PRACTICAL NO

Page no: Date:

AIM: - [A] Identification of reactions of carbohydrates.
REFERENCE: -
“Praticals in Biochemistry and clinical pathology” by Dr. Ramesh K. Goyal, B.S Shah
Prakash

1. Molisch test:
It is general test for carbohydrates. It is positive for any compound which can be dehydrated
to furfural or substituted furfural by concentrated sulphuric acid, the color is due to
condensation products of furfural or its derivatives with various phenolic substance e.g.
naphthol

2. Anthoron test
In this test keto form of 9-hydroxyanthracene, reacts with carbohydrates to produce a green
to blue green colour.

3. Iodine test
Except for polysaccharides all carbohydrates are oxidized by iodine solution and do not give
color. Different polysaccharides form different complex, with different colors which are
important for the identification.

4. Benedict’s test / Fehling’s test

Because of reactive carbonyl group, carbohydrates readily reduce metallic oxides
particularly that of copper and cupric solution, all reducing sugars precipitate Cu2O.

5. Barfoed’s test
In acid solution only, monosaccharides reduce cupric salts. This fact is the basis of barfoed’s
test. It cannot be used for testing urine or similar liquid containing chlorides.

6. Seliwonoff’s test
Due to the conversion of fructose by hot hydrochloric acid to levulinic acid and
hydroxymethyl furfural. Later compound is condensed with resorcinol to give red color.
Sucrose, which is readily hydrolysed to glucose and fructose, give red color with
 Seliwonoff’s reagent. Aldoses do not give this test positive but upon continued boiling, due
to gradual conversion to ketoses by hydrochloric acid, they give red color with reagent.

7. Osazone test
All reducing sugar react with phenylhydrazine to form crystalline Osazone. They have
characteristic crystalline forms depending upon the sugar from which they are derived,
AIM: - [B] chemical test for carbohydrates
SR TEST OBSERVATION INTERFERENCE
NO:
1 Molisch’s test Purple color ring at Carbohydrates
3ml sugar solution + 2 drops Molisch’s junction of two present
reagent mix in the test tube + 3 ml conc. liquids
H2SO4 from the side of the test tube so
as to form two layers.
2 Anthron test Green to the blue Carbohydrates
0.2 ml sugar solution + 2 ml Anthron green color present
reagent
3 Iodine test Blue color Starch present
3 ml sugar solution + dil iodine solution Purple -red color Dextrin present
dropwise. Red brown color Glycogen present
Colourless Mono/di
saccharides are
present
4 Reduction of methylene blue Blue color Reducing sugar
1 ml sugar solution + 1 drop of disappears due to present
methylene blue + 0.5 % dil sodium formation of leuco -
hydroxide and boil. methylene blue
5 Reduction of alk. Ferricyanide Yellow color Reducing sugar
3 ml alk. ferricyanide solution + 1 ml disappears present
dil NaOH, boil + dropwise sugar
solution
6 Tommer’s test Clear blue solution. Reducing sugar
2 ml CuSO4 + 2 ml glucose solution +2 After boiling present
ml dil. NaOH, boil it. yellow or red ppt of
Cu2O
7 Fehling’s test Red to yellow ppt Reducing sugar
1 ml sugar solution + 2 ml Fehling present
solution A + 2 ml Fehling solution B +
Boil
8 Benedict’s test Green, yellow or Reducing sugar
0.5 ml sugar solution + 0.5 ml sugar red ppts of Cu2 O. present
solution., boil for 2 min, cool.
9 Barfoed’s test Red ppts. Settle Monosaccharides
3 ml sugar solution + 3 ml Barfoed’s down at the bottom. sugar present
reagent. boil for 3 min, and cool
10 Picric acid test Red color due to Reducing sugar
3 ml sugar solution + 1 ml picric acid picramic acid present
solution + 1 ml dil. NaOH solution. +
boil
11 Rapid furfural test Deep purple color Fructose present
1 ml sugar solution + 6 drops of develops as soon as
Molisch’s reagent + 5 ml conc. HCl and the mixture begins
boil. to boil.

12 Seliwonoff’s test Red color after Fructose present

1 ml sugar solution + 3 ml seliwonoff’s further boiling
reagent, boil for 2 min. brown red ppts

13 Osazone test Needle shape Glucose /fructose

5 ml sugar solution + 10 drops of Small needle Lactose
glacial acetic acid + phenyl hydrazine appears like a ball
mixture of prickles
Plate like crystal Maltose
appears like sun
flower.
AIM: [C] GENERAL SCHEME FOR THE IDENTIFICATION OF CARBOHYDRATES