Angela Beatriz C.

Mamauag

Reaction Paper on Qualitative Analysis of Amino Acids

In the short video titled “Qualitative Analysis of Amino acids by Amrita University” different tests were
conducted to determine the presence of amino acids in certain solutions. Amino acids play many critical
roles in our body. They are vital for processes such as synthesis of proteins, hormones, and
neurotransmitter. Amino acids contain an amine group, a carboxylic group, and a side chain which
differs between various amino acids. Amino acids in a solution can be identified in the laboratory by
conducting tests by presence of color, precipitation, and more. Ten chemical tests namely Ninhydrin
test, Xanthoproteic acid test, Pauly’s Diazo Test, Millon’s test, Histidine test, Hopkins cole test, Sakaguchi
test, Lead Sulphide test, Follins McCarthy Sullivan’s test, and Isatin test

Aa are molecules containing an amine group, a carboxylic group and a side chain that ranges between
different amino acids. They are amphoteric in nature. In the laboratory, presence of amino acids can be
indentifieed in a soulution by color, precipitation and more. Ten chemical tests namely ninhydrin test,
xanthoproteic acid test, pauly’s diazo test, millon test, histidine test, Hopkins cole test, sakaguchi test,
lead sulphide test, follins mccarthy sullivan’s test, isatin test

Ninhydrin test

Transferred few drops of an unknown solution to a test tube and added a few drops of Ninhydrin
reagent. The test tube was placed in boiling water bath for five minutes. After the water bath, the color
of the soulution was observed. A blue colored solution t indicates that there is a presence of alpha
amino acids while a yellow colored solution indicates that there is a presence of immunoacids in it.

Xanthoproteic acid test

One milliliter of an unkn”own solution was transerred to a test tube and was added a few drops of
concentrated nitric acid solution in a fume hood. Marble chips were added in the solution to avoid
bumping of the solution while boiling. The test tube with solution was heated on the Bunsen burner and
was cooled by placing under running water. A few drops of 40% NaOH was added in the soulution. Color
of soulution changed to an orange color which indicates that there is presence of aromatic amino acids.

Pauly diazo test

A test tube containing an unknown soulution was placed in an ice bucket. A few drops of sulphanilic acid
gathered from the ice bucket and sodium nitrate were added in the unknown solution. Then, a few
drops of sodium bicarbonate solution were also added. The appearance of a red colored solution in
addition of sodium bicarbonate solution indicates the presence of histadine and thyroxine

Hopkins Cole Test

One ml of an unknown solution was placed in a test tube. 1 ml of acetic acid and glyoxylic acid reagent
were added to the same test tube. The solution was shook carefully. A few drops of concentrated
sulfuric acid were added on the side of the test tube. A purple violet ring was observed which indicates
the presence of tryptophan.

Lead sulphide test

A few drops of 40% NaOH were added in a test tube containing one milliliter of an unknown solution.
Marble chips were also added in it to prevent bumping of the solution while heating in the Bunsen
burner for 5-10 minutes. After heating, the contents were allowed to cool by placing the test tube under
running water. A few drops of 10% Lead acetate were in the solution. Black precipitate was observed
which indicates the presence of cysteine.

ISatin test

A drop of an unknown solution was placed on a filter paper strip using a dropper. The filter paper was
dried using a hair blower. A drop of isatin ragent was applied on the same spot and was heated again. A
blue colored spot was observed on the filter paper which indicates the presence of amino acids.

FOllins Mccarthy Sullivan’s test

A few drops of 40% ml NaOH followed by a few drops of glycine and sodium nitroprausside were added
in a test tube containing one ml of an unknwon solution. The soluton was placed in a hot water bath
with temperature of 40 degrees celsuis in 15 minutes. 0.5 mL of 6 N HCL in the solution taken from
water bath. Upon adding HCl, the color of the solution changed to a red color which indicates the
presence of methionine.

Sakaguchi test

A few drops of an unknown solution were placed on a test tube. A few drops of 40% NaOH, alpha-
naphthol solution, 5% urea solution, hypo bromic solution were added respectively in the solution. The
color of the solution changed to red which indicates the presence of alanine.

Histidine test

A few drops of 5% Bromine in 33% acetic acid solution were added in a test tube containing 2 mL of
unknown solution. The test tube containing solution was kept in room temperature for 10 minutes. 2 ml
of ammonium carbonate was added in the solution and was placed in boiling water bath for 5 minutes.
After the water bath, the color of the solution changed to blue indicating the presence of histidine.

Millon’s test

Millon’s reagent was added in a test tube containing solution and was shook. A few drops of
concentrated nitric acid solution was added. The color of the solution changed into red indicating the
presenc of tyrosine.

Building blocks of all proteins, alpha amino acids are known to react with excess ninhydrin to from a
product