LC-MS/MS法测定人血浆中达沙替尼的浓度及两种片剂的生物等效性研究

摘要

目的：建立测定人血浆中达沙替尼浓度的方法，并研究两种片剂的生物等效性。方法：24名男性健康受试者随机分成两组，采用两制剂两周期双交叉试验设计，先后空腹及餐后口服受试制剂或参比制剂100 mg，采用液相色谱-串联质谱（LC-MS/MS）法测定人血浆中达沙替尼的浓度。以甲磺酸伊马替尼为内标，色谱柱为Welchrom C18，流动相为乙腈-0.1％甲酸水溶液（70∶30，V/V）。以多反应监测方式进行正离子扫描，采用电喷雾离子源，用于定量分析的离子对分别为m/z 488.5→401.2（达沙替尼）、m/z 494.6→394.2（内标）。用DAS 3.2.8软件进行数据处理，采用方差分析考察两种片剂的生物等效性。结果：达沙替尼血药浓度在1～300 ng/ml范围内线性关系良好。空腹口服受试制剂和参比制剂的cmax分别为（165.599±67.592）、（164.533±77.960）ng/ml，tmax分别为（1.145±0.504）、（1.080±0.467）h，t1/2分别为（5.080±2.262）、（3.771±1.596）h，AUC0-36 h分别为（550.487±256.494）、（585.986±324.885）ng·h/ml；餐后口服受试制剂和参比制剂的cmax分别为（163.058±47.533）、（165.440±53.012）ng/ml，tmax分别为（1.630±1.066）、（1.576±0.530）h，t1/2分别为（4.720±2.677）、（4.311±2.610）h，AUC0-36 h分别为（568.036±192.521）、（601.100±216.855） ng·h/ml。空腹、餐后AUC0-36 h的相对生物利用度分别为（100.2±7.5）％、（99.2±3.8）％。方差分析表明，两制剂的主要药动学参数在药物间、周期间差异均无统计学意义（P＞0.05），但在受试者个体间差异有统计学意义（P＜0.05）。结论：LC-MS/MS法能够快速测定人体血浆中达沙替尼的浓度；两种片剂生物等效。%OBJECTIVE:To establish a method for the determination of dasatinib concentration in human plasma and study the bioequivalence of 2 kinds of tablets. METHODS:In a randomized two-way crossover study,24 healthy male volunteers were divid-ed into two groups,and were administered respectively with test and reference preparations 100 mg under fasting and fed condi-tions. The plasma concentration of dasatinib was determined by LC-MS/MS. Using imatinib mesylate as internal standard,the deter-mination was performed on Welchrom C18 column with mobile phase consisted of acetonitrile-0.1% formic acid(70∶30,V/V). Posi-tive ion scanning was conducted under MRM mode,ESI,and ion pair for quantitative analysis were m/z 488.5→401.2 (dasatinib) and m/z 494.6→394.2(internal standard). DAS 3.2.8 software was used for data processing and variance analysis was adopted to in-vestigate the bioequivalence of 2 kinds of tablets. RESULTS:The linear rang of dasatinib was 1-300 ng/ml. The pharmacokinetic pa-rameters of test and reference preparations under fasting conditions were as follows:cmax were (165.599 ± 67.592) and (164.533 ± 77.960)ng/ml;tmax were(1.145±0.504)and(1.080±0.467)h;t1/2 were(5.080±2.262)and(3.771±1.596)h;AUC0-36 h were (550.487 ± 256.494) and (585.986 ± 324.885) ng·h/ml. The pharmacokinetic parameters of test and reference preparations under fed conditions were as follows:cmax were(163.058±47.533)and(165.440±53.012)ng/ml;tmax were(1.630±1.066)and(1.576± 0.530)h;t1/2 were(4.720±2.677)and(4.311±2.610)h;AUC0-36 h were(568.036±192.521)and(601.100±216.855)ng·h/ml. Relative bioavailability of AUC0-36 h under fasting and fed conditions were(100.2±7.5)% and(99.2±3.8)%. Analysis of variance showed there were no significant difference in the pharmacokinetic parameters of between 2 preparations and cyde(P>0.05),there was statistical significance among subjects(P<0.05). CONCLUSIONS:LC-MS/MS method can determine the concentration of da-satinib in human plasma rapidly;and the two preparations are bioequivalent.