PEPTIDE SEPARATION BY GEL-FILTRATION HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY USING A GRADIENT ELUTION SYSTEM

Peptides were separated by gel filtration high-performance liquid chromatography with gradient elution. Two gradient elution systems were applied: (1) 0 to 20% acetonitrile in 30 mM phosphate buffer, pH 8.0 and (2) 30 mM phosphate buffer, pH 8.0 to water; with the polymer-type gel filtration column we used, excellent separation of tryptic peptides from hen egg-white lysozyme was achieved within 50 min. System 2 was applied to a separation of V8 protease peptide from yam tuber chitinase and yielded two to three times higher amounts of peptides than reversed-phase high-performance liquid chromatography.