Systematic separation of medium-sized biologically active peptides by high-performance liquid chromatography.

The systematic separation of medium-sized biologically active peptides by high-performance liquid chromatography (HPLC) is described. Three steps are involved: first, high-performance sodium dodecyl sulphate (SDS) gel chromatography on a newly developed column. TSK-GEL 2000SW; secondly, ion-pair reversed-phase HPLC using stepwise elution mobile phases containing SDS and tetrabutylammonium phosphate; thirdly, high-performance cation-exchange chromatography on a Partisil SCX column for purification, using stepwise gradient elution with volatile buffers. Removal of SDS was possible in the final step. This systematic method is fast, reproducible and gives excellent separations and recoveries.