Guru Nanak Institute of Pharmaceutical Science and Technology

Sub: Lab on Genetic Modification in Agriculture and Medicine Paper code: DSEBT-691A
Experiment No: 2 Date:

AIM – DETERMINATION OF AMINO ACIDS USING XANTHOPROTEIC TEST

OBJECTIVE:

• To detect the presence of aromatic groups-containing amino acids like tyrosine and
tryptophan.
• To differentiate tyrosine and tryptophan from other amino acids.

PRETEXT QUESTIONS:

1. Xanthoproteic test is performed to identify which of the following amino acid?

a) Tryptophan b) Glycine c) Arginine d) Histidine

2. What is the percentage of NaOH which is used in xanthoproteic test?

a) 20% b) 30% c) 40% d) 50%

PRINCIPLE:

Xanthoproteic test is a biochemical test for the detection of amino acids containing phenolic
or indolic groups like phenylalanine, tyrosine, and tryptophan (aromatic amino acids). The
test is named Xanthoproteic test due to the formation of a yellow precipitate of xanthoproteic
acid. The term ‘Xantho’ refers to ‘yellow’, so the test is often termed as the Yellow Protein
Test. The test gives a positive result for amino acids containing benzene rings or other
aromatic groups. The test is a qualitative test that provides information only on the presence
or absence of the amino acids.

The Xanthoproteic test is based on the fact that aromatic groups in the amino acids are
nitrated by heating with concentrated HNO3 to yield intensely yellow-colored nitro
derivative. On the addition of alkali, however, the residue turns orange due to the formation
of a salt of the tautomeric form of the nitro compound. Benzene ring-containing amino acids
like phenylalanine don’t give a positive test to this test because the phenyl group in
phenylalanine is very stable, which doesn’t react with nitric acid in the conditions of this test.
However, phenylalanine might give a positive result after an extended period of heating.
 Guru Nanak Institute of Pharmaceutical Science and Technology
Sub: Lab on Genetic Modification in Agriculture and Medicine Paper code: DSEBT-691A
MATERIALS REQUIRED:

Reagent
• Concentrated Nitric acid
• 40% NaOH
• Test solution

Material Required
• Test tubes
• Test tube stand
• Pipettes

METHODOLOGY:

1. About 1 ml of the sample solution is taken in a test tube. To this, the same amount of
concentrated nitric acid is added.
2. The test tube is allowed to cool down to room temperature. If the sample is a protein
solution, a white precipitate might develop due to the denaturation of proteins.
3. Then, 1 ml of 40% NaOH solution is added to the test tube and observed for color change.

RESULT AND OBSERVATION:

• Positive result: The appearance of a dark yellow or orange-colored solution represents a

positive test. This indicates the presence of aromatic groups in the proteins and amino
acids.
• Negative result: The absence of a dark yellow or orange-colored solution represents a
negative test. This indicates the absence of aromatic groups in proteins and amino acids.
 Guru Nanak Institute of Pharmaceutical Science and Technology
Sub: Lab on Genetic Modification in Agriculture and Medicine Paper code: DSEBT-691A

CRITICAL THINKING QUESTIONS:

1. What is the colour of protein in Xanthoproteic test?

2. What is the mechanism of Xanthoproteic test?

ATTENDANC VIVA PERFORMANCE TOTAL

E(10) (10) (20) (40)

CLEANLINESS CONDUCT PRACTICAL CRITICAL

& MANUAL & ACCURACY THINKING
MAINTENANCE INVOLVEMENT (4) (8)
(4)
(4)

Signature of the Teacher