whey protein

1. WHEY PROTEIN RESEARCH
Supervisor’s: /Du Ming/
/D.Bolormaa/
Student: Borkhuu Nomin-Erdene
2020

2. Title
Rationale and novelty of the research work
The purpose and objectives of the research
Chapter1 . Studied
1.1 Composition and technological properties of whey
1.2 Whey protein
1.3 Significance of whey protein
1.4 Membrane-technology milk processing
1.5 Whey protein concentrate
1.6 atomic absorption spectroscopy backing
Chapter 2. Research methodology
2.1 Methodology for analysis of whey
2.1.1 Weight method for determination of moisture
2.1.2 Determination of ash by weight
2.1.3 Total obligations to determine Kiyeldaliin way
2.1.4 Determination of total oil by Soxhlet method
2.1.5 Determine the content of macro and micro elements by AAC
2.2 Methodology for analysis of whey protein
2.2.1 Separation of whey protein
2.2.2 Dialysis of protein
2.2.3 Refractometric method for the determination of dry matter
2.2.4 Determination of total hydrocarbons by phenol-sulfuric acid reaction
2.2.5 Infrared spectroscopic determination ( IRS )
2.2.6 Determination of protein amino acids by porosity
2.2.7 Method for microbiological analysis to determine the number of bacteria
2.2.8 Detection of Salmonella pathogen in the sample
Assumptions about the outcome
Schedule
Used by the union PP im ag average

3. Rationale for research work
Our country is a nomadic pastoral country and has an ancient tradition of producing the widest
variety of sour milk products from its milk. Ministry of Food and Agriculture reported that
demand for Mongolian population of dairy lit 117.0 million in 2015, milk 33475tn are
product technique and develop Brother 857 kg of soy milk refuse to be is news that there [1] .
The production of curds, cheese, curds, food and technical casein produces large amounts of
whey, which is a valuable raw material containing 5-7% dry matter, such as lacto globulin, lacto
albumin, milk sugar, minerals and water-soluble vitamins. that is. Soy milk contains high quality
proteins in the immune system, IDD c provided, antioxidants floor. Whey protein contains
immune-stimulating substances -globulin, -lacto albumin, glycol macro peptide and immune
globulins. They activate the immune system and improve the body's resistance.
In Mongolia, whey is not used for health purposes or for 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.
Novelty of the research:
The novelty of the research is the processing of whey, which is rich in minerals, proteins and
vitamins, during the processing of milk, into biologically active feed additives.
Purpose of the research: To separate protein from whey to obtain biologically active food
supplements.
Objective:
1. Determine the composition of whey
2. Separation and drying of whey protein
3. Determining the chemical composition of proteins
4. Determine the number of bacteria

4. Introduction
Cheese from soy milk with lactic acid, enzyme or enzyme-rich but well-Chan, a protein
contains the hood. It is also a high-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 have shown that this drink can prevent high
blood pressure, stroke and heart attack, as well as balance cholesterol, increase muscle mass,
prevent osteoporosis and improve the immune system, reduce stress and increase energy .
The technology of gelfiltration 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 lactoglobulin and lactalbumin, are introduced, which can separate whey
protein from milk sugar and minerals. The technology for the production of cheese and cheese
products with whey 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.
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, special parts are separated from whey for use in the
pharmaceutical industry and for medical purposes .

5. CHAPTER1.
STUDIED
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 from 15-22 all its protein % and 0.7-1.3 milk protein to calculate the total % of
serum albumin and 7-12 % β-globulin, 2-5 % of lacto albumin, 1.9-3.3 % immunoglobulin, 2-
6 % protease-peptone. Casein- precipitated whey contains glycol macro peptides due to the
action of renin (Ana R. Madureira , Pereira, Gomes, Pintado , & Xavier Malcata ,
2007).
Goat milk yield, composition, properties, goat color and milk relationship were
studied. The study also compared black and white goat's milk with cow's milk ( M. Narangerel, a
lecturer at the Agricultural University ) .
Research on the use of whey protein in food production . A comparison was
made between Mongolian cheese, curd, lemon, whey protein and other ingredients derived from
industrial whey (Khuree Institute of Information and Communication Technology, O.Maral-
Erdene, Ko Jae Hyun) .
Casein and whey protein enzyme cleavage into peptides. The possibility of extracting
whey protein and enzymatic action to produce food supplements containing biologically active
peptides was studied ( NUM Nerguiin Uranchimeg , B.Tuyaagerel, S.Munkhchimeg ).
1.1 Milk of ingredients and technological characteristics
One of the by-products of the dairy industry is whey. Soy milk is 5-7% of the dry ingredients and
substances , such as milk solids, 50% narrower than the IDS cream, milk , nitrogen compounds,
minerals, protein, lactose, and a variety of vitamins, particularly water-soluble B 1, B 2
and C vitamins, is a valuable raw material containing enzymes, trace elements and organic
acids [2] .

