P2. Laboratory Exam. Part 1: Test For Amino Acids

Biochemistry Laboratory
P2. Laboratory Exam. Part 1

Instructions: Research and complete the table below with information that are essential for further understanding
of Carbohydrates and Proteins. This will be the part one of your examination in laboratory, the second half or
practical exam will be an oral recitation regarding this topic so make sure that you are familiar with these tests.
The oral examination will be on April 25, 2021 at 1 PM.
Note: This is a group examination, indicate which group member is assigned for each tests. Deadline for this
research activity will be on April 25, 2021 at 5 PM.
TEST FOR CARBOHYDRATES

I. General test for carbohydrates
Test Principle Reagent End-product Interfence

Molish test Concentrated sulphuric acid Molish reagent 5- hydroxymethyl Positiveresult purple
hydrolyses glycosidic bonds - alphanaphthol furfural (bottom ring on the junction
Use/ Purpose: to give the in 95% ethanol reaction) and furfural
-specific for monosaccharaides, which (top reaction) Negative result: No
carbohydrates are then dehydrated to H2so4 purple ring on the
furfural and its derivatives. junction
Anthrone test
II. Test For Carbohydrates Based On Their Ability To Form Furfural And Its Derivatives
Test Principle Reagent End-product Interference

Seliwanoff’s test
Bial’s orcinol test
Mucic acid test
1|Page
Biochemistry Laboratory
III. Test For Carbohydrates Based On The Reducing Property Of Sugars Define:
a. Reducing sugars
b. Non- Reducing Sugars
Test Principle Reagents End-product Interferences

Benedict’s test
Barfoed’s test
Nylander’s test
Tollen’s test
TEST FOR AMINO ACIDS

General Tests for Amino acid
BRIONES, CHERUBIN FAITH T.
Test Principle Reagent Results
Biuret Test  Biuret test is a general  Copper sulfate (CuSO40 Biuret test positive:
Use/Purpose: test for compounds  Sodium hydroxide  color changes to purple
A Biuret test is a chemical test having a peptide bond. (NaOH)  all peptides and protein
used to determine the presence Biuret is a compound give the test positive
of a peptide bond in a formed by heating urea  Sodium potassium
substance. to 180° C. When biuret tartarate (commonly  Histidine is the only
is treated with dilute known as Rochelle salt) amino acid that give
copper sulfate in biuret test positive.
alkaline condition, a
purple colored Biuret test negative:
compound is formed.
This is the basis of  No color change
biuret test widely used
for identification of
proteins and amino
acids.
 The principle of biuret

test is conveniently
used to detect the
presence of proteins in
biological fluids.
Xanthoproteic Test Xanthoproteic test is used to  Concentrated Nitric When yellow colour appears, it
Use/Purpose: detect amino acids containing acid means aromatic amino acid is
 this test is used to an aromatic nucleus (tyrosine,  Original solution present in the solution.
differentiate aromatic tryptophan and  Aqueous ammonia
amino acids which give phenylalanine) in a protein
positive result from solution which gives yellow color
other amino acids nitro derivatives on heating with
conc. HNO3. The aromatic
benzene ring undergoes nitration
to give yellow colored product.
Phenylalanine gives negative or
weakly positive reaction though
this amino acid contains aromatic
nucleus because it is difficult to
nitrate under normal condition.
On adding alkali to these nitro
derivative salts, the color change
from yellow to orange
Ninhydrin The amino group belonging to a  Dissolve 0.35g of  The presence of a
Use/Purpose: free amino acid undergoes a ninhydrin in 100 ml ethanol purple-colored complex in
The ninhydrin test is a chemical chemical reaction with ninhydrin, (iso-propanol or 1:1 the tube represents a
test which is used to check which behaves as an oxidizing mixture of acetone/butanol positive result and indicates
whether a given analyte contains agent. When exposed to may be used instead of the presence of amino acid
amines or α-amino acids. ninhydrin, the amino acid ethanol). in the sample.
undergoes oxidative  Diluent solvent (for the  The absence of the
deamination, resulting in the quantitative test): Mix complex in the tube
liberation of CO2, NH3, and an equal volumes of water and represents a negative result
aldehyde along with hydrindantin n-propanol. and indicates the lack of
(which is a reduced form of  Standard solution (1% amino acids in the sample.
ninhydrin). protein solution)
 Sample solution
Now, the ammonia goes on to
react with another ninhydrin
molecule to form diketohydrin
(which is also known as
Ruhemann’s complex). This
complex is responsible for the
deep blue colour. When the
analyte contains Imino-acids like
proline, a yellow coloured
complex is formed. When
asparagine is used, the colour of
the resulting complex is brown.
Test for Amino Acids

Test Principle Reagent Results
Millon’s test
Lead Acetate Test or

Sulfur Test
Sakaguchi Test
2|Page

P2. Laboratory Exam. Part 1: Test For Amino Acids

Uploaded by

Copyright:

Available Formats

P2. Laboratory Exam. Part 1: Test For Amino Acids

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

P2. Laboratory Exam. Part 1: Test For Amino Acids

Uploaded by

Copyright:

Available Formats

Biochemistry Laboratory

P2. Laboratory Exam. Part 1

TEST FOR CARBOHYDRATES

Test Principle Reagent End-product Interfence

Test Principle Reagent End-product Interference

Bial’s orcinol test

Mucic acid test

b. Non- Reducing Sugars

Test Principle Reagents End-product Interferences

TEST FOR AMINO ACIDS

BRIONES, CHERUBIN FAITH T.

Test Principle Reagent Results

 The principle of biuret

Test for Amino Acids

Lead Acetate Test or

You might also like