HPLC-MS/MS determination of febuxostat in human plasma and its application to pharmacokinetic study / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop a sensitive and rapid HPLC-MS/MS method for the determination of febuxostat in human plasma. METHODS: The plasma samples were extracted with ethyl acetate after addition of internal standard and 0.1% formic acid solution, the extract was evaporated and the dry residue was reconstituted with 1 mL of methanol and 5 μL was injected to the API3000 HPLC-MS/MS system for analysis. The analytical column was SHISEIDO, CAPCELLPAK C18(2.0 mm × 50 mm, 5 μm). The mobile phase was composed of 0.1% formic acid in water-0.1% formic acid in acetonitrile (15 85) and the flow rate was 0. 25 mL · min-1. Detection was performed with multiple reactions monitoring (MRM) using positive electrospray ionization (ESI). The pharmacokinetic characteristics of the febuxostat tablet(febuxostat 40 mg) was investigated in 12 healthy volunteers after single oral dose administration of 40 or 80 mg febuxostat and multiple oral dose administration of 40 mg febuxostat. The serial blood samples were collected and after centrifugation, the plasma was separated and analysed by HPLC-MS/MS established. RESULTS: The calibration curves were linear over the concentration ranges of 10.02-8020 ng · mL-1. The lower limit of quantifications was 10.02 ng · mL-1. Inter- and intra-day precisions were less than 12.40% and accuracy was within 85.75%-105.91%. Extraction recoveries were around 87% and the ana-lytes were proved to be stable under all the required conditions. Total runtime of an analyte was only 2.0 min. Results of the statistical analysis indicated that febuxostat followed linear kinetics in healthy Chinese volunteers at the investigated dose range of 40 to 80 mg. No significant drug accumulation was found after multiple dose administration of 40 mg febuxostat tablets. Furthermore, there was no significant difference of pharmacokinetics between males and females in Chinese population. CONCLUSION: This method is rapid, sensitive, specific and applicable to the pharmacokinetic study in human of febuxostat.