“AMOUNT OF CAESIN

IN MILK”

Submitted in partial fulfillment of the requirements for the award

of the Class XII Board Practical in Senior Secondary Education
Submitted By
MITHRESH M

REGISTER NO:
Under the Guidance of
Mr. KAMACHIYAPPAN P M. Sc., B.Ed., PhD

SRI CHAITHANYA TECHNO

SCHOOL
A Senior Secondary School Affiliated to Central Board of
Secondary Education (CBSE), New Delhi.
Affiliation No: 1931222
(2023-2024)
 Certificate

This is to certify that this report on “AMOUNT OF CAESIN IN MILK” is the bonafide

work of MITHRESH M (Register No: ) who carried out the project under my

supervision. Certified further, that to the best of my knowledge the work reported herein does

not form part of any other project report or assertation on the basis of which a degree or

award as conferred on an earlier occasion on this or any other certificates.

Mr. KAMACHIYAPPAN P Mr. RAUVURI SANJEEVA REDDY

Subject Teacher Principal

Submitted for the Project Work and viva-voce Examination held on

………………………

INTERNAL EXAMINER PRINCIPAL EXTERNAL EXAMINER

i
 DECLARATION

I hereby declare that the project entitled “AMOUNT OF CAESIN IN MILK” submitted to
Sri Chaitanya Techno School, Erode, in partial fulfilment of the requirements for the award of the
class XII Board Practical’s in Senior Secondary Education, is a record of original project work
done by me under the supervision and guidance of Mr. P. KAMACHIYAPPAN., Sri Chaitanya
Techno School, Erode.

PLACE: ERODE SIGNATURE OF THE STUDENT

DATE: ………………

ii
 ACKNOWLWDGEMENT

It is new elevation in my schooling history to undertake this project entitled -

“AMOUNT OF CAESIN IN MILK” which helped our project team to gain more knowledge.

I would like to express our deep gratitude and heartfelt thanks to our Principal Mrs. S.
Mr. RAUVURI SANJEEVA REDDY M.Sc., B.Ed., Sri Chaitanya Techno School, Erode, for
providing the facilities to complete this project work.

I would like to thank our Senior Lecturer Mr. GADDI ASHWINI KUMAR M.Sc.,
B.Ed., & Mr. GAJULA SATHISH KUMAR M.Sc., B.Ed., Sri Chaitanya Techno School,
Erode for his prolonged support, encouragement and motivation.
I owe my immense gratitude and highest respect with heartfelt and sincere thanks to our
project guide Mr. P. KAMACHIYAPPAN M.Sc., B.Ed., PhD and Mrs. C. Priya B. Tech,
who took keen interest on my project and guided me to complete my project work.

This project would not have completed without their enormous help and worthy
experience. Whenever I was in need, they were behind me.

Although, this report has been prepared with utmost care and deep routed interest.
Even then I accept respondent and imperfection.

I express my sincere thanks to my parents and friends who helped me to complete this
project successfully.

iii

iii
 TABLE OF CONTENTS

S.NO. CHAPTERS PAGE NO.

ABSTRACT 01

1. INTRODUCTION
02

2. AIM 03

MATERIALS REQUIRED
3. 03

4. THEORY
04

PROCEDURE
5. 05

6. OBSERVATION
06

7. PROPERTIES AND COMPARISION 07

8. RESULT
10

9. PRECAUTIONS
11

10. BIBILIOGRAPHY
11

iv
 ABSTRACT

Casein is a family of related phosphoproteins that are commonly found in mammalian milk.

Casein has a wide variety of uses, from being a major component of cheese to use as a food additive.

A small fraction of the population is allergic to casein. Casein intolerance also known as “milk

protein intolerance” is experienced when the body cannot break down the proteins of casein.

Supplementation of protease enzyme has been shown to help casein intolerant individuals digest the

protein with minimal adverse reaction. Casein protein provides the body with all of the amino acids

necessary to help build body muscle.

