Reactions of Monosaccharides

REACTIONS OF GLUCOSE

S.NO EXPERIMENTS OBSERVATION INFERENCE

1 MOLISCH’S TEST: A violet colour ring was Carbohydrate +

5% solution of α- naphthol in got at the junction of two conc.Sulphuric acid + furfural
alcohol. liquid. furfural + α naphthol gives
To 5.0 ml of glucose solution, violet color. The reaction is
added two drops of Molisch’s due to the condensation of
reagent and 3.0ml of hydroxy methyl furfural with
concentrated sulphuric acid sulphonated naphthol.
along the sides of the test tubes
without shaking, so as to form a
layer of acid beneath the glucose
solution.
2 IODINE’S TEST: No characteristic change Glucose does not answer this
To a small amount of glucose was noticed. test, as it is not a
solution, added a few drops of polysaccharides.
dilute iodine solution.
3 FEHLING’S SOLUTION:
PREPARATION OF FEHLING’S REACTION
Fehling’s reagent is a mixture of copper sulphate and alkaline tartaric acid. About 34.65 gm
of copper sulphate was dissolved in distilled water and made up to 500ml- 125gm of
 potassium hydroxide and Rochell’s salt were dissolved in distilled water and made upto
500ml. The solution are mixed.
To 5.0ml of Fehling’s reagent in A reddish brown Cu2+ + reducing sugar NaoH
a test tube added a small amount precipitate was obtained. Cupric hydroxide (CuoH) →
of glucose solution and heated in Cuprous hydroxide (Cu2o).
a boiling water bath for three The red precipitate is cuprous
minutes. oxide. This shows that
glucose is reducing sugar.
4 BENEDICT’S TEST:
PREPARATION OF BENEDICT’S REAGENT
Dissolved 173gm of sodium citrate and 100gm of sodium carbonate in 500ml of distilled
water by warming. Filtered through a filter paper and made up of volume of the filtrate
(carbonate titrate solution) to 250ml of distilled water. Dissolved 173gm of copper sulphate
with volume with distilled water to one liter.

To 2.0ml of glucose solution First a greenish yellow and Cu2++ reducing sugar Na2Co3
added 8 drops of benedict’s then a reddish brown cuprous hydroxide(cu2o). This
reagent and heated in boiling precipitate was obtained. shows the reducing action of
water bath for 3 min and then glucose.
allowed to cool spontaneously.
5 BARFOED’S TEST:
PREPARATION OF BARFOED’S REAGENT:
Dissolved about 13.3gm of neutral crystalline copper acetate in 200ml of distilled water.
Filtered if necessary then added 1.8ml of glacial acetic acid.
 To 5.0ml of Barfoed’s reagent, A brick red precipitate was This indicates that glucose is
added 5.0 ml of glucose solution got at the bottom and at the a monosaccharides.
and heated in boiling water bath side of the test tube.
for three mintues.
6 PICRIC ACID TEST:
PREAPARTION OF PICRIC ACID:
This is prepared by allowing distilled water to stand over picric acid with occasional shaking,
making 1 to 2% solution.
To 5.0 ml of glucose solution A mahagony red colour This is due to the reduction of
added 2,0ml of saturated picric solution was got. picric acid to picramic acid
acid of 1.0ml of 10% saturated and it indicates that glucose is
sodium carbonate solution and a reducing sugar.
heated in a boiling water for
three minutes.
7 SELIVANOFF’S TEST:
PREPARATION OF SELIVANOFF’S TEST:
Dissolved 0.5gm of resacinal in 100ml of 10% hydrochloric acid.
To 5.0 ml of glucose solution No characteristic change This shows the absent of keto
added 5.0ml of selivanoff’s was noticed. group in glucose.
reagent and heated in a dilute
flame for few minutes.
8 BIAL’S TEST: A bluish color solution This shows that glucose in a
To 5.0ml of glucose added, was obtained. pentose sugar.
5.0ml of bial’s reagent and
boiled for few minutes in water
bath.
9 PHENYL HYDRAZINE TEST: A yellow precipitate was Glucosazone is formed as
To a small amount of phenyl obtained, yellow needle needle shaped crystals in the
hydrazine mixture, added a few shaped crystals in the form form of “sheaves of corn’.
drops of glacial acetic acid and of “sheaves of corn” were
equal volume of glucose seen under the microscope.
solution and heated in boiling
water bath for half an hour,
Allowed to cool at room
temperature and viewed the
crystals under microscope.

