Simultaneous determination of ifosfamide and its metabolite ifosforamide mustard in human plasmy by high performance liquid chromatography

摘要

Because ifosforamide mustard (IFM) is the active alkylating metabolite of ifosfamide (IFO) it is of particular interest in the pharmacokinetic analysis of patients undergoing IFO treatment. This paper presents an assay for the simultaneous determination of IFM andIFO after deri.vatization with diethyldithiocarbamate (DDTC), subsequent liquid-Iiquid extraction of the plasma with acetonitrile (AcN) and using reversed phase high performance liquid chromatography (RP-HPLC) with ultra-violet (UV) detec- tion at 276 nm. Structural confirmation of the analytes was accomplished using mass spectrometry (MS). Reaction conditions such as incubation duration, temperature, and concentration of dcrivatization agent were investigated; 30 min at 70°C with 100 mg/mL DDTC was optimal. The presented analytical method proved to be accurate, precise, and linear for IFM andIFO concentrations, ranging from 0.1 00~50.0 and 0.100- 100 #mu#g/mL, respectively, and with lower limits of quantitation of 0.100#mu#g/mL for both analytes. A typical patient pharmacokinetic profile is presented to demosnstrate the applicability ofthe assay in clinical samples.The analytical method could be employed in high-throughput clinical analysis of IFM and IFO patient samples.