HPLC-MS研究盐酸恩丹西酮缓释胶囊人体药动学及生物等效性

Pharmacokinetics and Bioequivalence of Ondansetron Extended-Release Capsule by HPLC-MS

目的首次建立人血浆中盐酸恩丹西酮浓度的HPLC-MS分析方法,用以测定健康受试者口服盐酸恩丹西酮缓释胶囊后的血药浓度,估算受试制剂和参比制剂的药动学参数,评价两种制剂的生物等效性和相对生物利用度。方法血浆中加入内标米氮平后,乙酸乙酯提取,HPLC-MS分离、分析。色谱系统:ODS柱(4.6mm×150mm,5μm),甲醇-乙腈-醋酸铵溶液(0.01mol.L-1)(35∶30∶35)为流动相,流速1.0mL.min-1,柱温35℃。质谱检测方式:SIM。结果盐酸恩丹西酮的线性范围为0.1～80μg.L-1(r=0.9999),最低检测质量浓度达0.1μg.L-1,提取回收率≥85%。口服单剂量和多剂量的盐酸恩丹西酮缓释胶囊后的相对生物利用度分别为(94.1±15.8)%和(94.8±14.3)%。结论本法简便,准确,灵敏度高。统计学结果表明,口服单剂量和多剂量的缓释胶囊与普通参比胶囊相比,达峰浓度低,达峰时间延长,稳态的波动度小,缓释特征明显。经方差分析和双单侧t检验表明,缓释胶囊和普通参比胶囊生物等效。

OBJECTIVE To build a sensitive and specific LC-ESI-MS method for the determination of ondansetron in human plasma. The method was then applied successfully to the pharmacokinetic and bioequivalence study of an extended-release capsule of ondansetron. METHODS Mirtazapine was used as the internal standard. After adding mirtazapine, the plasma samples were extracted with ethyl acetate and determined by HPLC-MS, The separation was carried out on a Shimadzu VP-ODS column（4. 6 mm × 150 mm,5 μm） using a mobile phase consisting of methanol-acetonitrile： 10 mmol · L^-1 ammonium acetate （35：30： 35） delivered at 1 mL · min^-1. HPLC-MS was performed with the selected ion monitoring （SIM） mode using target ions at m/z 294. 5 for ondansetron and m/z 265.5 for mirtazapine, A randomized crossover design was performed in 20 healthy volunteers. RESULTS The linear calibration curve was obtained in the concentration range of 0. 1-80 μg · L^- 1 （ r = 0. 999 9 ）. The detection limit of ondansetron in plasma was 0. 1μg · L^-1. The average recovery was more than 85%, The relative bioavailability of the test formulation in single-dose and multipledose pharmacokinetic study were （94. 1±15.8）% and （94. 8 ± 14. 3 ）%, respectively. CONCLUSION The assay was proved to be sensitive, accurate and convenient. The results of statistic analysis showed that the test ondansetron extended-release capsule and the reference ondansetron tablet were bioequivalent, and the test capsule had the characteristics of extended-release formulation.