Experiment 2

Analysis of Carbohydrates

Carbohydrates are defined chemically as polyhydroxy aldehydes or ketones and yield these
products after hydrolysis in a slightly acidic solution. Also known as saccharides, they are
represented by the general formula (CH2O)n. Monosaccharides, such as glucose and fructose,
are the smallest units of carbohydrates. On the other hand, maltose, lactose, and sucrose are
classified as disaccharides. Starch and cellulose are the common examples of polysaccharides.
Various chemical tests are used to determine the properties and differentiate carbohydrates
from one another.

Materials
Test tubes, 250 mL beaker, graduated cylinder, cheesecloth, marbles, cotton, water bath setup,
droppers, mortar and pestle, potato/cassava

Reagents
0.1 M sugar solutions (glucose, fructose, xylose, galactose, lactose, sucrose), 0.01 M iodine
solution, Molisch reagent, Bial’ls orcinol reagent, Seliwanoff’s reagent, Tollen’s reagent,
concentrated H2SO4, Fehling’s reagents (A and B), Benedict’s reagent, distilled water,
CH3COONa

*Masks and gloves are required for this experiment

Procedure

Extraction of Starch from Potato/Cassava

1. Cut a small part of the root crop and grind it.
2. Transfer into a small beaker, add 100 mL of distilled water and mix.
3. Strain the mixture using a cheesecloth and allow the starch to settle.
4. Separate samples for use in general tests for saccharides and for hydrolysis.

Acid Hydrolysis
1. Add 5 drops of conc. HCl to every 5 mL of extract.
2. Cover with a marble and boil in a water bath for 30 minutes.
Tests for Saccharides
A. Molisch’s Test – Add a few drops of Molisch’s reagent to 1 mL of samples (extract,
hydrolysate, and 0.1 M solutions). Carefully place H2SO4 down the sides of the tube.
Observe changes that will occur at the interface of the acid and sample.
B. Iodine test – Add a few drops of 0.01 M iodine solution into a small amount of sample.
Heat sample in a water bath and observe change.

Barfoed’s, Benedict’s, Bial’s Orcinol, Tollen’s, and Seliwanoff’s Tests

*Perform one test at a time
1. In separate tubes, mix 5-10 drops of your samples and 1 mL of required reagent.
2. Place the tubes in a boiling water bath.
3. Note the time it takes to produce visible results and take out the sample once a result
is seen.

Fehling’s Test
1. Add a few drops of sample to equal amounts of Fehling’s A and B.
2. Heat in a water bath until it produces visible results.

Phenylhydrazone Test
1. Mix 4 grams of phenylhydrazine HCl, 6 grams of CH3COONa, and 20 mL of distilled
water. This should be enough for the entire class.
2. Please the reagent in a warm water bath, stirring until the solution is clear.
3. Mix 2 drops of sample with 4 drops of phenylhydrazine reagent. Mix and cover with
cotton.
4. Heat in a boiling water bath for 30 minutes. Note the time when crystals first appear
5. Allow tubes to cool.
6. Place the solution in a glass slide and view under the microscope. Draw
observations.
Name: _______________________________ Date: ___________________
Section:______________________________ Instructor: _______________

Analysis of Carbohydrates

Results
Sample
Molisch’s Test Iodine Test Barfoed’s Test Benedict’s Test

Fructose

Glucose

Xylose

Lactose

Sucrose

Starch Extract

Hydrolysate

Results
Sample
Bial’s Test Tollen’s Test Seliwanoff’s Test Fehling’s Test
Fructose

Glucose

Xylose

Lactose

Sucrose

Starch Extract

Hydrolysate
Name: _______________________________ Date: ___________________
Section: ______________________________ Instructor: _______________

Mono-/Oligo- Reducing/Non-
Sample Hydrolysable? Aldose/Ketose?
/Polysaccharide? reducing?

Fructose

Glucose

Xylose

Lactose

Sucrose

Starch Extract

Hydrolysate

Draw the osazones observed.

Name: _______________________________ Date: ___________________
Section: ______________________________ Instructor: _______________

1. Explain how the hydrolysis of starch happens. What is/are the product/s of starch
hydrolysis?

2. What does a “reducing sugar” mean? Which of the following tests is/are specific to reducing
sugars?

3. What is the observed brick-red precipitate in the experiments?

4. Which test can be used to differentiate the following? Explain

a. Ribose and glucose

b. Fructose and galactose

Name: _______________________________ Date: ___________________
Section: ______________________________ Instructor: _______________

c. Glucose and maltose

5. Explain the principle involved for Tollen’s test.

6. Explain why the iodine test is specific to a certain class of sugars.

7. What is the difference between starch and cellulose that makes one digestible and the
other not digestible to humans? Explain AND illustrate.