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 illus trated with an example. The aim is to find the mobile phase system. i. e. buffer and organic modifier used for gradient formation, that admit s the steepest gradient with sufficient resolution. The result is a ra pid separation where the detection limits are low and the loading capa city is high. This strategy is a hybrid between experimental design us ed for optimizing the selection of the mobile phase system and gradien t theory used for gradient predictions.