Ex.

no:

REACTIONS OF CYSTINE

Date:

S.no
1.

Experiment
Solubility Test
A small amount of amino
acid is dissolved in
a. Cold water
b. Hot water
c. Dilute acid
d. Dilute alkali
e. Alcohol

Observation

Inference

Insoluble
soluble
soluble
Insoluble
Insoluble

2.

Ninnhydrin Test
To 1ml of cystine solution A violet coloured It is due to the
added 0.5ml of 1% ninhydrin solution
was formation
of
and heated for 2min and obtained
Ruhemanns purple by
allowed to cool
the
amino
acid
containing
-NH2
group.

3.

Ferric Chloride Test

To the cystine solution
added dilute solution of Fecl 3
and a drop of copper sulphate
solution.

does
No
transitory Cystine
violet or blue answer this test.
colouration
appears on the
addition of copper
sulphate.

not

4.

Nitoprusside Test
To 1-2ml of cystine No Red
solution added dilute sodium appears .
nitroprusside solution and a
drop of dilute sodium
hydroxide solution.

5.

Lead Acetate Test

To the cystine solution A black precipitate It is due to the
formation of lead
added a small quantity of 10% was obtained.
lead acetate and 4% of NaOH
sulphide.
solution and boil for few
minute.

6.

Cyanide Test:
To the cystine solution A
Beet
added a 2 drops of liquid coloration
ammonia and
5 % sodium obtained.
cyanide and sodium nitroprusside solution.

colour Cystine
does
answer this test.

not

red Cystine answers this

was test.

Ex.no:

REACTIONS OF CYSTEINE

Date:

S.no

Experiment

Observation

1.

Solubility Test
A small amount of
amino acid was dissolved in
a. Cold water
b. Hot water
c. Dilute acid
d. Dilute alkali
e. Alcohol

Insoluble
soluble
soluble
soluble
Insoluble

Inference

2.

Ninnhydrin Test
To 1ml of cysteine A violet coloured It is due to the formation
solution added 0.5ml of 1% solution
was of Ruhemanns purple
ninhydrin and heated for obtained.
by the amino acid
2min and allowed to cool
containing
-NH2
group.

3.

Ferric Chloride Test

To the cysteine solution
added dilute solution of Fecl3
and a drop of copper
sulphate solution.

The
transitory It is due to the
violet
colour sulfhydryl group of
appears on the cysteine.
addition of copper
sulphate.

4.

Nitoprusside Test
colour It is due to the presence
To 1-2ml of cysteine Red
solution added dilute sodium appears and faded of sulfhydryl group of
nitroprusside solution and a after 2minutes.
cysteine.
drop of dilute sodium
hydroxide solution.

5.

Lead Acetate Test

To the cysteine solution
added a small quantity of
10% lead acetate and 4% of
NaoH solution and boil for
few minute.

6.

Cyanide Test:
To the cysteine solution
added a 2 drops of liquid
ammonia and 5 % sodium
cyanide and sodium nitroprusside solution.

A black precipitate It is due to the formation

was obtained.
of lead sulphide.

A beet red
coloration was
obtained.

Cysteine answers this

test.

Ex.no:

REACTIONS OF METHIONINE

Date:
S.no

Experiment

Observation

1.

Solubility Test
The given amino acid was
dissolved in small amount of ,
a. Cold water
b. Hot water
c. Dilute acid
d. Dilute alkali
e. Alcohol

Insoluble
Insoluble
Insoluble
Insoluble
Insoluble

2.

Ninnhydrin Test
To 1ml of methionine A violet coloured
solution, added 0.5ml of 1% solution
was
ninhydrin and heated for 2min obtained.
and allowed to cool.

3.

Bollins Test
(Modification of mecrothy A red coloration
Sullivans test)
was got.
To 1 ml of methionine
solution the following reagents
are added in order & mixed
after each addition
1. 1.5 ml of 1% glycine.
2. 1.5 ml of 5N NaOH.
3. 0.3 ml of 10% sodium
nitroprusside .
Placed in a water
bath at 37-40 C for 15 min.
Placed in a ice bath for 5-7
min & then added 6N Hcl &
left to stand at
room
temperature for 15 min.

Inference

It is due to the
formation
of
Ruhemanns
purple.

