High Pressure Liquid Chromatography

Louvain-La-Neuve

High Pressure Liquid Chromatography allows the separation or the purification of one or more compounds of a mixture with a view to their identification and their quantification.

This technique makes it possible to carry out a large part of the analyzes made in Gas Chromatography but especially those which are impossible with these techniques such as the analysis of thermosensitive compounds or of molecular masses which are both very large and even polar.The compounds to be separated are extracted in a solvent.

This mixture is introduced into the liquid mobile phase called eluent. Depending on the nature of the compounds, these interact more or less with the stationary phase in a column.The mixture to be analyzed is injected and then transported through the chromatographic system where the eluent is pushed by a pump under high pressure.

The compounds in solution are then distributed according to their affinity between the mobile phase and the stationary phase.The different separate solutes are characterized by a peak at the column outlet by means of an appropriate detector. The set of recorded peaks is called a chromatogram.

Various detectors are used on the platform.

Applications of this technique:

  • Amino acid determination

  • Determination of water-soluble vitamins (Vit. B and Vit. C) and fat-soluble vitamins (Vit. A and Vit. E)

  • Sugar dosage

  • Determination of the GSH/GSSG ratio (oxidative stress indicator)

  • Determination of volatile fatty acids (from C2 to C6)

  • Assay of cytotoxics

  • Determination of chlorophylls and xanthophylls

  • Determination of polyamines

  • Identification of lipopeptides by LC-MS