CHEM123

BIOCHEMISTRY FOR MEDICAL LABORATORY SCIENCE

LABORATORY ACTIVITY #1
DETERMINATION OF CARBOHYDRATES

Name: JOHN LLOYD H. DANO

Section: MEDTECH 2-YA-1
Group No. GROUP 1
Date:

I. Tests Based on the Formation of Furfural and Its Derivatives

1. Molisch’s Test
a. Mix 4 ml of distilled water and two drops of the Molisch’s reagent in a test tube. This tube
will serve as the control.
b. Place 4 ml of 3% solution of glucose in a second test tube. Add two drops of the Molisch’s
reagent and mix the contents by gently shaking the test tube.
c. Incline the test tube and cautiously add about 5 ml of concentrated sulfuric acid, allowing
the acid to run down the side of the tube. Sulfuric acid is denser than water and will form
a lower layer. Note the color of the ring formed at the junction if the two liquids.
d. In the same manner of adding acid, add sulfuric acid to the control tube. What do you
observe?
e. Repeat the above test with 3% sample solutions of glucose, xylose, lactose, and starch.
f. Record all results.

Substance Tested Results Interpretation

Control

Glucose

Xylose

Lactose

Starch

1
2. Bial’s Orcinol Test
a. Place 1 ml of each 3% solution of xylose, glucose, and starch in separately labeled test
tubes.
b. Add 3 ml of Bial’s reagent to each test tube.
c. Carefully heat each tube over a Bunsen flame until the solution begins to boil. Add one to
two drops of 10% FeCl3 solution.
d. Note the color of the product formed.
e. Record your results in the table.

Substance Tested Results Interpretation

Xylose

Glucose

Starch

3. Seliwanoff’s Test
a. Place 1 ml each of 3% xylose, glucose, fructose, and sucrose in separately labeled test
tubes.
b. Add 4 ml of Seliwanoff’s reagent to each test tube.
c. Place the test tubes in a water bath filled with boiling water and allow them to stay there
for exactly 1 minute.
d. Note the changes and record which test tube gives a positive result in the shortest time.
e. Continue heating and observe the color changes at 1-minute intervals.
f. Record the time required for a positive test for each sample.

Substance Tested Time Result Interpretation

Xylose 3
CLEAR NEGATIVE
CLEAR NEGATIVE
Glucose 3 REDPI
CLEAR
NK

Fructose 1 RED POSITIVE

POSITIVE
2 PINK
Sucrose

2
II. Tests Based on the Reducing Property of Sugars
1. Benedict’s Test
a. Place 1 mL each of 3% solutions of glucose, xylose, fructose, lactose, and sucrose in
separately labeled test tubes. Add 5 mL of Benedict’s reagent in each test tube.
b. Place all the tubes in boiling water bath for 2 to 3 minutes.
c. Observe the color of the solution and note whether a precipitate was formed. A change
in color of the solution is not considered a positive reaction.
d. Avoid prolonged heating.
e. Record your results.

Substance Tested Results Interpretation

2. Barfoed’s Test
a. Place 1 mL each of 3% solutions of glucose, xylose, fructose, lactose, and starch in
separately labeled test tubes.
b. Add 3 mL of Barfoed’s reagent in each test tube.
c. Prepare a control tube using distilled water instead of sugar solution.
d. Place all the tubes in a boiling water bath for 10 minutes. Record your observations.

Substance Tested Results Interpretation

Glucose CLEAR BLUE NEGATIVE

Xylose CLEARBLUE
CLEAR NEGATIVE
BLUECLEAR NEGATIVE
BLUE BLUE
CLEAR
Fructose NEGATIVE

3
 Lactose
CLEAR
CLEAR BLUE NEGATIVE
BLUECLOUDY NEGATIVE
Starch CLOUDY
BLUE BLUE

3. Tollen’s Test
a. Place 5 drops of 3% solutions of glucose, xylose, and sucrose in separate test tubes.
b. Add 2 mL of Tollen’s reagent into each tube.
c. Boil for about 5 minutes. Note and record your observations.

Substance Tested Results Interpretation

GLUCOSE CREAM POSITIVE

XYLOSE YELLOW-BROWN POSITIVE

SUCROSE GRAY POSITIVE