The methyl group

in split off to form
homocystine which
gives red coloration
with
sodium
nitroprusside
solution.

Ex.no:

REACTIONS OF ARGININE

Date:

S.no
1.

2.

3.

Experiment
Solubility test
A small amount of arginine
was dissolved in
(a) Hot water
(b) Cold water
(c) Dilute hydrochloric acid
(d) Dilute potassium
hydroxide
(e) Alcohol

Observation

Inference

Soluble
Soluble
Soluble
Soluble
Soluble

Ninhydrin test
To 1ml of arginine solution A purple coloration It is due to the
added 0.5 ml of 1% ninhydrin was obtained.
formation
of
and heated for 2 min.
Ruhemanns
purple
Sakaguchi test
To 5 ml of arginine solution A caramel red This reaction is
added 5 drops of 15% NaOH and colour
solution due
to
the
4 drops of freshly prepared was obtained
guanidine group
-napthol in alcohol and bromine
of arginine
water

Ex.no:

REACTIONS OF HISTIDINE

Date:
S.no

Experiment

1.

Solubility Test
The given amino acid is
dissolved in
(a) Cold water
(b) Hot water
(c) Dilute acid
(d) Dilute alkali
(e) Alcohol
Ninhydrin test
To 1ml of histidine solution
added 0.5 ml of 1% ninhydrin
and heated for 2 min and
allowed to cool.

2.

3.

4.

Test for Histidine

To 2ml of histidine solution
added 1% bromine in 33%
acetic acid until a yellow colour
develops.
Ehrlischs Diazo test
To 2ml of histidine solution,
add 1ml of 1% sulphanilic acid
in 10% Hcl followed by 1ml of
5% sodium nitrite. Heated and
cool and then add 1ml of 10%
sodium carbonate and few drops
of ethanol.

Observation

Inference

Soluble
Soluble
Soluble
Soluble
Insoluble
A violet coloured It indicates the
solution
was formation
of
obtained.
Ruhemanns
purple by amino
acid
containing
-NH2 group.
A dark blue or Histidine answers
violet
coloured this test and the
solution
was presence
is
obtained.
confirmed.

Appearance of red Histidine answers

colouration.
this test and the
presence
is
confirmed.

Ex. No:

REACTIONS OF TRYPTOPHAN

Date:

s. Experiment
no
1.

Observation

Inference

Solubility test
A Small amount of tryptophan
was dissolved in
a. Cold water
b. Hot water

Soluble
Soluble

c. Dilute Potassium hydroxide

Soluble
solution
d.
Ammonium
hydroxide Soluble
solution

Soluble

e. Alcohol

Soluble

f. Hydrochloric acid.
2.

Ninhydrin Test
To 1ml of tryptophan solution
added 0.5ml 1% ninhydrin and
heated for 2 min and allowed to
cool.

3.

A
purple It is due to the
coloration was got formation
of
Ruhemanns purple.

Hopkins Cole Test

To 1ml of trptophan solution A Violet coloured It

is

due

to

the

added 2ml of hopkins cole ring was got.

reagent and mixed thoroughly
and added 5ml of conc. H2SO4
along the sides of the test tube
so that the acid layer is formed
beneath the solution.

condensation
of
aldehyde with indole
group in presence of
sulphuric acid.

4.

Xanthoproteic test

5.

To 2ml of tryptophan solution A red coloration It is due to the nitration

of phenyl group.
added 1ml of conc. HNO3 was got.
boiled, cooled and added 40%
NaOH in drops.
Voisnet Rhode
Treated 5ml of the solution with A violet coloured It is due to the
paradimethyl
amino ring was got.
condensation of CHO
benzaldehyde and conc.H2SO4.
group with the indole
group in presence of
H2SO4.

6.

Aldehyde Test
Treated 1ml of tryptophan A Violet coloured
solution with 1 drop of 15% ring was got.
mercuric sulphate in 10%
H2SO4, added 3-4 drops of conc.
H2SO4.

7.

It is due to the
condensation of CHO
group with the indole
group in presence of
H2SO4

Ehrlischs Test
Treated 5ml of the solution with A pink coloured
paradimethyl
amino solution was got.
benzaldehyde and a few drops
of conc.Hcl.

It is due to the
condensation of CHO
group with the indole
ring with the presence
of Hcl.

