Reversed-phase paired-ion high-performance liquid chromatographic method for the separation and quantification of multiple bleomycin congeners.

A rapid, linear gradient chromatographic technique for separating and quantifying copper(II)-chelated bleomycin congeners is described. This method is also capable of separating divalent from trivalent metal chelates; determining the purity of many chemically modified bleomycins; and assaying bleomycin hydrolase activity on complex mixtures. Quantification at 280 nm is sensitive to 50 pmol and is linear at least up to 10 nmol per injection.