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Exercise no. 3
TEST FOR THE PRESENCE OF CARBOHYDRATES: MOLISCH TEST
I. INTRODUCTION

The most important carbohydrate is glucose, a simple sugar (monosaccharide)

that is metabolized by nearly all known organisms. Glucose and other carbohydrates are
part of a wide variety of metabolic pathways across species: plants synthesize
carbohydrates from carbon dioxide and water by photosynthesis storing the absorbed
energy internally, often in the form of starch or lipids. Energy obtained from metabolism
(e.g. oxidation of glucose) is usually stored temporarily within cells in the form of ATP.
Organisms capable of aerobic respiration metabolized glucose and oxygen to release
energy with carbon dioxide and water as byproducts.
Carbohydrates can be chemically into complex and simple. Simple carbohydrates
consist of single or double sugar units (monosaccharide and disaccharides, respectively).
Sucrose or table sugar (a disaccharide) is a common example of a simple carbohydrate.
Complex carbohydrates contain three or more sugar units linked in a chain and are called
polysaccharides. They are digested by enzymes to release the simple sugars. Starch, for
example, is a polymer of glucose units and is typically broken down to glucose. Simple
and complex carbohydrates are digested at similar rates. Carbohydrates are typically
stored as long polymer of glucose molecules with glycosidic bond for structural support
(e.g. chitin, cellulose) or for energy storage (e.g. glycogen, starch).
All carbohydrate shares a general formula of approximately CnH2nOn; glucose is
C6H12O6. Monosaccharides may be chemically bonded together to form disaccharides
such as sucrose and longer polysaccharides such as starch and cellulose.
Carbohydrates metabolism begins with digestion in the small intestine where
monosaccharides are absorbed into the blood stream. Blood sugar concentrations are
controlled by three hormones: insulin, glucagon, and epinephrine. If the concentration of
glucose in the blood is too high, insulin is secreted by the pancreas. Insulin stimulates the
transfer of glucose in the blood into the cells, especially in the liver and muscles.
In the experiment, you will investigate how to detect the presence of carbohydrates
using the Molisch Test and or Barfoed Reagent.
II. OBJECTIVES:
At the end of the experiment, each student will be able to:

1. ___________________________________________________
___________________________________________________
 2. ___________________________________________________
___________________________________________________

III. MATERIALS:
2pcs medium size fresh potato, 2-500ml beaker, 6 test tube, test tube rack,
test tube bruch, dropper, cheese/plain white cloth,2ml sucrose soln, 2ml
albumin soln, 2ml oil, 2ml Molisch reagent (5%alpha naphthol in 95%
alcohol) and/ or Barfoed’s Reagent ( 0.33M neutral copper acetate and 1%
acetic acid soln), 2ml sulfuric acid (conc)

IV. PROCEDURE

A. Preparation of Starch
Prepare a small potato. Grate into fine pulp and replace the pulp in a
beaker. Mix well with water. Strain the pulp using a cheese cloth,
catching the liquid in a beaker. Allow the starch to settle and discard the
liquid using decantation. Allow the starch to drain thoroughly and set
aside to dry. Reserved the prepared starch for the next procedure.

B. Molisch Test
1. In four clean dry test tubes, place separately 1 ml of sucrose
solution, 1 ml of starch solution (from A), 1 ml albumin solution
and 1 ml oil. Add two drops of the Molisch reagent (5% alpha-
naphthol in 95% alcohol) in each test tube.

2. Carefully pour 2 ml of concentrated sulfuric acid down the sides

of the test tubes as so to form a layer. Observed the color at the
junction of the liquids. Note the result and compare the result in
each test tube. Give an explanation for the results.
DATA:
Sucrose Potato Albumin Oil

Color Change

Presence of
Carbohydrates
(Yes/No)
 V. RESULTS AND OBSERVATION

1. What is the role of concentrated sulfuric acid in Molisch Reagent test?

Ans: The Molisch test is for the presence of carbohydrates. It is based on

the dehydration of the carbohydrate (the reason for the presence of the
sulfuric acid in the Molisch reagent test) to produce an aldehyde which
then condenses with two molecules of alpha-naphthol (or other phenol like
resorcinol or thymol) to give colored products. With alpha-naphthol, the
product is violet in color. The reactions involved are shown below.

2. What does Molisch Test tell you in conducting the experiment?

Ans: Molisch's test is a sensitive chemical test, named after Austrian

botanist Hans Molisch, for the presence of carbohydrates, based on the
dehydration of the carbohydrate by sulfuric acid or hydrochloric acid to
produce an aldehyde, which condenses with two molecules of a phenol
(usually α-naphthol, though other phenols.

3. What is the composition of Molisch Reagent?

Ans: The test solution is combined with a small amount of Molisch's

reagent (α-naphthol dissolved in ethanol) in a test tube. After mixing, a
small amount of concentrated sulfuric acid is slowly added down the sides
of the sloping test-tube, without mixing, to form a layer.

VI. CONCLUSION

Molisch's test is a chemical test which is used to check for the presence of
carbohydrates in a given analyte. This test is named after Czech-Austrian botanist
Hans Molisch, who is credited with its discovery. A test for carbohydrate (as
sugar) in which a reddish violet color is formed by reaction with alpha-naphthol
in the presence of concentrated sulfuric acid.

LIBRARY WORK
 In an A4 bond paper research at least two (2) journals / articles on
carbohydrates (Times New Roman 13, landscape)

Author(s) Year Title Methods Results

published