LC-MS/MS法测定人血浆中雷沙吉兰的浓度及其生物等效性评价  

Determination of rasagiline concentration in human plasma by LC-MS/MS and application to bioequivalence study

作　　者：于婧 冯晓晖 祁琪 王菊香 董菁 李黎 张鹏[1] YU Jing;FENG Xiao-hui;QI Qi;WANG Ju-xiang;DONG Jing;LI Li;ZHANG Peng(Shenyang Pharmaceutical University Wuya College of Innovation,Shenyang 110016,China;Changzhou Siyao Pharmaceuticals Co.,Ltd.,Changzhou 213000,China;United-Power Pharma Technology Co.,Ltd.,Beijing 102206,China)

机构地区：[1]沈阳药科大学无涯创新学院,沈阳110016 [2]常州四药制药有限公司,常州213000 [3]军科正源(北京)药物研究有限公司,北京102206

出　　处：《药物分析杂志》2021年第11期1914-1922,共9页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立快速灵敏的液相色谱-质谱联用(LC-MS/MS)方法,测定人血浆中雷沙吉兰的浓度;评价2种甲磺酸雷沙吉兰片在中国健康受试者体内的生物等效性。方法:以雷沙吉兰-^(13)C_(3)为内标,采用液液萃取法进行生物样品前处理,使用反相ACE C_(18)-300?(2.1 mm×100 mm,5μm)色谱柱,以含0.1%甲酸的5 mmol·L^(-1)乙酸铵水溶液(A)和含0.1%甲酸的乙腈溶液(B)为流动相,进行色谱分离,在正离子模式下,采用多反应监测分别检测m/z 172.1→117.0(雷沙吉兰)和m/z 175.1→117.0(雷沙吉兰-^(13)C_(3))。结果:该方法仅使用150μL血浆即可达到0.020 ng·mL^(-1)的定量下限,在0.020~20.0 ng·mL^(-1)浓度范围内具有良好的线性关系,批内、批间精密度分别为1.0%~10.6%和2.6%~12.8%,准确度分别为-3.0%~4.7%和-3.0%~1.7%。应用该方法测定雷沙吉兰浓度,受试制剂(T)和参比制剂(R)在空腹试验中的主要药动学参数:C_(max)分别为(8.528±3.595)ng·mL^(-1)和(8.873±4.128)ng·mL^(-1);AUC_(0-t)分别为(5.838±1.660)ng·h·mL^(-1)和(5.911±1.797)ng·h·mL^(-1);AUC_(0-∞)分别为(5.990±1.716)ng·h·mL^(-1)和(6.104±1.895)ng·h·mL^(-1);Cmax、AUC_(0-t)和AUC_(0-∞)的T-R比值的90%置信区间分别为88.26%~108.46%、94.16%~105.35%和93.55%~105.01%。受试制剂和参比制剂在餐后试验中的主要药动学参数:C_(max)分别为(4.333±2.434)ng·mL^(-1)和(4.359±2.980)ng·mL^(-1);AUC_(0-t)分别为(4.931±1.361)ng·h·mL^(-1)和(4.807±1.382)ng·h·mL^(-1);AUC_(0-∞)分别为(5.011±1.386)ng·h·mL^(-1)和(4.893±1.403)ng·h·mL^(-1);C_(max)、AUC_(0-t)和AUC_(0-∞)的T-R比值的90%置信区间分别为89.54%~118.23%、99.88%~107.07%和99.59%~107.05%,均符合生物等效性的等效范围要求(80.00%~125.00%),判断2种制剂具有生物等效性。结论:该方法灵敏度高,前处理简单,适用于雷沙吉兰生物样本的高通量分析,并成功应用于雷沙吉兰片的临床一致性评价,为雷沙吉兰片仿制药的临床用药提供数据支�Objective:To establish a sensitive and rapid liquid chromatography-mass spectrometry(LC-MS/MS)method for the determination of rasagiline in human plasma and apply to bioequivalence evaluation of two rasagiline mesylate tablets in healthy Chinese subjects.Methods:Liquid-liquid extraction was used to separate rasagiline from plasma in 96-well plate.Separation was performed on ACE C_(18)(2.1 mm×100 mm,5μm)column with the mobile phase of 5 mmol·L^(-1) ammonium acetate aqueous solution and acetonitrile contained 0.1%formic acid at a flow rate of 0.5 mL·min-1.Under the positive electrospray ionization interface,the multiple reaction monitoring transitions of m/z 172.1→117.0 and m/z 175.1→117.0 were used to measure rasagiline and rasagiline-^(13)C_(3),respectively.Results:The lower limit of quantification(LOQ)was identifiable and reproducible at 0.020 ng·mL^(-1),using 150μL plasma sample.The intra-run and inter-run precision were less than 12.8%,and the accuracy was within±4.7%.The main pharmacokinetic parameters of rasagiline after a single oral dose of rasagiline tablet under fasting condition for T and R were as follows:Cmax were(8.528±3.595)ng·mL^(-1) and(8.873±4.128)ng·mL^(-1);AUC_(0-t) were(5.838±1.660)ng·h·mL^(-1) and(5.911±1.797)ng·h·mL^(-1),and AUC_(0-∞)were(5.990±1.716)ng·h·mL^(-1) and(6.104±1.895)ng·h·mL^(-1),respectively.The 90%CIs for the geometric mean ratios of Cmax,AUC_(0-t) and AUC_(0-∞)of T and R under fasting condition were 88.26%-108.46%,94.16%-105.35%and 93.55%-105.01%,respectively.The main pharmacokinetic parameters of rasagiline under fed condition for T and R were as follows:C_(max) were(4.333±2.434)ng·mL^(-1) and(4.359±2.980)ng·mL^(-1),AUC_(0-t)(4.931±1.361)ng·h·mL^(-1) and(4.807±1.382)ng·h·mL^(-1),and AUC_(0-∞)(5.011±1.386)ng·h·mL^(-1) and(4.893±1.403)ng·h·mL^(-1).The 90%CIs for the geometric mean ratios of C_(max),AUC_(0-t) and AUC_(0-∞)of T and R under fed condition were 89.54%-118.23%,99.88%-107.07%and 99.59%-107.05%,respectively.The results

关 键 词：雷沙吉兰 液相色谱质谱联用 血药浓度 生物等效性 高灵敏度 高通量分析 

分 类 号：R917[医药卫生—药物分析学]