REACTIONS OF GLUCOSE

1. Molisch Test
1. It is general Reaction for all Carbohydrates.
2. All carbohydrates that have 5 carbons answer
the test.

Reagent: α-naphthol in absolute alcohol

Principle
• Carbohydrates undergo dehydration when
treated with Conc. H2SO4 and form
FURFURALS
• Furfural is a Heterocyclic aldehyde with a ring
structure.
• Pentoses – Furfurals
• Hexoses – Hydroxymethyl Furfurals.
Experiment Observation Inference

Take 2ml of glucose A PURPLE COLOR Glucose undergoes

Solution in a test ring is seen at the dehydration when
tube and add 2 junction of two treated with Conc.
drops of MOLISCH layers H2SO4 and forms
REAGNT. Then hydroxymethyl
incline the Test furfural which on
tube is slightly and condensation with
add 2ml of Conc.
H2SO4 α-naphthol gives a
Slowly along the purple color
sides of the test complex thus
tube. confirming it as a
carbohydrate.
 2. Benedict’s Test
- This is specific test for reducing
carbohydrates.
- Differentiates between Reducing and Non
reducing carbohydrate.
- Reducing Carbohydrate: A carbohydrate
with a free aldehyde or free keto group of
anomeric carbon.
Reagent
a) Copper Sulphate: Provides Cu2+ ions.
b) Sodium Carbonate: Provides Alkaline
Medium.
c) Sodium Citrate:
- Prevents precipitation of cupric ions by
forming cupric sodium citrate complex and
ensures continuous supply of Cupric ions.
- Also improves the shelf life of reagent by
preventing interaction between sodium
carbonate and copper sulphate.
Principle
• Reducing sugars in alkaline medium undergo
TAUTOMERIZATION to form ENEDIOLS, which
are powerful reducing agents.
• Enediols reduce Cupric ions to Cuprous ions
which forms a yellow precipitate of cuprous
hydroxide.
• This cuprous hydroxide on further heating,
forms a RED precipitate of Cuprous Oxide.
Experiment Observation Inference

5ml of Benedict’s GREEN to BRICK RED Glucose undergoes

reagent is taken in a PRECIPITATE is tautomerisation in
test tube and add 8 observed (Color alkaline medium to
drops of glucose depends on the form enediol, a
solution & boil for 1 – amount of sugar powerful reducing
2 min present) agent which reduces
Cu++ ions to Cu+ ions
which forms a yellow
precipitate of CuOH
which on further
heating, forms a RED
precipitate of CuO
confirming it as a
reducing
carbohydrate.
• Benedicts is also called as SEMIQUANTITATIVE
test.
Based on the color of the precipitate we can
approximately tell the amount of sugar present in
the given solution

GREEN PPT  0.1 - 0.5 gm / 100 ml (+)

YELLOW PPT  0.5 -1.0 gm / 100 ml (++)
ORANGE PPT  1.0 - 1.5 gm / 100 ml (+++)
RED PPT  1.5 - 2.0 gm / 100 ml (++++)
BRICK RED PPT  ≥ 2.0 gm / 100 ml (+++++)

- BED SIDE TEST FOR GLYCOSURIA

 3.Barfoed’s Test
A specific reducing test to identify
MONOSACCHARIDES.
This test uses copper ions to detect
reducing sugars in an mild ACIDIC medium
BARFOED’S REAGENT :
a) 13.3 gm of Copper acetate.
b) 1.8 ml GLACIAL ACETIC ACID
c) 200 ml Distilled Water
 Principle:
- Monosaccharides form enediols in mild acidic
medium which can reduce Cu2+ to Cu+ ions
which up on heating form a scanty reddish
precipitate of CUPROUS OXIDE.

- Monosaccharides are powerful reducing

agents even in an unfavourable mild acidic
medium, unlike disaccharides which have an
hinderance from the second monosaccharide
component.
Experiment Observation Inference

Take 3ml of Barfoed’s A RED SCUM at the Glucose undergoes

Reagent in a Test bottom and the sides tautomerisation in a
tube and add 1 ml of of the test tube wall mild acidic medium
Glucose solution. is formed due to to form enediol, a
Then boil for 30 cuprous oxide. powerful reducing
seconds to 1 minute. agent which reduces
Cu++ ions to Cu+ ions
which up on heating
form a scanty reddish
precipitate of CuO
confirming it as a
monosaccharide.
 Seliwanoff’s Test

- This test is specific for KETOHEXOSES.

