MIDLANDS STATE UNIVERSITY

MANICALANDS COLLEGE OF APPLIED SCIENCES

DEPARTMENT OF CHEMICAL AND PROCESSING SYSTEMS
ENGINEERING
BSc (Hons) Chemical and Processing Engineering
HCHE 315 BIOCHEMICAL ENGINEERING

Name: Panashe Muduzu

Reg #: R14966Z
Level: 3.1
Title: TESTS FOR CARBOHYDRATES
Practical Number:
Partner(s):

Tanaka Makoena
Chiedza Mudimu
Blessed Zhou

Lecturer:

Dr Nyamunda

Date: 28 November 2016

Aim:
To test for carbohydrates.
Objective(s):

To determine the identity of an unknown carbohydrates by carrying out a

series of chemical reactions

Abstract
A carbohydrate is an organic compound with the general formula Cm(H2O)n, that is,
consists only of carbon, hydrogen and oxygen, with the last two in the 2:1 atom ratio.
Carbohydrates make up the bulk of organic substances on earth and perform numerous roles
in living things. The carbohydrates (saccharides) are divided into four chemical groups:
monosaccharides, disaccharides, oligosaccharides and polysaccharides. Monosaccharide
is the smallest possible sugar unit. Examples include glucose and fructose. When
monosaccharides merge together in linked groups they are known as polysaccharides.
Disaccharide is two monosaccharide molecules bonded together. Examples of disaccharides
include sucrose and maltose whereas polysaccharides include starch and glycogen. In this lab,
we will be concerned with the nature and activities of the carbohydrates and with their
structure.

Introduction
Simple sugars, starches and cellulose are organic compounds that have the approximate
formula C(H2O)n, which accounts for the name carbohydrate (or hydrate of carbon) that is
usually applied to this group of compounds. They are not truly hydrates of carbon but are
polyhydroxy (alcohol) compounds that contain an aldehyde or ketone functional group. These
functional groups give the carbohydrates some of their chemical properties that will be
studied in this lab. Simple sugars are called monosaccharides (one sugar), or disaccharides (2
sugars). Some monosaccharides are glucose, fructose and galactose. Note fructose is a ketose.

Two common disaccharides are sucrose (table sugar) and lactose (milk sugar); sucrose is a
combination of glucose and fructose linked together by their anomeric carbons to produce a
nonreducing sugar (it does not reduce Cu 2+), whereas lactose is a combination of galactose
and glucose linked together by a -1, 4-glycosidic bond to produce a reducing disaccharide.

When many sugar molecules are linked together into a polymer, the resulting compound is
called a polysaccharide. Starches and celluloses are polysaccharides. Amylose is a linear
chain polymer of glucose, whereas amylopectin (a plant starch) and glycogen (an animal
starch) are branched polymers of glucose.

Qualitative Tests for Carbohydrates

Reducing sugars are usually detected with Benedict's reagent, which contains Cu2+ ions in
alkaline solution with sodium citrate added to keep the cupric ions in solution. The alkaline
conditions of this test causes isomeric transformation of ketoses to aldoses, resulting in all
monosaccharides and most disaccharides reducing the blue Cu 2+ ion to cuprous oxide (Cu2O),
a brick red-orange precipitate. This solution has been used in clinical laboratories for testing
urine.
Iodine forms a deep blue colour in the presence of starch. Potassium iodide is added to the
reagent solution in order to make the iodine more soluble in water. Some forms of starch may
yield a greenish colour. Simple carbohydrates (mono- and disaccharides) and cellulose do not
cause any change in the orange-brown colour of the iodine reagent.
Since carbohydrates are readily identified by colour change in specific reactions, we will
explore some of these methods of identification as we carry out specific tests for particular
carbohydrates. Solutions of the following mono-, di-, and polysaccharides are available: (a)
glucose, (b) fructose, (c) galactose, (d) sucrose, (e) starch and (f) unknown. These solutions
are available at 2% concentrations (percent solutions are by weight/volume, thus 2% is 2
gram in 100 ml of solvent).

PRECAUTIONS AND SAFETY

Benedicts reagent is caustic, rinse thoroughly with water if you get this solution
on your skin or clothing.
Be sure to heat the tubes slowly in a water bath, tilting it slightly and
heating along the entire side of the tube
Do not point the tube at your fellow co-workers
DO NOT place your pipettes in reagent bottles as this leads to contamination.

APPARATUS
Test tube, 400 mL beaker, droppers, hot plate, 5-10 mL graduated cylinder, watch glass or
spot plate.

Reagents
Chemical name

Chemical formula Manufacturer

Mass/ concentration

Benedict's reagent

SKYCHEM

50 mL

Iodine reagent

ALPHA CHEMIKA

50 mL

glucose

C6H12O6

ASSOCIATED CHEMICAL 2%
ENTERPRISES

fructose

C6H12O6

ASSOCIATED CHEMICAL 2%
ENTERPRISES

sucrose

C12H22O11

ASSOCIATED CHEMICAL 2%
ENTERPRISES

galactose

C6H12O6

ASSOCIATED CHEMICAL 2%
ENTERPRISES

starch

(C 6H 10O 5) n

ASSOCIATED CHEMICAL 2%
ENTERPRISES

Red litmus paper

RIEDEL DE HAN

Sodium hydroxid

NaOH

ASSOCIATED CHEMICAL 10%

ENTERPRISES

Hydrochloric acid

HCl

ASSOCIATED CHEMICAL 10%

ENTERPRISES

Instrumentation
Name

Model

Manufacturer

Use in the practical

Hot plate

JJ224BC

G&G

To heat the contents of

the test tubes

PROCEDURE
i.

ii.

