Experiment Title: Carbohydrates Materials:

Experiment Objectives: At the end of the experiment, Laboratory Manual

students are expected to: References:
1. identify common carbohydrates in nature; Bettelheim, Frederick., et., al.
2. classify the different carbohydrates and; (2010) Laboratory Experiments
for Introduction to General,
3. perform simple qualitative tests for carbohydrates. Organic, and Biochemistry, 7th
ed., ISBN-13: 978-0-495-
39196-8
Voet, Donald., et., al. (2008)
Fundamentals of
Biochemistry: LIFE AT THE
MOLECULAR LEVEL, 2nd ed.,
ISBN-13 978-0470-12930-2.

Productivity Tip:

Isolate yourself from distractions!

Introduction

Carbohydrates are most abundant organic compounds found in living organisms and are
composed of carbon, hydrogen and oxygen. Carbohydrates act as the primary source to provide
energy for functioning of living organisms. These are called carbohydrates because they can be
considered as hydrates of carbon. Carbohydrates, or saccharides, are divided into
monosaccharides, disaccharides, and polysaccharides.

A monosaccharide consists of a single carbohydrate molecule, containing between 3 and

7 carbons. Glucose and fructose are examples of monosaccharides. A disaccharide consists of
two monosaccharides that are linked together. Sucrose and lactose are disaccharides. A
polysaccharide consists of many monosaccharides linked together. Starch, pectin, glycogen, and
cellulose are examples of polysaccharides.

PRE-LABORATORY ACTIVITY

A. Answer the following questions

1. What is carbohydrates? What is the structural characteristic of carbohydrates?

2. How are carbohydrates classified? Give examples for each classification.

3. Discuss the principles behind each of the following test.

a) Fehling’s test
b) Seliwanoff’s test
c) Iodine test

4. What are reducing sugars? Why is sucrose non-reducing while lactose also a
disaccharide, is reducing? Why are monosaccharides reducing sugars?

5. How do starch and glycogen differ structurally?

Materials/Reagents

▪ Test tubes (9 pcs) ▪ Test tube rack (1 pc)

MC2, Biochemistry Laboratory

 ▪ Test tube brush (1 pc) ▪ Glucose
▪ Beaker (1 pc 500 ml for boiling) ▪ Maltose
▪ Test tube holder (1 pc) ▪ Lactose
▪ Pipette (4 pcs, 1 ml) ▪ Fructose
▪ Aspirator (1 pc) ▪ Galactose
▪ Fehlings A&B ▪ Sucrose
▪ Seliwanoff’s Reagent ▪ Starch
▪ I2KI solution ▪ Distilled water
▪ Arabinose
Procedure

A. General Tests

Fehling’s Test
General Principle: Fehling's test is an indicating reaction for reducing groups like Aldehyde
functions. The principle of this test is based on the fact that the aldehyde group of sugar is oxidized
to form acids by complex Copper ions. The Red Copper (I) oxide then precipitates.

Procedure:

1. In nine labeled test tube, place 2 ml of the following:

Test tube No. Sugar Solutions
1 5% fructose
2 5% galactose
3 5% arabinose
4 5% glucose
5 5% maltose
6 5% sucrose
7 5% lactose
8 5% starch
9 water

2. Add 1 ml of Fehling's solution A and Fehling's solution B to it.

3. Place the solution in a boiling water bath for about 10 minutes. Observe

*The formation of red precipitate indicates the presence of carbohydrate.

B. Specific Test for some Carbohydrates

Seliwanoff’s Test – Resorcinol – HCL Test (Test for Ketoses)

Principle: Ketoses, particularly fructose, form hydroxymethyl furfural by the action of HCL and
condenses with resorcinol to form a red color.

Procedure:
1. In nine labeled test tube, place 1 ml of the following:
Test tube No. Sugar Solutions
1 5% fructose
2 5% galactose
3 5% arabinose
4 5% glucose
5 5% maltose
6 5% sucrose
7 5% lactose
8 5% starch
9 water

2. Add 2 ml of Seliwanoff’s reagent in a labelled test tube, mix and heat the mixture to boiling for
10 minutes.
3. Observe

MC2, Biochemistry Laboratory

*The appearance of cherry red color with or without the separation of a brown – red precipitate
indicates the presence of a ketose.

Note: the time of appearance of s red color.

Aldoses, like glucose, may give a positive reaction upon prolonged heating. This is due to catalytic
transformation of glucose to fructose by HCL.

Iodine test
General principle: The basic principle involved in the iodine test is that Amylose interacts with starch
to form a blue-black colored complex with the iodine. The helical structure of Amylose forms a
charge transfer (CT) complex with iodine, wherein iodine is present inside the spiral or helical
structure of the Amylose.

Procedure:
1. In nine labeled test tube, place 1 ml of the following:
Test tube No. Sugar Solutions
1 5% fructose
2 5% galactose
3 5% arabinose
4 5% glucose
5 5% maltose
6 5% sucrose
7 5% lactose
8 5% starch
9 water

2. Pipette 1 ml of 0.1 M I2KI solution and Add to the solution.

3. observe for a change in color from reddish brown to blue.

MC2, Biochemistry Laboratory