利巴韦林血药浓度测定方法的改进及生物等效性研究

目的:改进利巴韦林血药浓度测定方法,研究2种国产利巴韦林片的生物等效性。方法:血样用甲醇沉淀蛋白后离心,取上清液30μL以液-质联用法进样分析。20名男性健康受试者随机交叉双周期给药,分别单剂量口服利巴韦林受试制剂及参比制剂,DAS2.0软件计算二者的药动学参数及相对生物利用度,并评价二者的生物等效性。结果:利巴韦林检测浓度在1～1500ng.mL-1范围内线性关系良好(r=0.9993),定量下限为1ng.mL-1;萃取回收率>88%;日内、日间RSD均<6%;准确度在92%～108%之间。口服利巴韦林300mg受试制剂及参比制剂的主要药动学参数,t1/2ke分别为(42.5±16.9)、(46.8±26.3)h,tmax分别为(1.5±0.8)、(1.6±0.8)h,Cmax分别为(251.9±111.5)、(237.7±86.5)ng.mL-1,AUC0～72分别为(5003±1192)、(4891±1138)ng.h.mL-1,AUC0～∞分别为(7142±2337)、(7434±2869)ng.h.mL-1。受试制剂相对于参比制剂的生物利用度为(105.3±28.7)%。结论:本方法特异性强、准确可靠,所需样本量少,适合利巴韦林药动学研究。二者生物等效。

Improvement in Determination of Human Plasma Ribavirin Concentration and Its Bioequiavailability Study

OBJECTIVE： To improve the method for the determination of human plasma ribavirin concentration, and to study the relative bioequiavailability of two kinds of domestic ribavirin tablets. METHODS： Blood sample was extracted with methanol and centrifuged, then 30μL supernatant liquid was injected into column and analyzed by high - performance liquid chromatography - tandem mass spectrometry. A single oral dose test and reference formulations （300 mg） was given to 20 healthy male volunteers in a randomized cross over study. DAS2.0 was used to assess the pharmacokinetic parameters, relative bioavailability and bioequivalence of two kinds of ribavirin tablets. RESULTS： The linear range of the calibrationcurves （r = 0.999 3） was 1-1500 ng·mL^-1 for ribavrin with a lower limit of quantitation at 1ng·mL^-1. The recovery rate was above 88% ; the intra - day and inter day precision were all less than 6%. The accuracy ranged from 92% - 108%. The main pharmacokinetic parameters of two formulations （test and reference） were as follows： t 1/2ke were （42.5±16.9） h and （46.8 ±26.3） h; tmax were （1.5±0.8） h and （1.6±0.8） h; Cmax were （251.9±111.5）ng·mL^-1 and （237.7＋86.5） ng·mL^-1; AUC0-72 ewere （5 003±1192） ng·mL^-1 and （4 891±1138） ng·mL^-1 ;AUC0-72 were （7 142±2337） ng·mL^-1 and （7 434±2 869） ng ng·mL^-1, respectively. The bioavailability of test ribavrin as against reference ribavrin was （105.3 ± 28.7） % . CONCLUSION： The method is specific, accurate and reliable, and applicable for the pharmacokinetic study of ribavirin, and in which the sample size needed is small. The test and reference ribavirin tablets were bioequivalent.