You are on page 1of 5



What are proteins

Milk (non-fat vs fat)
Isoelectric point and pH electrophoresis
Acid and Bases Neutralization

Biomolecules are organic compounds that compose all living cells thus being
essential to every living organism. There are four types of biomolecules that is
naturally produced by the body, and these are the proteins, carbohydrates, lipids
and nucleic acids. Among the four, only proteins, carbohydrates and lipids are
polymeric or linked by polymers.
Protein is one of the most common biomolecules that is naturally occurring and
can be found throughout the body such as in hair, skin, bone and muscles. It is
made up unbranched polymers, with monomers called amino acids and that are
linked together by peptide bonds. There are two types of protein fibrous and
globular. A fibrous protein tends to form long and strong repeated sets of amino
acid residues and usually forms intermolecular interactions between the side
chains of the residues while a globular protein has no systematic structure,
relatively spherical in shape, tends to fold back on themselves into compact units
and do not form intermolecular interactions between protein units.
The amino acids that make up the protein in the body is either made from
scratch, undergo a posttranslational modification or derived from the food we eat.
Amino acids that are derived from food or protein supplements are called the
essential amino acids. These essential amino acids can then be derived from
meat, fish, dairies, poultry and eggs, and must be eaten every day so that more
amino acids can make the protein that is needed by the body to function.
In order for the body function properly, nutrients and vitamins are needed and are
supplemented by our daily food and drink intake. Among the variety of food and
drinks, milk is one of the drinks that should be consumed daily as it contains
vitamins like histamine, riboflavin, pantothenic acid, A, D and K, minerals like
calcium, potassium, sodium, phosphorus, trace metals, proteins, carbohydrate
(lactose) and lipids. According to USDA (2013), there are about 23, 913 million
people consume milk everyday, and 36% of this population can consume up to
three glasses.



Retrieved from: Minard, R. (2000). Isolation of

Casein, Lactose and Albumin from Milk.

A. Isolation of Casein from Milk
In a 100-milliliter beaker, five grams of powdered non-fat milk was
added and was dissolved by adding 20 milliliters of warm distilled water.
After the powdered milk dissolved in warm water, the solution placed in
the beaker, was heated to 55C on a hot plate and was removed when the
temperature reached 55C. Once the solution had been prepared, its initial
pH was recorded using a pH meter. After the initial pH was recorded, ten
percent (10%) acetic acid (CH3COOH) was added to the solution,
dropwise method, while stirring with a stirring rod. The acid solution was
continuously dropped until the pH reached 4.6, and the volume of the acid
(based on the number of drops) was recorded. After letting the solution

stand until a large amorphous mass was formed, the solution was
decanted to separate the large mass from the solution. One the isolated
casein was collected; it was dried in between filter papers and was
weighed and the percent yield was determined. The percent yield of the
isolated casein was computed for by dividing the actual yield over the
theoretical yield and was multiplied to 100. After determining the percent
yield, the isolated casein was divided into two portions the first portion
used for the acid and base hydrolysis and the second portion used for the
experiment on Color Reaction of Intact protein and Hydrolyzate.
B. Alkaline Hydrolysis and Neutralization
For the alkaline hydrolysis of the casein, the collected intact protein
was isolated and cut into very small pieces and was placed into a 50milliliter Erlenmeyer flask. After adding the protein isolate into the flask,
five milliliters of boiling water and two grams of Barium Hydroxide
Ba(OH)2 was added into the flask, and was plugged with a cotton plug
covered with aluminum foil and was labeled. After the preparation, the
appearance of the sample was noted. The flask was then autoclaved at
fifteen psi for five hours, and after the flask was autoclaved, the
appearance of the hydrolyzate was noted. The hydrolyzate was diluted
with 15 milliliters of distilled water and was transferred to a 250-milliliter
beaker. Once diluted, the hydrolyzate was neutralized by adding one
milliliter of 16N sulphuric acid (H2SO4), and using the litmus paper, the pH
was checked. Once placed in the hydrolyzate, the color of the litmus paper
changed from blue to purple, which indicated that the pH was 7. Once
neutralized, the precipitate was filtered and the filtrate was collected.

Results and Discussion

Table 1.
Weight of Milk
5.0997 g
Initial pH
Final pH
Volume 10% Acetic acid (drops of 28 drops
acetic acid)
Weight of Casein
2.5709 g
Percent yield
50.41 %