药学学报, 2022, 57(5): 1465-1470
引用本文:
瞿向阳, 刘倩, 张思敏, 朱建雄, 黄民, 毕惠嫦. 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[J]. 药学学报, 2022, 57(5): 1465-1470.
QU Xiang-yang, LIU Qian, ZHANG Si-min, ZHU Jian-xiong, HUANG Min, BI Hui-chang. Application of micro-pen electrospray ionization tandem mass spectrometry in rapid detection of metabolic stability of testosterone and dextromethorphan[J]. Acta Pharmaceutica Sinica, 2022, 57(5): 1465-1470.

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
瞿向阳1,2, 刘倩1,3, 张思敏1, 朱建雄3, 黄民1, 毕惠嫦1,2*
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
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。
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     
Application of micro-pen electrospray ionization tandem mass spectrometry in rapid detection of metabolic stability of testosterone and dextromethorphan
QU Xiang-yang1,2, LIU Qian1,3, ZHANG Si-min1, ZHU Jian-xiong3, HUANG Min1, BI Hui-chang1,2*
1. Lab of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China;
2. School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China;
3. Guangdong RangerBio Technologies Co., Ltd., Dongguan 523000, China
Abstract:
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, a μPen-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. A μPen-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 the μPen-ESI-MS/MS had high efficiency with 0.3 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 sensitive μPen-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.
Key words:    ambient ionization mass spectrometry    probe electrospray ionization    micro-pen electrospray ionization tandem mass spectrometry    drug metabolism stability evaluation    cytochrome P450   
收稿日期: 2021-11-17
DOI: 10.16438/j.0513-4870.2021-1650
基金项目: 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).
通讯作者: 毕惠嫦,Tel:86-20-39943470,Fax:86-20-39943000,E-mail:bihchang@mail.sysu.edu.cn
Email: bihchang@mail.sysu.edu.cn
相关功能
PDF(399KB) Free
打印本文
0
作者相关文章
瞿向阳  在本刊中的所有文章
刘倩  在本刊中的所有文章
张思敏  在本刊中的所有文章
朱建雄  在本刊中的所有文章
黄民  在本刊中的所有文章
毕惠嫦  在本刊中的所有文章

参考文献:
[1] Noor Z, Ahn SB, Baker MS, et al. Mass spectrometry-based protein identification in proteomics-a review[J]. Brief Bioinform, 2021, 22:1620-1638.
[2] Goodwin RJA, Takats Z, Bunch J. A critical and concise review of mass spectrometry applied to imaging in drug discovery[J]. Slas Discov, 2020, 25:963-976.
[3] Zhang X, Li Q, Xu Z, et al. Mass spectrometry-based metabolomics in health and medical science:a systematic review[J]. RSC Adv, 2020, 10:3092-3104.
[4] Gachumi G, Purves RW, Hopf C, et al. Fast quantification without conventional chromatography, the growing power of mass spectrometry[J]. Anal Chem, 2020, 92:8628-8637.
[5] Feider CL, Krieger A, DeHoog RJ, et al. Ambient ionization mass spectrometry:recent developments and applications[J]. Anal Chem, 2019, 91:4266-4290.
[6] Hiraoka K, Nishidate K, Mori K, et al. Development of probe electrospray using a solid needle[J]. Rapid Commun Mass Spectrom, 2007, 21:3139-3144.
[7] Yoshimura K, Chen LC, Asakawa D, et al. Physical properties of the probe electrospray ionization (PESI) needle applied to the biological samples[J]. J Mass Spectrom, 2009, 44:978-985.
[8] Thompson TN. Early adme in support of drug discovery:the role of metabolic stability studies[J]. Curr Drug Metab, 2000, 1:215-241.
[9] Asha S, Vidyavathi M. Role of human liver microsomes in in vitro metabolism of drugs-a review[J]. Appl Biochem Biotech, 2010, 160:1699-1722.
[10] Ackley DC, Rockich KT, Baker TR. Metabolic stability assessed by liver microsomes and hepatocytes[M]//Yan Z, Caldwell GW, Eds.Optimization in Drug Discovery:in vitro Methods. Totowa:Humana Press, 2004:151-162.
[11] Kuo CP, Shiea J. Application of direct electrospray probe to analyze biological compounds and to couple to solid-phase microextraction to detect trace surfactants in aqueous solution[J]. Anal Chem, 1999, 71:4413-4417.
[12] Mandal MK, Chen LC, Hashimoto Y, et al. Detection of biomolecules from solutions with high concentration of salts using probe electrospray and nano-electrospray ionization mass spectrometry[J]. Anal Methods, 2010, 2:1905-1912.
[13] Mandal MK, Yoshimura K, Saha S, et al. Biomolecular analysis and biological tissue diagnostics by electrospray ionization with a metal wire inserted gel-loading tip[J]. Anal Chem, 2014, 86:987-992.
[14] Mandal MK, Saha S, Yoshimura K, et al. Biomolecular analysis and cancer diagnostics by negative mode probe electrospray ionization[J]. Analyst, 2013, 138:1682-1688.
[15] Danielson PB. The cytochrome p450 superfamily:biochemistry, evolution and drug metabolism in humans[J]. Curr Drug Metab, 2002, 3:561-597.
[16] Wang J, Liu H. Lead compound optimization strategy (1)——changing metabolic pathways and optimizing metabolism stability[J]. Acta Pharm Sin (药学学报), 2013, 48:1521-1531.
[17] Clarke SE, Jeffrey P. Utility of metabolic stability screening:comparison of in vitro and in vivo clearance[J]. Xenobiotica, 2001, 31:591-598.
[18] Kola I, Landis J. Can the pharmaceutical industry reduce attrition rates?[J]. Nat Rev Drug Discov, 2004, 3:711-715.
[19] Shang F, Feng S, Chen Q, et al. In vitro inhibitory effects of jiawei foshou san capsule on activity of cytochrome p450 enzymes in rat and human liver microsomes[J]. Acta Pharm Sin (药学学报), 2016, 51:926-930.
相关文献:
1.张思敏, 刘倩, 瞿向阳, 朱建雄, 黄民, 毕惠嫦.微探针电喷雾串联质谱与液相色谱-电喷雾串联质谱的基质效应对比研究[J]. 药学学报, 2022,57(4): 1123-1129