Abstract
Reversed-phase high-performance liquid chromatography (RP-HPLC) with a pyridine formate-1-propanol solvents system and fluorometric detection has been used to separate complex peptide mixtures. Excellent separation wre obtained with mixtures containing a large number of relatively small peptides such as those generated by trypic cleavage of proteins. RP-HPLC was also suitable for the separation of larger peptides such as those obtained, for example, by treating proteins with cyanogen bromide. The proposed method is rapid and simple, and it gives good peptide recovery. In addition, high detection sensitivity and large sample capacity make this technique suitable at both the analytical (1 nmol) and the preparative (1 μmol) level. The method is generally applicable to such problems as peptide mapping and protein sequencing.
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