Ex. No:
Date:

REACTIONS OF TYROSINE

S . No

Experiment

1.

Solubility test

Observation

Inference

A Small amount of tyrosine

was dissolved in
a. Hot water

Insoluble

b. Cold water
c.
Dilute
sulphate
d. Potassium
solution

2.

Insoluble
ammonium

Soluble

hydroxide
Soluble

e. Alcohol

Insoluble

f. Hydrochloric acid.

Soluble

Ninhydrin Test
To 1ml of tyrosine solution A purple
added 0.5ml 1% ninhydrin coloration was
and heated for 2 min and got.
allowed to cool.

3.

Mormers test
To small amount of A green
tyrosine solution added 3ml colouration was
of mormers reagent and got.
gently raise the temperature
to boiling point and boil for
5minutes.

4.

It is due to the
formation
of
Ruhemanns purple.

It is due to the
presence
of
phenolics hydroxyl
group.

Sulphuric acid test

Warm a little of the tyrosine A solution was
solution in a water both for got.
20 minutes with 3-5 drops
of concentrated H2SO4, cool
and wash into a beaker with
water.
Add
calcium
carbonate with constant
stirring until the solution is
no longer acidic filter,
concentrate the filterate and

Formation of ferric
salt of tyrosine with
sulphur group.

REACTIONS OF UNKNOWN AMINO ACID

Ex.no:
Date:
S.no

Experiment

1..

Ninhydrin test
To 1ml of histidine solution added
0.5 ml of 1% ninhydrin and heated
for 2 min and allowed to cool.

2.

Hopkins Collin Test

To 1ml of trptophan solution added
2ml of hopkins collin reagent and
mixed thoroughly and added 5ml
of conc. H2SO4 along the sides of
the test tube so that the acid layer
is formed beneath the solution.

3.

Aldehyde Test

Observation

Inference

Treated 1ml of tryptophan solution

with 1 drop of 15% mercuric
sulphate in 10% H2SO4, added 3-4
drops of conc. H2SO4.
4.

Ehrlischs Test
Treated 5ml of the solution with
paradimethyl amino benzaldehyde
and a few drops of conc.Hcl.

5.

Mormers test
To small amount of tyrosine
solution added 3ml of mormers
reagent and gently raise the
temperature to boiling point and
boil for 5minutes.

6.

Millon-naisse Test
To a few drops of tyrosine
solution add 3ml of millon nasse
reagent, heat and cool.then add 1ml
of 1% sodium nitrite solution.

7.

Sakaguchi test
To 5 ml of arginine solution added
5 drops of 15% NaOH and 4 drops
of freshly prepared
-napthol
in alcohol and bromine water

8.

Test for Histidine

To 2ml of histidine solution added
1% bromine in 33% acetic acid
until a yellow colour develops.
After 10 minutes added 2ml of 5%
NH4CO3 and placed in waterbath
for 10 minutes.

9.

Ehrlischs Diazo test

To 2ml of histidine solution, add
1ml of 1% sulphanilic acid in 10%
Hcl followed by 1ml of 5% sodium
nitrite. Heated and cool and then
add 1ml of 10% sodium carbonate
and few drops of ethanol.

10.

Lead Acetate Test

To the cysteine solution added a
small quantity of 10% lead acetate
and 4% of NaoH solution and boil
for few minute.

11.

Cyanide Test:
To the cysteine solution added a 2
drops of liquid ammonia and 5
% sodium cyanide and sodium
nitro-prusside solution.

12.

Nitoprusside Test
To 1-2ml of cystine solution added
dilute
sodium
nitroprusside
solution and a drop of dilute
sodium hydroxide solution

13.

Ferric Chloride Test

To the cystine solution added dilute
solution of Fecl3 and a drop of
copper sulphate solution.

14.

Bollings Test
(Modification of mecrothy

Sullivans test)
To 1 ml of methionine solution
the following reagents are added in
order & mixed after each addition
1. 1.5 ml of 1% glycine.
2. 1.5 ml of 5N NaOH.
3. 0.3 ml of 10% sodium
nitroprusside .
Placed in a water bath at
37-40 C for 15 min. Placed in a ice
bath for 5-7 min & then added 6N
Hcl & left to stand at room
temperature for 15 min.