CHEMISTRY

INVESTIGATORY PROJECT
Submitted for
ANNUAL EXAMINATION

2021-2022

INVESTIGATION OF AMOUNT OF CASEIN PRESENT

IN DIFFERENT SAMPLES OF MILK

Done By
M. SANJAY LAKSHMINARAYAN
XI-E
SCIENCE STREAM
SUBMITTED TO THE DEPARTMENT
OF CHEMISTRY
f

CERTIFICATE

This is to certify that, M. Sanjay Lakshminarayan

studying in class 11-E (science stream) during the

academic year 2021-2022 has satisfactorily completed the

investigatory project in chemistry a s s i g n e d to him

within the given time frame.

SUBJECT TEACHER VICE PRINCIPAL

SCHOOL SEAL PRINCIPAL

 ACKNOWLEDGEMENT

I would like to express my special thanks of gratitude to my chemistry teacher

Mrs. Joy Hepshebah for her guidance and support throughout the duration of the

project. I was able to complete the project successfully by her motivation and their

extended support .

I would also like to thank our Correspondent Dr. R.Kishore Kumar, our

Principal Mrs. Shanthi Samuel and Vice Principal Mrs. Ramya Uday who gave

me the golden opportunity to do this investigatory project, which helped me in

doing a lot of research and made me aware of so many new things.

I would also like to thank my parents and friends who helped me a lot in

finalizing this project within the limited time frame.

Above all I would like to thank God Almighty, who has given me the strength and

courage to do this project efficiently.

 CONTENTS

S.NO. TITLE PAGE NUMBER

1. COVER PAGE 1

2. CERTTIFICATE 2

3. ACKNOWLEDGEMENT 3

4. CONTENTS 4

5. ABSTRACT 5

6. INTRODUCTION 6-7

7. EXPERIMENTAL SETUP 8-9

8. OBSERVATION 10

9. CONCLUSION 11

10. BIBLIOGRAPHY 12
 ABSTRACT

The main objective of this investigatory project is to determine the amount of

casein present in different samples of milk.

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 is a complete balanced
diet.

Milk is a multinutrient fluid and it is the primary source of nutrition for

human. It consists of 80% of proteins. The protein in the milk is classified
into casein and whey protein. Milk protein consists of 80% of casein and
20% whey protein. The function of casein is to provide energy to human
body. The name of casein is related to the family of phosphoproteins. These
proteins are commonly found in the mammalian milk. This study deals with
the precipitation of casein from the various milk samples such as cow milk,
goat milk, buffalo milk and also the samples that availed from the market.
The technique of precipitation of casein is used to predict the protein content
in the milk samples.
 INTRODUCTION

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 degree, it forms
semi-solid mass, called curd. Casein is present in milk as calcium caseinate in
the form of micelles. These micelles have negative charge and on adding acid
to milk, the negative charges are neutralized.

Pure casein is an amorphous white solid without taste or odour. Commercial

casein is slightly yellow, with a pleasant odour. Dry casein keeps well if
protected from insects and rodents; damp casein is quickly attacked by molds
and bacteria and acquires a disagreeable odour. The specific gravity is 1.25 to
1.31. Casein is a mixture of phosphoproteins of differing molecular weight.
Casein A1 milk gets its name from A1 beta-casein. This has to do mostly
with breeds of cows that originated in Northern Europe. Casein A2 milk is
the healthier choice between the two kinds of milk because casein A2 milk
doesn’t usually bring about the same digestive issues and health concerns. 

CASEIN :

Molecular
C81H125N22O39P
Formula

Casein
9000-71-9
Synonym Caseinogen
s Casein from bovine milk
Bovine beta-casein monophosphopeptide

Molecular
2062.0
Weight

CHEMICAL STRUCTURE OF CASEIN

 EXPERIMENTAL SETUP

AMOUNT OF CASEIN IN MILK

MATERIALS REQUIRED:

1. 250 ml beakers
2. Funnel
3. Glass rod
4. Chemical balances
5. Test tubes
6. Filtration flask
7. Bunsen burner
8. Different samples of milk
9. 1% acetic acid solution
10.Saturated ammonium sulphate solution

THEORY:

Casein, like proteins, are made up of many hundreds of individual amino

acids.   Each  may have a positive or a negative charge, depending on the pH
of the [milk] system. At some pH value, all the positive charges and all the
negative charges on the [casein] protein will be in balance, so that the net
charge on the protein will be zero. That pH value is known as the isoelectric
point (IEP) of the protein and is generally the pH at which the protein is least
soluble. For casein, the IEP is approximately 4.6 and it is the pH value at
which acid casein is precipitated. In milk, which has a pH of about 6.6, the
casein micelles have a net negative charge and are quite stable. During the
addition of acid to milk, the negative charges on the outer surface of the
micelle are neutralized (the phosphate groups are protonated), and the neutral
protein precipitates. 

Ca+2 (Casemated) + 2CH3 COOH(aq) Casein(s)+(CH3COO)2 Ca (aq)

PROCEDURE:

1. Take a clean dry beaker, put into it 20cc of cow’s milk and add 20 ml
of saturated ammonium sulphate solution slowly and with stirring. Fat
along with casein will precipitate out.
2. Filter the solution and transfer the precipitates in another beaker.
3. Add about 30 ml of water to the precipitate.
4. Only casein dissolves in water forming milky solution leaving fat
undissolved.
5. Heat the milky solution to about 40˚ C and add 1% acetic acid solution
drop wise, when casein gets precipitated.
6. Filter the precipitate, wash with water and let the precipitate dry.
7. Weigh the dry solid mass in a previously weighed watch glass.
8. Repeat the 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

Weight of casein
% of casein = ---------------------- X 100
Weight of milk

Sample Source Weight of milk Weight of casein % of casein

No. W1 g W2 g

1 Cow Milk 18.17 0.62 3.1 %

2 Goat Milk 17.86 0.56 2.8 %
3 Buffalo Milk 18.1 0.66 3.3 %
4 Market Milk 14.2 1.19 8.38 %
CONCLUSION:

Different samples of milk contain different percentage of Casein. Cow milk

contains the largest amount of casein protein than goat milk and buffalo milk.
Thus, the cow milk is suitable for the best muscle growth and basic body
building achievements. It was found that goat milk contains the small amount
of casein protein. For someone who has casein sensitivity, goat milk may
provide an alternative to which they do not react.

RESULT:

The amount of casein precipitated from the Cow’s milk was higher than goat
milk
and buffalo milk samples.

PRECAUTION:

1. Handle apparatus and chemicals carefully.

2. Add ammonium sulphate solution very slowly.
3. Stir milk while adding chemicals.
4. Do not disturb milk after adding ammonium sulphate solution and wait
some time for fat and casein to precipitate out.
5. Take the amount readings carefully with digital weighing machine
 only.

BIBLIOGRAPHY

1. https://www.alamy.com/stock-photo/casein.html

2. https://pubchem.ncbi.nlm.nih.gov/compound/Casein

3. https://en.wikipedia.org/wiki/Casein

4. http://www.milkfacts.info/Milk%20Composition/Milk%20Composition%20Page.htm

5. http://nem.org.uk/chem1.htm

6. https://www.britannica.com

7. https://www.webmd.com

8. https://www.iosrjournals.org

9. https://www.researchgate.net
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