Structural Biochemistry/Proteins/Purification/High pressure Liquid chromatography

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High Pressure Liquid Chromatography (HPLC for short or also called High Performance Liquid Chromatography) is a precise and hi-spec form of column chromatography that can separate and identify components in an unknown (or known) compound mixture.

Like all forms of chromatography, HPLC consists of a stationary phase and a mobile phase. The stationary phase is some solid material packed into the column, while the mobile phase is a liquid solvent that carries the analyzed compound through the column.

Chromatography operates on the principle of polarity where the compound interacts with the solid stationary phase based on their differences in polarity. In general, if the stationary phase is very polar, a non-polar component of the compound would exit the column faster than a polar component would.