INVESTIGATORY

PROJECT
TOPIC: Casein in different samples of
milk.

Philipose alexander
Harikrishnan KC
Vignesh bhaskar
Aswin saji
 CONTENTS

1. Objective
2. Introduction
3. Applications
4. Aim
5. Theory
6. Apparatus Required
7. Chemicals Required
8. Procedure
9. Observations
10. Result
11. Precaution
12. Bibliography
 OBJECTIVE
To study the quantity of Casein present in different
samples of milk.

Casein
 INTRODUCTION
Milk is a complete diet as it contains proteins,
carbohydrates, fats, minerals, vitamins and water. The
average composition of milk from different sources of
milk is given below:
SOURCE WATER MINERALS PROTEINS FATS CARBOHYDARTES
OF MILK (%) (%) (%) (%) (%)
COW 87.1 0.7 3.4 3.9 4.9
HUMAN 87.4 0.2 1.4 4.0 4.9
GOAT 87 0.7 3.3 4.2 4.8
SHEEP 82.6 0.9 5.5 6.5 4.5

Casein is the most predominant phosphoprotein found

in milk and cheese. When coagulated with rennet,
casein is sometimes called Paracasein.
Casein is not coagulated by heat. It is precipitated by
acids and by rennet enzymes, a proteolytic enzyme
typically obtained from the stomach of calves. The
enzyme trypsin can hydrolyse off a phosphate
containing peptone.
Casein consist of a fairly high number of praline
peptides, which do not interact. There are also no
disulphide bridges. As a result, it has relatively little
secondary structure or tertiary structure. Because of
this, it cannot denature. It is relatively hydrophobic,
making it poorly soluble in water. It is found in milk as
suspension of particles called casein micelles which
show some resemblance with surfactant-type micelle
in a sense that the hydrophobic parts reside at the
surface. The casein in the micelles are held together by
calcium ions hydrophobic interactions. These micelles
have negative charge on adding acid in milk the
negative charges are neutralised.
Ca2+-Caseinate + 2CH3COOH (aq) →Casein + (CH3COO)2Ca (aq)

The isoelectric point of casein is 4.7. The purified

protein is water insoluble. While it is also insoluble in
neutral salt solutions, it is readily dispersible in dilute
alkali and in salt solution such as sodium oxalate and
sodium acetate.
 APPLICATIONS
In addition to being consumed in milk, casein is used in
the manufacture of adhesive, binders, protective
coatings, plastics, fabrics, food additives and many
other products. It is commonly used by body builders
as a slow-digestive source of amino acids as opposed
to the fast-digestive when protein, and also as an
extremely high source of glutamine. Another reason it
is used in body building is because of its anti-catabolic
effect, meaning that casein consumption inhibits
protein breakdown in the body. Casein is frequently
found in otherwise non-dairy cheese substitutes to
improve consistency especially when melted.
 AIM
To study the quantity of casein in different samples of
milk.

THEORY
Milk contains 3% to 4% casein suspended in water in
the colloidal form. It is precipitated in a weekly acidic
medium.
 APPARATUS REQUIRED
Funnel
Funnel stand
Glass rod
Filter paper
Weight box
Test tubes

CHEMICALS REQUIRED
Different samples of milk.
Saturated ammonium sulphate solution.
1% acetic acid solution.
 PROCEDURE
1. Wash the beaker with the distilled water and dry it
2. Take 20ml of milk in the beaker and find its
weight.
3. Add 20 ml saturated solution of ammonium
sulphate slowly with stirring. Fat and casein will
separate out as precipitate.
4. Filter the above solution and transfer the
precipitate in another beaker.
5. Treat the above precipitate with 30ml distilled
water. Casein dissolves forming milky solution
whereas fat remains as such.
6. Warm the above contents of the beaker to 40-
45oC on a low flame. Now, add 1% acetic acid
solution drop wise with stirring when casein gets
precipitated.
7. Filter the precipitated casein and wash with
distilled water and dry it.
8. Find the weight of dry precipitate.
9. Repeat the whole experiment with other samples
of milk.
 OBSERVATION
Volume of milk taken in each case = 20 ml

Weight of milk taken = w1 g

Weight of casein isolated = w2 g

Percentage of casein = (Weight of Casein/Weight of
Milk) x100

Serial Type of Volume Weight Weight Percentage

No. Milk of Milk of Milk of of Casein
taken (w1 g) Casein
(ml) (w2 g)
1. 20
2. 20
3. 20
 THEORY
Milk contains 3% to 4% casein suspended in water in
the colloidal form. It is precipitated in a weekly acidic
medium.