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Presenter: Mr. Z. E. Clarke Originally Prepared by K.


Recall the first time you were introduced to colour in art class and you learnt of the three primary colours and the resulting secondary colours.

Chromatography is used to separate mixtures of substances into their components.

There are many types, however, you need to be familiar with the following 1. 2. 3. 4. Paper Column Thin layer (tlc) Gas-liquid (glc)

All types of chromatography have the following principles in common.

1. There are two phases:
stationary phase (a solid, or a liquid supported on a solid) mobile phase (a liquid or a gas)

All types of chromatography have the following principles in common.

2. The separation of the mixture is due to differing interaction of the components with the stationary phase 3. The two mechanisms which can occur are partitioning and adsorption 4. Different components travel at different rates.

This is the immovable phase and may either be a solid support:

Cellulose in the paper Silica or alumina

Liquid support:
Non-volatile, viscous liquid coated unto a solid surface
A long-chain alkane of high boiling point on a SiO2 support

The mobile phase is a fluid (gas or a liquid) which flows through or over the surface of the stationary phase and carries the components of the mixture with it.
In paper chromatography, the mobile phase is a suitable liquid solvent or mixture of solvents.

There are different types of solvents, ranging from polar (such as water) to non-polar (such as alkanes)
The mobile phase can be a mixture of solvents, in which case the ratio is quoted e.g. methanol:water (2:1)

Adsorption chromatography occurs when the components of the mixture (solute molecules) are bound to the surface of the stationary phase.

The stationary phase is a polar solid (such as silica or alumina) and the polar solute molecules are bound to this surface. The more polar the solute, the more strongly bound it is to the stationary phase
As the mobile phase passes over this, the solute molecules are eluted, from least polar to most polar in turn.

If both the stationary and the mobile phases are fluids, then the solute will be partitioned between the two.

1. Filter paper is used to aid in separation

2. Cellulose fibres contain water which acts as the stationary phase.

3. A small dot of mixture is placed on the paper, which is placed in a jar containing a shallow layer of solvent and is sealed pload/Chromatography.jpg

Methods of Analysis and Detection, Cambridge University Press

It is important that the solvent level is below the line with the spots on it.

It is important to cover the container so that the atmosphere in the beaker is saturated with solvent vapour
Saturating the atmosphere in the beaker with vapour stops the solvent from evaporating as it rises up the paper.

As the solvent moves up the paper, the different components are partitioned between the stationary and mobile phase.
Least polar molecules move ahead of the more polar molecules. Can be used to separate components of black pen ink

Retention factor (also called retardation factor) : Rf The movement of a solute relative to the movement of the solvent front

Stationary phase is a thin layer of silica (SiO2) or alumina (Al2O3) coated unto a plastic or glass support.
Setup is similar to that of paper chromatography, but this utilises adsorption and not partition. Useful for separating colourless components which can be detected by use of appropriate chemicals or techniques (visualising agents)


Department of Chemistry, UWI Mona Kamau Francis My Family A. Miller Mr. Campbell IT Department, MBCC My Pre-University Year 2 Chemistry Students (LOVE U ALL)