Principle:
- Acid in Seliwanoff reagent dehydrates
KETOHEXOSES to form 5 - Hydroxy Methyl
Furfurals which Condenses with RESORCINOL to
form CHERRY RED Color Precipitate.
- Aldohexoses will not show a significant color
change.
- Ketohexoses are more reactive in presence of
Hcl.
Experiment Observation Inference

3 ml of Cherry Red Color Glucose is not a

Seliwanoff’s Precipitate is not ketohexose, it is an
Reagent is taken in observed. aldohexose.
a test tube and 1
ml of glucose
solution is added
& boiled for 30 sec
– 1 min and cooled
immediately
 Foulger’s Test

- This test is specific for KETOHEXOSES.

Principle:
- Acid in Foulgers reagent dehydrates
KETOHEXOSES to form 5 - Hydroxy Methyl
Furfurals which Condenses with Stannous
Chloride to form DEEP BLUE Color Precipitate.
- REAGENT : a) 2% Stannous Chloride
b) 40% Urea
c) 40% H2SO4
Experiment Observation Inference

3ml Foulger’s DEEP BLUE Color Glucose is not a

Reagent is taken in Precipitate is not ketohexose it is an
a test tube and 1 observed. aldohexose.
ml glucose solution
is added. It is
boiled for 30 sec –
1 min and cooled
immediately.
 Rapid furfural test
• This is a specific test for KETOHEXOSES.

PRINCIPLE: Similar to Molischs test the only

difference is in place of Conc Sulphuric acid,
freshly prepared Conc. HCl is used to form
coloured complex.
Experiment Observation Inference

To 1ml of Conc. HCl A PURPLE COLOR Glucose is not

is taken in a test ring is not seen at Ketohexose
tube, to this add 1- the junction of two
2 drops of layers
(MOLISCH
REAGNT) and mix
it. To this add few
drops of
carbohydrate
solution.
 OSAZONE TEST
• Reducing sugars react with phenylhydrazine
to form phenylhydrazones, which further
react with excess of phenylhydrazine to from
Osazones .
• REAGENTS: 1) Phenylhydrazine hydrochloride.
2) Sodium Acetate
3) Glacial acetic acid.
Experiment Observation Inference
To 5ml of carbohydrate Characteristic
solution add 1 spatula crystals are
of mixture of observed:
phenylhydrazine a) Needle shaped – Glucosazone is formed
hydrochloride and Glucose, Fructosazone is formed.
sodium acetate. To this Fructose.
add 8 – 10 drops of b) Sunflower petal
glacial acetic acid and shaped – Maltosazone is formed.
place in boiling water Maltose
bath for 30 min and c) Powderpuff Lactosazone is formed.
then cool under tap shaped – Lactose
water . Then mount
crystals on a glass
slide, and observe
under microscope.
• Time of formation: Ketohexose form osazones rapidly
followed by aldohexoses then disaccharides.
• Solubility:
Monosaccharide osazone – Insoluble in hot
solution, so observed as precipitate.
Disacccharide osazone – Soluble in hot solution.
• Colour: Glucose, Fructose – Greenish yellow.
Maltose, Lactose – Yellow
• Individual Osazones:
Glucose, Fructose – Needle shaped.
Maltose – Petal, Lactose – Fiber
• Associations: Glucose, Fructose – haystack
Maltose – Sunflower Lactose – Powderpuff
Experiment Observation Inference
To 5ml of glucose Long Needle shaped Glucosazone is
solution add 1 spatula greenish yellow individual formed
of mixture of crystals are observed.
phenylhydrazine Group of crystals will appear
hydrochloride and as hay stack.
sodium acetate. To this
add 8 – 10 drops of
glacial acetic acid and
place in boiling water
bath for 30 min and
then cool under tap
water . Then mount
crystals on a glass
slide, and observe
under microscope.
 Glucose
• Monosaccharide, Reducing sugar, Aldohexose.
• Most important carbohydrate.
• Pyranose and Furanose forms are Structural
forms of glucose.
• Glucose circulates in the blood of animals
as blood sugar.
• Insulin and Glucagon are two hormones
important in regulation of Glucose levels in
the body.
• Epimers of Glucose: Mannose and
Galactose.
• Isomer of Glucose: Fructose.
• Disaccharides: Cellobiose, Lactose, Sucrose.

• Polysaccharides: Starch, Cellulose in Plants

Glycogen in Animals
 Sources of Glucose
• Bread, Rice, Potatoes, Sweet corn.
• Sugarcane, Fruits.
• Table sugar.
• Sugar syrups.