ABSTRACT

The application of high-performance liquid chromatography (HPLC) technology in biochemistry has been instrumental in the isolation and eventual structural elucidation of a plethora of new peptides and proteins. As new peptides were isolated and characterized, the need for their duplication by total synthesis became imperative, and it is that the power and usefulness of preparative reversed-phase chromatography (RPC) was unmistakably realized. Early work in the laboratory on the preparative purification of peptides dealt with the identification and application of solvent systems and conditions that would maximize purity. Once the isocratic analytical conditions have been determined, successive and rapid assessment of the identity and purity of the fractions obtained from the preparative HPLC purification can be carried out. Due to the strong elutropic characteristics of the triethylammonium phosphate buffer, preparative gradient conditions are generally started at about 10% lower acetonitrile concentration than the isocratic analytical conditions determined in a trifluoroacetic acid system.