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A high-performance liquid chromatographic method coupled with electrospray mass spectrometry was developed for the simultaneous determination of dolasetron and its major metabolite, hydrodolasetron, in human plasma. A new sample pretreatment method, i.e., salt induced phase separation extraction (SIPSE), was proposed and compared with four other methods, i.e., albumin precipitation, liquid-liquid extraction, hydrophobic solvent-induced phase separation extraction and subzero-temperature induced phase separation extraction. Among these methods, SIPSE showed the highest extraction efficiency and the lowest matrix interferences. The extraction recoveries obtained from the SIPSE method were all more than 96% for dolasetron, hydrodolasetron and ondansetron (internal standard). The SIPSE method is also very fast and easy because protein precipitation, analyte extraction and sample cleanup are combined into one simple process by mixing acetonitrile with plasma and partitioning with 2 mol/L sodium carbonate aqueous solution. The correlation coefficients of the calibration curves were all more than 0.997, in the range of 7.9-4750.0 ng/mL and 4.8-2855.1 ng/mL for dolasetron and hydrodolasetron, respectively. The limits of quantification were 7.9 and 4.8 ng/mL for dolasetron and hydrodolasetron, respectively. The intra-day and inter-day repeatability were all less than 10%. The method was successfully applied to the pharmacokinetic study of dolasetron.

Optimization of Sample Pretreatment Methods for Simultaneous Determination of Dolasetron and Hydrodolasetron in Human Plasma by HPLC-ESI-MS