SHEAVES OF CORN SHAPED GLUCOSAZONE:

 REACTIONS OF MONOSACCHARIDES
REACTION OF FRUCTOSE

S.NO EXPERIMENTS OBSERVATION INFERENCE

1 MOLISCH’S TEST
To 5.0 ml of fructose solution ,added A violet coloured ring was got at Carbohydrates
two drops of Molisch’s reagent and 3.0 the junction of two layers +con.H2SO4
ml of concentrated sulphuric acid along Furfural .furfural +alpha
sides of the test tube without shaking so naphthol
as to form the acid layer along the sugar Violet colour.This
solution reaction is due to the
condensation of
hydroxyl methyl furfural
with sulphonated
napthol
2 IODINES TEST
To a small amount of fructose solution, No characteristic change was Fructose does not
added a few drops of dilute iodine noticed answer this test as it is
solution not a polysaccharide
3 FEHLING’S TEST
To 5.0 ml of Fehling’s reagent in a test A reddish brown precipitate was Cu 2+ + reducing sugar
tube added of fructose solution and obtained
heated in boiling water bath for three Cupric hydroxide(CuOH
minutes )
Cuprous oxide
(cu2O).This shows that
lactose is a reducing
sugar
4 BENEDICT’S TEST
To 2.0 ml of fructose solution ,added8 First greenish yellow and then the Cu 2+ + reducing sugar
drops of Benedicts reagent and heated reddish brown precipitate was
in boiling water for three minutes and obtained Cupric hydroxide(CuOH
allowed to cool spontaneously )
Cuprous oxide (cu2O).
The red precipitate is
cuprous oxide .This
shows that lactose is a
reducing sugar
5 BARFOED’S TEST
To 5.0 ml of Barfoeds reagent added 5 A brick red precipitate was It indicates that fructose
ml of fructose solution and heated in obtained at the bottom and the is a monosaccharide
boiling water bath for 3 minutes sides of the test tube
6 PICRIC ACID TEST
To 5.0 ml of fructose solution ,added 2.0 A mahagony red colour was It indicates that the
ml saturated picric acid solution and 1.0 obtained reduction of picric acid
 ml of 10% sodium carbonate solution to picramic acid
and heated in boiling water bath for indicates fructose is a
three minutes reducing sugar
S.NO EXPERIMENTS OBSERVATION INFERENCE
7 SELIVANOFF’S REAGENT
To 5.0 ml of fructose solution, added A cherry red colour solution was This shows that fructose
selivanoff’s reagent and boiled for few got is a hetose
minutes in a direct flame
8 BIAL’S TEST
To 5.0 ml of fructose solution, added No characteristic change was This shows that absence
bial’s reagent and boiled for three noticed of pentose sugar
minutes in a water bath
9 PHENYL HYDRAZINE TEST
To a small amount of phenyl hydrazine A yellow precipitate was obtained Fructosazone is formed
mixture added a few drops of glacial .Yellow needle shaped crystal in as needle shaped
acetic acid and added equal volume of the form of “Sheaves of corn” crystals in the form of
lactose solution and heated in boiling were seen under the microscope “Sheaves of corn”.This
water bath for half an hour .Allowed to confirms the presence
cool at room temperature and viewed of fructose
the crystals under microscope
 REACTIONS OF XYLOSE
S.NO EXPERIMENTS OBSERVATION INFERENCE

01 MOLISCH’S TEST
To5.0 ml of xylose solution added A violet color ring was got Carbohydrate + conc.
two drops of Molisch’s reagent and at the junction of two Sulphuric acid → furfural
3.0 ml of concentrated sulphuric acid liquids. + α naphthol →violet
along the sides of the test tube color. The reaction is
without shaking so as to form a layer due to the condensation
of acid beneath the xylose solution. of hydroxyl methyl
furfural with
sulphonated naphthol.

Xylose does not answer

IODINE’S TEST this test, as it is not a
polysaccharide.
To a small amount of xylose solution, No characteristic change
02 added few drop of dilute iodine was noticed.
solution.

NaOH

FEHLING’S TEST Cu2+ + reducing sugar→

cupric hydroxide (CuOH)
To 0.5 ml of Fehling’s reagent in test
→ cuprous oxide. The
tube added a small amount of xylose
A reddish brown red precipitate is
03 solution and heated in boiling water
precipitate was obtained. cuprous oxide. This
bath for three minutes.
shows that the xylose is
a reducing sugar.
 NaOH

Cu2+ + reducing sugar→

BENEDICT’S TEST
Cupric hydroxide (CuOH)
To2.0ml of xylose solution added 8 →cuprous oxide. The red
drops of Benedict’s reagent and precipitate is cuprous
heated in boiling water bath for 3 oxide. This shows that
minutes and then allowed to cool First a greenish yellow and xylose is reducing sugar.
spontaneously. the a reddish orange
04
precipitated was got.

