LC-MS/MS法测定人血浆中氯吡格雷活性代谢物衍生物浓度的不确定度评定

目的:评定液相色谱串联质谱(LC-MS/MS)法测定人血浆中氯吡格雷活性代谢物衍生物(CAMD)浓度的不确定度。方法:对LC-MS/MS法测定人血浆中CAMD浓度的整个过程进行分析,对测定过程中的测量重复性、称量、工作液配制、生物样品制备、回收率、仪器允差和标准曲线拟合等因素引起的不确定度分别进行了评定,最后计算合成不确定度并进行扩展。结果:血浆CAMD低(0.107ng/ml)、中(4.467ng/ml)、高(155.667ng/ml)质量浓度质控的扩展不确定度分别为0.010 2、0.188、6.413ng/ml(k=2,P=95%)。结论:LC-MS/MS法测定人血浆中CAMD的不确定度在低浓度时主要由曲线拟合引入,在中浓度和高浓度时主要由生物样品配制、萃取过程、对照品配制和仪器允差引入。

氯吡格雷活性代谢物在人体内的血药浓度测定

目的:采用液相色谱-串联质谱(LC-MS/MS)建立人血浆中氯吡格雷活性代谢产物浓度测定的方法。方法:血浆经冷冻干燥处理,以Accucore C_(18)为色谱柱;乙腈(含0.01%甲酸)-水(含0.01%甲酸)为流动相,流速0.4 mL·min^(-1),柱温30℃,进样量2μL;通过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活性代谢产物衍生物(CAMD)m/z 504.2→155.1,内标氯雷他定m/z382.8→336.9。结果:血浆中氯吡格雷活性代谢产物衍生物在0.5~500 ng·mL^(-1)范围内线性关系良好;最低检测限为0.5 ng·mL^(-1);提取回收率为83.81%~97.65%;日内和日间精密度的RSD均小于9.86%。结论:本方法准确可靠,简便快速且灵敏度高,适用于人血浆中氯吡格雷活性代谢物的测定,可进一步用于研究氯吡格雷活性代谢产物的药代动力学特征。

液质联用法同时测定人血浆氯吡格雷活性和非活性代谢物浓度

目的建立快速测定人血浆中氯吡格雷活性和非活性代谢物浓度的LC-MS/MS法。方法乙醚液液萃取法,采用Acquity UPLC BEH C18色谱柱。以正离子多反应离子监测方式进行检测。结果氯吡格雷羧酸代谢物、卡马西平、氯吡格雷活性代谢物衍生物分离良好,氯吡格雷羧酸代谢物的线性范围良好。两种代谢产物日内、日间RSD小于12%。在不同贮存条件下代谢物稳定性良好。结论该方法操作简便快速,重复性好,灵敏度高,适用于氯吡格雷代谢产物血药浓度的测定。

冠心病患者氯吡格雷群体药动学-药效学模型研究

目的建立冠心病患者氯吡格雷群体药动学-药效学模型,为尽早诊断和干预氯吡格雷抵抗提供临床依据。方法前瞻性收集使用双联抗血小板治疗冠心病的患者101例,分别以患者体内氯吡格雷活性代谢产物(clopidogrel active metabolite,Clop-AM)浓度和血小板最大聚集率(maximum platelet aggregation rate,MAR)作为药动学和药效学指标,使用非线性混合效应模型定量考察患者性别、年龄、体质量指数(body mass index,BMI)、CYP2C19基因型、合并疾病等影响氯吡格雷抵抗的因素,建立冠心病患者氯吡格雷群体药动学-药效学模型。结果Clop-AM表观清除率的群体典型值为2910 L·h^(−1),BMI对Clop-AM的表观清除率有影响,最终模型公式为CL=2910×(BMI/25.09)×3.22;模型验证的平均预测误差(mean prediction error,MPE)和均方根预测误差(root mean square prediction error,RMSE)分别为药动学-药效学:MPE=0.14,RMSE=10.77;药动学:MPE=0.26,RMSE=2.91;药效学:MPE=–0.05,RMSE=17.10。结论初步建立了冠心病患者氯吡格雷群体药动学-药效学模型,BMI较高的人群,Clop-AM清除加快,对氯吡格雷个体化用药有一定的参考价值。

Effects of Rong Shuan capsule, Xue Zhi Kang capsule, Xin Yuan capsule and Songling Xue Mai Kang capsule on the pharmacokinetics of clopidogrel active metabolite in rats

In the present study, we aimed to examine the effects of Rong Shuan capsule, Xue Zhi Kang capsule, Xin Yuan capsule and Songling Xue Mai Kang capsule on the pharmacokinetics of clopidogrel active metabolite（CAM）. The traditional Chinese medicines（TCMs） Rong Shuan capsule, Xue Zhi Kang capsule, Xin Yuan capsule and Songling Xue Mai Kang capsule are widely used to treat cardiovascular disease in China. They are often prescribed in combination with clopidogrel, a common anti-platelet Western drug. We investigated the influence of the four TCMs on CAM pharmacokinetics following administration at human dose in rats. Pharmacokinetic parameters were determined following oral（PO） administration of clopidogrel（7.5 mg/kg） with or without Rong Shuan capsule（75 mg/kg, PO）, Xue Zhi Kang capsule（60 mg/kg, PO）, Xin Yuan capsule（120 mg/kg, PO）, or Songling Xue Mai Kang capsule（150 mg/kg, PO）. Compared with the animals in the control group, Xue Zhi Kang capsule significantly decreased the area under the plasma concentration-time curve（AUC_（0-t）） of the CAM derivative by 25.4%. However, the t1/2 and Vz/F of CAM derivative were significantly increased by 43.6% and 70.7%, respectively. It was also observed that the pharmacokinetic parameters were altered in groups pretreated with Rong shuan capsule, Xin yuan capsule or Songling Xue mai kang capsule compared with the control group, but not significant. This study indicated that Xue Zhi Kang capsule had an effect on the formation and metabolism of CAM. Therefore, in the beginning of co-administration of Xue Zhi Kang capsule and clopidogrel, the anti-platelet efficacy might be compromised because of the decreased formation of CAM. Otherwise, long-time co-administration might lead to side effects by the prolongation of the t1/2 and Vz/F increase of CAM.