Color Test

Biuret Test

Ninhydrin Test
Xanthoproteic Test

Millons Test

Hopkins-Cole Test

Sakaguchi Test

Nitroprusside Test
Fohls Test

Test for Amide

Pauly Test

Reagent

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
Five drops of 30%
NaOH
2 drops of
5%(CH3COO)2Pb
1 mL of 20% NaOH
(boiling water bath)

Diazo reagent (3-5

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

Visible Results
(GLUTEN)

Principle
Involved

Color
Reaction of
Intact
Protein
Purple
solution

Inference

Positive

To detect the presence

of peptide bonds

Blue-Violet
Solution

Positive

Test used for an

alpha-amino acid

Yellow
Solution

Positive

Detects the side

chains of aromatic
amino acids

Flesh

Negative
(means
gluten has
no tyrosine
residues)

Determines tyrosine
residues

Positive

Specific only for

tryptophan

Negative

Test for guanidines

(arginine and peptides
that contain it)

Positive

Determine the
presence of cysteine (SH)
Detect presence of
sulfur containing
amino acid

If tyrosine is
positive,
the color
must be
pinkish-red
Purple ring
at Interface
Yellowish
solution
If
guanidines
are
present,
the color
must be
bright-red
Red to Red
violet
colorization
Red to red
violet
colorization
Red -->
Blue Litmus
Paper
Red solution
(+)

Positive

Positive

Detect asparagines
and glutamine

Positive

This test answers

for tyrosine and
histidine residues

Color Test

Biuret Test

Ninhydrin Test
Xanthoproteic Test

Millons Test
Hopkins-Cole Test

Sakaguchi Test

Nitroprusside Test
Fohls Test

Test for Amide

Pauly Test

Reagent

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
Five drops of 30%
NaOH
2 drops of
5%(CH3COO)2Pb
1 mL of 20% NaOH
(boiling water bath)

Diazo reagent (3-5

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

Visible Results
(CASEIN)

Principle
Involved

Color
Reaction of
Intact
Protein
Purple
solution

Inference

Positive

To detect the presence

of peptide bonds

Blue-Violet
Solution

Positive

Test used for an

alpha-amino acid

Yellow
Solution

Positive

Detects the side

chains of aromatic
amino acids

Pinkish-red

Positive

Purple ring
at Interface

Positive

Determines tyrosine
residues
Specific only for
tryptophan

Bright-red

Positive

Test for guanidines

(arginine and peptides
that contain it)

Red to Red
violet
colorization
Red to red
violet
colorization

Positive

Determine the
presence of cysteine (SH)
Detect presence of
sulfur containing
amino acid

Red -->
Blue Litmus
Paper
Red solution
(more like
Red-orange)
(+)

Positive

Detect asparagines
and glutamine

Positive

This test answers

for tyrosine and
histidine residues

Positive

Color Test

Biuret Test

Ninhydrin Test
Xanthoproteic Test

Millons Test

Hopkins-Cole Test

Sakaguchi Test

Nitroprusside Test

Fohls Test

Test for Amide

Pauly Test

Reagent

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 Hopkins-Cole
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

Five drops of 30% NaOH

2 drops of
5%(CH3COO)2Pb

1 mL of 20% NaOH
(boiling water bath)
Diazo reagent (3-5 drops
of 1% sulfanilic acid with
3 drops of 5% NaNO2
soulution)
3-5 drops of 10% Na2CO3

Visible Results (MYOGLOBIN)

Principle Involved

Color
Reaction of
Intact Protein
Purple solution

Inference
Positive

To detect the presence of

peptide bonds

Blue-Violet
Solution

Positive

Test used for an alphaamino acid

Yellow Solution

Positive

Detects the side chains of

aromatic amino acids

Colorless
solution with
white
precipitate

Negative

Determines tyrosine
residues

Negative

Specific only for

tryptophan

Negative

Test for guanidines

(arginine and peptides
that contain it)

Negative

Determine the presence

of cysteine (-SH)

Positive

Detect presence of sulfur

containing amino acid

Positive

Detect asparagines and

glutamine
This test answers for
tyrosine and histidine
residues

If positive:
Pinkish-red
Dark flesh to
colorless
solution
If positive:
Purple ring at
Interface
No change
If positive:
Bright-red
Yellow with
small amount
of precipitate
If positive:
Red to Red
violet
colorization
Light yellow
with dark
brown
sediments
If positive:
Red to red
violet
colorization
Red --> Blue
Litmus Paper
Red solution
(more like Redorange)
(+)

Positive

https://www.scribd.com/doc/82512711/Qualitative-Color-Reaction-for-Casein
https://www.scribd.com/doc/63643592/Protein-Isolation-and-Characterization
https://www.scribd.com/doc/82850370/Formal-Report-for-Biochem
http://www.scribd.com/doc/48484530/Isolation-Qualitative-Color-Reaction-andAlkaline-Hydrolysis-of-Gluten-from-Yeast#scribd
https://www.scribd.com/doc/34761561/FR2-Isolation-of-Proteins-and-Color-Reaction
https://www.scribd.com/doc/49052097/Isolation-and-Characterization-of-Proteins