CEBU DOCTORS’ UNIVERSITY

COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

Laboratory Activity Sheet No. 2.1

Analysis of Unknown Carbohydrates

Group Members: Camacho, Jianshe Laboratory Schedule: T/F – 3:00pm - 6:00pm

Horteza, Justinne
Laurente, Ashley Jade
Milan, Jasmine
Santisas, Sciarra Ann

Program and Year: BSN - 1L 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 A purplish-red orange-red brick red colorless yellow

ring precipitate precipitate solution

Unknown B purplish-red brick red blue solution colorless yellow

ring precipitate solution

Unknown C dark purple brick red brick red cherry red yellow
ring precipitate precipitate complex solution

Unknown D no ring blue solution blue solution colorless yellow

solution

Unknown E purple ring blue solution blue solution cherry red yellow
complex solution

Unknown F purplish-red blue solution blue solution colorless blue-black

ring solution
 CEBU DOCTORS’ UNIVERSITY
COLLEGE OF ARTS AND SCIENCES

Physical Sciences Department

2. Discuss below the interpretation of the results.

The Molisch’s test is a general test for the presence of carbohydrates. The emergence of a
purple ring denotes a positive outcome. In this condition, Unknowns A, B, C, E, and F were
capable of producing purple rings when tested for the Molisch’s Test, therefore, the solutions
contained carbohydrates. On the other hand, Unknown D was unable to produce a purple ring
indicating that Unknown D was water. Benedict's Test was the next test conducted; it is utilized in
the identification of simple carbohydrates; detecting whether they are reducing or non-reducing
sugars. The color of the precipitate can vary from red to orange to green depending on the amount
of sugar that is present in a solution. Unknown A produced an orange-red precipitate indicating a
moderate amount of reducing sugar was present. Unknowns B and C produced brick red
precipitates indicating large amounts of reducing sugars in the solutions. And, Unknowns E and F
produced blue solutions, indicating that the solutions were non-reducing sugars. Additionally,
Unknown D also produced a negative result since it was water.
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 A and C were able to produce brick-red precipitates indicating that these solutions were
monosaccharides, whereas Unknowns B, D, E, and F 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). The formation of a cherry red
solution is a positive result. Unknowns C and E gave positive results, forming cherry red solutions,
while Unknowns A, B, D, and F produced colorless solutions indicating that they were negative
when tested with Seliwanoff’s Test. Lastly, the Iodine Test is a test used to identify the presence
of polysaccharides, such as starch. When a few drops of potassium iodide solution are poured on
the sample, it turns blue-black in color. In this case, Unknown F gave a positive result with the
Iodine Test suggesting that Unknown F was a starch.
In conclusion, Unknown A is glucose, as it is a reducing sugar with an aldehyde group;
Unknown B is lactose, as it is a reducing disaccharide; Unknown C is fructose, as it is a reducing
sugar with a ketone group; Unknown D is water, as it has no presence of carbohydrates; Unknown
E is sucrose, as it is a disaccharide with a ketone group; and lastly, Unknown F is starch, as it is a
polysaccharide.

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

Unknown 1 - glucose
Unknown 2 - lactose
Unknown 3 - fructose
Unknown 4 - water
Unknown 5 - sucrose
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 A, B, C, D, E, and F

Molisch’s Test
with purple/purplish-red ring without purple/purplish-red ring
A, B, C, E, F D

Benedict’s Test
brick-red solution blue solution
A, B, C D, E, F

Barfoed’s Test
brick-red
blue solution solution
B A, C

Seliwanoff’s Test
colorless solution cherry red complex

A, D, F C, E

Iodine Test
yellow solution blue-black solution
A, D F