BIOCHEMISTRY LAB REPORT

NO. OF EXPERIMENT : 2
TITLE OF EXPERIMENT : QUALITATIVE TEST FOR AMINO ACIDS
DATE OF EXPERIMENT : 4 APRIL 2023
GROUP : AS1142A2
PROGRAM : DIPLOMA MICROBIOLOGY
CODE SUBJECT : MIC 180

STUDENT’S NAME MATRIX NUMBER

NURUL INSYIRAH BINTI SEDEK 2022661492
PUTERI AILHANNA NAJEEHAH BINTI ZAIDI 2022619254
ARNIEZA SHAFIQA BINTI SUZELI 2022675976
NURUL AQILAH BINTI MOHAMAD YAZID 2022822636
NISRINA NABIHAH RIDWANA BINTI BADRUL SHAHRI 2022483426
NOR AIDA NATASHA BINTI HASMAWI 2022475904
OBJECTIVE
To conduct a series of qualitative tests for amino acids.

INTRODUCTION

Qualitative test of amino acids is also defined as the analytical method to

determine the content of amino acid, peptides and protein amount present in the
sample. Amino acids are the basic building blocks of protein. The structure
consists of an amino group (-NH2), a carboxylic group (-COOH) and R-group
or side chain. The reaction between the functional groups of amino acids and
the chemical reagents will gives a characteristic colour to the sample. There are
few tests that can be carried out depending on the type of R-group of amino
acid. Qualitative test of amino acids includes Ninhydrin Test, Millon’s Test,
Hopkin’s Cole Test, Xanthoproteic Test, Sakaguchi Test and Lead Sulphide
Test. All these tests are able to tell us the function of protein and the effects of
their presence, absence and modification.

MATERIALS
 Test tubes  Phenylalanine
 Test tube rack  Proline
 Pipette  Millon reagent
 Boiling water bath  Natrium Nitric
 Dropper  Concentrated Sulphuric Acid
 Ice box  Cysteine
 Glycine  40% NaOH
 Ninhydrin  Sodium Plumbate (Na2O3Pb)
 Tyrosine  Methionine
 Tryptophan
METHOD

Test 1 : Ninhydrin Test

a) Pipette 1 ml of glycine in a test tube.

b) Add 5 drops of ninhydrin.
c) Heated the test tube mixture in boiling water bath for 2 minutes.
d) Recorded changes.
e) Repeated using stock solutions of tyrosine, tryptophan, phenylylalanine,
and proline.

Test 3 : Millon Test

a) Pipette 1 ml of glycine in a test tube.

b) Added 5 drops of Millon reagent.
c) Heated the test mixture in a boiling water bath for 10 minutes.
d) Cool it down to room temperature.
e) Added 5 drops of natrium nitric.
f) Recorded changes.
g) Repeated using stock solutions of tyrosine, and phenylalanine.
Test 4 : Hopkins-Cole Test

a) Mixed 1 ml of glycine with 1 ml of glacial acetic acid in a test tube.

b) Incline the test tube and slowly and 1 ml of concentrated sulfuric but do
not mixed.
c) Two layers should form.
d) Let the layers stand and note the colour at the interface sulfuric but do not
mixed.
e) Repeat using stock solutions of tyrosine and tryphtophan.

Test 5 : Lead Sulpide Test

a) Pipette 1 ml of cysteine in a test tube.

b) Added 1 ml 40% NaOH.
c) Swirled the test tubes.
d) Heat the test tube in cold water to cool them down.
e) Added 1 ml of sodium plumbate (Molecular Formula: Na2O3Pb)
f) Recorded any changes.
g) Repeated using stock solutions of cystine and methionine.
 RESULTS

NINHDYRIN TEST

SAMPLE OBSERVATION CONCLUSION

GLYCINE COLOURLESS → DARK PURPLE POSITIVE
TYROSINE COLOURLESS → VIOLET POSITIVE
TRYPTOPHAN COLOURLESS → SLIGHTY POSITIVE
PURPLE
PHENYLALANINE COLOURLESS → VIOLET POSITIVE
PROLINE COLOURLESS → YELLOW POSITIVE

Figure 1 : before adding nynhdyrin and heating process

 Figure 2 : after heating process

MILLON TEST

SAMPLE OBSERVATION CONCLUSION

GLYCINE COLOURLESS → COLOURLESS NEGETIVE
TYROSINE COLOURLESS → BROWN POSITIVE?
PHENYLALANINE COLOURLESS → COLOURLESS NEGETIVE

Figure 3 : glycine, tyrosine and phenylalanine after heating process

HOPKINS-COLE TEST

SAMPLE OBSERVATION CONCLUSION

GLYCINE COLOURLESS → COLOURLESS NEGETIVE
TYROSINE COLOURLESS → COLOURLESS NEGETIVE
TRYPTOPHAN COLOURLESS → A HUE OF POSITIVE
PURPLE RING CAN BE SEEN

Figure 4 : tryptophan after heating process

LEAD SULPHIDE TEST

SAMPLE OBSERVATION CONCLUSION

CYSTEINE COLOURLESS → BLACK PRECIPITATE IS POSITIVE
FORMED
METHIONINE COLOURLESS → COLOURLESS NEGETIVE

Figure 5 : cystein and methionine after heating process