Experiment # 2

GENERAL TESTS FOR CARBOHYDRATES

Group Number: 1                                                         Year/Section: BSN-1B

Date performed:  August 31, 2020                                            
Date Submitted: September 7, 2020
Group Leader: Jamiel James Arceno
        Members: Star Arricivita
Seth Basio
OBJECTIVE:  To study and perform the different tests that is used to establish the
presence of carbohydrate in a material.

APPARATUS AND MATERIALS:

 Test tube rack  5% solutions of glucose
 Test tubes  5% solutions of sucrose
 Graduated cylinder  5% solutions of starch
 Beaker  Molisch reagent
 Tripod  Sulfuric Acid
 Wire gauze  Sodium Hydroxide
 Tongs  Anthrone reagent

PROCEDURE:
MOLISCH TEST
a) In each of the three test tubes, place separately 2 mL of 5% solutions of
glucose, sucrose, and starch.

b) To each test tube, add 2 mL of Molisch reagent. Mix well. Incline each tube
and carefully pour down the side 2 mL of conc. H2SO4, taking care that the
concentrated acid forms a layer at the bottom of the test tube.

c) Note the color formed at the junction of the two layers.

MOORE'S TEST
a) In each of three test tubes, place 1 mL of conc. NaOH.

b) To the first, add 1 mL of 5% glucose solution. To the second, add 1 mL of 5%

sucrose solution. And to the third, 5% starch solution.

c) Boil the solutions and observe the change in color produced in each tube.
ANTHRONE TEST
a) To each of three test tubes, place separately 1 mL of 5% glucose solution, 1
mL of 5% sucrose solution, and 1 mL of 5% starch solution.

b) To each, add 1 mL of anthrone reagent and mix.

c) Let stand for some time and note the color produced immediately and after
one hour.

d) If the solution is milky, dilute with 50% H2SO4 or glacial acetic acid.

RESULTS AND OBSERVATIONS:

TEST RESULTS AND OBSERVATIONS

After the 2mL of molisch reagent (a solution of α-
MOLISCH napthol in ethanol) and 2 mL of Sulfuric acid were
added succesively to the 5 mL glucose, sucrose, and
starch, a positive reaction did happen to each of the
samples. Hence, indicating that carbohydrates are
indeed present in each one of them. The pentoses
that were dehydrated by the test reagent caused
them to form the furfural and the reagent also
dehydrated the hexoses that caused the
hydroxymethylfurfural. The α-napthol present in
the reagent, made the furfurals react further
 causing the violet ring in the middle.
The Glucose solution became significantly darker in
color compared to the Sucrose and Starch
solutions. Indicating that Glucose had a positive
MOORE'S
reaction based on the liberation of aldehydes which
subsequently polymerizes to form “caramel”. While
Sucrose and Starch gave a negative reaction.
After 1 mL of anthrone reagent were added
separately into 1 mL of 5% glucose solution, 1 mL
of 5% sucrose solution, and 1 mL of 5% starch
solution, all carbohydrates exhibited a dark
bluish/greenish color.In this test, the carbohydrates
ANTHRONE get dehydrated when they reacted with conc.
H2SO4 to form the furfural. This furfural reacted
with anthrone and gave the darkbluish green
colored complex.Since the results showed this type
of color, that means that all carbohydrates give test
positive.

CONCLUSION:
This experiment was made to study and perform the different tests that are
used to establish the presence of carbohydrate in given solutions namely, Glucose,
Sucrose, and Starch. In the Molisch test, there are presences of furfurals
(hydroxymethylfurfural and furfural) and the violet ring indicator is visible in all of the
test tubes, which means that all of the solutions have a positive result and
carbohydrates are present. Next, in the Moore test, Glucose is the only solution to
get a positive result while, Sucrose and Starch had a negative result, since both are
non reducing sugars, they do not have free aldehyde or ketone groups and therefore
there are no liberations of aldehydes and they do not undergo the polymerization
reaction. Finally, the Antrone test, all of the solutions also yielded a positive result
due to the change of color of the solutions to a dark blue/green after the anthrone
reagent was added into the test tubes and mixed with the solutions;the color change
was a result of the dehydration of carbohydrateswhen they reacted with sulfuric acid
to form a furfural and then reacted with the anthrone.

QUESTIONS:

1. Explain reasons for using Molisch Test, Moore's Test, & Anthrone Test in the
conduct of General Tests for Carbohydrates experiment.
 The main reason for using Molisch’s test is to find out if a particular substance
has carbohydrates. Same as for the Anthrone test, both of these tests are used for
the detection of carbohydrates in the given solution. For the Moore’s test, it is a test
used for reducing of sugars except for sucrose. All of these different tests are used to
establish the presence of carbohydrate in a material.
2. Describe the evidences for a positive test for the three tests.
       The factors that indicate that the samples are positive in the Molisch test are the
presence of the furfurals (hydroxymethylfurfural and furfural) along with a violet ring
in the middle, caused by the α-napthol in the reagent. The factors that indicate that
the samples are positive in Moore’s test are the residue substance, caramel. It is
identified by the golden-brown color and the smell the final result of the sample
emits. The positive results of the samples in Anthrone test is identified by the color
of the final result of the sample.