CARBOHYDRATES: TESTS BASED ON THE FORMATION OF FURFURAL

AND ITS DERIVATIVES

Introduction
 A carbohydrate is a polyhydroxy aldehyde, a
polyhydroxy ketone, or a compound that yields
polyhydroxy aldehydes or polyhydroxy ketones
upon hydrolysis.

Tests for Carbohydrates:

1. General Tests for Carbohydrates – Tests
based on the formation of furfural and its
derivatives
2. Reducing tests for Carbohydrates

General Tests for Carbohydrates

Classification of Carbohydrates MOLISCH TEST

 Monosaccharides - “ simple sugars” , are highly
soluble in water, less soluble in ethanol, and
insoluble in ether.
- They are either aldoses or ketoses
- They may also be classified into tetroses,
pentoses, or hexoses
- Free monosaccharides are all reducing
sugars
- They also exhibit muta-rotation
 is the general test for carbohydrates.
 The sugars are mixed with a-naphthol.
 The test tube is inclined, and concentrated
H2SO4 is added along the side of the tube,
causing the formation of a lower layer of acid.
 The concentrated H2SO4 will dehydrate the
sugar allowing it to react with the alcohol
forming furfural or hydroxymethyl-furfural.

 Disaccharides – are formed by two molecules

of monosaccharides. Examples are maltose,
which are abundant in germinating barley;
sucrose, also known as cane sugar or beet
sugar; and lactose or milk sugar, which does not
taste very sweet and is not fermented by yeast.

 Polysaccharides – found in nature function

either as structural units (e.g. cellulose), or for
storage such as starch, dextrin, glycogen, and
inulin.
GLYCANS

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 CARBOHYDRATES: Tests based on the Formation of Furfural and its Derivatives

PROCEDURE:
1. Mix 4 ml of distilled water and two drops of
the Molisch’s reagent in a test tube. This tube
will serve as the control.
2. Place 4 ml of 3% solution of glucose in a
second test tube. Add two drops of the
Molisch’s reagent and mix the contents by
gently shaking the test tube.
3. Incline the test tube and cautiously add about
5 ml of concentrated sulfuric acid, allowing
the acid to run down the side of the tube.
Sulfuric acid is denser than water and will
form a lower layer. Note the color of the ring
formed at the junction if the two liquids.
4. In the same manner of adding acid, add
sulfuric acid to the control tube. What do you
observe?
5. •Repeat the above test with 3% sample
solutions of fructose, lactose, and sucrose
6. Record all results.
Procedure:
1. Place 1 ml of each 3% solution of xylose, and
glucose in separately labeled test tubes
2. Add 3 ml of Bial’s reagent to each test tube
3. Carefully heat each tube over a Bunsen
flasme until the solution begins to boil.
4. Note the color of the product formed
5. Record your results in the table
SELIWANOFF‘S TEST
 This test is used to differentiate ketohexoses
from aldohexoses. Ketohexoses react faster
with the solution containing hydrochloric acid
and resorcinol than aldohexoses.
 The dehydrated ketohexoses form a bright
cherry red condensation product, while the
aldohexose yields only a pale pink coloration,
a negative result.
 In this test, prolonged heating of samples
should be avoided.

BIAL’S ORCINOL TEST

Concentrated HCl Ferric Chloride 5-
PROCEDURE
hydroxymethyl furfural
1. Place 1 ml each of 3% glucose, fructose,
 Used to determine the presence of pentoses
lactose and sucrose in separately labeled test
and nucleotides and nucleotides that contain
tubes.
pentose sugars.
2. Add 4 ml of Seliwanoff’s reagent to each test
 When pentoses are treated with orcinol,
tube
furfurals are formed and they will yield a
3. Place the test tubes in a water bath filled with
blue-green compound in the presence of
boiling water and allow them to stay there for
ferric ions.
exactly 1 minute
 The reaction is not specific for pentoses
4. Note the changes and record which test tube
because other compounds like trioses, uronic
gives a positive result in the shortest time.
acids, and certain heptoses will also give blue
5. Continue heating and observe the color
or green products.
changes at 1 minute intervals
 Hydroxymethyl furfural is formed from
6. Record the time required for a positive test
hexoses to give yellow-brown condensation
for each sample
products.

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