Identification Scheme for carbohydrates in given unknown sample:

Experiment test solution

Perform Molisch’s Test

if +ve if -ve

carbohydrate is present carbohydrate is absent

Perform Iodine test

if +ve if -ve

Polysaccharides is present Mono-or Di- Saccharide is present

Starch- Blue

Dextrin – Purple perform Benedict’s test

Glycogen- Reddish brown if +ve if -ve

Perform Half saturation Test Reducing sugar is present Non -reducing sugar is present
if +ve if –ve

Starch is present Dextrin, glycogen Perform Inversion

Test is present if +ve

Perform Barfoed’s Test Sucrose is

present
if +ve if -ve

Monosaccharide is present Reducing disaccharide is present

Perform Osazone
Perform Osazone Test
Test

Sunflower shape Puff shape

Fructose
Fructose is Glucose is crystal are formed crystal are
formed
present present

Maltose is Lactose is
Perform Needle shape present present
confirmatory crystal
test
SR TEST OBSERVATION INFERENCE
NO.
1 MOLISCH’S TEST : Reddish violet or purple Carbohydrate is
Take 3 ml of sugar solution in a test tube + 5 color ring is observed at confirmed
(CT) drops Molish’s reagent and mix + 3 ml junction of two liquid
con.H2SO4 along with the wall of test tube [NOTE: A green ring or
such that two layer are formed. Make the green color is
tube upright and rotate it gently between considered as negative
the palms. test]
2 ANTHRONE TEST : Green to blue-green Carbohydrate is present
(CT) Take 0.5 ml sugar solution in a test tube + 2 color is observed
ml Anthrone reagent , mix thoroughly.
3 IODINE TEST : Blue color is formed on Starch is present
(CT) Take 3 ml of sugar solution in a test tube + addition of iodine which
one drop of iodine solution. Mix, heat and disappear on heating
then cool. and reappears on
cooling
Purple –red color is Dextrin is present
formed on addition of
iodine which disappear
on heating and do not
appear on cooling.
Red-brown color is Glycogen is present
formed on addition of
iodine which disappear
on heating and
reappears on cooling
4 HALF SATURATION TEST : White precipitates are Starch is present
(CT) Take 5 ml of carbohydrate sample solution observed
in test tube + equal volume of saturated
solution of ammonium sulphate. Shake
vigorously and allow standing the mixture
for atleast 10 minute.
5 FULL SATURATION TEST : White precipitates are Starch or dextrin or
Take 5 ml of carbohydrate sample solution observed glycogen is present
in a test tube. Saturate it with solid
ammonium sulphate. Shake vigorously and
allow standing the mixture for atleast 10
minute.
6 REDUCTION OF METHYLENE BLUE : Blue color disappears Reducing sugar is
Take 1 ml sugar solution in a test tube + one present
drop of Methylene blue solution (1%)
+ 0.5 l dil NaOH and boil.
7 REDUCTION OF ALKALINE FERRICYANIDE Yellow color disappear Reduction sugar is
SOLUTION : present
 Take 3 ml of alkaline [K3Fe(CN)6] solution
(1%) in a test tube + 1 ml dil. NaOH, boil +
drop wise sugar solution
8 FEHLING’S TEST : Brick red precipitates Reducing sugar is
Take 2 ml of fehling sol0ution A + 2ml of are observed present
fehling solution B in a test tube. Heat to boil
+ 1ml sugar solution and boil for 2 min
9 BENEDICT’S TEST : Appearance of green, Reducing sugar is
Take 5 ml of benedict’s reagent in test tube yellow or red present
+ 8 drop of sugar solution, boil for 2 min, precipitates
cool
10 PIRIC ACID TEST : Blood red color is Reducing sugar is
Take 3 ml sugar solution in a test tube + 1ml observed present
picric acid solution (1%) +1 ml dil.NaOH
solution and boil.
11 INVERSION TEST : Benedict’s test and Sucrose is present
(CT) Take 5 ml of sugar solution in test tube + seliwanoff’s test comes
one drop of conc.HCl and boil for one positive
minute. Cool and divide the solution into
two test tube. Neutralize portion of one test
tube by adding a pinch of sodium
bicarbonate. Perform Benedict’s test with
this portion. Perform Seliwanoff’s test with
the second portion
12 BARFOED’S TEST : Red color due to Reducing agent
3 ml sugar solution solution + 3 ml Barfoed’s picramic acid
reagent , boil for 3 min. and cool.
13 OSAZONE TEST : Needle shape crystal Glucose or fructose is
(CT) 5 ml sugar solution + 10 drop of glacial lying singly or sheaves present
acetic acid + 100 mg phyenlhydrazine
mixture [knife point of phynelhydrazine HCl
and twice the amount of anhydrous sodium Small needle shape Lactose is present
acetate]. Shake well and heat on boiling crystal appear like ball
water bath for 60 min. Allow the test tube or prickles or puff shape
to cool down slowly to room temperature.
Spread a suspension on a clean, dry Crystal appear like sun Maltose is present
microscope slide and observe crystal under flower shape
low power of microscope.

14 RAPID FURFURAL TEST : Deep purple color Fructose present

(CT) Take 1 ml carbohydrate solution in test tube develop as soon as the
+ 6 drop of Molisch’s reagent + 5 ml mixture begins to boil
conc.HCL and boil.
15 SELIWANOFF’S TEST : Deep red color is Fructose is confirmed
(CT) Take 3 ml of Seliwanoff’s reagent in test observed
tube + 1 ml of carbohydrate solution. Boil
for 30 second
16 BIAL’S TEST : Blue-green color is Pentose sugar is
To 2 ml of Bail’s reagent , add 4-5 drops of obtained confirmed
test solution in a test tube and heat the
solution in boiling water bath.
17 MUCIC ACID TEST : Crystal of mucic acid is Galactose is present
TAKE 25 ml of carbohydrate solution in a formed
beaker + 5 ml of concentrated nitric acid.
Heat until the volume is reduced to about 5
ml. Allow the solution to cool gradually.
Observe the crystal so formed in the
microscope.