6. Table1. Whey composition and basic parameters
Specifications
Types of whey Nutrient
conversion rate,%
Cheese Cream Cases
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
Minerals - - - 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 -
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 soy milk, dairy core component of their Mo lyekul weight and kollo not in p is defined as the
quantity felt.
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 .

7. 1.2 Whey protein
Casein-derived milk contains about 0.6 % protein. This is called whey protein. Multi-
protein milk protein fraction established that 15-22 % and 0.7-1.3 total milk protein to calculate
the percent albumin serum of 7-12 % β-globulin, 2-5 % of lacto albumin, 1.9-
3.3 % immunoglobulin, 2-6 % protozoa-peptone. Casein- precipitated whey contains glycol
macro peptides due to the action of rennin. -immunoglobulin irreplaceable uric acid, sulfur-
containing uric acid, which is more nutritious than casein protein, can be filtered by gel filter
and used as food protein. Whey protein is small in size , but more biologically
important than casein . Whey protein can be separated at different pH values using
MgSO 4 or (NH 4 ) 2 SO 4 , and β-lacto globulin and –lacto albumin precipitate when 3% acetic acid
is used . Whey proteins are separated for research purposes. In practice, whey 90-95 together
with all protein fractions 0
C- including denaturation of separated. Sugar is also extracted from
whey. Although not enough whey protein is used for food, it is more nutritious than casein
because of its high 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
residue. 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 quickly absorbed, so it is more
important for an active adult. Total milk protein about 20 percent of whey protein in
blood cholesterol play an important role in reducing the concentration and the immune
system. The content of essential amino acids in whey protein is higher than that of meat
protein. Whey protein supplements slow down the cancer process. Protein concentrate is
produced from whey and used for treatment and fitness [ 3 ] .
Table 2. Whey protein and curd composition
№ Components Ingredients,%
Whey protein Curds
1 Us 75-78 77-80
2 Dry matter 22-25 20-23
3 Protein 18.0 15.0
4 Milk sugar 2.0 3.0
5 Ash 1.2 0.5
6 Lactic acid 0.8 1.0

8. 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 3 . Whey protein uric acid,% [ 4 ]
Uric acid Immune globulin -lacto albumin -lacto globulin
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
Valens 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
Histamines 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
-Lacto albumin: 18% whey protein, does not contain phosphorus in the molecular composition ,
is not digested by gastric enzymes. Albumin is a bubble-like protein that dissolves in water and
dilute saline. This protein coagulates with heating. The second type of milk protein is lacto
albumin.
-Lacto globulin: The molecule has one thiol group, which is a natural protein inside the
molecule and is shielded . -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 [ 3 ] .

9. 1.3 Significance of whey protein
Certain properties of whey protein are interesting. They affect the quality of the product
in different technological ways. On the other hand, changes in the whey protein fraction can be
inferred from milk processing. An important and observable 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 immune globulins, which are denatured at 60 degrees, followed by
serum albumin and beta lacto globulin, and the most resistant to temperature are alpha-lacto
globulin. The protease-peptone fraction begins to denature at -110 ° C [ 3 ] .
Whey contains high-quality proteins that nourish the immune system and provide
antioxidants. Whey protein contains immune-stimulating substances -globulin, -lacto albumin,
glycol macro peptide and immune globulins. 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 good effect on blood
clotting. 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 influence of temperature , the ratio
between –lacto albumin –lacto globulin 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 with temperature, and precipitates with casein when salt is added. To distinguish
between renin and whey, sialic acid is determined from renin. The content of whey can be
determined by the content of sialic acid in a mixture of powdered dried milk . I used to be
interested in the fat content of milk, but now I'm interested in the protein content. There is a
growing interest in low-fat, 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 male [ 5 ] .

