Color Test

Reagent

Visible Results
Color
Reaction
of Intact
Protein

Principle
Involved

20 drops of 2.5 M
NaOH
2-3 drops of 0.1 M
CuSO4 solution
6-10 drops of 0.1%
ninhydrin solution
(boiling water bath)
10 drops of
concentrated HNO3
10 drops
of concentrated NaOH
5 drops of Millons
reagent
20 drops of HopkinsCole reagent
20 drops of
concentrated H2SO4
10 drops of 10%
NaOH
10 drops of 0.02%
naphthol solution
0.5 ml of 3 M NaOH
0.25 ml of 2%
Nitroprusside solution

Purple
solution
(+)

Color
Reaction of
Acid
Hydrolysat
e
Greenish
Brown
Solution (-)

Blue-Violet
Solution
(+)
Yellow
Solution
(+)

OrangeBrown
Solution (-)
Yellow
Orange
Solution (+)

Test used for an

alpha-amino acid

Flesh (+)

Yellow
Solution (-)
Clear Brown
Solution (-)

Determines tyrosine
residues
Specific only for
tryptophan

Light Brown
Solution (-)

Test for guanidines

(arginine and
peptides that contain
it)
Determine the
presence of cysteine

Fohls Test

Five drops of 30%

NaOH
2 drops of
5%(CH3COO)2Pb

Black/
Brown
Solution
(+)

Test for Amide

1 mL of 20% NaOH
(boiling water bath)

Pauly Test

Diazo reagent (3-5

Red -->
Blue
Litmus
Paper (+)
Red
solution
(+)

Biuret Test

Ninhydrin Test

Xanthoproteic Test

Millons Test
Hopkins-Cole Test

Sakaguchi Test

Nitroprusside Test

drops of 1% sulfanilic
acid with 3 drops of
5% NaNO2 soulution)
3-5 drops of 10%
Na2CO3

Purple ring
at
Interface
(+)
White
precipitate
(-)
Yellow
solution
(-)

OrangeBrown
Solution with
Precipitate
(-)
Amorphous
Black
precipitate
(Clear
Solution)
(+)
Red --> Blue
Litmus Paper
(+)
Yellow
Solution with
Black
precipitate
(-)

To detect the
presence of peptide
bonds

Detects the side

chains of aromatic
amino acids

Detect presence of
sulfur containing
amino acid

Detect asparagines
and glutamine

This test answers

for tyrosine and
histidine residues