QUALITATIVE
ANALYSIS AND
BIOLOGICAL ROLE
OF
CARBOHYDRATES
Presented by:
Priyanka Boruah
Assistant Professor,
School of Pharmacy,
Kaziranga University
QUALITATIVE TESTS FOR CARBIOHYDRATE
1. Molisch's Test
Purpose: Detects any carbohydrate.
Procedure:
Add 2-3 drops of Molisch's reagent (alpha-
naphthol solution) to 2 ml of the sample solution.
Carefully add 1 ml of concentrated sulfuric acid
(H2SO4 ) down the side of the test tube without
mixing.
Positive Result: A violet ring at the interface
indicates the presence of carbohydrates.
2. Benedict's Test
Purpose: Detects reducing sugars.
Procedure:
Add 2 ml of Benedict's reagent to 1 ml of the
sample solution.
Heat the mixture in a boiling water bath for 2-5
minutes.
Positive Result: Color change from blue to
green, yellow, orange, or red, indicating the
presence of reducing sugars.
3. Fehling's Test
Purpose: Detects reducing sugars.
Procedure:
Mix equal parts of Fehling's solution A (Cu2SO4 ) and
Fehling's solution B (alkaline tartrate) to prepare
Fehling's reagent.
Add 2 ml of Fehling's reagent to 1 ml of the sample
solution.
Heat the mixture in a boiling water bath for 5-10
minutes.
Positive Result: Formation of a brick-red precipitate
indicates the presence of reducing sugars.
4. Barfoed's Test
Purpose: Differentiates monosaccharides from
disaccharides.
Procedure:
Add 1 ml of Barfoed's reagent to 1 ml of the sample
solution.
Heat the mixture in a boiling water bath for 1-2 minutes.
Positive Result for Monosaccharides: Red precipitate
within a few minutes.
Positive Result for Disaccharides: Red precipitate
after a longer time (usually 7-10 minutes).
5. Iodine Test
Purpose: Detects polysaccharides like starch.
Procedure:
Add 1-2 drops of iodine solution to 2 ml of the sample
solution.
Mix the solution gently.
Positive Result: A blue-black color indicates the
presence of starch. Heat the solution , and the blue
colour will disappear and on cooling the colour
reappears.
6. Seliwanoff's Test
Purpose: Differentiates between aldoses and
ketoses (especially fructose).
Procedure:
Add 2 ml of Seliwanoff's reagent (resorcinol and
hydrochloric acid) to 1 ml of the sample solution.
Heat the mixture in a boiling water bath for 1-2
minutes.
Positive Result for Ketoses (e.g., fructose): Red
color appears quickly.
Positive Result for Aldoses: Red color appears
slowly.
7. Osazone Test
Purpose: Identifies specific reducing sugars
based on crystal formation.
Procedure:
Add 5ml of sample solution, 0.5g phenylhydrazine
hydrochloride, and 0.1g sodium acetate and 5 drops of
glacial acetic acid.
Heat the mixture in a boiling water bath for 15-30 minutes.
Cool the solution to room temperature.
Positive Result: Formation of characteristic yellow
crystals (osazones), whose shapes can help identify specific
sugars (e.g., needle-shaped crystals for glucose and
fructose).
8. Anthrone Test
Purpose: Detects carbohydrates, especially
polysaccharides.
Procedure:
Add 2 ml of anthrone reagent (anthrone in sulfuric acid) to
1 ml of the sample solution.
Heat the mixture in a boiling water bath for 10 minutes.
Cool the solution to room temperature.
Positive Result: Formation of a blue-green color indicates
the presence of carbohydrates.
7. Bial's Test
Purpose: Detects pentoses (five-
carbon sugars).
Procedure:
Add 2 ml of Bial's reagent (orcinol,
hydrochloric acid, and ferric chloride) to 1 ml
of the sample solution.
Heat the mixture in a boiling water bath for 1-
2 minutes.
Positive Result for Pentoses: Formation of
a green or blue-green color.
BIOLOGICAL ROLE OF
CARBOHYDRATES
1. Energy Source
Glucose: The primary source of energy for cells.
Glycogen and Starch: Storage forms of glucose in animals and plants,
respectively. They can be broken down into glucose molecules when
energy is needed.
2. Structural Components
Cellulose: A major component of plant cell walls, providing structural
support.
Chitin: Found in the exoskeletons of arthropods and the cell walls of
fungi, providing strength and protection.
3. DNA and RNA Structure
Ribose and Deoxyribose: Sugars that are part of the backbone of RNA
and DNA, respectively. Ribose is a component of RNA, while deoxyribose is
found in DNA.
4. Metabolic Intermediates
Intermediates in Metabolic Pathways: Carbohydrates are involved in
various metabolic pathways, including glycolysis, the citric acid cycle, and
the pentose phosphate pathway. These pathways are essential for energy
production and the synthesis of nucleotides and amino acids.
5. Osmotic Balance
Maintaining Osmotic Pressure: Carbohydrates,
particularly polysaccharides, help maintain osmotic
balance in cells and tissues.
6. Digestive Health: Indigestible carbohydrates, such
as dietary fiber. They aid in bowel movements, prevent
constipation,
7. Storage and Transport of Energy
Starch in Plants: Plants store excess glucose as starch,
which can be broken down when the plant requires
energy.
Glycogen in Animals: Animals store glucose as
glycogen in liver and muscle tissues for rapid mobilization
during physical activity or between meals.
8. Component of Coenzymes
NAD+, NADP+, and FAD: Carbohydrate-derived
molecules that are crucial coenzymes in redox reactions,
playing key roles in metabolic pathways.
