Test for Carbohydrates

Molisch’s Test
It is a screening test for confirming the presence or absence of carbohydrates in a given
solution. It is a highly sensitive test for carbohydrates. Monosaccharides,
oligosaccharides, and polysaccharides all give positive Molisch’s test. 

Principle

This test is based on the reaction of the alpha-naphthol with carbohydrate in the presence
of sulfuric acid. The sugars react with alpha-naphthol in an acidic environment to form
purple-colored furfural or hydroxymethylfurfural derivatives. The intensity of the color is
directly proportional to the amount of carbohydrate present in the solution. 

Reagent

 Molisch’s reagent: 5% alpha-naphthol solution in ethyl alcohol

Observations

A violet-colored ring is formed at the junction of the two liquids i.e. solution with
Molisch’s reagent and the sulfuric acid. 

Result

The violet ring indicates the presence of carbohydrates in the solution. 

Points to Remember

 To give a positive test, the carbohydrate must have at least five carbons. (It
is so because it involves the formation of furfural derivatives that contain
five carbon atoms.)
 Impurities in the reagent give green color indicating a false-negative test.
 Oligosaccharides and polysaccharides are first broken down to
monosaccharides by acid which then give the Molisch’s test positive. 
 Proteins and lipids having an attached carbohydrate can also give this test
positive. 
Seliwanoff’s Test
This test is used to detect monosaccharides with a ketonic functional group. It is widely
used to differentiate fructose, a keto sugar, from glucose and galactose

Principle

This test involves the formation of furfural derivatives by monosaccharides with

hydrochloric acid. The furfural derivatives formed by a sugar with ketonic functional
group condense with resorcinol to form a chromogen having cherry-red color. 

Reagents

Seliwanoff’s reagent is used that contains in 100 ml of water;

 50  mg resorcinol
 33 ml of concentrated HCL
Observations

A cherry red color forms in the test tube upon cooling.

Results

The given solution contains a keto-sugar. 

Points to Remember

 Seliwanoff’s test is specific only for hexoses having a ketonic functional

group 
  Sucrose also gives a positive test because it is hydrolyzed to glucose and
fructose
 It is highly sensitive for sucrose even at .1% concentration
Phloroglucinol test
This test is specifically to detect galactose and lactose in a solution.

Principle

This test also involves the formation of furfural derivates in the presence of concentrated
HCL. The furfural derivatives formed by galactose then condense with the phloroglucinol
to form a red-colored compound. 

Reagents

Tollen’s reagent is used that contains;

 10 ml concentrated HCl mixed with 8 ml of 0.5% phloroglucinol

Observations

When the solution is boiled and allowed to cool, it turns yellow to red. 

Results

The change of color to red indicates the presence of galactose in the solution.

Points to Remember

 Lactose also gives this test positive as it is hydrolyzed by acid to yield

glucose and galactose. To differentiate between the two, perform Barfoed’s
test. 
 The furfural compounds formed by pentoses having a keto group also form
a similar red-colored compound with phloroglucinol.  

Fehling’s test
Fehling’s test is a chemical test used to differentiate between reducing and
non-reducing sugars. This test can also be used to distinguish ketone
functional group carbohydrates and water-soluble carbohydrates.

Principle of Fehling’s Test

 The carbohydrates having free or potentially free carbonyl groups
(aldehyde or ketone) can act as reducing sugars.
 The Fehling’s solution appears deep blue in color and consists of
copper sulfate mixed with potassium sodium tartrate and strong
alkali, which is usually sodium hydroxide.
Result and Interpretation of Fehling’s Test

 The appearance of a reddish-brown precipitate indicates a positive

result and the presence of reducing sugars.
 The absence of the reddish precipitate or the appearance of deep
blue color indicates a negative result and lack of reducing sugars.