DEQUINA, FERNANDEZ, GONZALES

GENERAL TESTS FOR

CARBOHYDRATES
MATERIALS

 Glycogen isolate sample, 1% aqueous solution

 Plant starch isolate sample, 1% aqueous solution
 Molisch Reagent
 Anthrone Solution
 Iodine Solution
IODINE TEST
WHAT IS IODINE TEST?

 The Iodine Test is used distinguish between

polysaccharides and monosaccharides.
 Also, it is to detect the presence of starch in the natural
materials used in the experiment.
PROCEDURE

1. Add one drop of Iodine solution to 1 ml. of aqueous

isolate solution.
2. Observe the color developed.
3. Warm in water bath and note any change in color.
4. Cool the solution. Is there any change in color?
5. Record all solution.
WHAT IS LUGOL’S SOLUTION?

 It is also known as the aqueous iodine and strong iodine

solution.
 Chemical formula: KI3
STARCH (CORN)
GLYCOGEN (HALAAN)
POSITIVE RESULTS

SAMPLE POSITIVE RESULTS

STARCH BLUE-BLACK COLOR

GLYCOGEN BROWN-BLUE COLOR

PRINCIPLE

 Starch will form a deep blue color with the Iodine solution.
It is because starch contains amylose.
 The iodine is able to form a large complex with the
amylose helix. Due to this, it will then absorb the light and
reflect the blue light only.
 Iodine Clock Reaction
 Amylopectin (and glycogen) are unable to assume a
stable helical conformation because of the branching. •
ANTHRONE TEST
WHAT IS ANTRONE TEST?

 It is a test used to determine the amount of carbohydrates

from the natural materials. The intensity of which is
directly proportional to amount of carbs present in given
solution.
PROCEDURE

1. Add one to two drops of Anthrone solution to 1 ml of

isolate.
2. Record the observation.
WHAT IS ANTHRONE SOLUTION?

 Anthrone is a tricyclic sweet-smelling ketone. It is utilized

for a well-known cellulose measure and in the colorimetric
determination of carbohydrates.
 The reagent is composed of 200 mg of anthrone reagent
in 100 ml. of concentrated Sulfuric acid (H2SO4)
STARCH (CORN)
GLYCOGEN (HALAAN)
POSITIVE RESULTS

SAMPLE POSITIVE RESULTS

GREEN TO BLUE GREEN COLOR

STARCH
COMPLEX
GREEN TO BLUE GREEN COLOR
GLYCOGEN
COMPLEX
PRINCIPLES

 Carbohydrates are dehydrated with concentrated H2SO4

to form “Furfural”, which condenses with anthrone to form
a green color complex. Anthrone react with dextrins,
monosaccharides, disaccharides, polysaccharides,
starch, gums and glycosides.
MOLISCH’S TEST
WHAT IS MOLISCH’S TEST?

 To detect the presence of carbohydrates, the solution is

first treated with a strong acid. This is for hydrolyzing the
carbohydrate to monosaccharide. A compound named
furfurol is then made when water is removed from
monosaccharides.
PROCEDURE

1. Add 2 drops of 5% alcoholic naphtol reagent to 1 ml. of

the aqueous isolate solution. Mix very well.
2. Incline the test tube and slowly and carefully pour 1 ml.
conc. Sulfuric acid down the side of the tube.
3. Note the color developed at the junction of the two
liquids.
WHAT IS MOLISCH’S REAGENT?

 A solution of alpha-naphthol in 95% ethanol

 Useful for identifying any compound which can be
dehydrated to furfural or hydroxymethylfurfural in the
presence of H2SO4
STARCH (CORN)
GLYCOGEN (HALAAN)
POSITIVE RESULTS

 The formation of a purple product at the interface of the

two layers.
 Monosaccharides give a rapid positive test while
disaccharides and polysaccharides react slower.
PRINCIPLES

 The carbohydrate gets dehydrated on contact with

concentrated sulfuric acid to form furfural derivatives
which on reaction with α-napthol (Molisch’s reagent)
results in a violet colored complex.
The test reagent dehydrates pentoses to form furfural.
The test reagent dehydrates hexoses to form 5-hydroxymethyl furfural.
POST LABORATORY QUESTIONS
WHY IS THERE A NEED TO USE SAND WHEN
GRINDING THE FRESH ANIMAL TISSUE SAMPLES FOR
THE EXTRACTION OF GLYCOGEN?

