Al-Mashreq University Name: ‫علي عبد األمير علي‬

First stage /B2 Experiment NO: 4

Experiment Title: Millon's test

Objective:

This test is specific for tyrosine. Because it is the only amino acid containing a phenol group, a hydroxyl
group attached to benzene ring.

Tools and Materials:

Tools

Tube , Dropper , spatula, flask

Materials

nitric acid, water, amino acid containing a phenol group, sodium nitrite

Procedure:

The phenol group of tyrosine is first nitrated by nitric acid in the test solution. Then the nitrated tyrosine
complexes mercury ions in the solution to form a brick-red solution or precipitate of nitrated tyrosine, in
all cases, appearance of red color is positive test.

Method:

1. About 2 ml of the sample solution or the tyrosine solution is taken

in a test tube.

2. Add about 2 ml of Millon’s reagent in the test tube and kept in the

water bath for about 2 minutes.

3. Cool and add 2 drops of 1% of sodium nitrite solution .

4. Formation of red colored precipitatered observed immediately

Results

1. Millon’s test:

+ve test -- > red color precipitate --- > the sample is nitrated tyrosine

Discussion:

Amino acid containing a phenol group reacts with Millon’s reagent which give positive results by
changing the color of the solution to red.

Conclusion: Millon’s test gives a satisfactory solution to distinguish the properties of Amino acid
containing a phenol group.