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Science@CSCB

Introduction to Chromatography
What is Chromatography?

Derived from the Greek word Chroma


meaning colour, chromatography provides a
way to identify unknown compounds and
separate mixtures
Applications of Chromatography

Forensics

Research
Pharmaceutical industry
Types of Chromatography…

Thin layer
Paper

HPLC Gas Column


Your mission should you choose
to accept is to……..
 Analyse coloured markers

Test whether they contain E133 (blue), E122


(pink), E124 (red) or E110 (yellow)
Thin Layer Chromatography

 Sample – marker
 Standard – food dyes
 Stationary phase – chromatography paper
 Mobile phase - water
Structures of E numbers…..

E122 pink
E110 yellow

E124 red E133 blue


So what will happen?

 Each dye will travel up the paper at different


speeds

 The speed depends on the solubility of the dye in


water and its interaction with the paper

 The dyes are all different molecules with different


characteristics
Calculation of results
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
Conclusions – writing up

 One of the key elements of all scientific


experiments is to write up your results

 At the end of this experiment we would like each


person to conclude from the Rf values as to which
E numbers are contained in the markers

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