Basics of HPLC

HPLC: high performance liquid chromatography

HPLC (or high performance or high pressure liquid chromatography) is a separation analysing technique, based on the progressive migration of compounds in a column that withstands high pressures.

The interactions that allow to retain and elute the compounds take place between:
  • The compound: C
  • The mobile phase: MP
  • The stationary phase: SP

Different separation modes exist in HPLC. The chosen mode depends on the characteristics of the molecules to be separated.

The different chromatographic types commonly used are:

  1. Partition chromatography (the most used)
  2. Ion-exchange chromatography
  3. Size-exclusion chromatography
  4. Adsorption chromatography (rarely used, intended for specific applications)

The below simplified diagram helps to identify the HPLC method best suited to the separation of the compounds according to their characteristics.

The quality of a chromatographic separation can be evaluated thanks to different chromatographic parameters that can be found in the drawer « Chromatographic parameters ».