 It is for the animal tissue sample to rupture so that

glycogen can be extracted. The grinding with sand cause
glycogen to precipitate. Also, it increases the surface area
of the animal tissue. The more area exposed in the
reaction, the more efficient the process is.
WHAT IS THE ROLE OF TCA IN THE PROCESS OF
GLYCOGEN EXTRACTION?

 Since TCA is a highly corrosive agent, it helps in breaking

the animal tissue.
THE PROCESS OF EXTRACTION OF PLANT STARCH
SEEMS TO BE SIMPLER THAN THAT OF GLYCOGEN;
CAN YOU GIVE AN EXPLANATION FOR SUCH
DIFFERENCE?
 It is because the plant starch is easier to extract than
glycogen. In the experiment, there is a need to
rupture/break the animal tissue to extract the glycogen.
Also, starch and glycogen are both polysaccharides but
branching is frequent in glycogen.
WHY IS IT IMPORTANT TO USE UNRIPE FRUITS
INSTEAD OF THE RIPE ONES AS THE SOURCE OF
PLANT STARCH?

 It is because more starch can be extracted from unripe

fruits. Since ethylene is present in ripe fruits, it will convert
starch to sugar.
DID THE IODINE TESTS FOR BOTH PLANT AND ANIMAL
STARCH GIVE THE SAME RESULTS?

 No. It is because of the presence of the amylose in the

starch that causes the blue-black color (Iodine Clock
Reaction) while glycogen has amylopectin.
WHAT IS THE NAME OF THE BLUE COLORED
COMPOUND FORMED BY THE PLANT STARCH WITH
IODINE SOLUTION?

 It is amylose.
EXPLAIN THE DIFFERENCE IN OBSERVATIONS NOTED
IN THE REACTION OF PLANT STARCH WITH IODINE
SOLUTION IN HOT TEMPERATURE AND AT ROOM
TEMPERATURE.

 The reaction in the Iodine Test took time for the Iodine
solution to react with the amylose for the starch and
amylopectin for the glycogen.
WHAT IS THE PRINCIPLE INVOLVED IN THE MOLISCH’S
AND ANTHRONE TEST?

 For Anthrone Test, the carbohydrates are dehydrated with

concentrated H2SO4 to form “Furfural”, which condenses
with anthrone to form a green color complex.
 For Molisch’s Test, the carbohydrate gets dehydrated on
contact with concentrated sulfuric acid to form furfural
derivatives which on reaction with α-napthol (Molisch’s
reagent) results in a violet colored complex.
GIVE THE ROLE OF THE COMPONENTS OF EACH
REAGENTS OF THE MOLISCH’S AND ANTHRONE
TESTS.

 For the Anthrone reagent, it helps in condensing the

furfural to form green color complex.
 For the Molisch’s reagent, it helps in the dehydration of
the reaction to produce a violet color complex.
RESOURCES

 http://allmedtests.com/iodine-test-starch/
 https://allmedicalstuff.com/anthrone-test-carbohydrates/
 http://allmedtests.com/anthrone-test/
 http://allmedtests.com/molischs-test-principle-reagent/
 http://www.biochemden.com/anthrone-method-carbohydrate-
determination/
 https://www.quora.com/What-is-the-principle-of-the-Molisch-test-
for-carbohydrates
 http://www.harpercollege.edu/tm-
ps/chm/100/dgodambe/thedisk/carbo/molisch/molisch.htm
 http://generalchemistrylab.blogspot.com/2011/12/molisch-test-for-
carbohydrates.html