Reagent Used
Test Purpose Principle/Purpose of reagent Result
A. Protein Denaturation
Effect of Heat Examine the effect of If the temperature is increased
heat on the solubility of slowly, a protein’s conformation
albumin generally remains intact until an
abrupt loss of structure and
function occurs over a narrow
temperature range.
Effect of 70% ethanol These compounds are capable
Alcohol 95% ethanol of engaging in intermolecular
hydrogen bonding with protein
molecules, disrupting
intramolecular hydrogen
bonding within the protein.
Effect of Heavy 1% AgNO3 These ion form strong bonds with
Metal 10% Pb(CH3COO)2 the carboxylate anions of the
acidic amino acids or SH groups
of cysteine, disrupting ionic
bonds and disulfide linkages.
Experiment 2: Carbohydrates
A. General Test for Carbohydrates
Molisch Test Used to test for All Molisch Reagent The test reagent dehydrates The
carbohydrates. Concentrated sulfuric acid pentoses to form furfural (top formation of
Monosaccharides give a reaction) and dehydrates a purple
rapid positive test. hexoses to form 5-hydroxymethyl product at
Disaccharides and furfural (bottom reaction). The the interface
polysaccharides react furfurals further react with - of the two
slower. naphthol present in the test layers.
reagent to produce a purple
product (reaction not shown).
Iodine Test Test for the presence of Iodine Solution The starch-iodide complex is The
starch. formed as charge - recall formation of
electrons are charged particles - a blue-black
is transferred between the starch complex.
and iodide ions - tri-iodide or
pentaiodide.
B. Comparative Test Reactions of Carbohydrates
Fehling’s Test The test is commonly used Fehling’s A solution Fehling’s test is one of the + Reddish
for reducing sugars but is Fehling’s B solution sensitive test for detection of brown ppt (
known to be NOT specific reducing sugars. Fehling’s glucose,
for aldehydes. reagents comprises of two fructose,
solution Fehling’s solution A and lactose)
solution B. Fehling’s solution A is
aqueous copper sulphate and
Fehling’s solution B is alkaline
sodium potassium tartarate (
Rochelle salt). Rochelle salts
(sodium potassium tartarate)
present in the reagent acts as
the chelating agent in this
reaction.These two solution are
mixed in equal amount before
test.
On heating an aldehyde or
reducing sugar with Fehling’s
solution give reddish brown
precipitate. Formation of red
precipitate of cuprous oxide
denotes the presence of
reducing sugar.
Benedict Test Benedict’s Test is used to Benedict’s Reagent The Reducing sugar under Formation of
test for simple alkaline condition form enediols. a reddish
carbohydrates. The Benedict’s solution contains precipitate
Benedict’s test identifies milder alkali Na2CO3. Enediols within three
reducing sugars are powerful reducing agents. minutes.
(monosaccharide’s and They can reduce cupric ions to Reducing
some disaccharides), cuprous ions which is the basis sugars
which have free ketone for Benedict’s reaction. The present.
or aldehyde functional cuprous hydroxide during the
groups. Benedict’s process of heating is converted
solution can be used to to red cuprous oxide.
test for the presence of
glucose in urine.
C. Specific Tests For Carbohydrates
Barfoed’s Test Detect the presence of Barfoed’s Reagent Reducing monosaccharides are The
monosaccharide oxidized by the copper ion in formation of
(reducing) sugars in solution to form a carboxylic a reddish
solution acid and a reddish precipitate precipitate
of copper (I) oxide within three within three
minutes. Reducing disaccharides minutes.
undergo the same reaction, but
do so at a slower rate.
Seliwanoff’s Test for the Ketoses. Seliwanoff’s Reagent The test reagent dehydrates Formation of
Test ketohexoses to form 5- a red
hydroxymethylfurfural. 5- product.
hydroxymethylfurfural further
reacts with resorcinol present in
the test reagent to produce a
red product within two minutes
(reaction not shown).
Aldohexoses react to form the
same product, but do so more
slowly.
Bial’s Orcinol Test for the Pentoses. Bial’s Orcinol reagent The test reagent dehydrates Formation of
Tets pentoses to form furfural. Furfural a bluish
further reacts with orcinol and product.
the iron ion present in the test
reagent to produce a bluish
product (reaction not shown).
Mucic Acid Test to identify Gluocse, Conc, Nitric Acid • The nitric acid oxidizes Presence of
Test Galactose and Lactose galactose to an isomer of Crystals
tetrahydroxyadipic
(tetrahydroxyhexanedioic)
acid that crystallizes out from
water under the conditions of
the test.
• Lactose is a disaccharide,
galactosyl(1- >4)glucose. The
nitric acid first catalyzes
hydrolysis of lactose to glucose
and galactose. Then the nitric
acid oxidizes both of those
sugars to the tetrahydroxy acids.
Mucic acid crystallizes out,
which is a positive test. The acid
from glucose remains in solution.
• This test will give positive results
if there is a
presence of crystal precipitate
