INVESTIGATORY

PROJECT
Study of the Quantity of
Casein Present in Different
Samples of Milk
Student Name: SHRUTI.T
Class: XII-“C”
Roll No.:12333
School Name: Jain Vidyaashram
Submitted To:
Subject: Chemistry
Date:

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Study the quantity of Casein in present in Different
sample of milk
AIM-
To determine the quantity of casein present in different milk samples (e.g., cow
milk, buffalo milk, goat milk, toned milk)

INTRODUCTION-
MILK-
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 are able to digest other types of food.
Early-lactation milk contains colostrums, which carries the mother's antibodies to
its young and can reduce the risk of many diseases. 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. Dairy farms produced about
730 million tones 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 as a
whole contains water, minerals (Ca, K, Na and trace metals), vitamins (A, D, K),
carbohydrates, proteins and fats. Average composition of milk from different
sources is given ahead:-

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CASEIN-
Casein (from Latin caseus "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. The j 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 Quantity of Casein, in different Sample of Milk
casein supplies amino acids, carbohydrates, and two essential elements, calcium
and phosphorus. Casein contains a high number of proline residues,which do not
interact. There are also no disulfide 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 sub micelles, the periphery consisting of micro vellosities 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 iso
electric point of casein is 4.6. Since milk's pH is6.6, casein has a negative charge in

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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. The enzyme trypsin can hydrolyze a
phosphate-containing peptone. It is used to form a type of organic adhesive.

Ca2+-Caseinate + 2CH3COOH(aq.)→Casein(s)+ (CH3COOH)2Ca

Types of Casein Proteins

There are four main types of casein, each with specific functions:

Approx. % in
Type Role
Milk

α_s1-
40%–45% Major structural component
casein

α_s2-
8%–10% Calcium-binding
casein
β-casein 25%–35% Surface-active, stabilizes micelles
Stabilizes micelles; prevents
κ-casein 8%–15%
aggregation

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Biological Importance of Casein

1. Nutritional Source of Protein

 Casein is a complete protein, meaning it contains all 9 essential amino

acids.
 It is the main protein in milk, making it a vital nutrient for infants and
growing children.
 It supports muscle growth, tissue repair, and immune function.

2. Efficient Calcium & Phosphorus Carrier

 Casein forms complexes with calcium and phosphate ions via its
phosphoserine residues.
 These complexes are stable and help in the transport and absorption of
calcium and phosphate in the intestines.
 This is essential for bone development and maintenance.

3. Milk Micelle Formation

 Casein forms micelles in milk, which:

o Keep calcium phosphate suspended.
o Maintain the colloidal stability of milk.
o Make milk white and smooth in texture.
 These micelles are easily digested by enzymes in the stomach.

4. Slow Digestibility

 Casein clots in the stomach into a gel-like substance.

 This leads to slower digestion, providing a sustained release of amino
acids.
 This makes it ideal for:
o Night-time consumption (slow muscle feeding)
o Long-term satiety in meal replacement products

5. Supports Infant Growth

 In human and animal milk, casein provides bulk protein nutrition during
early development.

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  Plays a role in developing the gut microbiome and immunity in infants.

6. Bioactive Peptides from Casein

 When digested, casein releases bioactive peptides with beneficial effects:

o Opioid peptides (e.g., casomorphins) – may help regulate digestion
and stress responses.
o Antihypertensive peptides – may help lower blood pressure.
o Antimicrobial peptides – support immune defense.

7. Used in Nutraceuticals and Supplements

 Due to its slow-digesting nature, casein is used in:

o Protein supplements
o Medical nutrition for patients with wasting diseases (e.g., cancer,
HIV)
o Infant formulas

Applications of Casein
 Casein has a wide range of applications across various fields due to its
nutritional, chemical, and physical properties. In the food industry, casein
plays a crucial role in the production of dairy products such as cheese,
paneer, and yogurt, where it coagulates to form the solid structure of these
foods. It is also added to infant formulas as a major protein source to mimic
the nutritional value of human milk. Casein is commonly used in protein
supplements, especially for athletes, because of its slow-digesting nature,
which provides a sustained release of amino acids.

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 In industrial applications, casein has been historically used in the
production of adhesives and glues, such as those used in woodworking and
bottle labeling. It is also a component of casein-based paints, which are
eco-friendly and known for their durability and matte finish. Before
synthetic plastics became widespread, casein was used to manufacture
plastic-like materials such as galalith, used in buttons, combs, and pens.
Additionally, it is used in paper coatings and textile sizing to improve
surface properties and strength.
 In the pharmaceutical field, casein is used in nutritional supplements for
patients with protein deficiencies or chronic illnesses. It also serves as a
carrier in controlled drug delivery systems due to its biocompatibility and
slow-release properties. Furthermore, bioactive peptides derived from
casein digestion have antimicrobial, antihypertensive, and immune-boosting
effects. In scientific research, casein is used as a model protein in enzyme
studies and as a blocking agent in immunoassays like ELISA and Western
blotting to prevent non-specific binding.
 Overall, the diverse applications of casein in food, industry, medicine, and
science highlight its significance as more than just a milk protein — it is a
valuable multifunctional biomaterial.

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PRINCIPLE-
The unique property of casein is that it remains suspended in milk at
normal pH (~6.6) but becomes insoluble when the pH is adjusted to its isoelectric
point (around pH 4.6). At this specific pH, casein molecules lose their net
charge, which causes the micelles to destabilize and precipitate out of solution.
This is achieved by carefully adding a dilute acid like acetic acid (vinegar) or
citric acid, which lowers the pH of milk without denaturing other components
significantly.

