Introduction

• Paper chromatography (PC) is a type

of planar chromatography whereby
chromatography procedures are run
on a specialized paper.

• Paper chromatography is one of the

types of chromatography procedures
that runs on a piece of specialized paper.
• It is a planar chromatography system
wherein a cellulose filter paper acts as a
stationary phase where compounds are
separated. It is a very inexpensive way to
separate different dissolved chemical
substances, and thus a powerful
analytical tool requiring nominal
amounts of material. 1
 Principle of Paper Chromatography

• The principle involved is partition

chromatography wherein the substances are
distributed or partitioned between liquid phases.
• One phase is the water held in the pores of the
filter paper used, and the other is the mobile
phase, which moves over the paper.
• The compounds in the mixture get separated due
to differences in their affinity towards water (in
the stationary phase) and mobile phase solvents
during the movement of the mobile phase. This
mobile phase moves under the capillary action of
pores in the paper.
• The principle can also be adsorption
chromatography between solid and liquid
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phases, wherein the stationary phase is the solid
surface of the paper, and the liquid phase is the
mobile phase. But most of the applications of
paper chromatography work on the principle of
partition chromatography (i.e., partitioned
between two liquid phases).
 Uses and Applications of Paper
Chromatography

1.Paper chromatography is specially used to

separate a mixture having polar and non-
polar compounds.
2.For separation of amino acids.
3.It is used to determine organic
compounds, biochemicals in urine, etc.
4.In the pharma sector, it is used to
determine hormones, drugs, etc.
5.Sometimes it is also used to evaluate
inorganic compounds like salts and
3
complexes.
6.It is often used to inspect cosmetics and
detect contaminants in food and drink.
Ascending Paper Chromatography

As the name indicates, the chromatogram

ascends. Here, the development of paper
occurs due to the solvent movement or
upward travel on the paper.
The solvent reservoir is at the bottom of
the beaker. The paper tip with sample
spots just dips into the solvent at the
bottom so that spots remain well above
the solvent.

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 Radial Paper Chromatography

Here, the solvent moves from the center (mid-

point) toward the periphery of the circular
chromatography paper. The entire system is kept in
a covered Petri dish for the development of the
chromatogram.
The wick at the center of the paper dips into the
mobile phase in a petri dish, by which the solvent
drains onto the paper and moves the sample
radially to form the sample spots of different
compounds as concentric rings.

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 Paper Chromatography Experiment
Method

The experimental method involves the following

steps:

1.Selection of suitable type of development

This depends on the complexity of the mixture,
solvent, paper, etc.
But in general ascending type or radial type
chromatography are used as they are easy to
perform, handle, less time-consuming, and give
chromatogram faster.

2. Selection of suitable filter paper

Filter paper is selected based on pore size, the
quality of the sample to be separated, and the 6

mode of development.
 Paper Chromatography Experiment
Method
The experimental method involves the following
steps:

5. Development of the chromatogram

Sample spotted paper is subjected to

development by immersing it in the mobile
phase.
The mobile phase moves over the sample on the
paper under the capillary action of paper.

6. Drying off the paper and detecting the

compounds
7

Once the chromatogram development is over, the

paper is held carefully at the borders to avoid
touching the sample spots and dried using an air
drier.
Sometimes the detecting solution is sprayed on
the developed paper and dried to identify the
sample chromatogram spots.