Experiment – 18
Aim: - To separate the color components present in the mixture of red and blue
ink by ascending paper chromatography and find their Rf values.
Materials required: - Boiling test tube, glass rod, Whatman filter paper strip,
capillary tube, alcohol, distilled water and a Mixture of inks.
Theory: -
1. Chromatography is a technique for rapid separation of colored components
of a mixture of compounds.
2. The principle of chromatography is based on differential migration of an
individual component of a mixture through a stationary phase under the influence
of a moving phase.
3. The stationary phase may be a porous solid supported in the container for
example - filter paper.
4. The moving phase may be any solvent or water. It is called eluent.
5. Different types of chromatography
i. Column Chromatography
(1) Adsorption chromatography
(2) Partition chromatography
ii. Thin layer chromatography
iii. Gas chromatography
iv. Paper chromatography
Rf Value: - It represents Retention Factor or the ratio of the distance moved by
the component from the origin or point of application to the distance moved by
the solvent from the same point.
Rf =
Procedure:-
Cut a strip of filter paper of about 10 cm in length and 3 cm in width
tapering from one end.
Draw a line from the tapering end and mark the center on it.
On the center of the line, put a drop of ink mixture with the help of a fine
capillary.
Take a boiling test tube and put a little water in it.
With the help glass rod suspend the filter paper vertically in a boiling test
tube containing solvent (eluent).
Keep the boiling test tube undisturbed for 10 minutes. Notice the rising
solvent along with the mixture of ink.
Take the filter paper out of the test tube and mark the distance that the
solvent has risen on the paper with the pencil. This is called the solvent
front.
Dry the paper and measure the distance of the two spots from the original
line and the distance of the solvent from the original line.
Calculate the Rf values of the blue and red inks by using the formula.
Observation Table :
Color of the ink Distance travelled by Distance travelled by Rf Value
the different ink from the solvent
the original line (water)from the
original line
Red ink cm(x) cm(z) =
Blue ink cm(y) cm(z) =
Rf Value
Rf for Red ink cm(x) / cm(z) = -------
Rf for Blue ink cm(y) / cm (z) = --------
Results :-
The Rf value for red ink - --------
The Rf value for blue ink – ---------
Precautions:-
1. Use good quality pencil for drawing the reference line so that the mark does
not dissolve in the solvent in which the chromatography is carried out.
2. Always make use of a fine capillary tube.
3. Keep the test tube undisturbed during the experiment.
4. A spot should be small and rich in mixture.
5. Allow the spot to dry before putting the strip in the boiling test tube.
6. Keep the strip erect. Do not let it to be curled.
7. Do not allow the spot to dip in the solvent.
8. Do not remove the filter paper from water and dip it again.
