NAME:_______________________ DATE:__________________

RATING:________________

PHINMA UNIVERSITY OF PANGASINAN

COLLEGE OF ALLIED HEALTH SCIENCES
DEPARTMENT OF NURSING

ACTIVITY NO. 7
COLOR TEST FOR PROTEINS AND SPECIFIC AMINO ACIDS

I. DISCUSSION:
Classification and Structure of Amino Acids
Structure of Proteins (Peptide Bond)
Name of test, purpose of test, composition of reagent, positive results obtained.

II. PROCEDURE:

A. GENERAL PROTEIN COLOR TESTS (20 points)

(Perform the following tests on 1 ml. each of 1% solution of egg albumin, glycine and
phenylalanine)

1. 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.:

2. 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.

3. 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 on the next page.

Why does nitric acid stains a yellow color?

 Name of Test Importance of Composition of Positive Results/ Group
Test Reagents Color Produces Responsible

Biuret Test

Ninhydrin Test

Xanthoproteic
Test

B. COLOR TEST FOR SPECIFIE AMINO ACIDS (30 Points)

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 Composition of Positive Results/ Group

Test Reagents Color Produces Responsible

Millon’s Test for

Tyrosine

Hopkins-Cole
Test for
Tryptophan

Sakaguchi Test
for Arginine

Lead Acetate for

Labile Sulfur