10. 1.4 Membrane-technology milk processing
With the help of membrane filtration, milk components are separated into different molecular
weights and used for enrichment and production of various human products [ 6 ].
Figure 1. Membrane filter

11. Table 4 . The principle of membrane filtration
Type of judgment Judicial principle Purpose
Osmosis filtration -H 2 O
-Mineral
-Lactose
-Protein
Produces milk, whey and cotton dry matter,
increased protein, condensed milk, yogurt and
drinking milk.
Very fine judgment -H 2 O
-Mineral
-Lactose
-Protein
Milk, whey and cotton minerals are separated and
used in medical and children's nutrition.
Too narrow a
judgment
-H 2 O
-Mineral
-Lactose
-Protein
Produces milk, whey, cheese, concentrated protein,
milk and dairy products.
Micro-judgment -H 2 O
-Mineral
-Lactose
-Protein
"Nanny."
-Mechanical pollution
Milk microorganisms, their armored cells and
protein precipitates are filtered out.
The table above sets out the principles of the judiciary in relative terms. In real life, it is not
possible to completely separate high and low molecular weight compounds. For example, ultra-
fine filters can filter only 60% of milk protein. The principle of membrane filtration differs from
that of ordinary filtration. During simple filtration, solids or amorphous substances accumulate
on one side of the filter. However, membrane switch matter filter in the filter separated the two
liquids from one area to another ( diffusion ) went reactions, thereby Unlike mutual
concentrations of the fluids. For example, when whey is processed through a very fine filter, 18%
of the dry matter concentrate on one side of the filter and 5% of the dry matter serum accumulate
on the other side. Another feature of membrane filtration is that the filtration process is based on
the action of pressure. The pressure force to regulate the filtration process depends on the type of
alkali . In the dairy industry, whey processing technology using ultra-fine filters is the most
commonly used. In particular, the use of biologically and nutritionally valuable whey protein for
food and medical purposes by concentrating it with ultra-fine filters accounts for more than 50%
of the use of membrane technology. [ 7, 8 ].

12. Table 5 . The amount of milk components
Components diameter, nm Molecular weight
Oil bubbles 100..10000 10 12
.. 10 14
Casein micelles 10..300 10 7
..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
Membrane filters shall be made of a special material with a fine pore size of 0.3..600 nm. Use
natural ( biological ) and synthetic fabrics as filter materials . Synthetic filters are made of
organic polymers and inorganic ceramics.
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 specifics of the technology of whey protein
concentrate, it is used to stabilize, soften and foam foods . Based on the biological, nutritional
and technological properties of whey protein concentrate, it is widely used in the food industry in
developed countries [ 7 ] .
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 sports food for athletes, cheese,
yogurt and protein,
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 treatments
Emulsion formation
Mayonnaise, sausages, meat substitutes, health
products
Foaming ability
In the manufacture of ice cream, mayonnaise,
beverages, confectionery and cocktails

13. Whey for food use:
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, yellow
to introduce a comprehensive industry to find a better way for food and feed use as basic
chemicals in milk composition before being placed almost all countries around the world, and
scientists in this field, experts do great work and delivering results of a judgment is beyond
technology . In the milk industry, milk filters are used to filter out whey and extract valuable
compounds such as protein-soluble preparations and lactose. Grocery film filters
condensing a vacuum sets the evaporator used in 30% of the low cost and equipment
maintenance costs 20-25% less than that produced experimental studies. This technology, which
differs from the protein, amino acid, in the technological process of biologically active
substances such as vitamins chemical, physical and biological effects of what factors exempt
from the stores as full biological activity and un spoil. It has already been established in the
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 .
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
the pharmaceutical industry and for medical purposes [ 9 ].
More than 40 different substances are now used worldwide instead of sugar, with corn starch,
milk whey and fruit-based spices predominating. However, US scientists have recently found
that such substances can cause obesity. In the United States, 61% of the sugar needed by the
canning industry 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.
Some of the main technologies used in the use of whey components can be adapted to the
specific situation of our country, but firstly, there are insufficient raw material resources,
secondly, the equipment required for production is expensive, and thirdly, dairy production in
Mongolia is seasonal. 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, cotton, and whey are used extensively to improve the taste of bread [ 10 ] .

