Application of Reversed Phase High Performance Liquid Chromatography in Solid Phase Peptide Synthesis

Abstract The application of ion-pair partition reversed phase high performance liquid chromatography to the analysis and purification of peptides produced by solid-phase synthesis is described. This approach permits very rapid recognition of deletion or partial deprotection products which are often generated during the synthesis and, depending on their molecular characteristics, allows these components to be resolved. The use of hydrophobicity parameters has been examined and found useful in the prediction of the relative elution order of these closely related peptides. The separation of a series of synthetic preparations, including the angiotensins, leu-enkephalin amide, a fragment of β-TSH, and linear antamanide, illustrates the speed of the method which can be used to assess the homogeneity of the reaction mixtures and also provide, preparatively, highly purified peptides.