Once casein is precipitated, it can be separated from the rest of the liquid (whey)
by filtration or centrifugation. The solid casein residue is then washed, dried,
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and weighed. This mass represents the amount of casein present in the sample. By
applying this method to various milk samples—such as cow’s milk, buffalo’s
milk, toned milk, and packaged milk—a comparative analysis of the casein
content in different types of milk can be performed.

This simple but effective principle demonstrates how protein precipitation at the
isoelectric point can be used as a biochemical technique for protein isolation and
estimation. It is also the fundamental principle behind processes such as cheese
and paneer production, where casein forms the structural base.

MATERIALS REQUIRED:-
•Conical flask

• Beakers

• Funnel

• Measuring Cylinder (100 ML)

•Watch glass

•Filter paper

•1% acetic acid

•Different samples of milk

•Glass rod

Chemicals:

Different sample of milk

 Ammonium Sulphate solution

 1% Acetic acid solution

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PROCEDURE:-
1. A clean dry beaker has been taken follow by putting 20 ml of cow milk into
it on adding 20 ml of saturated ammonium sulphate solution slowly and with
string fat along with casein was precipitated out.
2. The solution was filtered and transfered 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 dropwise and when casein gets 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 occure in other animal milk.
Animal Name Quantity Acetic acid Ammonium
Sulphate

Cow 20ML 4grams 20ML

Buffalo 20ML 5grams 20ML

Goat 20ML 19grams 20ML

Warm 20ML 18grams 20ML

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Note: The goat milk this process not properly work in this project In this process
firstly add acetic acid mix it well and the heat them PPT get form

Calculation:
Let them ammonium sulphate present in powder form then convert in dilute
solution.

The conversion of powder to dilute solution

According to Google

800ml →761 gram

200ml→?

Then,

761×20/800=19.025 gram

20ml=19.025gram

OBSERVATION
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 Animal Name Wt of Casein Wt of Casein
(HEATED MILK) (Normal Milk)

Cow 1grams

Buffalo 0.8grams

Goat 1.4 grams 1.2grams

RESULT-
According to our analysis of various samples of milk, we conclude that:

Goat milk heated wt of casein 1.4 gram.

Goat milk (normal) wt of casein 1.2 gram.

CONCLUSION-
The experiment was successfully conducted to estimate the quantity of casein in
different samples of milk, including cow milk, buffalo milk, goat milk, and

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packaged milk. The results clearly showed that buffalo milk contains the highest
amount of casein, followed by cow milk, packaged milk, and goat milk. This
variation is due to the differences in nutritional composition and fat content of milk
from different sources. The simple acid precipitation method used proved to be
effective in isolating casein. This study highlights the nutritional value of milk
proteins and their importance in the human diet as well as industrial applications
such as cheese production, dietary supplements, and pharmaceuticals.

PRECAUTION-
1. Use fresh milk samples to ensure accurate results.
2. Always heat the milk gently; avoid boiling, as it may denature the proteins.
3. Add acetic acid slowly and with constant stirring to prevent splashing and
ensure even precipitation.
4. Use clean and dry apparatus to avoid contamination of samples.
5. Filter the precipitate carefully to avoid loss of casein during transfer.
6. Wash the precipitated casein properly with distilled water to remove
impurities.
7. Allow the casein to dry completely before weighing to get correct mass.
8. Handle acids with care; wear gloves or safety glasses if available.
9. Label each milk sample clearly to avoid confusion during analysis.
10.Perform the experiment in a well-ventilated and safe lab environment.

Viva Question
Q: What is casein?
A: Casein is the major protein found in milk, making up about 80% of the total
protein content. It is a phosphoprotein.

Q: What is the isoelectric point of casein?

A: The isoelectric point of casein is around pH 4.6, where it becomes insoluble
and precipitates out of solution.

Q: How does casein exist in milk?

A: Casein exists in milk in the form of micelles – spherical structures made of
multiple types of casein molecules stabilized by calcium phosphate.
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Q: Why does casein precipitate when acid is added to milk?
A: Acid lowers the pH to the isoelectric point of casein, causing it to lose its
charge and become insoluble, resulting in precipitation.

Q: Which acid is commonly used to precipitate casein from milk?

A: Acetic acid (vinegar) or citric acid is commonly used.

Q: Which type of milk contains the highest amount of casein?

A: Buffalo milk typically contains more casein compared to cow or goat milk.

Q: What are the types of casein found in milk?

A: Alpha-s1, alpha-s2, beta, and kappa casein.

Q: What is the role of kappa-casein in milk?

A: Kappa-casein stabilizes the micelle structure and prevents the aggregation of
casein micelles.

Q: How can you separate casein from milk?

A: By heating milk and adding acid to precipitate casein, then filtering, washing,
drying, and weighing it.

Q: Mention one industrial use of casein.

A: Casein is used in the production of cheese, adhesives, paints, and protein
supplements.

Bibliography

1. NCERT Chemistry/Biology Textbook – Class 12

2. Modern ABC Practical Book
3. Internet sources: Wikipedia, NCERT.nic.in, research articles
4. Lab manual references and teacher guidance
5. https://arihantbooks.com/laboratory-manual-chemistry-class-xii.
6. https://en.wikipedia.org/wiki/Casein.
7. https://en.wikipedia.org/wiki/Milk.
8. https://www.slideshare.net/dineshpol/amount-of-casein-in-milk.
9. https://www.slideshare.net/Neelanjyan/study-of-quantity-of-caesin-present-
in-different-samples-of-milk-54726663.

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