1
 INTRODUCTION
Milk is a complete diet as it contains proteins, carbohydrates, fats, minerals, vitamins and

water. The average composition of milk from different sources is given below:

SOURCE WATER MINERALS FATS CARBOHYDRATES

PROTIENS (%)
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 is found in milk as cheese. When coagulated
with rennet, casein is sometimes called Para casein. British terminology, on the other hand, uses the
term caseinogen for the uncoagulated protein and casein for coagulated protein. As it exists in milk, it
is a salt of calcium.
Casein is not coagulated by heat. It is precipitated by acids and by rennet enzymes, a
proteolytic enzyme typically obtained from the stomachs of calves. The enzyme trypsin can
hydrolyze off a phosphate containing peptone.
Casein consists of a fairly high number of praline peptides, which do not interact. 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 a suspension of
particles called as casein micelles which show some resemblance with surfactant type micellize in a
sense that hydrophilic parts resides at the surface. The caseins in the micelles are held together by
calcium ions and hydrophobic interactions. These micelles have negative charge and on adding acid
to milk the negative charges are neutralized.
Calcium caseinate + acetic acid → casein (s) + calcium acetate (aq)
The isoelectric point of casein is 4.7. The purified protein is water insoluble. While it is also
insoluble in neutral salt solution, it is readily dispersible in dilute alkalis and in salt solutions such as
sodium oxalate and sodium acetate.

2
AIM:

To study the quantity of CASEIN present in different samples of milk.

APPARATUS REQUIRED:

Funnel, funnel stand, glass rod, filter paper, weight box, test tubes, pestle and mortar.

CHEMICALS REQUIRED:

 Different samples of milk.

 Saturated ammonium sulphate solution.

 1% acetic acid solution

3
THEORY:

Milk contains 3 to 4% casein suspended in water in the colloidal form. It is precipitated in a

weakly acidic medium. Milk is a refreshing beverage and can be used as a base for other drinks such

as hot chocolate or specialty coffees. Milk is the main ingredient in puddings, custards and mousses

and is commonly included in recipes for soups, sauces and batters.

4
PROCEDURE:
1. Wash the beaker (250 ml) with the distilled water and dry it.

2. Take 20 ml of cow's milk in 250 ml 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 30 ml distilled water. Casein dissolves forming milky solution

whereas fat remains as such.

6. Warm the above contents of the beaker to 40 - 45°C 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 cow’s milk, goat’s milk and packet milk.

5
 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 x 100 / Weight of milk

Volume of Weight of Weight of Percentage

S. No Type of milk
milk taken milk taken casein of casein
1 Packet milk 20ml 46.35g 8.67g 18.70%
2 Cow’s milk 20ml 47.30g 10.8g 22.83%
3 Goat’s milk 20ml 46.55g 15.68g 33.68%

6
 PROPERTIES OF CASEIN

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 is a lyophilic colloid akin to albumin and gelatin. It is isoelectric at pH 4.6 where its

solubility in water is but 0.01 percent. It is amphoteric: below pH 4.6 casein forms moderately

soluble salts such as casein chloride; above pH 4.6 casein forms salts with bases. Sodium caseinate

and other alkali salts are soluble without limit, while calcium caseinate, other alkaline earth salts, and

heavy metal salts are nearly insoluble. Caseinates readily form gels when slowly coagulated from

concentrated solutions. Formaldehyde forms an insoluble compound with casein. Casein is insoluble

in most organic solvents. Paracasein is less lyophilic but otherwise identical with casein.

COMPOSITION OF CASEIN
Casein contains a high number of proline amino acids which hinder the formation of
common secondary structural motifs of proteins. 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
7
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. The enzyme trypsin can hydrolyze a phosphate-containing peptone. It is used to form a type
of organic adhesive.

8
 MANUFACTURE OF CASEIN

Casein is usually made from skim milk (rarely from buttermilk), by one of three methods:
(1) Naturally soured casein curdles when enough lactic acid develops from fermentation of milk
sugar by the ever present bacterium Streptococcus lactase
(2) Acid casein is precipitated by adding dilute hydrochloric acid or sulfuric acid
(3) For rennet casein, warm skim milk is set with rennet extract until the calcium
Para caseinate clots, after which the clot is cut into small pieces to allow the whey to drain.
In all three methods the whey is drawn off, the curd washed with water, drained or pressed, dried in
warm air, ground, and packed for sale. Rennet casein retains much of the calcium phosphate from the
milk.

9
 APPLICATIONS OF CASEIN

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 commonly
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.

RESULT

 Different samples of milk contain different percentage of casein.

 Highest percentage of casein is present in goat’s milk.

10
 PRECAUTIONS

1. Handle the 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 for some time for fat and
casein to precipitate out.
5. Take the amount readings carefully with digital weighing machine only.

BIBLIOGRAPHY

1. www.wikipedia.com
2. www.encyclopedia.com
3. www.casein-pro.com
4. www.sciencejournals.com
5. www.icar.nic.in
6. www.zetascience.com
7. www.scribd.com

11