Chromatography

Chromatography is a technique used for analysis and purification of mixtures.

After a reaction, it is very unlikely that you will only get the desired product. So, you need to be able to
analyse and identify how many different compounds you have in the reaction mixture.

A chemist might then use chromatography again on a larger scale to separate each of the compounds to
obtain the desired/pure product (purification).

almost all the focus is on analysis, which requires a very small amount of product. But larger scale
chromatography is a common technique that allows all the mixture to be separated.

You will need to know the general principles and a few techniques but they all work on the same
principle.

The Basic Principle

the basic principle is to separate compounds in a mixture based on differences in polarity

There are two phases in chromatography: the mobile phase (liquid or gas) and the stationary phase
(solid or a liquid supported on a solid).

You would put your sample "on" to the stationary phase and then pass the mobile phase over it. The
molecules bond to the stationary phase through intermolecular forces.

Lets say there are two compounds in a mixture. We are looking for one compound to like the
stationary phase and cling on to it. And the other compound to prefer the mobile phase and leave the
stationary phase quickly.

We would say that those compounds have a different affinity for either of the phases. This gives good
separation and we can then see how many components are in the mixture or possibly even identify each
compound.

In exams, to explain a compounds preference for either phase you can talk about solubility or the
strength of interaction with liquid/stationary phase or forces of attraction with either phase or affinity
for either phase.

There are several different techniques. The main difference between the various chromatography
techniques is simply different mobile and stationary phases.
Different techniques
Paper Chromatography
This is the most basic technique that you can use. You wont get asked about it specifically but it
demonstrates the principles very well. You probably used it in science in earlier years to separate out ink
from pens or different dyes in sweets.

You would have taken a piece of paper and a beaker with some water in it. You then dissolved up the
sweet or the dye and "spotted" it on to the paper. You also have to draw a line at the top of the paper, as
you need to remove the paper from the beaker when the solvent reaches this line.

The paper is the stationary phase. You would then place the paper standing upright in the beaker of
water. The water (the mobile phase) would then run up the paper. Once the water reached the top of
the paper you would remove and dry it.

The idea is that you end up with noticeable spots on the paper, each representing a different
compound in the initial mixture.

the spots nearer the top of the paper have a higher affinity for the mobile phase as they have moved
the furthest. Or alternatively, you could say that those compounds are more soluble in the mobile
phase than those near the bottom.

TLC (Thin Layer Chromatography)

Carried out exactly the same as paper chromatography but using a solid containing silica as the stationary
phase (instead of the paper).

You sometimes have to calculate Rf values:

the distance from the bottom of the silica plate to the centre of the spot/distance from the bottom
to the top (solvent front)

For example:

Solvent front
Spot that has
ran up the plate
3.06 cm 2.62 cm

Original sample

Rf = 2.62/3.06 = 0.86 (always a number less than 1)

HPLC (High Performance/Pressure Liquid Chromatography)
This is where it gets more modern and sophisticated, and is controlled via a computer. The end result is a
chromatogram (chart), which is discussed below.

The principle of how the compounds are separated is exactly the same as above.

It is a very powerful technique that can be combined with mass spectroscopy to give you a lot of
information about what is in your mixture.

The stationary phase is a long thin silica column and the mobile phase is a liquid, which varies depending
on the compounds in the mixture.

GC (Gas Chromatography)
Again works on the same basic principle but this time the stationary phase is a liquid supported on an
inert solid and the mobile phase is an unreactive gas such as nitrogen or argon.

They may ask why cant you use oxygen as the gas? Its just too reactive.

Analysis - The Chromatogram (chart)

This is the sort of chromatogram that you will get from HPLC or GC.
You can see that there are 4 or 5 main peaks that tell you that there must be 4 or 5 different compounds
in the mixture.

The x-axis is time, in minutes, and it shows how long it takes the various compounds to come off the
column.

The longer the time, the slower that compound has been removed from the column. So the peaks
furthest to the left come off first.
The y-axis is absorbance, which tells you how much of each compound there is i.e. a large peak means
there will be a lot of that compound.
A chemist would use this purely for analysis or use it to optimise conditions to then separate these
compounds by varying the solvents used in the mobile phase.

if they give you different compounds in an exam question and ask which would come off the column
first, you need to look at solubility and polarity.

If the molecule is polar and the solvent is polar then it will come off the column very quickly i.e. it will
be very soluble in that solvent. If the molecule is non-polar and the solvent is polar then it will not be
so soluble in that solvent and will stick to the column for a lot longer.