CEBU DOCTORS’ UNIVERSITY

COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

Laboratory Activity Sheet No. 2.1

Analysis of Unknown Carbohydrates

Group Members: Floro, Feodor Wayne Jaime Laboratory Schedule: T & F 1:30 PM – 6:00 PM
Gaviola, Averie Gail
Gonzalez, Ma. Luizza Paola
Gumapac, Claire
Imperial, Julia Emanuelle

Program and Year: BSN 1-J Score:

I. Intended Learning Outcomes:

1. Perform various chemical tests on given unknown solutions to identify the type of
carbohydrates present in them;
2. Organize and interpret obtained data from the chemical tests conducted; and
3. Illustrate the flow of analysis conducted to identify the unknown solutions through a
schematic diagram.

II. Instructions:

A. Analysis of Unknown Carbohydrates

1. On your lab desks, you are provided with all the necessary apparatus, chemicals, and
materials for the conduct of the following procedures below. Including in the display of
materials are the 6 unknown solutions that may contain any of the following:
a. Glucose
b. Fructose
c. Lactose
d. Sucrose
e. Starch
f. Water

2. Perform the following chemical tests on the unknown solutions to elucidate their
identities:
 Molisch’s Test
1. Add 2 mL each of the unknown solutions in 6 different test tubes. Label
them accordingly.
2. To each of the test tubes, add 1mL of Molisch’s reagent. Mix well.
3. Pour down concentrated H2SO4 slowly at the side of each test
tubewhile tilting it at an angle of 45o. DO NOT SHAKE.
4. Observe for the formation of a purple or purplish-red ring.
5. Record your results in your notebook.
 Benedict’s Test
1. Add 2 mL each of the unknown solutions in 6 different test tubes. Label
them accordingly.
2. To each of the test tubes, add 3 mL of Benedict’s reagent. Mix well.
3. Heat the mixtures in a boiling water bath for 2 minutes.
 CEBU DOCTORS’ UNIVERSITY
COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

4. Observe for the formation of yellow to brick red precipitate.

5. Record your results in your notebook.
 Barfoed’s Test
1. Add 2 mL each of the unknown solutions in 6 different test tubes. Label
them accordingly.
2. To each of the test tubes, add 3 mL of Barfoed’s reagent. Mix well.
3. Heat the mixtures in a boiling water bath for 5 minutes.
4. Observe for the formation of brick red precipitate. To see the
precipitate, it may be necessary to view the tube against a dark
background in good light.
5. Record your results in your notebook.
 Seliwanoff’s Test
1. Add 2 mL each of the unknown solutions in 6 different test tubes. Label
them accordingly.
2. To each of the test tubes, add 3 mL of Seliwanoff’s reagent. Mix well.
3. Heat the mixtures in a boiling water bath for 2 minutes.
4. Observe for the formation of cherry red solution.
5. Record your results in your notebook.
 Iodine Test
1. Add 2 mL each of the unknown solutions in 6 different test tubes. Label
them accordingly.
2. To each of the test tubes, add 1 drop of I2 in KI solution. Mix well.
3. Observe for changes in color.
4. Record your results in your notebook.

B. Data Organization and Analysis

1. Create a table to organize your results below.
Solution Molisch’s Benedict’s Barfoed’s Seliwanoff’s Iodine test
test test test test

Unknown 1 (+) (+) (+) (+) (-)

Unknown 2 (-) (-) (-) (-) (-)

Unknown 3 (+) (-) (-) (+) (-)

Unknown 4 (+) (+) (+) (-) (-)

Unknown 5 (+) (+) (-) (-) (-)

Unknown 6 (+) (-) (-) (-) (+)

 CEBU DOCTORS’ UNIVERSITY
COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

2. Discuss below the interpretation of the results.

Molisch’stest
Molisch’s test is very useful in detecting any presence of carbohydrates in a substance.
The formation of a purple ring indicates a positive outcome. However, during the testing
of the unknown carbohydrates, only one of the tests for the unknown carbohydrates
didn't react the same way as the other test. Unknowns 1,3,4,5, and 6 produce a purple
ring which means it contained carbohydrates. The only test that didn't react and unable
to produce a purple ring is Unknown 2 because it’s a water.

Benedict’s test
Benedict's Test, which is used in identifying simple carbs and determining whether or not
they reduce or do not reduce sugars. Depending on the amount of sugar in a solution,
the precipitate's color might range from red to orange to green. Unknowns 1, 4, and 5
generated a brick-red precipitate, indicating the presence of some reducing sugar.
Moreover, Unknowns 2, 3 and 6 generated blue solutions, suggesting non-reducing
sugars in the solutions.
.
Barfoed’s test
Barfoed’s Test is a chemical test used for detecting the presence of monosaccharides. A
positive test for monosaccharides is indicated by the formation of a brick red precipitate.
Unknowns 1 and 4 were able to produce brick-red precipitate indicating that these
solutions were monosaccharides, whereas Unknowns 2, 3, 5, and 6 were disaccharides
since they produced blue solutions. Moving on, Seliwanoff’s Test is used to distinguish
between sugars with a ketone group (ketose) and sugars with an aldehyde group
(aldoses).

Seliwanoff” s test
Seliwanoff’s Test is used to distinguish between sugars with a ketone group (ketose)
and sugars with an aldehyde group (aldoses). The formation of a cherry red solution
indicates a positive result. Unknowns 1 and 3 gave a positive result, producing a cherry
red solution while Unknowns 2, 4, 5, and 6 unable to produce a positive outcome since
the solution is colorless.

Iodine test
Iodine test is used to identify the presence of polysaccharides, such as starch. To
identify the starch, it reacts into blue-black color when in contact with iodine. Amylose, a
component of starch, creates helices in which iodine molecules gather to produce a
blue- black color. The blue-black color is not created when starch is digested or
hydrolyzed into smaller carbohydrate molecules. We noticed that Unknowns 1,2,3,4,5
had a negative reaction but Unknowns 6 had a positive reaction to it. Concluding that
Unknown 6 is the starch.
 CEBU DOCTORS’ UNIVERSITY
COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

3. Based from the results of your experiment, identify the different unknown solutions:

Unknown 1 - fructose
Unknown 2 – water
Unknown 3 - sucrose
Unknown 4 - glucose
Unknown 5 - lactose
Unknown 6 - starch
 CEBU DOCTORS’ UNIVERSITY
COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

4. Sketch a schematic diagram below showing a logical flow of the chemical tests that
would lead you to the analysis of the individual unknown solutions.

Unknowns 1, 2, 3, 4, 5, and 6

Molisch’s Test
with purple/purplish-red ring without purple/purplish-red ring
1, 3, 4, 5, 6 2

Benedict’s Test
brick-red solution blue solution

1, 4, 5 2, 3, 6

Barfoed’s Test
brick-red
blue solution solution
5 1, 4

Seliwanoff’s Test
colorless solution cherry red complex

2, 4, 6 1, 3

Iodine Test
yellow solution blue-black solution
2, 4 6