INDEX
S. No. CONTENT PAGE NUMBER
1. ABSTRACT 2
2. INTRODUCTION 3
3. CASEIN 4
4. STRUCTURE OF CASEIN 5
5. COMPOSITION AND USES OF CASEIN 6-9
6. APPLICATIONS 10-11
7. EXPERIMENT 12
8. PROCEDURE 13
9. OBSERVATION 14-15
10. PRECAUTION AND ANALYSIS 16
11. RESULT 17
12. CONCLUSION 18
13. BIBLIOGRAPHY 19
1
ABSTRACT
Analysis of different samples of
milk to determination amount of
casein, protein, minerals,
carbohydrates, fats and water
content. Casein is a major
constituent in milk and is a mixed
phosphorous protein. Casein is
present in milk as a caseinate in
the form of micelles. Micelle has
negative charge and adding acid
to milk the negative charges are
neutralized. These substances
serve a vital role in our day-to-day
life expenditures, food, industries,
etc.,
Ca²+ + caseinate + 2CH3COOH (aq) = Casein + (CH3COO)2Ca
Addition of saturated ammonium sulphate solution to the milk casein was
precipitated out. Then add 30ml water heat the precipitate at 400C then add 1% acetic
acid drop wise precipitate is obtained and filter the precipitate, dry weigh. The weight
of precipitate gives the amount of Casein present in a milk. This procedure is repeated
for different samples of milk.
For analysis of amount protein present in milk done by adding the small amount of
calcium oxide to 5 drops of milk in a test tube and 3 drops of water check the sample
with litmus then heated the test tube in flame precipitate is obtained and filter the
precipitate, dry weigh. The weight of precipitate gives the amount of protein present in
milk. This procedure is repeated for different samples of milk. For analysis of fat
Butyrometer is used. It shows amount of fat and water present in milk.
Likewise amount of casein, protein, minerals, carbohydrates, fats and water content
is determined in different samples of milk. Cow, Buffalo, goat, Sheep, Horse, Camel
and Donkey milk. Out of these samples Cow milk have good for nutrition.
2
INTRODUCTION
Casein is the name of related phosphor
proteins. These proteins are commonly
found in mammalian milk, making up 80%
of the proteins in cow milk and between
20% and 45% of the proteins in human
milk. As a food source, casein supplies for
amino acids, carbohydrates and two
inorganic elements, calcium and
phosphorus. It contains many other nutrients
including protein and lactose. Interspecies
consumption of milk is not uncommon,
particularly among humans, many of whom
consume the milk of other mammals. As an
agricultural product, milk, also called dairy
milk, is extracted from farm animals during
or soon after pregnancy. Milk is a nutrient-
rich white liquid food produced by the
mammary glands of mammals. It is the
primary source of nutrition for infant
mammals (including humans who are
breastfed) before they can digest other types
of food. Early-lactation milk contains colostrum, which carries the mother's antibodies
to its young and can reduce the risk of many diseases.
Dairy farms produced about 730 million tons of milk in 2011, from 260 million dairy
cows. India is the world's largest producer of milk, and is the leading exporter of
skimmed milk powder, yet it exports few other milk products.
The ever-increasing rise in domestic demand for dairy products and a large demand-
supply gap could lead to India being a net importer of dairy products in the future. The
United States, India, China and Brazil are the world's largest exporters of milk and
milk products. China and Russia were the world's largest importers of milk and milk
products until 2016 when both countries became self-sufficient, contributing to a
worldwide glut of milk.
Throughout the world, more than six billion people consume milk and milk products.
Over 750 million people live in dairy farming households.
Milk contains water, minerals (Ca, K, Na and trace metals), vitamins (A, D, K),
carbohydrates, proteins and fats.
3
CASEIN
Casein (from Latin cases "cheese") is a family of related phosphor proteins (αS1, αS2,
β, κ). These proteins are commonly found in mammalian milk, comprising c. 80% of
the proteins in cow's milk and between 20% and 45% of the proteins in human milk.
Casein has a wide variety of uses, from being a major component of cheese, to use as a
food additive. The most common form of casein is Sodium caseinate. As a food source,
casein supplies amino acids, carbohydrates, and two essential elements, calcium and
phosphorus.
