经皮冠状动脉介入治疗术后替格瑞洛及其代谢物浓度测定LC-MS方法的建立和应用

Establishment and application of an LC-MS method for the determination of the concentration of ticagrelor and its metabolites after percutaneous coronary intervention

目的建立测定经皮冠状动脉介入治疗术后患者血浆中替格瑞洛及其活性代谢物浓度的液相色谱串联质谱(LC-MS/MS)方法,并初步探索血浆药物浓度在此类患者中的分布特征。方法对2020年9月至2022年1月在东部战区总医院秦淮医疗区行经皮冠状动脉介入治疗术的120例急性冠脉综合征患者,口服替格瑞洛90 mg,1天2次,达到稳态后,在签署知情同意书情况下,于下次服药前1 h内采集静脉血5 mL于EP管中。采用C18色谱柱,以流动相A:0.1%甲酸水溶液;流动相B:乙腈为流动相进行梯度洗脱,使用液相色谱-三重四极杆串联质谱仪进行检测分析,并对结果进行统计分析。结果建立了血浆中2种药物的HPLC-MS/MS测定法,血浆中杂质不干扰样品的测定,线性关系良好;高、中、低三种浓度的批间和批内变异均小于15.0%,绝对回收率在89.03%~110.10%,相对回收率在91.0%~105.69%,样品稳定性良好,符合生物样品定量分析要求。同时分析发现替格瑞洛谷浓度在≤50岁与>60岁2组之间差异具有统计学意义(P=0.041)。结论建立了经皮冠状动脉介入治疗术后患者血浆中替格瑞洛及其活性代谢物HPLC-MS/MS测定法,方法稳定且符合生物样品定量分析要求,并成功应用于临床,为今后替格瑞洛在临床的安全及个体化应用打下基础。

Objective This study aimsto establish a liquid chromatography tandem mass spectrometry(LC-MS/MS)method for the determination of plasma concentrations of ticagrelor and its active metabolites in patients after percutaneous coronary intervention,and to preliminarily explore the distribution characteristics of plasma drug concentrations in such patients.Methods A total of 120 patients with acute coronary syndrome who underwent percutaneous coronary intervention in our hospital from September 2020 to January 2022 was enrolled.After reaching steady state with oral ticagrelor 90 mg BID,5 ml of venous blood was collected in an EP tube within 1 hour before the next dose.All patients signed informed consent.The results were analyzed using a C18 column with gradient elution using mobile phase A:0.1%formic acid aqueous solution;mobile phase B:acetonitrile as mobile phase,and detected and analyzed using liquid chromatography-triple quadrupole tandem mass spectrometer.Results In this study,an HPLC-MS/MS method for the determination of 2 drugs in plasma was established.Impurities in plasma did not interfere with the determination of the samples,and the linearity was good.the inter-and intra-batch variability was less than 15.0%for high,medium and low concentrations,and the absolute recoveries ranged from 89.03%to 110.10%,and the relative recoveries ranged from 91.0%to 105.69%.The sample stability was good and met the requirements for quantitative analysis of biological samples.The analysis also revealed a statistically significant difference in the distribution of ticagrelor Between groups≤50 years old group and>60 years old group(P=0.041).Conclusion A HPLC-MS/MS method for the determination of ticagrelor and its active metabolites in plasma of patients after percutaneous coronary intervention is established.The method is stable and meets the requirements for quantitative analysis of biological samples,and has been successfully applied in clinical practice,laying the foundation for the safe and individualized application of ticagrelor in clinical practice in the future.