Optimizing selectivity during reversed-phase high performance liquid chromatography method development: prioritizing experimental conditions

Abstract

Several different conditions can be varied to improve selectivity for reversed-phase chromatography (RPC). A reexamination of literature data suggests that changes in selectivity due to a change of column or mobile phase pH are largely replicated by changes in temperature or mobile phase composition (concentrations of acetonitrile and/or methanol). This suggests a reconsideration of the role of mobile phase pH and the column during method development.

Keywords: Experimental conditions; Experimental design; HPLC; Method development; Optimization; Selectivity.