14. 1.6. Atomic absorption spectroscopy method
The atomic spectroscopy method, which is based on measuring the intensity of light emitted
when the atoms of a test substance are exposed to a flame (thermal stimulation), is called the
atomic emission or atomic absorption photometry method. The scheme of atomic emission of the
reaction that takes place here can be shown as follows.
A + E → A x
+ hν
These are: A-determining atom E-absorbed energy,
A x is the
excited atom
hv is the quantum of emitted energy.
The general principle of this tool is shown in the figure :
2 - Fig . Flame - photometric block diagram
1. Research solution
2. Pulverize
3. Gas-Liquid Waste Tap
4. Convex mirror (reflector)
5 . Oh, the flames
6 . D diaphragm
7.8 . Guide lens
9. C search filter
10.D lens (guide)
11. X goggles
12. Photocell
13. Grower
14. Micrometer
15. Power supply block ( V = 220V)

15. Consider a flame , which is an integral part of an instrument that excites an atomic molecule and
generates a radiation spectrum . A flame generator produces a mixture of various flammable
gases.
1. Bhutan - air 1300-1900 0
C
2. pr o r a n air - 1900-2000 0
C
3. Hydrogen - air 2000 -2 100 0
С
4. Gasoline - air 2000-2200 0
С
5. acetylene - air 2100-2400 0
C
6. Hydrogen - Oxygen 2500-2700 0
C
7. A cetylene - oxygen 3100-3200 0
C
Flames that provide such temperatures usually identify alkali and alkali metals with low
ionization potential . However, many other high- temperature ionizing metals can be identified in
this way if high- temperature flames occur . Let's take a special look at the process that takes
place in a flame .
Figure 3 . Flame structure
In this case, a mixture of test solution, air, and gas enters the flame retardant probe and forms an
aerosol with the initial thermal decomposition reaction. However, due to the low flame
temperature, CO and H 2 are not formed due to incomplete combustion . Instead, it enters the
oxidizing probe and is completely oxidized (CO 2 , H 2 O) as the temperature rises . In other words,
conditions are created for the formation of the radiation spectrum. Therefore, this zone is the
most important part of the analysis, and the process that takes place here is very
complex. However, the atomic molecules identified as a result of combustion form stable
monoxides, mono hydroxides, nitrides, and carbides. The concentration of a substance is
determined by measuring the photocurrent by converting the light signal generated here into
electrical energy through a photocell.

16. Analytical parameters of the flame photometric method
It is as simple and fast as conventional photometers, with high accuracy and automation,
and is a highly sensitive analytical method that can detect a large number of metals at very low
concentrations. Sodium ions (10 -3
mg / ml can determine when
content). Therefore, micronutrients contained in various objects and mineral water are widely
used in modern analysis.
The limits for determining the elements by this method depend on the design of the instrument,
its sensitivity, and the structure of the flame. For example, German flame photometers can
simultaneously detect many elements in a very small (0.3 ml) sample. This is essentially the
same as for the spectrophotometric method, since the intensity of the analytical signal is linearly
related to the concentration of the analyses, so the analytical methods used in spectrophotometric
analysis can be used.
It is possible to identify many elements in a mother at the same time. The analytical aggregates
used in spectrophotometric analysis can be used because the intensity of the analytical signal
is linearly related to the concentration of the analyses, similar to the spectrophotometric method .

17. CHAPTER 2.
RESEARCH METHODOLOGY
2.1 Yellow analytical methods for milk
2.1.1 Weight method for determination of moisture
Method roots Rank : Determination of moisture in the plant samples 105 0
is based on the
calculation by measuring the weight difference dehydrate drying C oven.
Process : 2d chopped airtight container well-sealed sample quality ingredients are kept
unchanged conditions. Stabilize the weight, weigh 2 g of the sample to a precision of 0.0002 g in
a pre-prepared package , place it at 105 °
C for 120 min, place in a desiccator, cool and weigh.
Calculate the results of the survey :
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 mother's weight, g
2.1.2 collation determination evident Gravimetric
Ash determination is a method of determining the amount of minerals in a food sample.
Calculate the difference between the weight of the sample before and after combustion. 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 of the studied plant samples in 4 repetitions, bake in a
muffle furnace at 450 °
C for 1 h, cool in a desiccator and weigh for 1.5 to 2 h. Repeat this
process until the difference in weight does not exceed 0.001 g. Calculate by weight difference.
Calculate the results of the survey :
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, g
c is the weight of the sample, g
2.1.3 Kjeldahl method for protein determination
The essence of the Kjeldahl method is that the oxidation of the product under analysis with high
concentrations of sulfuric acid produces nitrogen ammonium sulfate which is neutralized

