EXPERIMENT 8

Topic :
Separation Process - Chromatography

Purpose :
To separate and identify the colours found in a type of commercial food dye.

Theory :

Chromatography is a technique used to separate the different components in a

mixture. All chromatography techniques consists of two major components : the
stationary phase and the mobile phase. The method consists of dissolving the
mixture in a suitable solvent and which is then carried over the surface of the
stationary phase.

Separation is possible because different substances have different degrees of

absorption or solubility between the stationary and the mobile phase. The
component will move along the stationary phase at different speeds, and thus
separated. In paper chromatography, the stationary space is the strip of
chromatography paper and the mobile phase is suitable solvent.

By comparing the Rf value of the component with the R f value of known substances,
the component in the mixture can be identified. The value of R f for each of the
component in the mixture can be identified. The value of R f for each of the
component is calculated using the formula :

Rf = distance moved by the component

distance moved by the solvent
List of Apparatus :

Measuring cylinder
Chromatography paper
Beaker
Pencil
Ruler
Fine stick

List of Materials:

Food dye
Ethanol

Procedure:

1. 100 cm3 of ethanol and water solvent was prepared in the ratio of 3:1.
2. The solvent was poured in a 1 dm3 tall container.
3. The mouth of the container was covered with plastic and it was leaved to stand
for 30 minutes.
4. A line was lightly marked with a pencil about 2 cm from one end on a strip of
filter paper measuring about 4 cm wide and 38 cm long.
5. The solution of food dye was spotted onto a marked spot in the middle of the line
as shown in the diagram by using a fine glass capillary pipette.
(Refer to Figure 1)
6. The spot was dried with a hair dryer.
 Figure 1: Method of spotting the sample onto filter paper

7. The filter paper was suspended in the container with its lower end touching the
solvent.
8. The pencil line was ensured to be above the solvent level and the side of the
filter paper is not touching the walls of the high container as shown in the
diagram below. (Refer to Figure 2)

Figure 2 : Development chamber for paper chromatography

9. The apparatus was leaved to stand for 1 to 2 hours.

10. Then the filter paper was removed and the position of the solvent is marked
immediately. The paper was dried with a hair dryer.
11. The value of Rf was obtained for each of the spots separated. The distance
moved by the solvent front and the distance moved by the color components
from the starting line.

Figure 2 : Development chamber for paper chromatography

Data and Observation:

Substances Component Color of Distance of Distance of Rf

component movement of solvent
component / cm front/ cm

Ethanol Green food Yellow 9.0 13.0 9.0

dye 13.0
= 0.69

Blue 11.5 13.0 11.5

13.0
= 0.88
Discussion:

a) Using your result from the table above, identify the color components in
the sample of food dye provided.
Yellow and blue

b) In this paper chromatography experiment, state the substance that

function as the
i. Stationary phase : Chromatography paper
ii. Mobile phase : Ethanol

c) Explain how you would ensure that the drop of food dye at the starting line
is concentrated and small in size?

 Only a little amount of the food dye is spotted.

 The drop of the food dye is made once in a time.
 Let the first spot dried before the second spot is made.

d) Why the starting line containing the drops of food dye solution must be
placed above the solvent level at the beginning of the experiment?
 To ensure that the separation process occur easily

e) State the steps taken to ensure effective separation.

1. The mouth of the container is covered with plastic and it is leaved to stand for
30 minutes.
2. The pencil line is ensured to be above the solvent level.
3. The side of the filter paper is ensured not to touch the walls of the high
container.