Structure of Casein:
Casein molecular structure:
4
STRUCTURE OF CASEIN
❖ It is a phosphoprotein, which has phosphate groups attached to some of the
amino acid side chains. Mostly these amino acids are serine and threonine.
❖ Casein is a mixture of at least three similar proteins, which differ
primarily in molecular weight and amount of phosphorus they contain
(number of phosphate groups).
❖ Casein is made up of the main 3 types of proteins are α-casein, β-casein, and κ-
casein.
❖ All casein proteins have different hydrophobic and hydrophilic regions along
the protein chain.
❖ α-Caseins are the major casein proteins. Its containing 8-10 phosphate groups,
❖ β-casein contains about 5 phosphate residues,
❖ β-casein it is more hydrophobic than a-caseins and κ-casein
❖ Because α-caseins and β-caseins are highly phosphorylated, they are very
sensitive to the concentration of calcium salts, that is, they will precipitate with
excess Ca2+ ion.
❖ Unlike other caseins, κ-caseins are glycoproteins, and they have only 2
phosphate group.
❖ Hence, they are stable in the presence of calcium ions, and they play an
important role in protecting other caseins from precipitation and make casein
more soluble forming casein micelles.
❖ Neither a nor the ẞ-casein is soluble in milk, singly or in combination.
5
COMPOSITION AND USES
Casein contains a high number of proline residues, which do not interact. There are
also non-di-sulfide bridges. As a result, it has relatively little tertiary structure. It is
relatively hydrophobic, making it poorly soluble in water. It is found in milk as a
suspension of particles, called casein micelles, which show only limited resemblance
with surfactant-type micelles in a sense that the hydrophilic parts reside at the surface,
and they are spherical. However, in sharp contrast to surfactant micelles, the interior
of a casein micelle is highly hydrated. The caseins in the micelles are held together by
calcium ions and hydrophobic interactions. Any of several molecular models could
account for the special conformation of casein in the micelles. One of them proposes,
the micellar nucleus is formed by several submicelles, the periphery consisting of
microvellosities of κ-Casein. Another model suggests the nucleus is formed by casein-
interlinked fibrils. Finally, the most recent Model proposes a double link among the
caseins for gelling to take place. All three models consider micelles. As colloidal
particles formed by casein aggregates wrapped up in soluble κ-casein molecules. The
isoelectric point of casein is 4.6. Since milk’s pH is 6.6, casein has a negative charge
in milk. The Purified protein is water insoluble. While it is also insoluble in neutral
salt solutions, it is readily dispersible in dilute alkalis and in salt solutions such as
aqueous sodium oxalate and sodium acetate.
USES
❖ Glue
Casein-based glues were popular for woodworking, including for aircraft, as late as
the de Havilland Mosquito. Casein glue is also used in transformer manufacturing
(specially transformer board) due to its oil permeability.
6
❖ Paint
Casein paint is a fast drying, water soluble medium used by artists. Case in paint
has been used since ancient Egyptian times as a form of tempera paint and was widely
used by commercial illustrators as the material of choice until the late 1960s when,
with the advent of acrylic. Paint, casein became less popular. It is still widely used by
scene painters, although acrylic has made inroads in that field as well.
❖ Medical and dental uses
Casein-derived compounds are used in tooth remineralization products to stabilize
amorphous calcium phosphate (ACP) and release the ACP onto tooth surfaces, where
it can facilitate remineralization.
7
❖ Protein Supplements
An attractive property of the casein molecule. Is its ability to form a gel or clot in the
stomach, which makes it very efficient in nutrient supply. The clot can provide a
sustained slow release of amino acids into the blood stream, sometimes lasting for
several hours.
8
CONTROVERSIES
❖ AUTISM
Although research has shown high rates of use of complementary and
alternative therapies for children with autism, including gluten and/or casein
exclusion diets, as of 2008 there is a lack of evidence for the efficacy of these diets.
A 2006 review of seven studies indicated that, although all reported benefits of
exclusive diets in reducing autism symptoms, all suffered design flaws, and there
was not enough evidence overall to justify recommending exclusion diets to
patients.