18. by 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 are determined together, except for protein nitrogen. 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 CuSO 4 , K 2 SO 4 , and H 2 O 2 , a
strong oxidizer, is usually added as a catalyst .
(NH 4 ) 2 SO 4 Free NH 3 is decomposed by concentrated alkali in a special distillation device called
the Kjeldahl apparatus . NH 3 is expelled by water vapor and the titer combines with a specific
sulfuric acid in the receiving flask. Titrate the sulfuric acid that has not reacted with NH 3 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 all subsequent operations are
performed in the same way. The amount of ammonia-bound acid in the test and control samples
is determined, the total nitrogen content is calculated from the difference, and the protein content
is calculated.
Reaction equation:
a. Burning:
Nitrogen-containing organic matter (protein) CO 2 , H 2 O, NH 3
2NH 3 + H 2 SO 4 (NH 4 ) 2 SO 4
b. Distillation:
(NH 4 ) 2 SO 4 + NaOH Na 2 SO 4 + 2NH 3 + H 2 O
c. In the receiving flask:
NH 3 + H 2 SO 4 (NH 4 ) 2 SO 4 + excess H 2 SO 4 (in test and control samples) Excess H 2 SO 4 + 2
NaOH Na 2 SO 4 + H 2 O (specific titer 0.01 n NaOH)
Identification process:
1. Incineration of the sample: Take 0.1-0.5 samples of the Kjeldahl flask and add 2 g
of sulfuric acid to 1 g of catalyst (0.25 g of CuSO 4 + 0.75 K 2 SO 4 ). As a control, the same
amount of water is taken and burned instead of the sample. Combustion is carried out until
a clear green color is obtained. To completely evaporate the water, the flask is burned
without a lid, and then with a special glass lid.
Preparation of the distillation apparatus: the flask is filled 1 2 - 2 3 with 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

19. the second tube, which is formed at the moment in the flask , enters the steam
distributor. Place a few pieces of porcelain on the bottom of the steamer to boil
evenly. The steam distributor has three tubes. One tube comes out of the steam generator
and almost reaches the bottom. The reciprocating secondary tube is designed to remove
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 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. Concentrated alkaline burette is connected to this joint 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 3 . Kjeldahl method and distillation tools
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

20. 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 indicator, fill the refrigerator fork ( inner tube ) with acid and transfer it to the
distillation flask. Add 3 ml of concentrated alkali and start distilling. The released
ammonia goes to the receiving flask together with the water vapor. At this time, the nozzle
is strongly heated. After heating the top ball of the nozzle for 5 min, distill the frosts for 3
min 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 0.01 N
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 0.01 N H 2 SO 4 solution 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 ) .
2.1.4 Soxlet separation and determination of fat
Rationale for the method : The total amount of dried tissue lipids is continuously extracted with
an organic solvent to determine the difference in weight before and after extraction.
Tools and reagents: Soxhlet apparatus, drying oven, box, bag, filter paper, weighing instrument,
crucible holder, forceps, water bath with constant heat, thermometer up to 200 0
C
The Soxhlet apparatus consists of an evaporating flask, an extractor, and a condenser, and is
placed 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 returns to the extractor, and the narrow tube returns the ether
collected to the extractor to the evaporator flask.
Procedure : Hygroscopic moisture was determined by weighing 2-3 g of the sample in a filter
paper bag with a constant weight box to determine the oil content , placing it in the box

