摘要
目的：建立测定人血浆中氯吡格雷及其活性和非活性代谢产物浓度的超高效液相色谱-串联质谱检测方法,并将其应用于氯吡格雷在中国健康人群的药代动力学特征研究。方法：采用液液萃取法和蛋白沉淀法处理血浆样品,测定原药、活性代谢产物和非活性代谢产物浓度。液相色谱-质谱条件：采用Thermo C18（100 mm×2.1 mm,5μm）色谱柱,以乙腈（含0.1%甲酸）-水（含0.1%甲酸）（9∶1）为流动相,流速200μL·min^-1;正离子模式多反应监测扫描分析,离子通道分别为氯吡格雷m/z 321.7→211.9,活性代谢产物m/z 503.6→353.8,非活性代谢产物m/z 307.7→197.8,内标氯雷他定m/z382.8→336.9和卡马西平m/z 237.1→194.1。11例健康受试者口服300 mg负荷剂量的氯吡格雷,于不同时间点分别采集静脉血进行药代动力学分析。结果：血浆中氯吡格雷及其活性和非活性代谢产物线性范围分别为0.1-20、1-200和25-10 000 ng·m L^-1,定量下限分别为0.1、1和25 ng·m L^-1,回收率分别在49.2%-53.7%、50.9%-54.3%和95.5%-103.5%之间,批内、批间精密度RSD均小于11%,内标氯雷他定的提取回收率为58.9%,卡马西平的提取回收率为105.9%。稳定性试验中,在各种贮存条件下氯吡格雷、活性代谢产物、非活性代谢产物均稳定性良好。药动学结果显示,健康受试者口服300 mg氯吡格雷后,氯吡格雷、活性代谢产物及非活性代谢产物在人体内平均Cmax分别为（3.84±1.26）、（90.75±44.75）和（15 423.31±5 960.89）ng·m L^-1,平均tmax分别为（1.02±0.43）、（0.82±0.28）和（0.98±0.31）h,平均t1/2分别为（6.22±4.00）、（0.46±0.12）和（7.16±0.92）h,平均AUC0-∞分别为（12.88±4.72）、（102.47±38.13）和（52 797.00±17 854.92）ng·m L^-1·h。结论：该方法操作简便快速,重复性好,特异性强,灵敏度高,可用于氯吡格雷的药动学研究。
Objective： To establish a UPLC-MS / MS method for the determination of clopidogrel and its active and inactive metabolites in human plasma,and to investigate the pharmacokinetics profile in healthy Chinese subjects. Methods： The plasma treatment procedure involved single-step protein precipitation and liquid-liquid extraction to determine plasma concentrations of bulk drug,active and inactive metabolites. The samples were chromatographed on a Thermo C18column（ 100 mm × 2. 1 mm,5 μm） using a mobile phase consisting of acetonitrile（ 0. 1% formic acid） and water（ 0. 1% formic acid）（ 9 ∶ 1） at a flow rate of 200 μL·min^-1. The protonated ions of analytes were detected in positive ionization in multiple reaction monitoring mode（ MRM）. The mass transition pairs of m / z 321. 7 →211. 9,503. 6 →353. 8,307. 7 →197. 8,382. 8 →336. 9 and 237. 1 →194. 1 were used to detect clopidogrel,trans-clopidogrel-MP derivative（ MP-AM,active metabolite）,clopidogrel carboxylic acid metabolite（ CCAM,inactive metabolite）,loratadine（ internal standard） and carbamazepine（ internal standard）,respectively. Venous blood samples for pharmacokinetic measurements were taken at different time points from 11 healthy volunteers after receiving 300 mg loading dose of clopidogrel. Results： The calibration curves were linear over a concentration range of 0. 1-20 ng·m L^-1for clopidogrel,1-200 ng·m L^-1for MP-AM and 25-10 000ng·m L^-1for CCAM and the lower limits of quantitation were 0. 1 ng·m L^-1,1 ng·m L^-1and 25 ng·m L^-1,respectively. The recovery rates ranged from 49. 2 % to 53. 7 %,50. 9 % to 54. 3 % and 95. 5 % to 103. 5 % for the three compounds,and the relative standard deviations for intra-and inter-batch precision were all less than11 %. The extraction recovery rates for internal standards loratadine and carbamazepine were 58. 9 % and105. 9 %. In stability tests,clopidogrel,MP-AM and CCAM were found to be stable in plasma under various storage conditions. In 11 human subjects rec
出处
《药物分析杂志》
CAS
CSCD
北大核心
2015年第1期56-63,共8页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金(81173132)