❖ CASEIN INTOLERANCE
Casein intolerance, also known as “milk protein intolerance,” is experienced
when the body cannot break down the proteins of casein. The prevalence of
casein allergy or intolerance ranges from 0.25 to 4.9% of young children.
Numbers for older children and adults are not known.
❖ A1/A2 BETA CASEINS IN MILK
According to Food Standards Australia New Zealand (FSANZ), “Milk contains
many types of proteins can be quite different in the milk from different breeds of
cows and in the milk from other animals. Of the six major protein types in Cow’s
milk, four are casein proteins and the other two are whey proteins. The caseins
usually make up about 80% of the protein in Cow’s milk. One of the major caseins
is beta-Casein. There are different beta casein types, But the most common are beta
casein A1 (milk high in this type is known as A2 milk). Certain breeds of cows,
such as Friesians, produce mostly A1 milk, whereas other breeds, such as
Guernsey’s, as well as sheep and goat, produce mostly A2 milk. Milk produced in
Australia and New Zealand is normally a mix of A1 and A2 milks.
❖ CANCER
Colin Campbell’s The China Study (2005), a book, describes a direct correlation
between casein administered to rats and the promotion of cancer cell growth when
exposed to carcinogens. Alfa-toxin a potent carcinogen) was administered to these
rats over a 2-week dosing period. The rats were given a 1 week post dosing period
before beginning the test (promotion period). During the promotion. Period, one
group of rats was put on a 5% casein protein diet and another group on a 20%
casein protein diet. None of the rats. On 5% casein protein developed foci,
precursors to cancerous cell growth, and every rat on 20% casein protein developed
the pre cancer foci. It should be noted that all test groups were fed a 20% casein
diet for a total of 5 weeks (2-wk acclimation, 2-wk dosing, 1-wk post-dosing) prior
to the 12-week promotion period to survive the initial aflatoxin B1(AFB1) dosing,
regard less of whether they were in the 5% or 20% test groups. Campbell has
performed additional studies using a range of different carcinogens and other
experimental animals and claims to have found a consistent correlation between
cancer growth and the amount of casein protein in diet.
9
APPLICATIONS
Edible Casein can be used in Food, Beverage, Pharmaceutical, Health & Personal care
products, Agriculture/Animal Feed/Poultry. Edible Casein is used in cheese making
and protein supplement such as in cream-based soups, sherbet, pudding and custard.
Casein is the principal protein of cow’s milk. It is the curd that forms when milk is left
to sour. It is the most used milk protein in the food industry and contains 21 amino
acids.
Edible acid casein is highly nutritional, low in fat and cholesterol, and flavorful,
making it ideal for medical and nutritional applications. Edible acid is used in coffee
whiteners, infant formulas, processed cheese, and for use in pharmaceutical products.
Hydrolyzed casein is casein that has been broken down partially or completely to its
constituent amino acids.
EDIBLE CASEIN USES AS FOLLOWS:
❖ In Food:
Casein can be used as nutritional supplements, thickener, emulsifier and
texture stabilizer in food such as in cream-based soups, sherbet, pudding and
custard.
❖ In Beverage:
Casein can be used as nutritional supplements, thickener, emulsifier and
texture stabilizer in beverage.
❖ In Pharmaceutical:
Casein can be used as intermediate in Pharmaceuticals.
❖ In Health and Personal care:
Casein used as Hair Conditioning Agent; Skin-Conditioning Agent –
Miscellaneous; ANTISTATIC; SKIN CONDITIONING in Cosmetics and
personal care products.
❖ In Agriculture/Animal Feed/Poultry:
Casein can be used in Agriculture/Animal Feed/Poultry feed.
❖ In Other Industries:
Casein can be used in ointments, paper, paints, glues, textiles, varnishes.
10
Application of casein
S.NO PRODUCT FUNCTION
1
Bakery Products (Bread, biscuits, Nutritional, sensory, emulsifier,
breakfast cereals, cake mixes, frozen dough consistency, texture, volume.
cakes, pastries)
2
Dairy Products (Imitation cheese, Fat and water binding, texture
coffee creamer, yogurt, milk enhancing, shredding properties,
beverages, butter like spreads, milk emulsifier, increase gel firmness,
shakes) nutritional and foaming properties.
