ABSTRACT
The excellent resolving power of reversed-phase chromatography (RPC) has resulted in its becoming the predominant mode of high-performance liquid chromatography for peptide separations in recent years. This chapter describes a series of five commercially-available synthetic decapeptide standards and demonstrates their usefulness in monitoring the effects of column dimensions, column aging, organic modifier, and temperature on peptide separations in RPC. Peptide standards are best suited for monitoring peptide retention in RPC, since it is preferable to use standards that are structurally similar to the sample of interest and that presumably interact with the reversed-phase packing in a similar manner. RPC is often used during peptide mapping of proteins, and the 10-residue length of the standards was designed to reflect the average size of cleavage fragments from proteolytic digests of proteins. Reversed-phase peptide standards allow the researcher to: identify non-specific interactions of peptides with the reversed-phase packing, monitor column performance, and monitor run-to-run reproducibility of peptide separations.
