Experiment: 7

amino acids
Objectives: To detect the presence of aromatic groups-containing
like tyrosine and tryptophan by Xanthoproteic Test
Xanthoproteic Test *****"*****".********u************.****************************************. *********"

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Xanthoproteic test is biochemical test for the detection of amino

a

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
Protein Test. The test gives a
to 'yellow, so the test is often termed as the Yellow
or other aromatic groups.
positive result for amino acids containing benzene rings
on the presence or
The test is a qualitative test that provides information only
absence of the amino acids. ************

Principle of Xanthoproteic Test

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in the amino acids
The Xanthoproteic test is based on the fact that aromatic groups
are nitrated by heating with concentrated HNO, to yield intensely
yellow-colored
residue turns orange due to
nitro derivative. On the addition of alkali, however, the
the nitro compound. Benzene ring-
the formation of a salt of the tautomeric form of
test to this test
containing amino acids like phenylalanine don't give positive doesn't react with
a

which
because the phenyl group in phenylalanine is very stable,
nitric acid in the conditions of this test. However, phenylalanine might give
a

positive result after an extended period of heating.

OH
+HNO3 ON OH
- H20
NH2 HO
NH2
HO
Tyrosine dark yellow or orange color

Requirements
Reagent
Concentrated Nitric acid
40% NaOH
Test solution
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Procedure of Xanthoproteic Test

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1. About 1 ml ofthe 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.
 solution is added to the test tube and observed for
Then, 1 ml of 40% NaOH
3 color change.
of Xanthoproteic Test *e****************************************************"ireuss

Result
.
andInterpretation
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Principle, Procedure,
Result, Uses|
Xanthoproteic Test- Definition,
Xanthoproteic
Xanthoproteic Test Positive
Test Negative
Presence of aromatic
Absence of aromatic
amino acids (tyrosine
amino acids (tyrosine
and tryptophan)
and tryptophan)
Presense of the dark
Absence of the dark color
yellow or orange

yellow or orange color

solution
dark yellow or orange-colored
Positive result: The appearance of a aromatic groups in the
test. This indicates the presence of
represents a positive
acids.
proteins and amino orange-colored solution
The absence of a dark yellow or
Negative result: absence of aromatic groups
in
test. This indicates the
represents a negative
acids.
proteins and amino
Limitations
aromatic group-containing
amino acid, it
is an
Even though phenylalanine test due to the highly stable phenyl
doesn't give a positive Xanthoproteic

group.