Isolation and Characterization of Carbohydrates

ISOLATION AND
CHARACTERIZATION OF
CARBOHYDRATES
GROUP 4
PRE-
LAB
1
Define the following terms briefly as they relate to the
experiments. Cite an example for each using the Fischer/Haworth
projection formula.
a. Aldohexose
b. Reducing
Sugar
c. Hemiacetal
ALDOHEXOS
E
A monosaccharide
sugar having 6 carbons
atoms and aldehyde
group.
Reducing
Sugar
It is any sugar that is
capable of acting as a
reducing agent
because it has a free
aldehyde group or free
ketone group. It is a
carbohydrate that is
oxidized by weak
oxidizing agent in basic
aqueous solution.
Hemiacetal
Is a carbon connected
to two oxygen atoms,
where one oxygen is
an alcohol (OH) and
the other is an ether
(OR).
2
Complete the table below. Indicate what is observed if each test
is positive. State what this result implies about each of the
carbohydrates tested.
POSITI
VE
TESTS
Expected Observation Substance Indicated by
Test
for a Positive Test a Positive Test
I2 solution Blue Carbohydrates and

polysaccharides
Molisch’s Dark violet region Carbohydrates and

between junctions of polysaccharides
two layers
Benedict’s Red precipitate Reducing sugars
Seliwanoff’s Cherry red Ketones
Bial’s orcinol Blue-green product Pentoses
Barfoed’s With red in blue solution Monosaccharide
3
Which test can be used differentiate the following pairs of

carbohydrates?
a. Fructose and
Galactose
b. Ribose and Glucose
c. Glucose and Maltose
both can be distinguished by
Fructose and Galactose Barfoed’s test.
both can be distinguished by Bial’s

test which is a chemical test for the
Ribose and Glucose presence of pentose.
both can be distinguished by

Barfoed’s test. You can also use
Glucose and Maltose Seliwanoff’s test for it distinguishes
aldose and ketose sugar.
4
Describe the reaction that takes place when starch is hydrolyzed
by an enzyme. What is the product of starch hydrolysis?
The glycosidic bond is broken and an –
OH is added to one glucose and an –H
is added to the other glucose.
Whenever starch molecules undergo
hydrolysis, it forms monosaccharide,
disaccharide or oligosaccharide.
5
What enzyme is present in the saliva? What conclusions can be
made regarding the action of this enzyme on starch?
The enzyme present in the saliva is the
enzyme amylase, also called as ptyalin,
which is capable of broken-down starch
into simpler sugars such as maltose and
dextrin that can be further broken down
into the small intestine. We can say that
the action of this enzyme is strongly
affected by several factors, such as
temperature and pH.
DATA
SHEETS
ISOLATION AND GENERAL TEST FOR
POLYSACCHARIDES
DESCRIPTION MOLISCH’S TEST KI/I2
ISOLATE: POTATO A potato, or "spud", is Appearance of a Blue-black colored is
a solid vegetable that reddish violet colored observed.
is a native of the ring at the junction of
Peruvian Bolivian 2 liquids.
Andes. The potato is
made up of
potassium, vitamin C,
fiber, and starch.
ISOLATION AND GENERAL TEST FOR
POLYSACCHARIDES
DESCRIPTION MOLISCH’S TEST KI/I2
UNKNOWN: The liver is an organ An appearance of The brown colour of
GLYCOGEN only found in reddish violet or the aqueous solution
vertebrates which purple colored at the remains.
detoxifies various junction of two liquids
metabolites, is observed in the
synthesizes proteins solution. And
and produces therefore, yielded a
biochemicals positive result.
necessary for
digestion.
HYDROLYSIS OF POLYSACCHARIDES
HYDROLYSATE DESCRIPTION BENEDICT’S TEST
ACID HCl mixes with the isolate (glucose) When Benedict’s reagent solutions
made a slight viscosity and reducing sugar, are heated
together, the solution changes its
color to orange-red/brick red
UNKNOWN SAMPLE
QUALITATIVE TESTS FOR
CARBOHYDRATES
Carbohydrate Benedict’s test Barfoed’s test Seliwanoff’s Bial’s Test
Solution test
Glucose Brick red ppt. Scanty red ppt Cherry red yellow
at the bottom of
test tube
Fructose Brick red ppt. Scanty red ppt Cherry red Light brown
at the bottom of
test tube
Xylose Brick red ppt. scanty red ppt Dark yellow Blue-green
at the bottom of
test tube
QUALITATIVE TESTS FOR
CARBOHYDRATES
Carbohydrate Benedict’s test Barfoed’s test Seliwanoff’s Bial’s Test
Solution test
Lactose Brick red ppt. No change in brown
color
Sucrose No change in No change in Cherry red Dark brown
color color
Starch White Blue Yellow Gold Yellow
Hydrolysate
unknown
MUCIC ACID
CRYSTAL
OSAZONE
CRYSTALS
THIN-LAYER CHROMATOGRAPHY
Rf value
Dextrin <0.207
Maltose 0.14
glucose 0.35
Conclusion:
SUGAR HYDROLYSABLE OR MONOSACCHARIDE, REDUCING OR PENTOSE OR ALDOSE OR
NON-HYDROLYSABLE DISACCHARIDE, NON HEXOSE KETOSE
POLYSACCHARIDE REDUCING
GLUCOSE Non-hydrolysable Monosaccharide Reducing Hexose Aldose
FRUCTOSE Non-hydrolysable Monosaccharide Reducing Hexose Ketose
XYLOSE Non-hydrolysable Monosaccharide Reducing Pentose Aldose

LACTOSE Hydrolysable Disaccharide Reducing Hexose Aldose
SUCROSE Hydrolysable Disaccharide Non-reducing Hexose Ketose
STARCH Hydrolysable Polysaccharide Non-reducing Hexose Aldose
HYDROLYS Non-hydrolysable Monosaccharide Reducing Hexose Ketose
ATE
GLYCOGEN Hydrolysable Polysaccharide Non-reducing Hexose Ketose
(unknown
sample)
POST-
LAB
1
An unknown carbohydrate solution tested (+) with
Molisch's reagent formed a red precipitation with
Barfoed's reagent within 3 minutes and turned blue-green
when mixed with Bial's orcinol reagent. What is this
carbohydrate? Draw its Haworth structure.
The carbohydrate is a reducing
monosaccharide (positive Benedict’s
and Barfoed’s test) and since the Bial’s
test is a confirmatory test for pentoses
so the given carbohydrate could be a
pentose.
2
Erythrose is an aldotetrose. Describe the result when it is
subjected to each of the following tests:
a. Molisch’s Test
b. Barfoed’s Test
c. Benedict’s Test
d. Seliwanoff’s Test
Erythrose or tetroses does not
Molisch’s test give a positive reaction in this
test.
A scanty brick red precipitate is

Barfoed’s test
observed in a positive reaction.
Benedict’s test Red cuprous oxide
A cherry red color is observed in

Seliwanoff’s test
a positive reaction.

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ISOLATION AND

I2 solution Blue Carbohydrates and

Molisch’s Dark violet region Carbohydrates and

Which test can be used differentiate the following pairs of

both can be distinguished by Bial’s

both can be distinguished by

XYLOSE Non-hydrolysable Monosaccharide Reducing Pentose Aldose

A scanty brick red precipitate is

Benedict’s test Red cuprous oxide

A cherry red color is observed in

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