LC-MS/MS测定人血浆中利伐沙班浓度及生物等效性研究  

Determination of rivaroxaban in human plasma by LC-MS/MS and its application to a bioequivalence study

作　　者：刘瑞娟 李泽运 刘帅兵[1] 孙素珂[1] 王素云[1] 张晓坚[1] 田鑫[1] LIU Ruijuan;LI Zeyun;LIU Shuaibing;SUN Suke;WANG Suyun;ZHANG Xiaojian;TIAN Xin(Department of Pharmacy,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052

出　　处：《郑州大学学报（医学版）》2021年第3期302-308,共7页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家科技重大专项重大新药创制项目(2020ZX09201-009)。

摘　　要：目的:建立检测人血浆中利伐沙班质量浓度的液相色谱-串联质谱(LC-MS/MS)分析方法,研究利伐沙班片在中国健康受试者体内的药动学特征并评价生物等效性。方法:血浆样品经乙腈沉淀,采用C18色谱柱分离,梯度洗脱,质谱采用电喷雾离子源正离子检测。采用随机、两序列、四周期、全重复交叉试验设计,健康受试者分别空腹和餐后口服20 mg利伐沙班受试制剂与参比制剂,采集不同时间点的血样,利用建立的方法检测血药质量浓度,计算受试制剂和参比制剂的药动学参数并进行统计分析。结果:利伐沙班在1.0~600.0μg/L范围内线性良好,方法的批内精密度小于13.6%,批间精密度小于8.3%,准确度为86.7%~102.2%。空腹和餐后状态下利伐沙班受试制剂与参比制剂Cmax和AUC几何均值比的90%CI均在80.00%~125.00%,受试制剂和参比制剂药动参数的受试者个体内变异比值的90%CI的上限均小于2.5。结论:建立的LC-MS/MS方法灵敏、快速、准确,适用于人血浆中利伐沙班浓度的测定;本研究中利伐沙班片受试制剂和参比制剂在空腹和餐后状态下生物等效。Aim:To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for determination of rivaroxaban in human plasma,and to study its pharmacokinetic characteristics and evaluate the bioequivalence of rivaroxaban tablets in healthy Chinese subjects.Methods:After protein precipitation by acetonitrile,the rivaroxaban in human plasma sample was separated on a C18 column with gradient elution.A tandem mass spectrometric detection was conducted using an electrospray ionization source in positive mode.A randomized,two-sequence,four-period and fully replicated crossover clinical trial was carried out.Healthy Chinese subjects were administered 20 mg dose of test and reference rivaroxabanunder fasting and fed conditions.Blood samples were collected at different time points.The concentrations of rivaroxaban in human plasma were determined using the established method.The pharmacokinetic parameters of rivaroxaban for the test and reference preparations were calculated and statistical analysis was performed.Results:The calibration curves were linear over the concentration range of 1.0~600.0μg/L for rivaroxaban.The intra-and inter-day precision was lower than 13.6%and 8.3%respectively in terms of relative standard deviation.The accuracy was in range of 86.7%~102.2%in terms of relative error.The 90%CI of the geometric mean ratios of Cmax and AUC of the rivaroxaban test and reference preparations were within the range of 80.00%~125.00%.The upper limit of the 90%CI for the test-to-reference ratio of the within-subject variability was less than 2.5.Conclusions:The established LC-MS/MS method is sensitive,rapid,and accurate,which is suitable for the determination of rivaroxaban concentration in human plasma.In this study,the test and reference preparations of rivaroxaban tablet are bioequivalent under fasting and fed conditions.

关 键 词：利伐沙班 液相色谱-串联质谱 药动学 生物等效性 高脂饮食 

分 类 号：R969.1[医药卫生—药理学]