LC-MS/MS法测定人血浆中的替格瑞洛及其活性代谢物的质量浓度

Determination of ticagrelor and its active metabolite in human plasma by LC-MS/MS

目的建立一种快速、灵敏的液相色谱-串联质谱方法(LC-MS/MS)测定人血浆中的替格瑞洛及其主要活性代谢物AR-C124910XX的质量浓度,并研究其在健康受试者体内的药物动力学。方法血浆样品经过乙腈蛋白沉淀提取,以替格瑞洛-d7为内标,采用Dikma C18色谱柱(50 mm×4.6 mm,5μm),以乙腈:醋酸铵(5 mmol·L^-1为流动相进行梯度洗脱;质谱采用电喷雾离子化源,多反应监测(multiple reaction monitoring,MRM)检测模式,负离子方式检测。结果人血浆中替格瑞洛及其活性代谢物AR-C124910XX在质量浓度为0.781~800μg·L^-1内线性关系良好,定量下限为0.781μg·L^-1。替格瑞洛及AR-C124910XX的日内、日间的精密度(RSD,%)均小于12.61%,准确度(RE,%)均在±7.88%内;替格瑞洛的基质效应在99.53%~108.73%内;AR-C124910XX的基质效应在99.34%~105.96%内。结论该方法简单、快速、灵敏度较高,适用于替格瑞洛和AR-C124910XX在健康受试者中的临床药动学研究。

Objective A rapid,simple and sensitive LC-MS/MS method was established and validated for quantification of ticagrelor and AR-C124910 XX in human plasma and to study the pharmacokinetic characteristics in healthy human.Methods Ticagrelor-d7 was used as the internal standard.After plasma samples were deproteinized with acetonitrile,the post-treatment samples were chromatographed on a Dikma C18 column interfaced with a triple quadrupole tandem mass spectrometer.Electrospray negative ionization mode(ESI)and multiple reaction monitoring were adopted to assay ticagrelor.Acetonitrile and 5 mmol·L-1 ammonium acetate was used as the mobile phase with a gradient elution at a flow rate of 0.5 mL·min-1.Results The method was linear in the range of 0.781-800μg·L-1 for both ticagrelor AR-C124910 XX.The intra-and inter-day precision was within 12.61%in terms of relative standard deviation(RSD,%)and the accuracy within±7.88%in terms of relative error.The matrix effects were in the range from 99.53%to 108.73%for ticagrelor and 99.34%-105.96%for AR-C124910 XX.Conclusion The LC-MS/MS method was fully validated for its sensitivity,selectivity,stability,matrix effect and recovery.This convenient and specific LC-MS/MS method was successfully applied to the pharmacokinetic study of ticagrelor and AR-C124910 XX in healthy volunteers after an oral dose of 90 mg ticagrelor.