25ml of 1.

1(v/v) mixture of diethyl ether and ethanol were added to the residues
and themixture was stirred for 5minutes to dissolve additional fat.

The weight of solid and the percent yield (g per liter of milk ) and comparison with
thetheoretical value of 3.5% ,even the percentage recovery by comparing these two
valuesand analyze the amount of protein lost during the extraction process.

Lastly several test were performed the biuret test and the xanthoproteic acid test
The Biuret test;

A pea sized amount of prepared casein was added to a test tube and dissolved in
4ml of distilled water and mixed thoroughly with 4ml of 3M sodium hydroxide
solution .1%CUSO
4
solution while swirling one drop at a time and stop after 10drops
addition.Xanthoprotein Acid Test

A pea sized amount of prepared casein was added to a test tube and dissolved it in
4 ml of distilled water followed by addition of 1ml of concentrated nitric acid the
mixture waswarmed in hot water then cooled in a steam of cold water tap then
lastly 3M of sodiumhydroxide were added to the mixture. A positive test is when the
yellow color deppeningto orange is appearing.we have to make sure that the entire

tube does not turn to orange.we was also looking on pieces of precipitate on the
wall of the test tube.
5.RESULTS
Percentage yield=weight of casein sample yield x100%original weight of casein
sample=1g/100ml x100%104.7/100mlPercentage yield = 0.955

1.0%

Percentage recovery=theoretical value

calculated value= 3.5% -1.0%=2.5%Percentage recovery is =2.5%

Amount of protein lost obtained by=Percentage yield x weight of milk sample used=
2.5/100 x 104.7g=2.6175gAmount of protein(casein ) lost during extraction is equal
to 2.6175 g
Biuret test Xanthoprotein Test
violet (the complex color) yellow coloredThe appearance of these colors was
showing the positive reaction means the presence of Casein
6.DISCUSSION
Due to error made during vaccum filtration we backed with a very small amount of
casein 1%,according to the literature it`s normally between 3.0-3.5 % in milk Biuret
test

While performing the general test for the presence of protein by using the Biuret
Test. BiuretsReagent contains copper ions. These copper ions formed a complex

with the nitrogens andcarbons of the peptide bonds in an alkaline solution causing
the pale blue color of Cu
+2
tochange to violet (the complex color). The violet color was a positive reaction in a
Biurettest. Proteins give a strong biuret reaction because they contain a large
number of peptide bonds.Xanthoprotein TestProteins and amino acids that contain
phenyl rings formed a yellow colored compound whenconcentrated nitric acid was
used. The yellow colored product upon the addition of nitric acid is

the test for the presence of tyrosine and tryptophan (phenyl rings) in a protein. The
yellow stainson skin caused by nitric acid are the result of the xanthoprotein
reaction.
7. CONCLUSION
Caseins have an appropriate amino acid composition that is important for growth
and development of thenursing young. This high quality protein in cow milk is one of
the key reasons why milk is such animportant human food .Many people with an
allergy to milk are actually allergic to one of twocomponents of the milk: the casein
or the lactose. Removal of these products particularly thecasein allows individuals
with allergies to tolerate milk. The knowledge gained from this practical is
absolutely helpful people who can`t eat immoderately milk to be able to consume
itafter the isolation of casein. Casein-free dairy products are also used for children
with autism.caseins are highly digestible in the intestine and are high quality amino
acids .Most whey proteinsare relatively less digestible in the intestine, although all
of them are digested to some degree. When substantialwhey protein is not digested
fully in the intestine, some of the intact protein may stimulate alocalized intestinal
or a systemic immune response. This is sometimes referred to as milk protein
allergy andis most often thought to be caused by -lactoglobulin. Milk protein
allergy is only one type of food proteinallergy.
8. REFERENCEa)

www.ehow.com/fact743252-isolation_casein_html/accessed January,3,2013,12:34
PM
b)

laclone.blogspot.com/2011/12/isolation of casein from milk accessed

January,3,201312:45 PM
c)

redalyc.uaemex.mx/src/inicio/( isolation, molecular and biochemical characterization

of goat milk casein and its fraction) accessed January,3,2013 ,4:31PM
d)

http://facstaff.bloomu.edu/mpugh/Experiment% Isolation and characterization of

caseinfrom milk 2011.accessedJanuary,3,2013 accessed: January,3,2013
e)

McMaster University, Chem2o6 Lab Manual,

1997,http://www.chemistry.mcmaster.ca/

chem2o6/labmanual accessed /January,3,2013