TEST DESCRIPTION/ REAGENTS PROCEDURE PRODUCT POSITIVE

CHARACTERIS RESPONSIBLE RESULT

TICS FOR COLOR
BIURET TEST This test is used Biuret reagent Take 1 ml of test solutions in The Biuret test is violet colored
to detect the (Hydrated dry test tubes and in another based on the complex
presence of Copper sulphate tube take 1 ml distilled water ability of Cu (II)
peptide bond. + Potassium as control. ions to form
hydroxide Add 1 ml of biuret reagent to a violet-
solution + all test tubes, mix well. coloured chelate
Potassium Look for the development of complex with
sodium tartrate) blue colors peptide bonds (-
CONH- groups)
in alkaline
conditions.
The greater
the concentration
of peptide bonds,
the greater
the color
intensity. If the
concentration of
peptide bonds is
low – such as
when short-chain
peptides are
present - the
color change is
from blue to pink.
XANTHOPROTEIC It is an Concentrated nit To 1ml of the amino acid In the presence of Dark yellow
TEST identification ric acid solution taken in a test tube, concentrated
test of protein add few drops of nitric acid nitric acid, the
and it gives a and vortex the contents. Boil aromatic phenyl
positive result the contents over a Bunsen ring is nitrated to
with those flame, using a test tube holder, give yellow
proteins with for few minutes. Cool the test colored nitro-
amino acid derivatives. At
 carrying tube under running tap water alkaline pH, the
aromatic group. and add few drops of alkali. color changes to
orange due to the
ionization of the
phenolic group.
NINHYDRIN TEST This is a test for Ninhydrin To 1ml of amino acid Amines (including Deep blue or
amino acids and solution (0.2 g solution taken in a test tube, α-amino acids) purple color
proteins with Ninhydrin add few drops of ninhydrin react with
free – 0.5 ml Acetic reagent and vortex the ninhydrin to give
NH2 group. acid contents. Place the test tube a coloured
100 ml n- in a boiling water bath for 5 product
Butanol minutes and cool to room
4.5 ml Water) temperature.

MILLON’S TEST To detect amino Test solution: To 1ml of the amino acid Tyrosine when Brick red color
acid containing 1 % arginine, solution in a test tube, add reacted with
phenol group 1 % tyrosine, few drops of millon’s reagent acidified mercuric
phenol solution and vortex. Boil the contents sulphate solution
over a Bunsen flame for 3 – 5 gives yellow
Millon’s reagent minutes. Cool the contents precipitate of
(Acidified under running tap water and mercury-amino
mercuric add few drops of sodium acid complex. On
sulphate) nitrite solution. addition of
sodioum nitrate
1 % sodium solution and
nitrite heating, the
yellow complex of
mercury-amino
acid complex
converts to
mercury
phenolate which
is in red color.
HOPKINS COLE This test is Hopkins Cole Mix 1 ml of the amino acid The indole ring purple ring in the
TEST specific test for reagent solution with 1 ml acetic acid reacts with junction where
(Glyoxylic acid) – glyoxilic acid reagent, in a glyoxylic acid in
 detecting test tube, vortex. Then the presence of a the two liquids
tryptophan. carefully, add conc. Sulphuric strong acid to meet
acid along the side of the test form a violet
tube, keeping the tube in an cyclic product.
inclined position (do not
shake the test tube , while
adding acid)
SAKAGUCHI TEST Test used for Sakaguchi To 1 ml of prechilled amino Guanidinium Red
detecting the reagent (1- acid solution and few drops compound
presence of Naphthol and a of NaOH is mixed and 2
arginine in drop of sodium drops of alpha naphthol is
proteins. hypobromite) added. Mix thoroughly and
add 4-5 drops of hypobromite
reagent and observe.
LEAD SULPHIDE This test is for NaOH 40% To 1ml of the amino acid Cysteine or Brown or black
TEST the identification solution in H2O solution taken in a test tube, cystine respond ppts
of sulphur add few drops of sodium to this test but the
containing Lead Acetate hydroxide (40%) and boil the sulphur present in
amino acid solution contents for 5 – 10min over a the methionine
cystine and bunsen burner. Cool the doesn’t respond
cysteine. contents and add few drops of to this test.
10% Lead acetate solution and
observe.
FOLIN'S forms the basis Folin–Ciocalteu To 1ml of the amino acid Addition to Blue
MCCARTHY for reagent (FCR) solution taken in a test tube, sodium
SULLIVAN TEST the quantiative (mixture of add few drops of sodium nitroprusside[Na2
determinaton phosphomolybd hydroxide (5N), followed by Fe(CN)5NO] to
of methionine. ate and addition of few drops of an alkaline
phosphotungstat glycine (1%) and 10% solution of
e) sodium nitroprusside solution methionine
and vortex. Place the test followed by the
tube in a hot water bath, acidification of the
maintained at 40°C, for 15 reaction yields a
minutes. Cool the test tube in red colour.
ice cold water for 5 minutes
and add 0.5ml of 6N HCl.
 Vortex the contents and allow
to stand for 15 minutes at
room temperature.
NITROPRUSSIDE Rapid, simple, Sodium Take 5 ml of urine in a test Acetoacetic acid Purple color
and qualitative Nitroprusside tube and saturate it with or acetone reacts
TEST determination of ammonium sulphate. with nitroprusside
cystine Add a small crystal of sodium in alkaline
concentrations. nitroprusside. Mix well. solution to form a
Slowly run along the side of purple-colored
the test tube liquor ammonia complex
to form a layer.

ROSENHEIM TEST Test for indole F3Cl3 , CH2O Add 5 drops of CH2O and 3 FeCl3 gives the Purple ring
ring . Specific ( formaldehyde ) drops of FeCl3 Mix the these color to the
test for and H2SO4 two reagents together which solution
tryptophan, will form rosenheim reagent.
because Then add 30 drops of
tryptophan is the albumin and then shake .
only amino acid until here we doesn’t have
that contain any product or result . Add 20
indole test . drops of H2SO4 drop by drop
( DON’T SHAKE ) then
observe the formation of the
colored ring