Kendriya vidyalaya no.

1,PORT BLAIR

INTERNAL ASSESSMENT 2022-23

SUBJECT: CHEMISTRY

NAME : ISHA BASNET

CLASS : XII
SECTION : A
ROLL no. : 24

1
 declaration
I hereby declare that the project entitled
“DETERMINE THE AMOUNT OF CASEIN IN
DIFFERENT SAMPLES OF MILK” submitted by ISHA
BASNET for the internal assessment of subject
CHEMISTRY for AISSCE 2022-23, is the result of
original and independent research work carried out
under the guidance of
Mrs. ROSHNI SAMUEL
PGT (CHEMISTRY)

PRINCIPAL TEACHER

EXTERNAL EXAMINER

2
 Acknowledgement
I would like to take this opportunity to express special
gratitude to my biology teacher Mrs. ROSHNI SAMUEL
who gave me the wonderful opportunity to do this
investigatory project on the topic “DETERMINE THE
AMOUNT OF CASEIN IN DIFFERENT SAMPLES OF
MILK”.

The opportunity to participate in this project has helped

me improve my research skills and I am really greatful to
them.

I would also like to thank my family and friends for

constantly encouraging me during this project, which I
could not have completed without their support and
continuous encouragement.

3
 INDEX
S.NO. TITLE PAGE.NO.

1. INTRODUCTION 5-13

2. AIM 14

3. REQUIREMENTS 15

4. THEORY 16

5. PROCEDURE 17

6. OBSERVATION 18

7. CONCLUSION 19

8. BIBLIOGRAPHY 20

4
 INTRODUCTION
Casein is the main protein constituent of milk. It
constitutes about 80% of the total protein in cow’s
milk and about 3% of its weight. It group of protein
precipitated when the milk is slightly acidified. It
dissolves slightly in water, extensively in alkalis’ or
strong acids. Casein is a complete protein meaning
that it contains all of the essential amino acids,
which the body can not manufacture on its own.
When dried, it is a white, amorphous powder without
taste and odour. It is a mixed phosphoprotein and
occurs in milk as calcium salt (calcium caseinate) in
the form of micelle. The micelle has a negative
charge. When an acid is added to the milk, the
negative charges are neutralized.
Calcium caseinate + acetic acid → casein (s) +
calcium acetate (aq)
m in diameter. Milk composition varies with the stage
of location, age and breed. Milk is colloidal nature
due to the presence of proteins. The proteins are
heavy molecules; they form colloids when dispersed
in water medium. The primary function of protein in
living cells is to promote growth and maintenance.
The nitrogen content of milk is distributed among
casein 76%, when protein and non-protein nitrogen

5
is 6%.The structure of protein consist of a
polypeptide chain of amino acids joined together by
peptide linkages. Around the world, there are more
than six billion consumers of milk and milk products.
Over 750 million people live in dairy farming
households. It is used in paints for fast drying water-
soluble medium (Figure 1). Casein based glues are
formulated from the mixture of casein, water,
hydrated lime and sodium hydroxideThe quantity,
quality and fat-content from the various milk samples
differ with the type of particular mammals and their
fodder. The composition of milk varies with
according to the animals from which it comes,
providing the correct growth rate and development
for the young of that species. Casein is a slow
digesting protein and it was suspended in the milk in
a complex called micelle. The micelles are spherical
and are 0.04 to 0.03

6
 ▪ COMPOSITION OF CASEIN
Casein contains a fairly high number of praline
residues, which do not interact. There are also no
disulphide bridges. As a result hydrophobic, making
it purely soluble in water. It is found in milk as a
suspension of particles called “casein micelles”
which show only limited resemblance with
surfactant-type micelae 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. Several models 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 consists of
microvallosites of K-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 partcals
formed by casein aggregates wrapped up in soluble
K-casein molecules.

7
 ▪ USES
• PAINT: casein pain is a fast drying, water soluble
medium used by artists. Casein 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.
• GLUE: Casein based glues were popular for
woodworking, including for aircraft, as late as the
de Hacilland Mosquito. Casein glue is also used in
transformer manufacturing (specially transformer
board) due to its oil permeability. While largely
replaced by synthetic resins , casein-based glues
still have a use in certain niche applications, such
as laminating fireproof doors and the labeling of
bottles.

