Biochemistry

Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

Lesson title: Proteins and Amino acids
Learning Targets:
At the end of the module, students will be able to:
1. Define what is protein, their functions in the body and their
building blocks;
2. Describe the general characteristics of proteins;
3. Classify the different amino acids;
4. Identify the presence of specific amino acids using chemical
confirmatory tests with specified positive results, and
5. Explain how proteins become denaturated and how it affects
the protein structure.
Materials: Pen, Paper, drawing materials
References:
Petrucci, et al. General Chemistry Principles &
Modern Applications. 9th ed. Upper Saddle
River, NJ: Pearson Prentice Hall, 2007.
Sadava, et al. Life: The Science of Biology. 8th
ed. New York, NY. W.H. Freeman and
Company, 2007.

A. LESSON PREVIEW/REVIEW
Video supplementary link:
The instructor will guide the students to watch the following links of informative clip regarding proteins.
 https://www.youtube.com/watch?v=HSCUAjZQhXI
 https://www.youtube.com/watch?v=wvTv8TqWC48

B. MAIN LESSON

MATERIALS:
 Powdered egg albumin  10% NaOH solution
 100 ml Egg albumin solution
 Merucric chloride solution
 Egg white
 Soda lime  0.1% Cupric sulfate solution
 Lead acetate  Dilute ammonium hydroxide
 3M HCl  Ninhydrin solution
 Water  Tryptophan
 0.1M NaOH  Water bath
 0.1M HCl  Stirring rod
 Chloroform
 Litmus paper
 Glycine
 Phenylalanine  Test tube
 Nitric acid  Beaker
 Ammonium sulphate solution  Crucible with cover
 95% ethanol  Alcohol lamp
 Conc. Nitric acid  Tongs
 Conc. Sulfuric acid  Watch glass
 Conc. HCl

PROCEDURE:

Before
1. Use equipment only for its designated purpose.
2. Make sure you have complete materials and equipment. Wash them before use and check if everything is in good
condition.

This document is the property of PHINMA EDUCATION

 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

3. To avoid mix-up and cross-contamination, organize and segregate the set of chemical reagents and equipment
needed in different procedure.
4. Make a label of the reagents or materials that you will use.
5. The acquired reagents should be put into an appropriate receiving and holding vessels such as test tube for liquids
and watch glass for solids.

During

A. GENERAL CHARACTERISTICS OF PROTEINS

I. ELEMENTARY COMPOSITION

1. Mix thoroughly in a mortar and pestle 0.5 g of powdered egg albumin and 0.5 g of soda lime. Heat the mixture strongly
while holding moist litmus paper to the vapors emerging from the mouth of the test tube.

Result:

What element is indicated? ______________________________________

2. Continue to heat the mixture strongly for several minutes. Allow the tube to cool to room temperature. Acidify the
contents of the tube with 3M HCI. The resulting solution must be distinctly acid (test with litmus paper). Place a small
strip of filter paper moistened with lead acetate at the mouth of the tube gently.

Result:

What element is indicated? ______________________________________

3. Note any odor at the mouth of the test tube.

Result:

What substance gives this odor? __________________________________

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 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

4. Observe the residue in the test tube.

What element is indicated? ______________________________________

5. Observe what is formed at the cooler end near the mouth of the test tube.

What is it? ___________________________________________________

What elements are indicated? ____________________________________

II. IGNITION TEST

1. Put powdered egg albumin in a crucible enough to cover the bottom of the crucible. Heat gently at first and then
more strongly until the bottom of the crucible becomes hot.

2. Did the sample melt? ______ bubble? ______ smoke? ______ catch fire? ______ change color? _____ give off
any odor? ______ leave any residue? _______

III. SOLUBILITY

Test the solubility of 1 ml of fresh raw egg white in 2 ml. of each of the solvents listed below. Indicate whether soluble
or insoluble.

a. Water _________________________________
b. 0.1M NaOH _________________________________
c. 0.1M HCl _________________________________
d. Chloroform _________________________________

B. GENERAL PROTEIN COLOR TESTS

I. BIURET TEST
 In separate test tubes containing 1 ml. of each of the above solutions, add 1 ml. of 10% NaOH solution and drops
of 0.1% cupric sulfate solution. To a fourth test tube containing 1 ml. of distilled water which serves as a blank,
add the same amount of NaOH and CuSO4. Describe any color change that occurs. Record your results in the
table on the next page.
 Write the formula of biuret: _____________________________________

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 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

II. NINHYDRIN TEST

 In separate test tubes of the above solutions, add 1 ml. of ninhydrin solution. To a fourth test tube containing 1
ml. of dilute ammonium hydroxide, add the same amount of ninhydrin solution. Heat the tubes in a boiling water
bath for 3 minutes and observe the color changes which occur in each tube. Record your results in the table on
the next page.
 Write your equation involved in the Ninhydrin reaction: ________________

III. XANTHOPROTEIC TEST

 In separate test tubes containing the above solutions, add an equal volume of conc. nitric acid (CAUTION!) To
a fourth test tube containing 1 ml of tryptophan, add the same amount of conc. nitric acid. Heat t for 2 minutes
in a boiling water bath. Write your results in the table below.
 Why does nitric acid stains a yellow color?

