CHROMATOGRAPHY

Applications of Chromatography

Forensics

Research
Pharmaceutical industry
Chromatography is a technique for
Separating mixtures of compounds
Identifying unknown compounds
Establishing the purity or concentration of compounds
Monitoring product formation in the pharmaceutical and biotechnology industries

Chromatography is widely used by forensic teams to analyse blood and urine samples for
drugs, for paint analysis and testing for the presence of explosives. Most chromatography
uses modern instrumentation and involves placing the sample to be analysed on a
support (paper or silica) and transporting it along a mobile phase. The mobile phase can
be a liquid (liquid chromatography) or a gas (gas chromatography).
Types of Chromatography…

Thin layer
Paper

Most chromatography uses modern instrumentation and involves placing the sample to be analysed on a support
(paper or silica) and transporting it along a mobile phase. The mobile phase can be a liquid (liquid
chromatography) or a gas (gas chromatography). Gas Column
 Thin Layer Chromatography
Sample – marker
Standard – food dyes
Stationary phase – chromatography paper
Mobile phase - water

The principle is that the inks are water soluble and travel up the paper. Each marker is composed of more than one
dye so each dye will separate as it moves up the paper with the water. The inks used in the dye are soluble in water
and so will travel up the paper.

The sample refers to what you want to test which in this case are colour markers.
The standard refers to the reference material you will compare the sample to. In this case it is Goodalls food dye.
Each food dye contains E numbers.
The stationery phase is the support you apply your sample to – in this case chromatography paper. You spot each
sample on the paper and allow it to dry.
The mobile phase is water and the paper is placed in the water. Since each E number is a different chemical with its
own characteristics and properties it will behave differently when you run it on the chromatography paper.
Calculation of results

Analysis
You must now calculate an Rf value for
each spot.

Rf = Distance from the start to the middle of a spot

Distance from start to finish point of the water

The Rf value is a measure of how far each spot has

moved relative to the solvent front.
Each dye will have its own Rf value so you can compare
Rf values and confirm whether food dyes were used in
making these markers.