OPTIMIZATION STRATEGY FOR REVERSED-PHASE LIQUID-CHROMATOGRAPHY OF PEPTIDES

An optimization strategy for the separation of peptides from a complex matrix by reversed-phase liquid chromatography is presented and illustrated with an example. The aim is to find the mobile phase system. i.e. buffer and organic modifier used for gradient formation, that admits the steepest gradient with sufficient resolution. The result is a rapid separation where the detection limits are low and the loading capacity is high. This strategy is a hybrid between experimental design used for optimizing the selection of the mobile phase system and gradient theory used for gradient predictions.