Abstract
A simple restricted-access media (RAM) HPLC method for simultaneous determination of lactone and carboxylate forms of topotecan (TPT) in human serum was established. Using a RAM analytical column, serum samples were directly injected into the HPLC system. The eluted peaks of two forms of TPT were monitored with a fluorescence detector. The separation was completed in 18 min. The linear range was 15–1000 ng/mL, intra-day and inter-day variations being less than 7%. The kinetic equation was constructed according to the analytical results. The equation shows that the course of TPT lactone form converting to carboxylate form in human serum at 37°C follows a first-order kinetics. Concentration of each form at the moment of sampling was calculated by extrapolation.
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