It indicates that xylose is

BARFOED’S TEST a monosaccharide.
To 0.5 ml of Barfoed’s reagent added
5.0ml of xylose solution and heated
in boiling water bath for three
minutes.
A brick precipitate was
obtained at the bottom
PICRIC ACID TEST and the sides of the test It is due to the reduction
05
tube. of picric acid to picramic
To 5.0ml of xylose solution, added
2.0ml of saturated picric acid acid and it indicates
solution and 1.0ml of sodium xylose is a reducing
carbonate solution and heated water sugar.
in boiling water bath for three A mahogany red color
minutes. solution was obtained.

This shows the absence

06 SELIVANOFF’S TEST of keto group in xylose.
 To 5.0ml of xylose solution added
Selivanoff’s reagent and boiled for
few minutes in direct flame.

No characteristic change This shows that xylose is

BIAL’S TEST
was noticed. a pentose sugar.
To 5.0ml of xylose solution added
Bial’s reagent and boiled in water
bath for few minutes.
07

PHENYL HYDRAZINE TEST A greenish color solution Xylosazone is formed as

was obtained. needle shaped crystals in
To a small amount of phenyl
the form of “sheaves of
hydrazine mixture, added a few
corn”.
drops of glacial acetic acid, added
equal volume of xylose solution and
08
heated in a boiling water bath for
half an hour. Allowed to cool at
Yellow precipitate was
room temperature and viewed the
obtained. Yellow needle
crystal under microscope.
shaped crystal in the form
of “SHEAVES OF CORN”
Yellow needle shaped
were seen under the
crystals of xylosazone
microscope.
09
 REACTIONS OF DISACCHARIDES
REACTIONS OF SUCORSE
S.NO EXPERIMENT OBSERVATION INFERENCE
1. A violet colour ring Carbohydrate +
MOLISCH’S TEST: was formed at the concentrated sulphuric
junction of the layers. acid furfural.
To 5.0 ml of sucrose solution, added Furfural+α naphthol
two drops of Molisch’s reagent and 3.0 violet colour. this
ml of concentrated Sulphuric acid reaction is due to the
along the sides of the test tube without condensation of hydroxy
shaking so as to form an acid layer methyl furfural with
beneath the sucrose solution. sulphonated naphthol.
2. IODINE’S TEST: No characteristic Sucrose does not
change was noticed. answer this test as it is
To a small amount of sucrose solution not a poly saccharide.
added a few drops of dilute iodine
solution.

3. FEHLING’S TEST: No reddish brown This shows that sucrose

precipitate was got is a non reducing sugar.
To 5.0 ml of Fehling’s reagent in
a test tube added a small amount of
sucrose solution and heated in boiling
water bath for three minutes.

4. BENEDICT’S TEST: No reddish brown This shows that sucrose is

precipitated was got not a reducing sugar.
To 5.0 ml of sucrose solution
added 8 drops of Benedict’s reagent
and heated in boiling water bath for 3
minutes and then allowed to cool
spontaneously.

5. BARFOED’S TEST: No brick red This shows that sucrose

precipitated was got. is not a monosaccharide.
To 5.0 ml of sucrose solution,
added 5.0 ml of Barfoed’s reagent
heated in boiling water bath for 3
minutes.
6. PICRIC ACID TEST: No mahogany red This shows that sucrose
coloured solution was is a non reducing sugar.
To 5.0 ml of sucrose solution got.
added 2.0 ml of saturated picric acid
and 1.0 ml of sodium carbonate
solution and heated in boiling water
bath for three minutes.

7. SELIVANOFF’S TEST: A cherry red colour This shows that sucrose

solution was got is a ketose.
To 5.0 ml of sucrose solution,
added 5.0 ml of Selivanoff’s reagent
and boiled for few minutes in direct
flame.

8. BIAL’S TEST: No characteristic This shows that

change was noticed. absence of pentose
To 5.0 ml of sucrose solution added, sugar.
5.0 ml of Bial’s reagent and boiled for
few minutes in direct flame.

9. PHENYL HYDRAZINE TEST: No yellow Sucrose does not react

precipitated was with phenyl hydrazine.
To a small amount of phenyl obtained.
hydrazine mixture added a few drops of
glacial acetic acid added equal volume
of sucrose solution and heated in
boiling water bath for half an hour.
Allowed to cool at room temperature
and viewed the crystals under
microscope.
 HYDROLYSIS OF SUCROSE SOLUTION
To a 10.0 ml of sucrose solution added 10 drops of concentrated hydrochloric acid and heated
in boiling water bath for half an hour, cooled and neutralized with solid sodium carbonate and
filtered. The filtrate was used for following tests.

