UPLC-MS/MS同时测定血浆样品中他莫昔芬及代谢产物的浓度和应用

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (7): 773-778.

UPLC-MS/MS同时测定血浆样品中他莫昔芬及代谢产物的浓度和应用

胡东莉¹, 陈尧¹, 沈杰^1,2, 谢海棠², 钱荣华¹, 谭志荣¹, 王医成¹, 王丹¹, 周宏灏¹

¹中南大学临床药理研究所, 湖南省遗传药理学重点实验室,湖南 410078,长沙;
²皖南医学院弋矶山医院临床药学部,芜湖 241001,安徽

收稿日期:2012-04-20 修回日期:2012-05-23 发布日期:2012-07-17
通讯作者: 周宏灏,男,院士,博士生导师,研究方向:临床药理学和遗传药理学。Tel: 0731-84805379 E-mail: hhzhou2003@163.com
作者简介:胡东莉,女,硕士研究生,研究方向:临床药理学。Tel: 13787794503 E-mail: 23484261@qq.com;cbohua@163.com
基金资助:
湖南省自然科学基金重点项目(10JJ2008);中南大学中央高校基本科研业务费项目(201012200047)

UPLC-MS/MS determination of tamoxifen and its metabolites in human plasma and application

HU Dong-li¹,CHEN Yao¹, SHEN Jie^1,2, XIE Hai-tang², QIAN Rong-hua¹,TAN Zhi-rong¹, WANG Yi-cheng¹, WANG Dan¹, ZHOU Hong-hao¹

¹Institute of Clinical Pharmacology,Hunan Key Laboratory of Pharmacogenetics, Central South University,Changsha 410078, Hunan,China;
²Department of Clinical Parmacy, Yijishan Hospital, Wannan Medical College, Wuhu 241001, Anhui, China

Received:2012-04-20 Revised:2012-05-23 Published:2012-07-17

摘要/Abstract

摘要： 目的: 建立UPLC-MS/MS法测定人体血浆样品中他莫昔芬及代谢产物的浓度和应用。 方法: 采用Waters AcQuity型超高效液相色谱联用API4000型三重四级杆质谱仪,样品处理采用固相萃取的方法,采用Thermo hypurity C₁₈(150 mm×2.1 mm,5 μm)色谱分析柱, 流动相为 10 mmol/L 甲酸胺-乙腈(40∶60,V/V),流速为 0.3 mL/min,进样体积为 10 μL,柱温为 30 ℃, 样品室温度为 5 ℃,质谱应用ESI源、正离子模式、多反应监测(MRM)方式检测他莫昔芬及各代谢产物的浓度。 结果: 他莫昔芬、ND-Tam和TamNox线性范围为1~400 ng/mL,4-OH-Tam线性范围为 0.4~160 ng/mL,4-OHND-Tam线性范围为2~800 ng/mL,可以满足样本浓度定量检测的需要,方法稳定、特异性高,并成功地应用到用药后他莫昔芬及各代谢产物血浆浓度的检测。 结论: 该方法简便、准确、重复性好,可以准确地定量人体血浆他莫昔芬和代谢产物的浓度,除了为临床血药浓度监测及他莫昔芬药效学研究提供服务外,也可为遗传药理学科研提供技术方法。

关键词: 他莫昔芬, UPLC-MS/MS, CYP3A4

Abstract: AIM: To develop a UPLC-MS/MS method to determine the concentration of tamoxifen and its metabolites in human plasma and application. METHODS: The equipments were Waters AcQuity ultra-high performance liquid chromatography with API4000 type triple quadrupole mass spectrometer , Thermo Hypurity C₁₈ column (150 mm×2.1 mm, 5 μm) was used in the experiment.10 mmol/L ammonium formate-acetonitrile(40∶60, V/V) was used as mobile phase and the flow rate was 0.3 mL/min, the injection volume was 10 μL, the column temperature was 30 ℃ and the sample room temperature was set at 5 ℃.The concentrations of tamoxifen and its metabolites were monitored by Mass with ESI source in positive ion mode and multiple reaction monitoring (MRM) way. RESULTS: Solid phase extraction method was used for sample preparation, tamoxifen, ND-Tam and TamNox were linear range from 1-400 ng/mL, 4-OH-Tam and 4-OHND-Tam were linear range from 0.4-160 ng/mL and 2-800 ng/mL respectively, the limitation of tamoxifen and its metabolites were about 1-2 ng/mL, some even less than 0.4 ng/mL,the method was high sensitivity, stability and specificity and has already been used for determination of tamoxifen and its metabolites in human successfully. CONCLUSION: The method is simple, accurate, repetitive for the determination of tamoxifen and its metabolites in human plasma and suitable for the concentration monitoring in clinic, present study also provide technical method for pharmacogenetics research.

Key words: Tamoxifen, UPLC-MS/MS, CYP3A4

中图分类号:

R917

胡东莉, 陈尧, 沈杰, 谢海棠, 钱荣华, 谭志荣, 王医成, 王丹, 周宏灏. UPLC-MS/MS同时测定血浆样品中他莫昔芬及代谢产物的浓度和应用[J]. 中国临床药理学与治疗学, 2012, 17(7): 773-778.

HU Dong-li,CHEN Yao, SHEN Jie, XIE Hai-tang, QIAN Rong-hua,TAN Zhi-rong, WANG Yi-cheng, WANG Dan, ZHOU Hong-hao. UPLC-MS/MS determination of tamoxifen and its metabolites in human plasma and application[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(7): 773-778.

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