丙戊酸钠缓释片在健康人体内的药动学研究

Study on the pharmacokinetics of sodium valproate sustained-release tablets in healthy volunteers

目的:建立用于测定人血浆中丙戊酸的LC-MS/MS法,研究丙戊酸钠缓释片在健康人体内的药动学。方法:色谱条件:色谱柱为Waters YMC C18柱(150 mm×2.1 mm,5μm),流动相为乙腈和10 mmol/L醋酸铵(85∶15,V/V),流速为300μL/min,柱温为40℃,进样量为2μL;质谱条件:电喷雾负离子化,三重四极杆质谱多反应监测模式,丙戊酸和内标甘草次酸的选择性反应检测离子分别为m/z 143.0→143.0和m/z 469.4→425.5。24名中国成年健康男性作为受试者,按随机数表分为2组,采用自身双交叉试验设计,单剂量空腹口服0.5 g丙戊酸钠缓释片受试制剂或参比制剂,并于不同时间点采集肘静脉血(0~96h);随后进行多剂量试验,将另外24名受试者分成2组,连续8 d服用受试制剂或参比制剂(0.5 g,qd),于第8天服药后按单剂量试验中的采血方案在各时间点采集血样。对采集的所有血样进行血浆分离及处理,采用建立的LC-MS/MS法测定丙戊酸血药浓度,用DAS 2.0软件计算药动学参数。结果:分析方法学评价:丙戊酸血药浓度在0.087 5~89.6μg/m L范围内线性关系良好,方法精密度、准确度和回收率均良好,且无明显基质效应。药动学评价:受试制剂单剂量和多剂量给药后的主要药动学参数分别为Cmax(42.9±14.4)和C_(ss,max)(65.1±31.6)μg/m L,以及AUC_(0-96 h)(1 214.5±372.8)和AUC_(ss)(1 106.6±484.9)μg·h·m L^(-1),均与参比制剂无统计学差异(P>0.05)。结论:建立的LC-MS/MS法专属、灵敏、准确,可满足丙戊酸钠缓释片的药动学研究要求。

AIM：To establish a LC-MS/MS method for the determination of valproic acid in human plasma to investigate the pharmacokinetics of sodium valproate sustained-release tablets in healthy volunteers.METHODS：A Waters YMC C 1s column （150 mm×2.1 mm,5 μm） at a column tem-perature of 40 ℃ and a mobile phase consisting of acetonitrile-10 mmoL/L ammonium acetate （85：15,V/V） at a flow rate of 300 μL/min were used with an injection volume of 2 μL for LC.The negative ESI and MRM mode were used for triple quadrupole MS,in which the selected reaction monitoring ions were m/z 143.0→143.0 for valproic acid and m/z 469.4→425.5 for glycyrrhetic acid as the internal standard.In the randomized,crossover study,twenty four healthy male Chinese volunteers were divided into two groups and were given a single oral dose of 0.5 g sodium valproate sustained-release tablets （test or reference formulation） after an overnight fast,followed by collecting cubital venous blood samples at different time points （0-96 h）.Subsequently,another twenty four volunteers were given multiple oral doses （0.5 g,qd,for 8 d） of test or reference formulation and the blood samples were collected in the same way on day 8.All the blood samples after plasma separation and extraction were determined by the established LC-MS/MS method and the pharmacokinetic parameters of valproic acid were calculated by DAS2.0 software.RESULTS：The established LC-MS/MS method was evaluated as follows：the calibration curve of valproic acid in human plasma was linear in the range of 0.087 5-89.6 μg/mL,the precision,accuracy and recovery were good,and no significant matrix effect was found.The pharmacokinetics of sodium valproate sustained-release tablets were evaluated as follows：Cmax and AUC0-96 h,the main pharmacokinetic parameters,for test formulation were （42.9 ± 14.4） μg/mL and （1 214.5 ±372.8） μg· h · mL-1 for single oral dose and （65.1 ± 31.6） μg/mL and （1 106.6 ± 484.9） μg· h ＆· mL-1 for multiple oral doses（P 〉 0.05,vs reference formulation）.CONCLUSION：The LC-MS/MS method is proved to be specific,sensitive and accurate and it can meet the requirement for pharmacokinetic study of sodium valproate sustained-release tablets.