ABSTRACT
Synthetic peptides are a class of compounds with increasing therapeutic importance and this has led to a concomitant increase in the need for rapid and efficient peptide purification procedures. Although an efficient peptide synthesis should result in only a small number of synthetic impurities (deletion, terminated or chemically modified peptides), these impurities which are usually closely related structurally to the peptide of interest, often pose difficult purification problems. The excellent resolving power and separation time of reversed-phase chromatography (RPC), coupled with the availability of volatile mobile phases, has made this mode of high-performance liquid chromatography (HPLC) the favored method for both analytical and preparative separations of synthetic peptides. Preparative RPC, whether it is carried out in gradient elution or displacement mode, requires the optimization of several parameters in order to maximize separation and yield. However, the gradient elution mode of RPC is handicapped by relatively poor utilization of the stationary and mobile phases.
