Carbohydrates:

General and Specific Tests

Mario A. Tan, Ph.D.

Department of Chemistry
College of Science
 Carbohydrates
Carbohydrates, called sugars and starches, are
polyhydroxy aldehydes or ketones, or compounds
that can be hydrolyzed to them.
 Carbohydrates
Carbohydrates are synthesized in green plants
by photosynthesis, whereby energy from the sun is
stored as chemical energy in carbohydrates.

In the body, they are used for bursts of energy

needed during exercise in the form of glucose.
 Types of Carbohydrates
Monosaccharides, the simplest carbohydrates.
Disaccharides, which consist of two monosaccharides.
Polysaccharides, which contain many monosaccharides.
 Types of Carbohydrates -
Monosaccharides
Monosaccharides, the simplest carbohydrates,
generally have 3 to 6 C atoms in a chain with an
aldehyde or ketone ending and many OH groups.

Aldehyde monosaccharides are aldoses; ketone

monosaccharides are ketoses.
 Types of Carbohydrates -
Monosaccharides
The simplest aldose is glyceraldehyde.

The simplest ketose is dihydroxyacetone.

They are constitutional isomers of each other,

sharing the formula C3H6O3.
 Types of Carbohydrates -
Monosaccharides
A monosaccharide is characterized by the number
of C atoms in its chain:
A triose has 3 Cs. A tetrose has 4 Cs.

A pentose has 5 Cs. A hexose has 6 Cs.

The terms are then combined with the words aldose

and ketose:

Glyceraldehyde is an aldotriose.
Dihydroxyacetone is a ketotriose.
 Types of Carbohydrates
Disaccharides
Disaccharides are composed of two mono-
saccharide units joined together.
 Types of Carbohydrates
Polysaccharides

Polysaccharides are composed of three or more

monosaccharide units joined together.
Glycogen is the major form of polysaccharide
storage in animals, similar in structure to
amylopectin.
It is stored mainly in the liver and in muscle cells.
When glucose is needed for energy, glucose
units are hydrolyzed from the ends of the glycogen
polymer.
Because glycogen is highly branched, there
are many ends available for hydrolysis.
Cellulose is an unbranched polymer made up of
repeating glucose units joined by -(14)
glycosidic linkages.

Cellulose is found in the cell walls of all plants,

where it gives support and rigidity to wood,
plant stems, and grass.

Humans do not posses the enzyme to hydrolyze

cellulose (b-glycosidase) and cannot digest it.

Cellulose makes up the insoluble fiber in our

diets, which is important in adding bulk to waste
to help eliminate it more easily.
 General Tests for Carbohydrates
Molisch Test (-napthol reaction)
test for polysaccharides (sugars in general)
Reagents: -napthol, 95% EtOH, conc. H2SO4
(+) Result: Violet Ring/Interphase
Principle:
(1) hydrolysis of the glycosidic bond forming the
reduced sugar (monosaccharide)
(2) dehydration of the monosaccharide into
hydroxymethyl furfurals
(3) condensation of the furfural with -napthol
 General Tests for Carbohydrates
Anthrone Test
general test for hexoses (different colors for different
sugars)
Reagents: Anthrone in conc. H2SO4
(+) Result: Blue-green solution
Principle:
(1) hydrolysis of the glycosidic bond forming the
reduced sugar (monosaccharide)
(2) dehydration of the monosaccharide into
hydroxymethyl furfurals
(3) condensation of the furfural with anthrone reagent
 General Tests for Carbohydrates
Iodine Test
test to differentiate helical from non-helical
polysaccharides
Reagents: Iodine solution
(+) Result: Deep-Blue (Purple) Solution (Amylose)
Red-Violet Solution (Amylopectin)
Red Solution (Glycogen)
Principle: Complexation

* Cellulose yellow solution () due to the color of the

iodine solution
 Specific Tests for Carbohydrates
Based on Furfural Formation
Seliwanoffs Test
test for ketohexoses (fructose)
Reagents: Resorcinol, 6N HCl
(+) Result: Cherry-Red Solution
Principle:
(1) Dehydration of ketohexose to hydroxymethyl furfural
(2) Condensation with resorcinol
 Specific Tests for Carbohydrates
Based on Furfural Formation
Bials Orcinol Test
test for pentoses (xylose)
Reagents: Orcinol, 6N HCl
(+) Result: Blue-green Solution
Principle:
(1) Dehydration of pentose to furfuraldehyde
(2) Condensation with orcinol
 Specific Tests for Carbohydrates
Based on Oxidation of Sugars
Mucic Test
test for the presence of galactose
Reagents: conc. HNO3 (Oxidizing agent)
(+) Result: Crystal formation (galactaric acid)
Principle: Oxidation of galactose to galactaric acid
 Specific Tests for Carbohydrates
Based on Reducing Property of Sugars

Makes use of the Oxidizing Agent to test the reducing property of

the carbohydrate

Ability of the carbohydrate to reduce metal ions like Ag+ or

Cu2+ in alkaline medium
 Specific Tests for Carbohydrates
Based on Reducing Property of Sugars
BenedictsTest
test for reducing carbohydrates
Reagents: Na citrate, Na2CO3, CuSO4
(+) Result: Brick-red precipitate
* Disaccharides may be classified as Reducing or Non-reducing
depending on whether free or potentially free aldehyde or
ketone groups are present.
 Specific Tests for Carbohydrates
Based on Reducing Property of Sugars

BarfoedsTest
test for reducing monosaccharides
monosaccharides are more powerful RA than disaccharides
used to differentiate monosaccharides from disaccharides
Reagents: Cu acetate, glacial Hac (acidic)
(+) Result: Brick-red precipitate (Cu2O)