21. and drying it at 100-105 °
C for 5 hours to produce a constant weight. It is easy to continue to
determine the oil from the sample. To determine the oil content, place a sample of the moisture-
proof, filter paper bag in the Soxhlet apparatus extractor, add up to 2/3 of the flask, assemble the
apparatus, run the water in the refrigerator and hold the water bath at +80 °
C.
After distillation 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 to 105 °
C for 5
h, cooled for 15 min and weighed for 15 min .
Method of calculation: The weight of the sample, paper and box after extraction from the weight
of the box, paper sample without moisture, and the oil in grams. The total amount of fat is
calculated as a percentage by the following formula.
Including: X-total fat,%
m- bucks , paper weight before sample extraction,
m 1 - bucks and sample extraction post-weight paper,
c is the weight of the sample taken for analysis, d
100 - the conversion factor for
2.1.4 Determination of macro and micro elements by ACC curve
For example, to prepare standard solutions (serial) to build curve and their photo
current(i) measuring i (mkA) = f (c) pg / ml, linear calibration curve established and find the
calibration curve, the concentration of research solutions .
Figure 5 . It's too loose
2.2 Methods for testing whey protein
2.2.1 Separation of whey protein
Procedure : Filter the whey through a 10kDa membrane filter to separate the whey protein.

22. 2.2.2 Dialysis of protein
Principle : Semi-permeable membrane-quality natural and synthetic materials with ( colloid,
pragment ) 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 inherited 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 deposit o d
Procedure : 5 g of protein was placed in a dialysis cellophane bag in a glass beaker with water
for 48 hours . Calculate the yield by drying the protein solution .
2.2.3 Refractometric method for the determination of dry matter
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.
Reagents used: Refractor meter, distilled water, ethyl alcohol, filter paper
Procedure : To determine the dry matter, place a few drops of the sample on the prism for 1 to 2
minutes to make it similar to the temperature conditions being tested . Check that the refract
meter is working properly with pre-distilled water . Turn the refract meter screw and mark
the number of dashes on the instrument at the intersection of the black and white
lights . Measure one sample 3 to 5 times to obtain averages.
2.2.4 Determination of total hydrocarbons by phenol-sulfuric acid reaction
Principle : In hot acidic environments, glucose dehydrates to form hydro xymethyl furfural. This
form forms a brownish-yellow compound with phenol and measures light absorption at 490 nm.
Tools and reagents used: 5% phenol, 96% sulfuric acid, 1 mg / ml glucose , UV 1100
spectrophotometer
Procedure : Standard glucose: 20 mg of solution at a concentration of 1
mg / ml ; 40; 60; 80 ; Prepare working solutions at a concentration of 100 μg / 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 carbohydrates. Add 5 ml of

23. concentrated sulfuric acid solution, allow to stand at room temperature for 10 min and leave at
30 ° C for 20 min. Measure the absorbance at 490 nm.
2.2.5 Infrared spectroscopic determination ( IRS )
Principle of the method : Although many physical methods can be used to study the molecular
structure of organic matter, 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
the spectroscopy. The instrument used to determine the absorption spectrum of any compound is
called the NPT 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 better atmosphere
mortar pumped intake 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 .
2.2.6 Determination of protein amino acids by the hole method
Principle : to extract the protein hydrolysis used to suppress the coefficient of amino acids
stationary phase as a solution of peptide based distinguish the separation of the acid. In addition
to the use of hydrochloric acid, sulfur and other minerals, alkalis, and enzymes are often used to
break down proteins. By hydrolysis, 6 hydrochloric acids 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, cysteine 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 that loses the amino acids.
Reagents used: Adjustable thermostat, amino acids for protein hydrolysis, 6 n hydrochloric acid

24. 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
Standard amino acids : In addition to the amino acids present in the amino acid collection (a set
of amino acids produced in the USSR ) . Cystine, threonine, proline, and tyrosine are available in
pure form for analysis. Depending on the amino acid distribution coefficient , the preparation
was prepared for one of 0.1 N hydrochloric acid and 10% isopropyl alcohol, assuming a
concentration of 0.01 m in 2-3 mixtures .
Process : a thoroughly cleaned 50 mg protein ( 1-2mg ) of the test weight on the measure took
sealing tubes ( ampule ) 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 ) 100-105 and put in 24
hours 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 and evaporate to dissolve the dry residue in 4 ml of 10% trace propyl
alcohol . To dissolve, first rinse with 1-2 ml of alcohol, 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 upper clear solution is used for analysis , as the hemic substance settles on the
bottom of the fingers .
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 hand and the handle, and mark the dots at a distance of 2-2.5 cm from the starting
line. At these points, 20 ml of test solution and 5-40 μl of standard solutions containing 0.05-0.4
μl of amino acids are instilled and dried . 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 dihydric, heated on a wet surface and clarified the stain.
Analyze the amino acids by calculating the Rf .
Rf =
𝑡ℎ𝑒 𝑝𝑎𝑡ℎ 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑎𝑚𝑝𝑙𝑒
𝑡ℎ𝑒 𝑝𝑎𝑡ℎ 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑜𝑙𝑣𝑒𝑛𝑡

