Microsequence analysis of peptides and proteins : I. Preparation of samples by reverse-phase liquid chromatography

Reverse-phase supports for the separation of peptides and proteins are compared in two high-performance liquid chromatographic systems. One uses a trifluoroacetic acid-acetonitrile solvent system with a 206-nm detector, and the other uses pyridine-formate or pyridine-acetate and 1-propanol with a postcolumn fluorescence detector. Each system was examined with RP8, RP18, and alkylphenyl supports. In most applications, the trifluoroacetic acid-acetonitrile solvent system used in conjunction with an alkylphenyl column performed best. The use of this system for the preparation of low-microgram amounts of samples for microsequence analysis is illustrated.