Experiment: 6

Objective: To detect amino acid containing phenol group by Millon's test

Principle of Millon's test:

with Millon's reagent to form red

Compounds containing hydroxybenzene radical react
is tyrosine. Thus, this test is
complexes. The only amino acid having hydroxybenzene ring
this amino acid. Tyrosine when
specific for the amino acid tyrosine and the protein containing
reacted with acidified mercuric sulphate solution gives yellow
precipitate of mercury-amino acid
of mercury-
solution and heating. the yellow complex
complex. On addition of sodioum nitrate
which is in red color.
amino acid complex converts to mereury phenolate

Reagents:
1% tyrosine, phenol solution
Test solution: 1 % arginine.
Millon's reagent (Acidified mercuric sulphate)
1%sodium nitrite

Procedure of Millon's test:

test solution in dry test tube.

1. Take 1ml
mix well.
2. Add 1ml of Millon's reagent and
3. Boil gently for I minute.

Cool under tap water.

4.
nitrite.
5. Now add 5 drops of 1 % sodium
6. Heat the solution slightly.
of brick red precipitate.
7. Look for the development

Result interpretation:
and phenol solution)
Brick red color (Tyrosine
Positive Millon's test:
Millon's test: no red
color ( arginine)
Negative
Millon's test
**all phenol give positive