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Hopkins-Cole Test
20 drops of Hopkins-Cole reagent was
slowly added to the sample, and then
mixed. While the test tube was inclined,
concentrated sulfuric acid of about 20
drops was slowly added, without shaking,
and the color at the interface was taken
note of.
hydrolyzed samples.
Biuret Test
The group added 2-3 drops of 0.1 M
CuSO4 solution to a mixture of the sample
and 20 drops of 2.5M NaOH. The test tube
was shaken and the color changes in the
solution were observed.
Ninhydrin Test
For this test, 6-10 drops of 0.1% ninhydrin
solution was placed into the sample and
then it was heated in a boiling water bath.
After that, appearance of blue-violet
coloration was taken note of.
Xanthoproteic Test
10 drops of concentrated nitric acid and
concentrated sodium hydroxide was
slowly added to the diluted sample and
then mixed. Color changes after each
addition was observed. After that, 10 drops
of concentrated NaOH was added, then it
was to be observed whether there is any
color change.
Millons Test
5 drops of Millons reagent was added to
the diluted sample, and the color changes
were noted.
Sakaguchi Test
10 drops of 10% NaOH and 10 drops of
are present.
Biuret Test
Purple solution
Positive
Ninhydrin
Blue-Violet Solution
Positive
Ninhydrin test
This is done to know whether there is an
alpha amino acid present. It reacts with
ammonia, a primary amine, or a secondary
amine. Free amino acids can react with
ninhydrin reagent and it will yield a deep
purple solution upon heating with
ninhydrin. The ninhydrin test is positive
for amino acid and some proteins. Most of
Positive
Inference
Protein
Test
Xanthoprot
eic Test
(excess NaOH)
Millons
Negative
Positive
Negative
Yellow ppt
Positive
Fohls Test
Brown ppt
Positive
Test
Red
Test
Hopkins-Co
le Test
Sakaguchi
Test
Nitroprussi
de Test
for
-->
Blue
Amide
Paper
Pauly Test
Red solution
Litmus
Positive
Positive
Biuret test
This is used to look the presence
of peptide bonds. The Biuret test is a
positive test for proteins but not for amino
acid. The evidence for the test consists of
formation of a violet-pink complex when
cupric ion, in basic solution is added to
any polymer such as protein which
contains multiple amide bonds. A
blue-colored solution indicates a negative
test or those fewer than two peptide bonds
Xanthoproteic test
This is used to test for an aromatic
sidechain. The test depends upon a
reaction with a specific type of amino acid
chain. Aromatic rings have the ability to
undergo nitration reaction, which is the
addition of NO2 group to the ring that is
why it is a positive test for side chains in
tyrosine and tryptophan. With the addition
of nitric acid and heat, the reaction will
result to a yellowish-colored solution. The
intensity of the yellow color deepens when
the reaction occurs in basic solution (color
will change to orange). Gluten is positive
of aromatic side chain.
Millons test
This is a test for phenolic group containing
side chain or a test specific for tyrosine,
the only amino acid containing phenol
group. In this test, the phenol group
of tyrosine is first nitrated by nitric acid in the test
solution. Then the nitrated tyrosine
complexes Mercury (I) and Mercury
The Hopkins-Cole
This test is specific for trypthopan, the
only amino acid containing an indole
group. The clear violet ring produced is
due to the formation of a compound from
the glyoxylic acid in the reagent and the
Fohls test
This is a test for sulfur containing amino acids.
It also indicates the presence of methionine
and cysteine because those two amino acids
have sulfur in their structures. Its principle is
fusion followed by ionic interaction. A positive
result for this test is the formation of dark
brown or black precipitate from lead sulfide
trace of formaldehyde.
Gluten showed a positive result for the
presence of tryptophan.
Sakaguchi test
This test is for identifying the presence of
the guanido group of arginine. Since basic
hydrolysis destroys arginine and produces
Pauly Test
This test answers for the presence of
tyrosine and histidine residues. Imidazole
group reacts with diazotized aulphanilic
acid
to
form
highly
colored
azocompouds. The diazonium salt formed
couples with either tyrosine or histidine in
Nitroprusside test
This is for indicating the presence of
cysteine or free thiol groups. In this test,
cysteine is partially destroyed is evident in the
results of the experiment because it produces
a yellow precipitate. Its principle is
REFERENCES
Amrita University. (2015). Qualitative
Analysis of Amino Acid. Retrieved from
http://amrita.vlab.co.in/?sub=3&brch=63&
sim=1094&cnt=1
Retrieved
from
http://www.academia.edu/4836795/Qualita
tive_analysis_of_Amino_acids