BIOCHEMISTRY LABORATORY

EXPERIMENT #7

MILK

GROUP 7

MALREDDY

MENCHAVEZ

MONTALBO

MONTESEVEN

INSTRUCTOR

SIR RANEL LARINO

 MILK

I.

Milk is a fluid secreted by the mammary gland as food for the young mammal.
Consequently, milk is considered from the standpoint; first as a secretion and second as
a food. Milk is an emulsion of lipids in a solution of proteins, lactose and inorganic salts.
There are some organic acids or their salts, vitamins, enzymes, and some undetermined
constituents.

It is a white to yellowish-white fluid. The color is partly due to the calcium salt of casein,
and emulsified fat. The color is derived from pigments in the food which is dissolved in
fat. These are carotene and xanthophyll.

OBJECTIVES:

At the end of the activity, the students should be able to:

1. distinguish the physical and chemical properties of milk;

2. prepare casein from milk, and;

3. appreciate the importance of milk for the proper nourishment of the body.

II. Results and Discussion

A. Reaction with Acid-Base indicators

Indicators Color
Blue litmus paper Red
Red litmus paper Red
Congo red Blue
Phenolphthalein Colorless

How would you account for the different reactions of milk with the different indicators?

The color exhibited by both the litmus papers tells us that the sample was acidic, but not
acidic enough to invoke a change in Congo Red since as an indicator, Congo Red will
only change in color approx. at a pH of 3.0-5.0 and since milk ranges from 6.5-6.7 which
is only slightly acidic the change is not to be expected, and with the Phenolphthalein
being colorless, it only confirms that the sample is acidic.
B. Coagulation Test

Describe the result and explain why such result occurred.

Formation of white coagulum, it is due to the casein's unfolding which is brought about
by the lowering of the pH caused by the addition of acetic acid.

C. Film Formation

1.What was produced after heating few mL of milk in a small evaporating dish? To what
was the result due?

A film of protein was formed and it was caused by the denaturation of protein structures
and is accompanied with coagulation.

2. Give the composition of the film.

Clotted Proteins, Carbohydrates fused by Milk fats.

3. Enumerate the factors facilitating the formation of the surface film.

Stirring, Type of Milk and Temperature

4. What is milk?

White liquid produced by the lactation of mammals for nutrition and growth of their
young.

D. Action of Hot Alkali

Questions:

1.What was the resulting color after heating few mL of milk with drops of NaOH?

Yellowish Brown.

2. How do you account for the change in color?

The presence of lactose is indicated with the change in color.

3. What test in the study of carbohydrates involves the same principle?

Moore's Test.
E. Preparation of Casein

Questions:

1. Why should the addition of excess dilute HCL be avoided in the preparation of casein?

To reach Casein's isoelectric point, AN excess amount of acid will cause it's dissolution.

2. What is the purpose of treating the casein precipitate with alcohol and then with
ether?

To remove moisture with alcohol and to remove milk fat with ether.

3. Why is casein precipitate heated in a warm water bath without flame after the
addition of ether?

Ether is flammable and heating it would only mean a safety hazard.

4. How would you dry the casein precipitate?

The casein is dried by enclosing it between multiple sheets of Filter paper.

F. Tests for Casein

1. Solubility Test

Solubility Test Casein

Water Insoluble
Dilute HCL Insoluble
Dilute NaOH Insoluble
10% NaCl Slightly Soluble

2. Millon's Test

Describe the result of this test. What is the indication of the result?

Tyrosine was formed by a denatured protein and was proven true by it's positive result
with Millon's test and the formation of red ppt. Casein is conjugated phosphorylation

3. Biuret Test

The positive result for Biuret's Test is a violet solution, which indicates the presence of
proteins thus proving the presence of proteins in milk.
G. Detection of Lactose

Test reagent Result

Fehling's A & B solutions Formation of red ppt.
Benedict's test Formation of red ppt.

H. Detection of Milk Fat

1.Explain why the filtrate used in detecting the presence of milk fat is evaporated
spontaneously and not to be heated over a flame.

If milk is heated, it will be at risk of denaturation thus it is not advised so it is allowed to

evaporate spontaneously.

2. Describe the residue left after evaporation

There is a thick residue which consists of fats and proteins.

Conclusion:
Milk is one of the many essential substances for growth and development, it is slightly
acidic and is composed mainly of water and the rest of dry substance, it contains fat
globules and more essential proteins that are well required to aid in the rapid
development of growing organisms (Mammals). Milk also contains casein one of the
many essential amino acids.

References:
Brody T. (1999) "Calcium and phosphate". pp. 76194 in Nutritional biochemistry, 2nd ed. Boston: Academic
Press

Fox, P.F. Advanced Dairy Chemistry: Vol 2 Lipids. 2nd Ed. Chapman and Hall: New York, 1995.

Fox, P. F. Advanced Dairy Chemistry, Vol. 3: Lactose, Water, Salts and Vitamins. 2nd ed. Chapman and Hall:
New York, 1995.