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. To determine the color change of the given test samples in Molisch test.
2. To identify whether the provided test sample contains carbs.
3. To understand the different reagents used in the Molisch test, as well as their composition.

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
Purple with Dark Blue Blue Blue
Color Change black

Presence of Yes No No No
Carbohydrates
(Yes/No) Because Because Because
there’s no there’s no there’s no
purple/reddish purple/reddish purple/reddish
purple tint purple tint purple tint
 arises and the arises and the arises and the
provided provided provided
analyte does analyte does analyte does
not contain not contain not contain
carbohydrate. carbohydrate. carbohydrate.

V. RESULTS AND OBSERVATION

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

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

the dehydration of the carbohydrate. Concentrated Sulfuric acid is a strong
dehydrating agent. In Molisch’s test it dehydrates sugar. When adding
a concentrating sulfuric acid in Molisch Reagent, a bottom layer forms and
color forms between the two liquids (Molisch Reagent and carbohydrate). In
Molisch's test, concentrated sulfuric acid serves as a dehydrating agent.

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

The common test for all the carbohydrates is the Molisch test. This test will
evaluate whether the sample or substance contains a specific sugars or
carbohydrates. 2ml of each carbohydrate solution is placed in the four test tubes.
After that, 10 drops of Molisch solution are added in the four-test tube. 10-15 drops
of sulfuric acid were also put after that. It must not be stirred. In the bottom layer,
there is the sulfuric acid. After 2 minutes, the glucose, lactose, and sucrose has
become light color red on the upper part and in the lower is dark violet or purple
which is the sulfuric acid. Fructose become dark violet or purple. The indicator for
Molisch test is the formation of purple ring wherein we can observe it in the
experiment

3. What is the composition of Molisch Reagent?

The composition of Molisch reagent is made up of alpha-naphthanol

(C10H8OH), which is dissolved in ethanol (C2H5OH). It is used to test the presence
of carbohydrates.
VI. CONCLUSION

Molisch's test is a sensitive and specific method for determining the concentration
of carbohydrates. The test is named by Austrian botanist Hans Molisch. Based on the
video presented, the dehydration of the carbohydrate by sulfuric acid forms at the bottom
and causes a color difference between the added liquid. Molisch's test is used to detect if
a material contains carbs or sugars. The most practical application of this test is that if a
food product is labeled as sugar free, you can simply test the product using Molisch's test
to see if it includes carbohydrate.