Name: Hoang Le Minh 12MRLV

Topic: Protein denaturation

Date: 4th March 2014

Time: 1 hour

Denaturing albumin

I. Introduction
Research question
How does a change in heat, pH, the addition of 95% ethanol, and the addition of lead (II) nitrate/silver
nitrate affect the solubility of albumin in eggs?
Hypothesis
I postulate that:

In test tube 1 (investigating temperature), the albumin would undergo a colour change from

clear to white and become water-insoluble.

In test tube 2a (investigating acidic pH), the albumin would become water-insoluble.
In test tube 2b (investigating alkaline pH), the albumin would become water-insoluble.
In test tube 3 (investigating the effect of ethanol), the albumin would become water-insoluble
In test tube 4 (investigating the effect of lead (II) nitrate), the albumin would become waterinsoluble.

Variables
Controlled variables: Same starting concentration of albumin stock solution

Test tube 1: Variables kept the same: pH, absence of ethanol, absence of lead (II) nitrate/silver

nitrate
Test tube 2: Variables kept the same: temperature, absence of ethanol, absence of lead (II)

nitrate/silver nitrate
Test tube 3: Variables kept the same: pH, temperature, absence of lead (II) nitrate/silver nitrate
Test tube 4: Variables kept the same: pH, temperature, absence of ethanol

Dependent variable: Solubility of albumin

Independent variable:

Test tube 1: Temperature

Test tube 2: pH
Test tube 3: Presence of ethanol
Test tube 4: Presence of lead (II) nitrate or silver nitrate

II. Procedures and materials

Apparatus

6 test tubes
Test tube rack
Pipettes
Measuring cylinders
Beaker

Safety precautions

Eye protection

Procedures
1. Prepare a stock albumen solution by adding the white of one egg to 100 ml of distilled water in a
beaker. Stir until the albumin is in solution.
2. Place 5 ml of albumen solution into each of six test tubes. Label the tubes Control, A, B1, B2, C, and
D.
3. Treat the four solutions as follows:

Control: Let stand in room temperature. Measure the temperature and pH with pH paper.
A: Heat in a hot water bath for a few minutes.
B1: Add 2 ml of 1M HCl. Record the pH using pH paper.
B2: Add 2ml of 10% NaOH. Record the pH using pH paper.
C: Add 1 ml of 95% ethanol solution.
D: Add 5 10 drops of 2% lead (II) nitrate.

4. Record observations.

III. Data collection and presentation

Results

Denaturing agents

Observation

Control test tube (pH 8,

No change

240C)
A Heat, 650C

Turns slightly cloudy.

B1 1M HCl, pH 1
B2 1M NaOH, pH 11

Viscosity increases. Solution becomes thicker and runnier.

However, no perceptible colour change is observed.

C 95% Ethanol

Turns cloudy, but not to the extent of test tube D.

Turns cloudy at the top of the solution first, then eventually permeates to the
entire body of the solution.

D Lead (II) nitrate

Coagulates into a white solid immediately when coming into contact with
lead (II) nitrate.
Produces a white precipitate, soft when touched.

IV. Data processing

No quantitative data available.

V. Conclusion and evaluation

1) What was the purpose of the controlled test tube?

2) What common result occurred in all experimental tubes? Why?