S.NO EXPERIMENTS OBSERVATION INFERENCE

1. FEHLING’S TEST:
A reddish brown This is due to the reaction
To 2.0 ml of Fehling’s precipitated was of copper sulphate in
solution added 2.0 ml of obtained. Fehling’s reagent to
hydrolysed sucrose filtrate and cuprous oxide by glucose
heated in boiling water bath for 3 and fructose formed on
minutes. hydrolysis of sucrose.

2. BENEDICT’S TEST: Benedict’s reagent is

reduced to cuprous oxide
To 3.0 ml of the hydrolysed First greenish yellow by glucose and fructose
sucrose filtrate added 5.0 ml of and then a reddish formed on hydrolysis of
Benedict’s reagent and heated in orange precipitate sucrose.
boiling water bath for 5 minutes. was obtained.
and cooled.

3. PHENYL HYDRAZINE TEST: A yellow precipitate Glucose and fructose

was got. yellow formed on hydrolysis of
To a small amount of needle shaped sucrose solution reacts with
Phenyl hydrazine mixture, added crystals in the form phenyl hydrazine to form
5.0 ml of the hydrolysed sucrose of “sheaves of corn” glucosazone and
filtrate and a few drops of glacial were seen under the fructosazone crystals.
acetic acid, heated in boiling microscope.
water bath for half an hour.
Allowed to cool and viewed the
crystal under microscope.
 REACTIONS OF DISACCHARIDES
REACTION OF LACTOSE

S.NO EXPERIMENTS OBSERVATION INFERENCE

1 MOLISCH’S TEST
To 5.0 ml of lactose solution ,added two A violet coloured ring was got at Carbohydrates
drops of Molisch’s reagent and 3.0 ml of the junction of two layers +con.H2SO4
concentrated sulphuric acid along sides Furfural .furfural +alpha
of the test tube without shaking so as to naphthol
form the acid layer along the sugar Violet colour.This
solution reaction is due to the
condensation of
hydroxyl methyl furfural
with sulphonated
napthol
2 IODINES TEST
To a small amount of lactose solution, No characteristic change was Lactose does not
added a few drops of dilute iodine noticed answer this test as it is
solution not a polysaccharide
3 FEHLING’S TEST
To 5.0 ml of Fehling’s reagent in a test A reddish brown precipitate was Cu 2+ + reducing sugar
tube added of lactose solution and obtained
heated in boiling water bath for three Cupric hydroxide(CuOH
minutes )
Cuprous oxide
(cu2O).This shows that
lactose is a reducing
sugar
4 BENEDICT’S TEST
To 2.0 ml of lactose solution ,added8 First greenish yellow and then the Cu 2+ + reducing sugar
drops of Benedicts reagent and heated reddish brown precipitate was
in boiling water for three minutes and obtained Cupric hydroxide(CuOH
allowed to cool spontaneously )
Cuprous oxide (cu2O).
The red precipitate is
cuprous oxide .This
shows that lactose is a
reducing sugar
5 BARFOED’S TEST
To 5.0 ml of Barfoeds reagent added 5 No brick red precipitate was got It indicates that lactose
ml of lactose solution and heated in is not a monosaccharide
boiling water bath for 3 minutes
6 PICRIC ACID TEST
To 5.0 ml of lactose solution ,added 2.0 A mahagony red colour was It indicates that the
ml saturated picric acid solution and 1.0 obtained reduction of picric
ml of 10% sodium carbonate solution acidnto picramic acid
and heated in boiling water bath for indicates lactose is a
three minutes reducing sugar
S.NO EXPERIMENTS OBSERVATION INFERENCE
7 SELIVANOFF’S REAGENT
To 5.0 ml of lactose solution added No cheery red colour This shows that
selivanoff’s reagent and boiled for few solution was got absence of keto group
minutes in a direct flame in the lactose
8 BIAL’S TEST
To 5.0 ml of lactose solution added No characteristic change This shows that
bial’s reagent and boiled for three was noticed absence of pentose
minutes in a water bath sugar
9 PHENYL HYDRAZINE TEST
To a small amount of phenyl hydrazine A yellow precipitate was Lactosazone is formed
mixture added a few drops of glacial obtained .Yellow puff’s in the form of puff’s
acetic acid and added equal volume of shaped crystal were seen shaped crystal.This
lactose solution and heated in boiling under the microscope confirms the presence
water bath for half an hour .Allowed to of lactose
cool at room temperature and viewed
the crystalic under microscope
 REACTIONS OF POLYSACCHARIDES
REACTIONS OF STARCH
S.N EXPERIMENT OBSERVATIO INFERENCE
O
S N
MOLISCH’S
1 TEST
To 5.0ml A violet Carbohydrate
of starch coloured ring +coc.H2SO4→furfur
solution was got at the al .furfural+α
added two junction of napthol→violet
drop’s of two liquids. colour. This
Molisch’s reaction gives the
reagent condensation of
and3.0ml of hydroxy methyl
concentrated furfural with
sulphuric sulphonated α
acid along the napthol.
sides of the
test tube
without
shanking so
as to form an
acid layer
beneath the
starch
solution.