Name of Test Importance of Composition of Color of Chemical Group Responsible

Test Reagent Positive Result for Positive Test

This document is the property of PHINMA EDUCATION

 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

C. COLOR TESTS FOR SPECIFIC AMINO ACIDS

1. Millon’s Test for Tyrosine

In 3 separate test tube, place 2ml. each of 1% egg albumin, 1% gelatin and 0.02% salicylic acid solutions. Add 3
drops of Millon’s reagent and heat the tubes in boiling water bath for 2 minutes. Note the color produced. Record
your results in the table below.
2. Hopkins-Cole Test for Tryptophan
In 3 separate test tubes, place 2 ml. each of 1% egg albumin, 1% gelatin and e few granules of casein suspended
n 2 ml. o water. Add 2 ml. of Hopkins-Cole reagent to each tube and mix thoroughly. Hold each test tube in an
inclined position and carefully allow 3 ml. conc. sulfuric acid (CAUTION!) to slide down the side of each tube. Let
stand and observe the color at the zone of the two fluids. Record your results in the table below.
3. Sakaguchi Test for Arginine
To 5 ml. of gelatin solution, add 1 ml. of 10% NaOH and 1 ml. of 0.02% alpha-naphthol solution. After 3 minutes,
add 3 drops of sodium hypobromite solution. A strong red color develops but fades quickly. The color can be
stabilized by adding urea to destroy the excess hypobromite. Write your result in the table below. Repeat using
Arginine.
4. Lead Acetate Test for Labile Sulfur
To 2 ml. of 1% egg albumin in test, add 5 ml. of 5% NaOH solution and a few crystals of lead acetate. Heat in a
boiling water bat for 5-10 minutes, with occasional mixing of the contents of the tube. Note the color change.
Record your result in the table below.

Name of Test Importance of Test Composition of Color of Positive Group Responsible

Reagents Results

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 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

D. COAGULATION OF PROTEINS

1. By Heat
Place 3 ml. of egg albumin solution in a test tube and boil gently for a few minutes. Describe the change in the
albumin solution.
Result:

2. Heller’s Ring Test

Place 1 ml. of conc. nitric acid in a test tube (CAUTION!). Dilute 1 ml. of egg albumin solution with q ml. of water and
mix well. Incline the tube containing the nitric acid and carefully pour the 3 ml. of diluted albumin solution down the
side of the tube so that it forms a layer above the acid solution. What do you observe at the juncture of the two
liquids?
Result:

3. By Alcohol
Place 1 ml. of egg albumin in a test tube and add 1 ml. of 95% ethyl alcohol. What changes occurs in the albumin
solution?
Result:

E. PRECIPITATION OF PROTEINS
1. By Strong Acids (CAUTION!)
In each of 3 test tubes containing 3 ml. of clear 1% egg white solution, add 1 ml. of conc. HCI, conc. nitric acid and
conc. sulfuric acid. Note and describe the changes that occur.
Result:

2. By Heavy Metal Cations

In each of 3 test tubes containing 2 ml. of egg white solution, ad several drops of 5% cupric sulphate solution, 5%
lead acetate solution and mercuric chloride solution.
Result:

This document is the property of PHINMA EDUCATION

 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

3. Salting Out
To 10 ml. of 1% egg white in a small beaker, add 5g of solid ammonium sulfate and stir until the ammonium sulfate
has dissolved. Filter and test 2 ml of the filtrate for protein by the biuret test, adding a slight excess of sodium
hydroxide to destroy the ammonium sulfate.
Result:

4. Seyage Test
To 1 ml. of egg white solution, add 1 ml. chloroform and shake. Allow to stand and observe the boundary between
the two liquids.
Result:

After
1. Dispose all waste materials into the laboratory waste bin. Observe proper segregation of hazardous and non-
hazardous chemicals in discarding them into the waste bin.
2. Clean all your laboratory apparatus and return them to the laboratory custodian or technician.
3. Clean your working area and sink.
4. Make sure to unplug all electronic devices used in the operation.

CHECK FOR UNDERSTANDING

Guide Questions:
1. Define proteins.

2. What are the functions of proteins?

3. What are the elements present in proteins?

4. Identify the essential and non-essential amino acids

This document is the property of PHINMA EDUCATION

 Biochemistry
Module #5 Student Activity Sheet

Name: _____________________________________________________ Class number: ____

Section: ____________ Schedule: _______________________________ Date: ___________

5. What is protein folding and briefly explain the four levels of protein folding mechanism.

6. What is Marasmus? Explain briefly the relationship of protein to this disease.

7. What is Kwashiorkor? Explain briefly the relationship of protein to this disease. (10 points)

8. What is meant by isoelectric point? What is its importance?

Reference/s used: APA Format 7th Edition

C. LESSON WRAP-UP

Active Learning Strategy: THE MUDDIEST POINT

THE MUDDIEST POINT

In today’s session, what was least clear to you?
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