TEST FOR

CARBOHYDRATES
Iodine Test
Significance: The test is done to differentiate between helical and non-helical
polysaccharides. It is also used to determine the presence of starch in biological materials.
Principle Involved: Iodine forms a coordinate complex between the helically coiled
polysaccharide chain and iodine centrally located within the helix due to adsorption. The
color obtained depends upon the length of the unbranched or linear chain available for
complex formation.
Positive Result: Amylose – a linear chain component of starch gives a deep blue color.
Amylopectin- a branched chain component of starch gives a purple color.
Glycogen- gives a reddish brown color.
 TEST FOR
CARBOHYDRATES
Molisch Test
Significance: a chemical test which is used to check for the presence of
carbohydrates in a given analyte. 
Principle Involved: arbohydrates when treated with concentrated sulphuric acid
undergo dehydration to give furfural derivatives. These compounds condense
with Alpha naphthol to form colored products. Pentoses yield furfural while
Hexoses yield 5-Hydroxy methyl furfurals. 
Positive Result: A deep violet   coloration  is produced at the junction of two
layers
 TEST FOR
CARBOHYDRATES
Osazone Test
Significance: Reducing sugars
Principle Involved: A solution of reducing sugar when heated with phenyl hydrazine,
characteristic yellow crystalline compounds called Osazone are formed. These crystals have
definite crystalline structure, precipitation time and melting point for different reducing
sugars.
Positive Result: Needle shaped glucosazone crystals as viewed under the microscope.
Galactosazone crystals as viewed under the microscope(Rhombic plates). Sun flower shaped
Maltosazone crystals as viewed under the microscope. Powder puff/hedge hog shaped
crystals of lactose as viewed under the microscope.
 TEST FOR
CARBOHYDRATES
Benedict’s Test
Significance: Presence of reducing sugars
Principle Involved: Carbohydrates with free aldehyde or ketone groups have the ability to
reduce solutions of various metallic ions. Reducing sugars under alkaline conditions
tautomerise and form enediols. Enediols are powerful reducing agents. They reduce cupric
ions to cuprous form and are themselves converted to sugar acids. The cuprous ions combine
with OH- ions to form yellow cuprous hydroxide which upon heating is converted to red
cuprous oxide
Positive Result: The color of the precipitate gives a rough estimate of a reducing sugar
present in the sample. Green color - Up to 0.5 G% (+), Green precipitate - 0.5-1.0 G% (++),
Yellow precipitate -1.0-1.5 G% (+++), Orange precipitate- 1.5-2.0 G% (++++) and Brick red
precipitate- > 2.0 G %(+++++) 
 TEST FOR
CARBOHYDRATES
Anthrone Test
Significance: General test for carbohydrates.
Principle Involved: Carbohydrates are dehydrated by concentrated sulfuric
acid to form furfural. Active form of the reagent is anthranol, the enol tautomer
of anthrone, which reacts by condensing with the carbohydrate furfural
derivative to give a green colour in dilute and a blue colour in concentrated
solutions, which is determined colorimetrically.
Positive Result: bluish-green color
 TEST FOR
CARBOHYDRATES
Barfoed’s Test
Significance: Presence of reducing sugars.
Principle Involved: Aldoses and ketoses can reduce cupric ions even in acidic
conditions. This test is used to distinguish reducing mono saccharides from
disaccharides by controlling pH and time of heating. Mono saccharides react
very fast whereas disaccharides react very slowly.
Positive Result: The positive reaction indicates the presence of a reducing
mono saccharide. On prolonged heating disaccharides can also give this test
positive. Hence, the solution should be boiled for 3 minutes only. 
 TEST FOR
CARBOHYDRATES
Seliwanoff’s Test
Significance: Presence of ketoses and Presence of aldoses.
Principle Involved: Keto hexoses on treatment with hydrochloric acid form
5-hydroxy methyl furfural which on condensation with resorcinol gives a
cherry red colored complex. 
Positive Result:
Ketose: A cherry red colored precipitate within 5 minutes is obtained.
Aldose: A faint red colour  produced.
 TEST FOR
CARBOHYDRATES
Bial’s Test
Significance: Presence of pentoses and presence of hexoses. 
Principle Involved: The test reagent dehydrates pentoses to form furfural.
Furfural further reacts with orcinol and the iron ion present in the test reagent to
produce a bluish product. 
Positive Result:
Pentose: A blue-green product.
Hexose: A muddy brown to gray product
 TEST FOR
CARBOHYDRATES
Tollen’s Phloroglucinol Test
Significance: This reaction is positive for pentoses.
Principle Involved: This reaction is positive for pentoses, but not a specific indication of
them since hexoses and glycuronic acid also react. The distinction between the groups is in a
carefully conducted test. If the cherry red color develops very promptly, and if this is soluble
in amyl alcohol and alcoholic solution shows a specific absorbance band consistent with
resource data, them it can be considered a fairly reliable test for pentoses and glycuronic acid.
The colored compounds formed are condensation products of the various intermediate and
final acid decomposition products from carbohydrates with the phenolic compound
phloroglucinol. 
Positive Result: Cherry red color
 TEST FOR
CARBOHYDRATES
Mucic Acid Test
Significance: Specific test for galactose.
Principle Involved: This test is one in which concentrated nitric acid is heated
along with an aldose sugar to give a dicarboxylic acid. Nitric acid is able to
oxidize the terminal groups of aldoses, but leaves the secondary hydroxyl
groups unchanged. The dicarboxylic acid formed from galactose is
called mucic acid and is insoluble in cold aqueous solution. Those acids
formed from the other common sugars are soluble in water. Thus the formation
of the insoluble precipitate is an indication of the presence of galactose. 
Positive Result: White precipitate
MUCIC ACID TEST