Benedicts Test for Reducing Sugars

Unknown solutions of carbohydrates/sugars labelled A, B, C, D, E and F were
provided which could be glucose, fructose, galactose, sucrose, starch or water. 10
drops of solutions of the above carbohydrates were placed in separate test tubes and
each test tube is labelled. 2 mL of Benedicts reagent was added to each sample. The
test tubes were placed in boiling water bath for 3-4 minutes.
Iodine Test for Polysaccharides

5 drops of each solution of glucose, galactose, sucrose, starch and water were placed
using a spot plate. 1 drop of iodine solution is added to each sample.
iii.

Hydrolysis of Disaccharides and Polysaccharides

3 mL of each 2% carbohydrate solutions are placed in two test tubes. To one sample
20 drops of 10% HCl was added and to the other samples 20 drops of H 2O were
added. The test tube were labelled and heated in a boiling water bath for 10 minutes.

Tests for Carbohydrates

The test tubes were removed from the water bath and allowed to cool. To the samples
containing HCl, about 20 drops of 10% NaOH was added until one drop of the mixture
turned red litmus paper blue indicating neutralization of HCl. The samples were tested for
hydrolysis as follows:
Iodine test 5 drops of each solution was placed on a spot plate and 1 drop of iodine reagent
was added to each. The observations were recorded and the occurrence of hydrolysis in each
sample was determined.
Benedicts test 2 mL of Benedicts reagent were added to each of the samples and heated
for 3-4 minutes in a boiling water bath.
Results and Analysis
Table 1: Benedict and Iodine tests
Sample
A
B
C
D
E
F

Benedicts Test
Reddish orange ppt
Greenish brown
Brown
No change
No change
No change

Observations
Iodine Test
No change
No change
No change
Blue-black
No change
No change

Table 2: Hydrolysis tests

Sample
A
B
C
D
E
F

Benedicts Test
Water
Reddish orange
Brown
Reddish orange
No change
No change
No change

HCl
Reddish orange
Reddish orange
Reddish orange
No change
No change

Iodine Test
Water
No change
No change
No change
Blue-black
No change
No change

HCl
No change
No change
No change
Blue-black
No change
No change

From the observations recorded in the above table the samples were identified as follows:

A glucose, B fructose, C galactose, D starch, E sucrose, F water

Reducing sugars: A, B, C
Non-reducing sugars: E
Ketoses: B
Positive fermentation test: A, B, C
Positive Iodine test: D
E undergone hydrolysis

Discussion
The aim and objectives of the experiment were achieved. The carbohydrates present in an
unknown solution were characterized qualitatively and quantitatively on the basis of various
chemical assays. The results obtained were in accordance with the theoretical ones. The
variation in colour changes for positive Benedicts test was due variation in amount of
reducing sugar present in a sample. A gave a reddish orange precipitate showing it has the
greatest of a reducing sugar followed by B which gave a brown precipitate, C has the least
amount since it gave a greenish-brown precipitate. Analysis on sample F reviewed that it was
water since it did not give any positive result with the reagents given. Sample D gave positive
when it was treated with iodine solution in both iodine test and hydrolysis test showing that it
is a starch solution. E gave a negative result when it was treated with Benedicts reagent at
first but when it was hydrolysed and treated with Benedicts reagent again it gave a reddish
orange colour. This showed that sample E was a sucrose solution since sucrose a disaccharide
can be hydrolysed to give a monosaccharide.

Conclusion
By performing this experiment it was possible to identify various carbohydrate groups by the
virtue of their chemical properties. Glucose, fructose, sucrose, galactose, starch and water
were all identified from the given samples. Although glucose, fructose and galactose gave a
positive result with Benedicts reagent they did not give the same colour change. The
difference in colour changes was attributed to the variation in amount present of reducing
sugar present in each sample. The experiment was a success.
Answers to Questions
1. Cu2+ is blue, it forms a red precipitate, Cu 2O, a Cu+ compound. Copper is reduced from
+2 to +1. That means the sugar was oxidized. The aldehyde oxidized to a carboxyl group.
2. Fructose will form a red colour more rapidly since it is a ketose. Seliwanoff's agent is
most sensitive to ketose. Glucose, which is an aldose will develop slowly giving off a
pink colour.
3. Fermentation is due to enzymes. Yeast does not contain all the specific enzymes to
breakdown all the disaccharides.

4. Amylose is a type of starch stored in plant cells, while glycogen is used to store energy in
animal cells. The iodine test is used to see if a substance contains starch or not.

References
i. https://msu.edu/course/lbs/145/luckie/Lab1.html accessed on 26/11/16 at
12.15pm
ii. http://www.namrata.co/category/chemistry-of-carbohydrates/subjectivequestions-chemistry-of-carbohydrates/ accessed on 26/11/16 at 1.20pm
iii. http://studymoose.com/carbohydrates-lab-report-essay accessed on 26/11/16 at
3.05pm
iv. https://www.scribd.com/doc/48016350/Carbohydrates-test-report accessed on
26/11/16 at 3.15pm
v. http://www.slideshare.net/namarta28/qualitative-tests-for-carbohydrates35884145?from_action=save accessed on 26/11/16 at 4.50pm