ANGELES UNIVERSITY FOUNDATION

Angeles City

College of Allied Medical Professions MTChem2 Laboratory

Department of Medical Technology Biochemistry for Medical Laboratory Science

EXPERIMENT 2: Monosaccharides and Disaccharides

INTRODUCTION
Monosaccharides are the simplest carbohydrates, which contain only a single aldehyde
of ketone functional group. They are colorless, crystalline, and frequently possess a sweet taste.
A monosaccharide is classified either as aldose or a ketose depending on whether it contains
an aldehyde or a ketone functional group respectively.
Glucose, C6H12O6, also called dextrose, is the most abundant organic compound in
nature. It is the component of polysaccharides cellulose, starch and glycogen. In medicine,
sugar is often called blood sugar.
The most important disaccharide is sucrose or cane sugar. Hydrolysis of sucrose,
C12H22O11, yields a 50:50 mixture of monosaccharide fructose and glucose, a mixture often
called invert sugar which is a major component of honey. Lactose, or milk sugar, is a
disaccharide which contains alpha-D-glucopyranose ring and alpha-D-galactopyranose ring.

OBJECTIVES
At the end of the experiment, students should be able to:
1. Identify the presence of carbohydrates
2. Identify the presence of aldoses
3. Differentiate the properties of monosaccharides from disaccharides

MATERIALS/APPARATUS
Test tubes, test tube rack, test tube brush, test tube holder, mortar and pestle water
bath, tripod, hot plate, graduated cylinder, watch glass, stirring rod, and medicine dropper.

WASTE DISPOSAL
● Flush acid solutions with plenty of water
● All solid materials are to be thrown in the trash can
● Discard organic solutions and reaction products appropriately in labeled waste bottles

EXPERIMENTAL PROCEDURE
Note: The following test solutions for each test will be used: GLUCOSE SOLUTION,
LACTOSE SOLUTION, SUCROSE SOLUTION

A. Molisch’s test:
1. Prepare 3 test tubes containing 3 mL of the solutions
2. Add 3 drops of Molisch’s reagent to each of these test tubes. Shake gently to mix
the solution well. Incline each test tube and slowly and carefully pour down the
side 3 mL of concentrated sulfuric acid. DO NOT SHAKE. Carefully put the tubes
upright and note the color of the ring formed at the junction of the two liquids.
ANGELES UNIVERSITY FOUNDATION
Angeles City

College of Allied Medical Professions MTChem2 Laboratory

Department of Medical Technology Biochemistry for Medical Laboratory Science

3. Compare the results in each test tube.

B. Schiff’s test:
1. Place 3 mL of the following test solutions in three separate test tubes.
2. Add 1 mL of Schiff’s reagent into each of these test tubes and note if any
changes in color occur.
3. Record your results.

C. Reducing properties:
a. Tollen’s test
Using 2 mL of the test solutions in three separate test tubes, place 3 mL
of ammoniacal silver nitrate in each. Mix by shaking the tubes gently. Then,
warm the test tubes in a water bath. Describe the changes observed.

b. Fehling’s test
Place 2 mL of the test solutions in three separate test tubes. Add in each
tube 3 mL of Fehling’s solution (Fehling’s A and B combined in equal ratio) and
heat the test tubes in a water bath. Note the reaction and color changes, giving
attention to the precipitate formed at the bottom.

c. Benedict’s test
Place 3 mL of the test solutions in three separate test tubes. Add 3 mL of
Benedict’s solution in each of these test tubes and warm in a water bath.
Observe the reactions and record your results.

d. Moore’s test
Place 2 mL of the test solutions in three separate test tubes, add in each
3 mL of strong NaOH solution and boil for 3 to 5 minutes. Note for changes which
will appear in each of the test tubes.

e. Picric acid test

Prepare three test tubes each containing 2 mL of picric acid and 0.5 gram
of sodium carbonate. Add in each test tube 2 mL of the test solutions. Warm the
test tubes in a water bath and observe the color changes.

f. Nylander’s test
To 2 mL of the test solutions in three separate test tubes, add 1 mL of
Nylander’s reagent in each test tube and heat the test tubes in a water bath. Note
the changes in the test tube.
ANGELES UNIVERSITY FOUNDATION
Angeles City

College of Allied Medical Professions MTChem2 Laboratory

Department of Medical Technology Biochemistry for Medical Laboratory Science

EXPERIMENT 2: Monosaccharides and Disaccharides

Names: __________________________ Date: ________________________

_________________________________ Instructors: ________________________
_________________________________ ________________________
_________________________________ SCORE: ________________________
_________________________________

Section: _________________________________

DATA & RESULTS

A. General test for carbohydrates

MOLISCH’S TEST with: Visible result Interpretation

Glucose solution

Lactose solution

Sucrose solution

B. General test for aldehydes

SCHIFF’S TEST with: Visible result Interpretation

Glucose solution

Lactose solution

Sucrose solution
ANGELES UNIVERSITY FOUNDATION
Angeles City

College of Allied Medical Professions MTChem2 Laboratory

Department of Medical Technology Biochemistry for Medical Laboratory Science

C. Reducing property

Visible result obtained with:

Test performed
Glucose solution Lactose solution Sucrose solution

1) Tollen’s test

2) Fehling’s test

3) Benedict’s test

4) Moore’s test

5) Picric acid test

6) Nylander’s test
ANGELES UNIVERSITY FOUNDATION
Angeles City

College of Allied Medical Professions MTChem2 Laboratory

Department of Medical Technology Biochemistry for Medical Laboratory Science

QUESTIONS FOR RESEARCH

1. What is meant by General Test? When is it appropriately performed?

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2. When do carbohydrates become reducing sugars?

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3. How can Fehling’s test determine the reducing property of a sugar?

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4. Does sucrose exhibit a reducing property? Why?

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