NAME: GROUP NO.

: 6
BOLIVAR, Leonilyn SECTION: BSN1-A
LEGASPI, Ma. Lara Micaela INSTRUCTOR: SAMPOLLO, Leslie
SCORE:
OSUMO, Betiara Jazel
PELINGON, Robert Kris
PIO, Krea Joy
YSULAN, Cyky Mae DATE: December 11, 2020

Module 5: Precipitation, Denaturation and Coagulation

I. MATERIALS
PART A. Protein Denaturation

Test tubes

Beaker

Stirring rod

Water

Egg yolk

Funnel

Ring stand

Erlenmeyer flask

PART B. Methods of Protein Precipitation

Pipet Albumin solution

Acid
Test Tubes
 Protein solution
Test Tube Rack

 Precipitation of protein with heavy metals

Beaker Lead Acetate Solution
2 mL Acid Solution
Test Tubes

Pipet

 Albumin and Picric Acid

Pipet Albumin solution

Picric Acid
Test Tubes

 Precipitation of protein by organic solvents

Pipet Albumin solution

Ethanol
Test Tubes

 Addition of alcohol to protein solution displaces water molecules

associated in protein thus, decreasing solubility

Pipet Albumin solution

Acetone
Test Tubes

II. PROCEDURES
PART A. Protein Denaturation
 Separate in two 150 ml beakers an egg white from the yolk
 Add the same volume of water to each and sir it.
 Filtrate both solution and use the filtrate as the watery solution of protein to show
their denaturation.
  Take a 3 ml of an egg white protein solution placed in a test tube and heat it over
the flame.
 Repeat the experiment with the egg yolk solution
 Observe the calculation due to the heat.
 Concentrated nitric acid add it to an album of solution denaturates
 Discards all the solutions in the train. The remained protein solution and the
acidic ones have to be diluted first.
PART B. Methods of Protein Precipitation
 Albumin solution
 Test tube
 Piper
 Acid
 Protein solution
Precipitation of protein with heavy metals
 Using pipet get 2 ml of albumin solution and put it inside the test tube
 Add lead acetate solution.
 As you can see the addition of lead iron to the protein results in the precipitation
protein as a complex.
Albumin and Picric Acid
 Using pipet, get a 2 ml of albumin solution and put it into the test tube.
 Transfer 1 ml of picric acid using pipet into the test tube with an albumin solution
inside it.
 Observe.
Precipitation of protein by organic solvents
 Using pipet, get a 2 ml of protein/albumin solution and put it into the test tube.
 Add 2 ml of ethanol also into the test tube
 Observe.
Addition of alcohol to protein solution displaces water molecules associated in
protein thus, decreasing solubility
 Using pipet, get 2 ml of protein solution and put it into the test tube.
 Add 2 ml of acetone into the test tube.
 Observe.

III. RESULTS AND DISCUSSION

PART A. Protein Denaturation
 EGG WHITE DISCUSSION

Initially, the color was transparent but after

The egg white becomes
HEAT
firm when heated.
heating the egg firms and the color of the
solution turns white. Therefore,
denaturation does not alter the protein
content and only its secondary and tertiary
structure.

Initially, the color of the egg white was

The egg white turns white
but has a transparent
ACETONE
solution at the bottom part
of the test tube.
transparent but after putting the acetone it
turned into a white color in the top part but
the bottom part stays transparent.
Therefore, acetone is also altering the
secondary and tertiary structure of the egg
albumin protein.

Initially, the color of the egg white was

transparent but after putting the
The egg white turns white
concentrated nitric acid, it turned into a
NITRIC at the bottom part and has
white color on the bottom part but the top
ACID a transparent part at the
part stays transparent. Therefore,
top.
concentrated nitric acid also denature the
egg albumin.

PART B. Methods of Protein Precipitation

EGG ALBUMIN DISCUSSION

The egg albumin

turned from Since, the solution turned white or cloudy,
HCl transparent to white we can conclude that denaturation of protein
color after adding the is present.
HCL.
The solution turned
White from Since the solution shows a cloudy material,
LEAD
transparent after we can also conclude that denaturation was
ACETATE
adding the lead present.
acetate.
 The egg albumin
does not completely
mix with the picric
Even though the picric acid and egg albumin
acid. A yellow
does not mix totally we can still conclude
PICRIC ACID material was seen on
that picric acid denatures the protein
top and a white or
because a cloudy material is present.
cloudy material was
seen below the yellow
solution.
Same as with picric acid the egg albumin
and 95% ethanol does not mix completely
but we can conclude that denaturation of
Egg albumin also protein is present because a cloudy material
95% ETHANOL
turned cloudy. was seen in the solution. The solution does
not mix because the addition of alcohol to
the protein solution makes protein less
soluble.

POST-LAB QUESTIONS 
1. Give the rationale of the use of picric acid for burns and tannic acid for
diarrhea. 
 Picric acid is used to treat burns because it is astringent and an antiseptic. Picric
acid is a much stronger acid than phenol it decomposes carbonates and may be
titrated with bases. Taking a product containing tannic acid and gelatin “gelatin
tannate” might improve symptoms in children who have had diarrhea for no more
than 3 days. Tannic acid is a naturally occurring plant polyphenol and can be
found in practically all aerial plant tissues. Tannic acid was historically used for
the treatment of diarrhea, topically to dress skin burns and rectally for treatment
of unspecified rectal disorder.
2. Discuss the difference between coagulation and denaturation.
 Coagulation and denaturation are two processes that occur in molecules. Both
processes change the state of molecules from native state to a different state.
The difference between coagulation and denaturation is that denaturation is the
changing of the properties of a molecule while the coagulation is the action of
converting the liquid state molecules into the solid or semi-solid state by sticking
molecules together. These are the differences between coagulation and
denaturation.
 3. Explain the process of salting out.
 Salting out is a purification method that utilizes the reduced solubility of certain
molecules in a solution of very high ionic strength. It is typically, but not limited to,
the precipitation of large biomolecules such as proteins. It occurs in aqueous
solutions of high ionic strength that reduce the molecule’s solubility causing
certain proteins to precipitate. The process of salting out is when the salt
concentration is increased, some of the water molecules are attracted by salt
ions, which decreases the number of water molecules available to interact with
the charged part of the protein. A result of the increased demand for solvent
molecules, the protein-protein interactions are stronger than the solvent solute
interactions, the protein molecules coagulate by forming hydrophobic interactions
with each other.
4. Predict the results in the following tests

COLOR DENATURED COAGULATED

DISCUSSION
REACTION TEST PROTEINS PROTEINS
Initially, the egg
albumin was
colorless, after
  adding concentrate
Yellow to orange nitric acid there is
XANTHOPROTEIC Yellow to orange
coloration no change
TEST observed. After
heating the
solution’s color
turned yellow.
Initially, the egg
albumin was
colorless and after
adding the amount
of NaOH solution
and 1% CuSO4
solution there is no
change seen but
when the solution
BIURET TEST Purple color Purple color was heated the
color turned violet
  Violet ring In the solution, the
Purple-Violet ring
 Violet ring appears indicates
HOPKINS-COLE
the presence of
TEST
Tryptophan.