2 IODINE TEST
To a small A blue This shows
amount of colour that starch is an
starch solution was polysaccharide.
solution got.
added a few
drops of
 dilute iodine
solution.

3 FEHLING’S
TEST
To a No This shows
5.0ml of reddish that starch is a
Fehling’s brown non-reducing
reagent in a precipitate sugar.
test tube was got.
added a small
amount of
starch
solution and
heated in
boiling water
bath for three
minutes.

4 BENEDICT’S
TEST
To 5.0ml No This shows that
of starch reddish starch is non-
solution brown reducing sugar.
added 8 precipitate
drops of was got.
Benedict’s
reagent and
heated in
boiling water
bath for 3
minutes and
then allowed
to cool
spontaneousl
y
5 BARFOED’S
TEST
To 5ml of No brick This shows
starch red that starch is not a
solution precipitate monosaccharide.
added 5ml of was got.
Barfoed’s
reagent
heating in
boiling water
bath for 3
minutes.

6 PICRIC ACID
TEST
To 5.0 ml No This shows
starch mahagony that starch is a
solution red coloured non-reducing
added 2.0ml solution was sugar.
of saturated got.
picric acid
and 1.0ml of
10% sodium
carbonate
solution and
heated in
boiling water
bath for three
minutes.

7 SELIVANOFF’
S TEST
To 5.0ml No This shows that
of the starch characteristic starch is not a
solution ketose.
 added 5.0ml change was
of Selivanoff’s noticed.
reagent and
boiled for few
minutes in a
direct flame.

8 BIAL’S TEST
To 5.0ml No Starch is not a
of the starch characteristic pentose sugar.
solution change was
added 5.0ml noticed.
of Bial’s
reagent and
boiled in
water bath for
3 minutes.

9 PHENYL
HYDRAZINE
TEST
 To a No yellow Starch
small precipitate does not
amount of was react with
phenyl obtained. phenyl
hydrazine hydrazine.
mixture
added a few
drops of
glacial acetic
acid added
on equal
volume of
starch
solution and
heated in
boiling water
bath for half
an hour.
Allowed to
cool at room
temperature
and viewed
at crystal
microscope.

HYDROLYSIS OF STARCH SOLUTION

To a 10.0ml of starch solution added 10 drops of
concentrated hydrochloric acid and heated in boiling
water bath for half an hour ,cooled and neutralized with
solid medium carbonate and filtered. The filterate was
used for following tests.
FEHLING’S
3 TEST
 To 2.0ml A reddish CU2+
of Fehling’s brown precipitate +reducing sugar
solution was got. →through NAOH
added 2.0ml gives cupric
of hydroxide
hydrolyzed (CUOH)→cuprous
starch oxide (CU2O).The
fliterate and red precipitate is
heated in cuprous oxide
boiling water this shows that
bath for starch is a
three reducing
minutes. solution.

BEBEDICT’S
4 TEST
To a A reddish CU2+ +
3.0ml of orange precipitate reducing sugar
hydrolyzed was got. through
starch NA2CO3→cupric
filterate hydroxide→
added 5.0ml cuprous oxide.
of Benedict’s This shows the
reagent and reducing action
heated in of starch.
boiling water
bath for five
minutes and
cooled.

PHENYL
9 HYDRAZINE
TEST
To a A yellow This shows
small precipitate was got that starch on
amount of yellow needle hydrolysis form
phenyl shaped crystals in sheaves of corn
hydrazine the form of shaped
mixture , “Sheaves of corn” Glucosazone
added 5.0ml were seen under crystals.
of microscope.
hydrolyzed
starch
filterate and
a few drops
of glacial
acetic acid.
Heated in a
boiling water
bath for half
an half hour
.allowed to
cool and
viewed the
crystals
under
microscope.