High-performance liquid chromatography assay of bleomycin in human plasma and rat hepatocytes in culture

Abstract

A sensitive and rapid linear-gradient, ion-paired, reversed-phase high-performance liquid chromatography technique using fluorescence detection was developed to quantify bleomycin (BLM) metabolites in the plasma of patients undergoing BLM therapy and in rat hepatocytes that had previously been incubated with 5 x 10(-5) M BLM. We could detect about 70 ng/ml using this procedure. BLM metabolites were assayed in the supernatant fractions of precipitated human plasma and in pellets of rat hepatocytes. Metabolite concentrations were below the level of detection in human plasma samples. In hepatocyte pellets, metabolites such as deamido-BLM A2 and deamido-BLM B2 were detected, indicating that isolated rat hepatocytes in culture can metabolize BLM analogues to the corresponding deamido-BLMs. The high-performance liquid chromatography procedure developed during this work can be used to study the metabolism of BLM in cell culture systems.