Experiment 3:

CARBOHYDRATE
S
 To identify common carbohydrates in nature
 To classify the different carbohydrates
 To perform simple qualitative tests for
carbohydrates
 7 5 3 1
8 6 4 2

A. General Tests (Molisch test for all Carbohydrates)

1. In eight (8) labelled test tubes, pipette 1mL of the following:

Test tube No. Sugar solutions
1 5% fructose
2 5% galactose
3 5% ribose
4 5% glucose
5 5% sucrose
6 5% lactose
7 5% maltose
8 5% starch
 7 5 3 1
8 6 4 2

2. Add three drops of Molisch reagent and mix thoroughly

3. Incline each test tube and add 1mL of concentrated sulfuric acid
down the side of the tubes
 7 5 3 1
8 6 4 2

4. Do not shake to avoid mixing layers. Observe for the presence of a

violet ring at the interface of the two liquids.
 Bial’s [Orcinol] Test for Pentoses

1. In four labelled test tubes, place 1mL of the following solutions:

Test tube No. Sugar solutions
1 5% arabinose
2 5% glucose
3 5% maltose
4 Water
2. Add 2mL of the Orcinol reagent. Mix well and transfer to a boiling
water bath.
(Watch video on the right side)
3. Observe for the appearance of blue-colored solution after 15 minutes
of heating.
 B. Reduction Tests (Benedict’s test)
1. In nine labelled test tubes, place 2mL of Benedict’s reagent and 1mL
of the following sugar solution:
Test Tube No. Sugar Solution
1 5% fructose
2 5% galactose
3 5% arabinose
4 5% glucose
5 5% maltose
6 5% sucrose
7 5% lactose
8 5% starch
9 Water (control)
2. Place these test tubes with the carbohydrates in a boiling water bath
and note the time elapsed before the appearance of a brick red
precipitate is observed. If there is no brick red precipitate that appears
after 30 minutes, discontinue heating. (Watch video on the right side)
Watch video above
 Seliwanoff’s test
1. In nine labelled test tubes, place 2mL of Seliwanoff’s reagent and
1mL of the following sugar solution:
Test tube No. Sugar solutions
1 5% fructose
2 5% galactose
3 5% arabinose
4 5% glucose
5 5% maltose
6 5% sucrose
7 5% lactose
8 5% starch
9 Water (control)
2. Place these test tubes with the carbohydrates in a boiling water bath for 10 minutes
and note the time elapsed before a cherry red solution is observed. (Watch video on
the left side)
3. Compare the results of each sugar solutions. (Watch video on the right side)
 Fermentation Tests
1. Fill seven (7) labelled test tubes with 10 mL of the following sugar
solutions:
2. Add 2mL of the yeast suspension and mix well. Incubate the mixture
at 37oC .
3. Note the time required to produce a noticeable bubbling or gas as a
result of fermentation. (Watch video above)
4. If no gas has been formed after 3 hours, the test is considered
negative.
 Iodine test
1. In an evaporating dish, pipette 1mL of 1.0mL of I2/KI solution.
Test Tube No. Sugar solutions
1 5% Starch
2 5% glucose
3 5% sucrose
4 water
2. Add 2-3 drops of starch suspension and observe for a change in color
from reddish brown to blue. Repeat the test using the other carbohydrate
solutions.
 THANKS!
Does anyone have any questions?