8
• CHEESE MAKING: cheese consists of proteins
and fat from milk, usually the milk of cows, buffalo,
goats or sheep. It is produced by coagulation of
casein. Typically the milk is acidified and then
coagulated by the addition of rennet, containing a
protrolytic enzyme, typically obtained from the
stomachs of calves. The solids are separated and
pressed into final form. Unlike many proteins,
casein is not coagulated by heat. During the
process of clothing, mik-clothing proteases act on
the soluble portion of the caseins, k-casein, thus
originating an unstable micellar state that results in
cloth formation. When coagulated with chymosin is
an aspartic protease that specially hydrolyses the
peptide bond in Phe105-met106 of k-casein, and
is considered to be the most efficient protease for
the chgeese-making industry. British technology,
on the other hand, uses the term caseinogens for
the uncoagulated protein and casein for the
coagulated protein. As it exists in milk, it is a salt
of calcium.
• PLASTIC AND FIBER- Some of the earliest
plastics were based on casein. In particular,
galalith was well known for use in buttons. Fiber

9
 can be made from extruded casein. Lanital, a
fabric made from casein fiber(known as Aralac in
the United States), was particularly popular in Italy
during the 1930s. Recent innovations such as
QMilch are offering a more refined use of the fiber
for modern fabrics.
• 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 is able to
provide a sustained slow release of amino acids
into the blood stream, sometimes lasting for
several hours.
• 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.

▪ 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

10
 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. 16
• 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
betacasein. There are different beta casein
types, but the most common are beta casein 17
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 Guernseys, as wall as sheep and goat,
produce mostly A2 milk. Milk produced in

11
 Australia and New Zealand is normally a mix of
A1 and A2 milks. The European Food Safety
Authority carried out a literature review in 2009
concluding “a cause and effect relationship is
not established between the dietary intake
BCM7, related peptides or their possible protein
18 precursors and non-communicable diseases”
. Studies supporting these claims have had
significant flaws, and the data are inadequate to
guide autism treatment recommendations.
• CANCER- T.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. Aflatoxin( a potent
carcinogen) was adminis- 19 tered to these rats
over a 2 week dosing period. The rats were
given a 1 week postdosing 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%

12
casein protein developed the precancer 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 dos- 20 ing, 1-wk post-
dosing)prior to the 12 week promotion period in
order to survive the initial aflatoxin B1(AFB1)
dosing, regardless 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. A 2001 study
suggests another milk protein, when protein,
may play a protective role against colon tumors
in rats. According to 21 a study from the
Australian Dairy Council, casein has
antimutagenic effects.

13
 AIM
To determine the
amount of casein
present in different
samples of milk.

14
 Requirements
APPARATUS
• 250ML BEAKERS
• FUNNEL, GLASS ROD
• PORCELAIN DISH
• CHEMICAL BALANCES
• TEST TUBES
• FILTRATION FLASK
• BURNER

CHEMICALS
• DIFFERENT SAMPLES OF MILK
• 1% OF ACETIC ACID SOLUTION
• SATURATED AMMONIUM SULPHATE
SOLUTION

15
 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 25 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.

16
 Procedure
• 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.
• Filter the solution and transfer the precipitates in
another beaker.
• Add about 30 ml of water to the precipitate.
• Only casein dissolves in water forming milky
solution leaving fat undissolved.
• Heat the milky solution to about 40˚ C and add 1%
acetic acid solution drop wise, when casein gets
precipitated.
• Filter the precipitate, wash with water and let the
precipitate dry.
• Weigh the dry solid mass in a previously weighed
watch glass.
• Repeat the experiment with other samples of milk

17
 OBSERVATION TABLE
Volume of milk taken in each case = 20 ml.

S.NO. NAME OF MILK WEIGHT OF

CASEIN PRESENT

1. COW 7.8

2. BUFFALO 4

3. GOAT 6.4

4. MARKET MILK- A 6.8

5. MARKET MILK-B 5.5

18
 CONCLUSION
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.
According to the research findings, cow milk
contains the largest amount of casein protein. 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. Although the mineral content of goat's milk
and cow's milk is generally similar, goat's milk
contains more calcium, potassium, iron, magnesium
and sodium. All milk has lots of casein but there are
different types of casein and for someone who has
casein sensitivity, goat milk may provide an
alternative to which they do not react.

19
 BIBLIOGRAPHY
[1] https://en.wikipedia.org/wiki/Casein
[2]http://www.milkfacts.info/Milk%20Composition/Mil
k%20Composition%20Page.htm
[3] Patricia Trueman, Nutritional Biochemistry, India,
2011.
[4] http://nem.org.uk/chem1.htm

20