微探针电喷雾串联质谱方法快速检测睾酮和右美沙芬的代谢稳定性  

Application of micro-pen electrospray ionization tandem mass spectrometry in rapid detection of metabolic stability of testosterone and dextromethorphan

作　　者：瞿向阳 刘倩 张思敏 朱建雄 黄民 毕惠嫦 QU Xiang-yang;LIU Qian;ZHANG Si-min;ZHU Jian-xiong;HUANG Min;BI Hui-chang(Lab of Drug Metabolism and Pharmacokinetics,School of Pharmaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China;School of Pharmaceutical Sciences,Southern Medical University,Guangzhou 510515,China;Guangdong RangerBio Technologies Co.,Ltd.,Dongguan 523000,China)

机构地区：[1]中山大学药学院药物代谢与药动学实验室,广东广州510006 [2]南方医科大学药学院,广东广州510515 [3]广东联捷生物科技有限公司,广东东莞523000

出　　处：《药学学报》2022年第5期1465-1470,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82025034,81973392);博士后科学基金资助项目(2020M683141);广东省东莞市研究生联合培养(实践)工作站项目(20201900300012)。

摘　　要：探针电喷雾电离(probe electrospray ionization,PESI)技术是原位电离技术的典型代表之一,但由于其载样稳定性差,该技术在定量分析中的应用受到很大制约。作者前期对现有PESI技术的定量分析性能进行改进,开发了新型微探针电喷雾串联质谱(micro-pen electrospray ionization tandem mass spectrometry,μPen-ESI-MS/MS)。本研究旨在探索其在药物代谢稳定性及CYP450酶活力评价中的应用。建立检测肝微粒体孵育体系中的CYP3A4底物睾酮与CYP2D6底物右美沙芬的μPen-ESI-MS/MS检测方法,并对分析方法的线性、精密度和准确度进行考察;并应用该分析方法检测睾酮和右美沙芬在肝微粒体孵育体系中的代谢稳定性。结果表明,该μPen-ESI-MS/MS方法分析效率高,一个样品的喷雾时长约为0.3 min。在设定的定量范围内,睾酮和右美沙芬分析方法的标准曲线线性良好(R2>0.99),睾酮的批内和批间准确度为95.9%~109.3%,批内和批间精密度(RSD)为2.4%~13.5%。右美沙芬的批内和批间准确度为90.5%~107.3%,批内和批间精密度(RSD)为3.4%~12.1%。睾酮和右美沙芬在肝微粒体孵育体系中快速代谢(浓度下降一半所需时间分别为12和14 min)。综上,本研究建立了睾酮及右美沙芬的代谢稳定性评价快速灵敏的μPen-ESI-MS/MS分析方法,为药物代谢酶CYP3A4和CYP2D6的活性检测提供了新方法和新策略。Probe electrospray ionization(PESI)is one of the typical types of ambient ionization technology,but its application in quantitative analysis is limited due to its poor sampling stability.Previously,we developed a new micro-pen electrospray ionization tandem mass spectrometry(μPen-ESI-MS/MS)method based on PESI.In this study,aμPen-ESI-MS/MS method to measure testosterone and dextromethorphan in liver microsome samples was developed and validated to further applicate in evaluating drug metabolism stability and CYP450 enzyme activity.AμPen-ESI-MS/MS method for detecting the CYP3A4 substrate testosterone and CYP2D6 substrate dextromethorphan in the liver microsome incubation system were developed,and the linearity,precision and accuracy of the method was validated.The validated method was further used to detect the metabolic stability of testosterone in the liver microsome incubation system.The results showed that theμPen-ESI-MS/MS had high efficiency with0.3 min spraying time of each sample.The standard curve of the testosterone and dextromethorphan has good linearity(R2>0.99),the intra-and inter-batch accuracy of testosterone and dextromethorphan was 95.9%-109.3%and 90.5%-107.3%,respectively;the intra-and inter-batch precision was acceptable with RSD values of 2.4%-13.5%and 3.4%-12.1%.The half-lives of testosterone and dextromethorphan in the liver microsome incubation system were 12 min and 14 min,respectively.This study provided a rapid and sensitiveμPen-ESI-MS/MS method for the assay of testosterone and dextromethorphan in liver microsome samples,and provided a new strategy for the evaluation of drug metabolism stability and CYP3A4/CYP2D6 activity.

关 键 词：原位电离质谱 探针电喷雾 微探针电喷雾串联质谱 药物代谢稳定性评价 CYP450酶 

分 类 号：R917[医药卫生—药物分析学]