3
Beverages (Drinking chocolates, Stabilizer, whipping and foaming
fruit beverages, cream liqueurs, properties, emulsifier, clarification,
wine and beer industry) reduce color and astringency
4
Dessert Products (Ice-cream, Whipping properties, impart body,
frozen desserts, mousses, instant texture, film former, emulsifier
puddings, whipped toppings)
5
Meat Products (Communicated Emulsifier, water binding, improves
meat products) consistency, releases meat proteins
for gel formation.
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EXPERIMENT 1
OBJECTIVE
To study the quantity of casein present in different samples of milk.
THEORY
❖ Natural milk is an opaque white fluid secreted by the mammary
glands of female mammal.
❖ The main constituents of natural milk are protein, carbohydrate,
mineral vitamins, fats and water and are a complete balanced diet.
Fresh milk is sweetish in taste.
❖ However, when it is kept for long time at a temperature of 5 degree it
become sour because of bacteria present in air. These bacteria convert
lactose of milk into lactic acid which is sour in taste. In acidic condition
casein of milk starts separating out as a precipitate. When the acidity in
milk is sufficient and temperature is around 36 degrees, it forms semi-
solid mass, called curd.
REQUIREMENTS
APPARATUS REQUIRED
❖ Conical flask
❖ Beakers
❖ Funnel
❖ Measuring cylinder (100 mL)
❖ Watch glass
❖ Filter paper
❖ 1% acetic acid
❖ Different samples of milk
❖ Glass rod
12
PROCEDURE
1.A clean dry beaker has been taken follow
by putting 20 ml of cow milk into it on adding
of saturated ammonium sulphate solution
slowly and with string fat along with casein
was precipitated out.
2. The solution was filtered and transferred
precipitate in another beaker. Adding about
30 ml water to precipitate. Only casein
dissolve in water forming milky solution
leaving fat undissolved.
3. The milky solution was heated to about 40
degrees Celsius and adds 1 % acetic acid
solution drop wise and when casein get
precipitated.
4. Filter and precipitate washed with water
and the precipitate was allowed to dry.
5.Weight the dry solid mass in previously
weight watch glasses.
6.Same process of occur in other animal
milk.
13
OBSERVATIONS
❖ Volume of milk taken in each case = 20 ml
❖ Weight of milk taken = W1g
❖ Weight of casein isolated = W2g
❖ Percentage of casein = Weight of casein/ Weight of milk x100
The yield of casein precipitated from the various milk samples of goat milk,
cow milk, buffalo, goat, sheep, horse and contains 7.8gm, 4gm, 6.4gm, 6.5gm, 3gm
respectively. Similarly, the milk samples availed from the market such as milk like
Arogya and Nandini was 1.19 gm and 1.088 gm respectively. This shows that the
casein precipitated from the cow milk contains more amount of casein protein
than the goat and buffalo milk samples. The lower amount of casein in the buffalo
milk may be due to the more fat content in it.
14
Amount of casein in different samples of milk has been studied systematically.
This shows that the casein precipitated from the cow milk contains more amount of
casein protein than the goat and buffalo milk samples etc.
15
PRECAUTIONS
❖ Handle apparatus and chemicals carefully.
❖ Add ammonium sulphate solution very slowly.
❖ Stir milk while adding chemicals.
❖ Do not disturb milk after adding ammonium sulphate solution and wait same
time for fat and casein to precipitate out.
ANALYSIS
❖ Buffalo milk provides high amount of calcium, a mineral needed for bone
development.
❖ Casein is a major protein found in milk, comprising about 89% of buffalo milks total
protein content.
❖ Cow milk helps in improving the metabolism of the body and therefore prevents
weight gain.
❖ Cow milk is a good source of protein and calcium, as well as nutrients including
vitamin, B12 and iodine.
❖ Goat milk is a powerhouse of protein, healthy fats, vitamins, iron and other essential
nutrients.
