, the research
methods and methodologies were
selected and used correctly.
6 Achieving research goals and
objectives, writing conclusions
and comments
20 20
The goals and objectives of the
research were achieved and the
conclusions were well written
Total 100 100
Evaluation of
Supervisor: /N.Pagmadulam /
/Du Ming/
Signature description:
DATE: April 19, 2020

44. Appendix 3. Review of the research work
Student's name: Borkhuu Nomin-Erdene
Thus cuts thesis: Research of whey protein
№ Criteria for criticism Critic
teacher review
Share a total of
100 review
points
percent
Given by the
critic teacher
percent
Edited according to
reviews
1 whether the purpose and
objectives of the
research are appropriate
From the goals, objectives and
conclusions of the work, it is
clear that the work has achieved
its goals and objectives.
10 10
2 Whether the situation is
true, correct and
sufficient
This effort has
been very well met ,
but before he compiled cited in
particular side on the bugs to fix
that.
20 18
Compilation and
refinement of previous
work on the topic.
3 Whether the
methodology was
chosen correctly to
achieve the objectives of
the research work
Research work for enough materi
al to soy milk to determine the
composition of
content, protein identification ,
it all ürdüülegch amino acids to
identify ways to optimize the
right choice is.
20 20
4 Whether the data used
in the research work are
accurate and consistent
with the chosen method
and model
Although all information used in
the research is true and
accurate, and compares
domestic and foreign articles, it is
advisable to re-evaluate the
latest information.
For example, in 2015, the
number of dairy products should
be updated as in recent years. ...
15 14
The information used in
the thesis has been
updated with the latest
information.
5 Theoretical and practical
significance of the thesis
This research is considered to be
of great theoretical and practical
importance, as it addresses the
issue of recycling waste, which is
a problem in modern waste-free
eco-industry, and producing
high-yield products.
20 20
6 The wording of the
research, the
grammatical errors, the
organization and
elaboration of the
chapters and sections
In the research work,
spectrophotometer,
spectroscopy, spectrophometer,
etc. are erroneous and written
differently. It is also a good idea
to correct the citations in the
theoretical and methodological
15 13
Corrected erroneous
words in the research
work, corrected citations
and bibliography,
corrected obscure terms
using Insert → Equation,
and corrected titles with

45. sections, such as [1], (author's
name), and so on.
Use Insert → Equation to make
some of the formulas in the text
blurry, and to edit the graphic
text in Excel with a different font
or title.
different backgrounds
into a single font.
7 Suggestions for further
improvement
This research work has been fundamental research to whey protein.
It is hoped that this work will be further enhanced by electrophoresis
to determine the composition of whey proteins, and by testing
enzymatic and chemical hydrolysis methods to improve the biological
activity of the protein.
8 Critic's evaluation of the
research work
This work is considered to
be a good work that meets
the requirements of a
bachelor's degree, which
can serve as a basis for
further detailed analysis of
the goals and objectives.
10 9.5
Review by teacher:
National University of Mongolia, University of Chemical Sciences, Department of
Chemical and Biological Engineering,
/ B.Tuyagerel/
Signature description:
DATE: 2020-04-28