25. 2.2.7 Method for microbiological analysis to determine the number of bacteria
1. Scope
The instructions for this test method can be used for microbiological analysis to
determine the total number of bacteria in all products.
Nutrients, media and reagents used
 Nacl
 Distilled water
 PCA media
Tools and equipment used
Use the basic equipment and tools used in a microbiology laboratory.
 Wash the flask
 Shaker
 Shake bag
 Alcohol den
 Pipit
 Electronic weight
 Peter's cup
 Spatula
 Vortex
 Tubes
 Dimensional cylinder 500 ml
 Thermostat
 Colon counter
Sample preparation
Dilute 25 g of the test substance in a 225 ml flask, dilute with sterile saline, place in a shaker bag,
place in a shaker and shake well for 10 s to prepare a homogeneous suspension.
Analysis process Scheme 1

26. 2.2.8 Detection of Salmonella pathogen in the sample
1. Scope
The standard method instructions for this test method include Salmonella in all
products . Used for microbiological analysis to detect ssp .
2. Characteristics of Salmonella bacterial culture
Gram-negative, non-spore-forming, rod-shaped, motile. On the surface of the selected
nutrient medium grows a convex column with a white flat edge with a black center.
It breaks down glucose, does not break down lactose and sucrose, and produces
hydrogen sulfide.
3. Medium and reagents used
 Buffer peptone water
 Salmonella Shigella agar, Xylose Lysine Dioxylate agar
 Rapport Vassiliadis soup
 Three iron sugar agar
 API 20E test
 Semi-liquid agar
 Gram paint

27. 4. Tools and equipment used
The basic equipment used in the microbiology laboratory will be used.
 Conical flask shaker
 Shake bag
 Alcohol den
 Pipette
 The electronic scale has an accuracy of 0.01 grams
 Peter's cup
 Vortex
 Tubes
 Dimensional cylinders
 Thermostat
 Colon counter
Test process Scheme 2
+
agar agar
utrient agar
Triple sugar iron agar
A 0E
1 hr
4 4 hr
4 hr
1 hr
1 hr

28. Assumptions about the outcome
№ Specifications Whey Whey protein
1 Ash
2 Moisture
3 Protein
4 Total fat
5 Macro and micro element
content
6 Antibacterial activity

29. Schedule
№ Period (in months )
Work to be done lV V 6-8
months
l ll lll lV V
1 Read press reviews
2 Approval of
methodology
3 Sample collection
and initial testing
4 Sample preparation
5 Analyze
6 Discuss theoretical
seminars
7 Attend scientific
conferences

30. Used press overview
[1] . http://www.mofa.gov.mn Ministry of Food and Agriculture
[2]. R.Indra, M.Narangerel " dairy 2016
[3]. D.Urtnasan "Milk Biochemistry" UB 2012
[4]. .Tsendsuren, Ts.Erdenebileg “Traditional and modern advanced technology of
biological resource use technology” UB 01
[5]. N.Tsevegsuren, D.Purev, B.Munkhjargal , B.Tuyaagerel, B.Myagmarsuren “Food
chemistry” UB 2005
[6]. “ Ecological and food security” 00
[7]. L.Damdinsuren “ Microbiological and technological research of Mongolian cheese”
[8]. S.Tsendsuren, Kh.Altangerel, Ts.Dorjsuren, Z.Erdene, B.Badamsuren
[9]. . ansalmaa “ Technology of processing raw materials of animal origin” 000
[10]. .Tsetsgee, E.Enkhtsetseg “ lant food chemistry” UB 00
[11]. Food microbiology and biotechnology ”UB 01
[12]. https://www.slideshare.net/noonii/biological-valueble-mil k
[13]. Daniel WDWest , Sydney Abou Sawan , Michael Mazzulla , Eric Williamson,
Whey Protein Supplementation Enhances Whole Body Protein Metabolism and
Performance Recovery after Resistance Exercise 2017
[14 ]. D.Purevee “ Innovative technology based on Mongolian tradition of milk and dairy
products ” UB 010