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目的建立超高效液相色谱-串联质谱法同时测定人血浆中的氯吡格雷(Clo)及其非活性代谢物(CCAM)和活性代谢物(CATM)的浓度。方法 5名健康受试者单剂量口服Clo片300 mg,分别经时采集肘静脉血样进行药动学分析。血浆样品沉淀蛋白后经WATERS ACQUITY UPLC HSS T3柱(2.1 mm×50 mm,1.8μm)分离,流动相为水(含0.1%甲酸)-乙腈(含0.1%甲酸),梯度洗脱。采用电喷雾电离源(ESI)正离子模式、多反应监测(MRM),用于定量分析的离子通道分别为m/z 322.1→211.8(Clo),m/z 356.0→154.9(CATM),m/z 308.3→198.0(CCAM),m/z 253.1→180.0(内标卡马西平)。结果血浆中Clo、CATM和CCAM线性关系良好(r>0.99),日内、日间精密度(RSD)小于12.6%,准确度(RE)在-3.6%~7.6%之间。药动学结果显示,健康受试者单剂量口服Clo片300 mg后,Clo、CATM和CCAM的ρmax分别为(26.57±24.06)、(38.12±22.80)和(8 128.00±1 624.58)ng·m L-1,tmax分别为(1.8±0.8)、(2.0±1.0)和(2.0±1.0)h,AUC0-∞分别为(67.74±48.44)、(212.16±122.58)和(46 982.31±14 496.05)ng·m L-1·h。结论本方法操作简便、灵敏度高、分析时间短,适用于Clo药动学研究和常规血药浓度监测。
Objective To establish a simultaneous determination of clopidogrel (Clo) and its inactive metabolites (CCAM) and active metabolites (CATM) in human plasma by ultra performance liquid chromatography-tandem mass spectrometry. Methods Five healthy volunteers were given a single oral dose of Clo tablets 300 mg. The elbow vein blood samples were collected for pharmacokinetic analysis. Plasma samples were precipitated and separated on a WATERS ACQUITY UPLC HSS T3 column (2.1 mm × 50 mm, 1.8 μm) using a mobile phase of water (containing 0.1% formic acid) -acetonitrile with 0.1% formic acid, gradient elution. The ion channels used for quantitative analysis were m / z 322.1 → 211.8 (Clo) and m / z 356.0 → 154.9 (CATM) respectively, using electrospray ionization source (ESI) z 308.3 → 198.0 (CCAM), m / z 253.1 → 180.0 (internal standard carbamazepine). Results The linearity of Clo, CATM and CCAM in plasma was good (r> 0.99). The intra-day and inter-day precision was less than 12.6% and the accuracy was between -3.6% and 7.6%. Pharmacokinetic results showed that the ρmax of Clo, CATM and CCAM were (26.57 ± 24.06), (38.12 ± 22.80) and (8 128.00 ± 1624.58) ng · m L-1 and tmax were (1.8 ± 0.8), (2.0 ± 1.0) and (2.0 ± 1.0) h, and the AUC0-∞ were 67.74 ± 48.44, 212.16 ± 122.58 and 46 982.31 ± 14 496.05, ng · m L-1 · h. Conclusion The method is simple, sensitive, and short in analysis time. It is suitable for Clo pharmacokinetics and conventional blood concentration monitoring.