High Performance Liquid Chromatography

Instrument: Dionex ICS1100 ion chromatography system; Dionex Ultimate 3000 system

High performance (or high pressure) liquid chromatography is a step up from standard LC or FPLC but functions in a similar manner. Chemically-functionalised beads are packed into a column and these bind molecules, and a liquid is pumped through the column to elute absorbed components.

HPLC is different from standard LC in that the beads are smaller and more regular. They pack more tightly into the column and require higher pressures to force the liquid flow through the column.

Because beads are more regular, the performance of the column is much improved and this allows high resolution separation of chemicals or biomolecules that have very similar properties.

Eluted substances can be detected by a range of different detectors, the most common being UV absorbance, but mass spectrometry is also widely used.

Columns packed with beads functionalised in different ways are available to allow selective binding of chemicals with a range of different properties.
Technical Detail
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