❖ Goat milk keeps your bones and teeth stronger, due to its high calcium content.
16
RESULTS
According to the literature, milk with high concentration of casein is more
beneficial as compared to low concentration. Before estimation, these samples were
stored in ice and filtered to remove suspended impurities. As reported, casein
concentration varies in milk samples ranging up to 80%. Casein in the form of
white amorphous solid was collected and dried in desiccator. The results showed
that sheep milk contains high amount of casein (g/100mL) 5.81 ± 0.01 with casein
percentage 5.64 ± 0.01 and 5.69 ± 0.021 casein (g/100mL) and 5.52 ± 0.021
casein percentage in un-boiled and boiled milk samples among natural milk.
Similarly, buffalo milk is also beneficial with casein concentration (g/100mL)
5.71 ± 0.03 and 5.62 ± 0.025 with casein percentage 5.54 ± 0.03 and 5.46 ±
0.025 in un- boiled and boiled milk samples respectively. Cow milk contains lowest
casein concentration (g/100mL) 4.91 ± 0.036 and 4.75 ± 0.025 with casein
percentage 4.77 ± 0.036 and 4.61 ± 0.25 among un-boiled and boiled milk
samples. In case of UHT treated milk (tetra pack) samples, Nestle milk pack
contains highest concentration of casein while Opler milk contain lowest
concentration
7.39 ± 0.021 and 5.36 ± 0.021 with casein percentage 7.18 ± 0.021 and
5.21 ± 0.026 respectively.
From the given results, it is confirmed that buffalo milk contains more casein as
compared to cow milk and sheep milk contain more casein as compared to goat
milk. Among all these natural milk samples, sheep milk is supposed to be more
beneficial both in boiled and un-boiled form. While both have low concentration of
casein as compared to UHT treated Nestle milk pack. Among tetra packs, Nestle milk
pack is supposed to be more beneficial as compared to Opler’s and Everyday milk.
Sheep milk > buffalo milk > goat milk > cow milk Nestle Milk pack > Everyday milk> Opler’s milk.
Casein concentration in all milk samples is simply expressed as
Nestle Milk pack >Everyday milk>Sheep milk>Buffalo milk >Goat milk>Opler’s milk>cow milk.
The results showed that sheep milk contains high amount of casein (g/100mL)
5.81 ± 0.01 with casein percentage 5.64 ± 0.01 and 5.69 ± 0.021 casein
(g/100mL) and 5.52 ± 0.021 casein percentage in un-boiled and boiled milk
samples among natural milk.
17
CONCLUSION
As casein is major protein present in milk, contains almost all essential amino acids
and important for digestion, growth and development.
From this study, it is concluded that sheep milk is more beneficial for humans as
compared to cow, buffalo and goat milk as it contains high amount of casein
(g/100mL) 5.81 ± 0.01 in un-boiled form and 5.69 ± 0.021 in boiled sample than
other natural milk samples.
The casein percentage in sheep milk is also highest 5.64 ± 0.01 & 5.52 ± 0.021 in
un-boiled and boiled milk samples respectively.
Similarly, Nestle Milk pack contain high casein amount (g/100ml) 7.39 ± 0.021 than
Opler’s (5.36 ± 0.02) and Everyday milk (6.4 ± 0.015) and supposed to be more
beneficial than other’s tetra packs available in market.
This study clearly indicated that the amount of casein precipitated from the cow milk
was higher than that of the other milk samples.
The quantitative analysis of casein precipitated from the various milk samples.
Provide the ample scope to the cottage cheese manufacture.
Thus, the cow milk is suitable for the best muscle. Growth and basic body building
achievements.
“Different samples of milk contain different percentage of casein.”
18
BIBLIOGRAPHY
• https://www.google.co.in/
• https://arihantbooks.com/laboratory-manual-chemistry-class-xii
• https://en.wikipedia.org/wiki/Casein
• https://www.slideshare.net/dineshpol/amount-of-casein-in-milk
• https://www.slideshare.net/Neelanjyan/study-of-quantity-of-caesin-present-in-
different-samples-of-milk-54726663
• http://chemiprojects.blogspot.com/?m=1
• https://www.scribd.com/
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