Page "High-performance liquid chromatography" Paragraph 46

HPLC separation have theoretical parameters and equations to describe the separation of components into signal peaks when detected by instrumentation such as by a UV detector or mass spectrometer.

This is analogous to the separation equation for a thin layer chromatography ( TLC ) separation, but describes how well HPLC separates a mixture into two or more components that are detected as peaks on a chromatagrams.

The HPLC parameters are the: efficiency factor ( N ), the retention factor ( kappa prime ), and the separation factor ( alpha ).

Together the factors are variables in a resolution equation, which describes how well two components ' peaks separated or overlapped each other.

These parameters are mostly only used for describing HPLC reversed phase and HPLC normal phase separations, since those separations tend to be more subtle than other HPLC modes ( e. g. ion exchange and size exclusion ).

The factors are described in terms of measurements of the peaks of a chromatagram produced by HPLC detector as it separates a mixture.