Lab Activity 4

Polysaccharides

Alaa S Baraka
Islamic university of
Gaza
March 2013

Content

Inversion of sucrose
Iodine Test for Polysaccharides
Hydrolysis of Starch
Mucic Acid Test For Galactose
UNKNOWN

sucrose

Non-reducing sugar: CHO- of D-glucosyl unit

and C=O of
D-fructosyl unit are in glysosidic bond, no
. reducing end

Inversion of sucrose

As a result, sucrose is easily hydrolyzed in very dilute acid

:or enzymes
+
H

Sucrose

Glu(+52.2) + Fru (-92)

invertase (yeast, bacteria)
invert sugar (equimolar
or intestinal sucrase
(mixture of D-glucose and D-fructose

= +66.5 [ ]

= -39.8 [ ]

Reagent:

1% sucrose solution
Conc. H2SO4
Bendict reagent
Barium hydroxide Ba(OH)2

phenolphthalein,

Procedure

Add 25ml of sucrose solution in a beaker

Add one drop of H2SO4
Boil one min
Cool and neutralize with saturated
Ba(OH)2
Use phenolphthalein,as indicator
Remove the precipitate of barium sulfate
by filtration
Test for Benedict

Iodine Test for

Polysaccharides

Iodine forms colored adsorption

complexes with polysaccharides
Blue color with starch
Red-blue or purple color with dextrin
Red color with glycogen
With cellulose it gives no color

Reagent

1%solutions of Starch, Dextrine

Iodine solution (2% KI containing
sufficient I2 to color it deep yellow)

Procedure
1. In a test tube add 4 drops of the tested
solution
2. Add 1 to 3 drops of a dilute solution of
Iodine solution.
3. Compare the color obtained with that of
water and iodine solution.

Hydrolysis of Starch

Reagents:
1% starch solution,
Benedicts solution
Conc. HCl

Procedure

1.

2.

3.

Place 25 ml of 1% starch solution in a

small beaker, add 10 drops of conc. HCl
& boil gently.
At the end of each min transfer 1 drop of
the solution to the test plate and make
the regular iodine test.
Also at the end of each min transfer 3
drops of mixture to 5 ml portions of
Benedicts solution in a series of test
tubes.
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4. As the test proceeds, the reaction with

iodine should become weaker & finally
be negative.
5. At this point place all test tubes
containing Benedicts solution in a
boiling water-bath for 3 min.
6. Remove the tubes then allow to cool.
7. Note the degree of reduction in each
case compare with the rate of the
disappearance of iodine reaction.
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Mucic Acid Test For Galactose

Galactose & any sugar gives

galactose upon hydrolysis such
as lactose, Raffinose and some
gums give mucic acid on
oxidation with HNO3
Conc. HNO3 oxidizes galactose to
mucic acid (a fine white gritty
crystals separate, insoluble in
water, but readily soluble in
alkaline or ammonium
carbonate solution & reprecipitated on the addition of
nitric acid.

Galactaric acid

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Reagents

2% solution of Galactose
Conc. HNO3

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Procedure
1.
2.
3.

4.

To 10 ml of 2% of galactose solt. In porcelain

evaporating dish add 5ml conc. HNO 3
Place the dish on top of a beaker filled with
2/3 distilled water.
Evaporate (under hood) the contents of the
dish on the boiling water bath to about a
volume of 2 ml (it takes about 45 min boiling
to reach the 2 ml).
Remove the hot dish with tong and allow to
cool.
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6. add 5 ml of distilled water to the

dish and mix well with a string rod.
7. Note the insoluble mucic acid
crystals from the oxidation of
galactose.
8. Transfer one drop of the liquid plus
crystals to a clean microscope slide,
examine with a low power microscope
(10X).
Note the gritty crystals of mucic acid.
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