ANALYTICAL CHEMISTERY

Muhammad Azhar
1056
BS CHEMISTRY 4th (M)
 Paper Chromatography

Introduction:

Paper chromatography is one of the types of chromatography procedures which runs on a

piece of specialized paper. It is a planar chromatography system wherein a cellulose filter paper
acts as a stationary phase on which the separation of compounds occurs.

History:

Paper chromatography is an analytical method used to separate coloured chemicals or

substances. Erwin Chargaff credits in Weintraub's history of the man the 1944 article by
Consden, Gordon and Martin with sparking his discovery of Chargaff's rules, an important
precursor to Watson and Crick's discovery of the double-helix structure of DNA, for which they
were awarded the Nobel Prize in Physiology or Medicine in 1962. It is now primarily used as a
teaching tool, having been replaced in the laboratory by other chromatography methods such
as thin-layer chromatography (TLC).
Principle:

The principle involved is partition chromatography wherein the substances are

distributed or partitioned between liquid phases. One phase is the water, which is held in the
pores of the filter paper used; and 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
stationary phase) and mobile phase solvents during the movement of mobile phase under the
capillary action of pores in the paper. The principle can also be adsorption chromatography
between solid and liquid phases, wherein the stationary phase is the solid surface of the paper
and the liquid phase is of the mobile phase.

Explanation:

A paper chromatography variant, two-dimensional chromatography involves using two

solvents mobile phase is a solution that travels up the stationary phase, due to capillary action.
The mobile phase is generally a mixture of non-polar organic solvent, while the stationary phase
is polar inorganic solvent water. Here paper is used to support the stationary phase, water. Polar
water molecules are held inside the void space of the cellulose network of the host paper. The
difference between TLC and paper chromatography is that the stationary phase in TLC is a layer
of adsorbent (usually silica gel, or aluminum), and the stationary phase in paper chromatography
is less absorbent paper. And rotating the paper 90° in between. This is useful for separating
complex mixtures of compounds having similar polarity, for example, amino acids. The setup
has three components.
Applications:

 Paper chromatography is specially used for the separation of a mixture having polar and
non-polar compounds.

 For separation of amino acids.

 It is used to determine organic compounds, biochemical in urine, etc.

 In the pharmacy sector, it is used for the determination of hormones, drugs, etc.

 Sometimes it is used for evaluation of inorganic compounds like salts and complexes.
Types:

1. Ascending 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.

2. Descending chromatography: Here, the development of paper occurs due to solvent travel
downwards on the paper. The solvent reservoir is at the top. The movement of the solvent is
assisted by gravity besides the capillary action.

3. Ascending- descending mode: Here solvent first travels upwards then downwards on paper.

4. Radial mode: Here, the solvent moves from the center (mid-point) towards the periphery of
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 paper dips into the mobile phase in
a petri dish, by which the solvent drains on to the paper and moves the sample radially to form
the sample spots of different compounds as concentric rings.

5. Two-dimensional chromatography: Here the chromatogram development occurs in two

directions at right angles. In this mode, the samples are spotted to one corner of rectangular paper
and allowed for first development. Then the paper is again immersed in the mobile phase at a
right angle to the previous development for the second chromatogram.