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题名: Determination of UDP-glucuronosyltransferase UGT2B7 activity in human liver microsomes by ultra-performance liquid chromatography with MS detection
作者: Liu, Hui-Xin1, 3;  He, Yu-Qi2;  Hu, Ying1, 3;  Liu, Yong1;  Zhang, Jiang-Wei1, 3;  Li, Wei1, 3;  Wang, Zheng-Tao2;  Yang, Ling1
通讯作者: 杨凌
关键词: ultra-performance liquid chromatography ;  UGT2B7 ;  human liver microsomes
刊名: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
发表日期: 2008-07-01
DOI: 10.1016/j.jchromb.2008.06.009
卷: 870, 期:1, 页:84-90
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1806
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: A rapid and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS) method was developed for the qualitative and quantitative determination of UGT2B7 activity using 3'-azido-3'-deoxythymidine (AZT) as probe substrate in human liver microsomes (HLMs). The method was validated for the determination of AZT glucuronidation (AZTG) with respect to specificity, linearity, detection limit, recovery, stability, precision and accuracy. The chromatographic separation was achieved on a UPLC BEH C-18 column (50 mm x 2.1 mm i.d., 1.7 mu m), with phase of acetonitrile-water (ratio 6:94). Selective ion reaction (SIR) monitor was specific for AZT, AZTG and I.S. The method was linear over the concentration range 0.5-500 mu M for AZTG in spiked HLMs. Good precision and accuracy were obtained for concentrations over the standard curve range. AZTG was stable at 4 degrees C for at least 72 h in spiked liver microsomes samples. The method was successfully used to determine the kinetics of UGT activities toward AZT in HLMs. In addition, the method could determine the effects of fluconazole, a known UGT2B7 selective inhibitor, on AZTG in HLMs. Therefore, this method is suitable for in vitro studies using AZTG formation as an index reaction for UGT2B7 activity. (C) 2008 Elsevier B.V. All rights reserved.
关键词[WOS]: DRUG-DRUG INTERACTIONS ;  FLIGHT MASS-SPECTROMETRY ;  SOLID-PHASE EXTRACTION ;  IN-VITRO ;  2B7 UGT2B7 ;  GLUCURONIDATION ;  ZIDOVUDINE ;  MORPHINE ;  ASSAY ;  FLUCONAZOLE
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000257835000014
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/100111
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Lab Pharmaceut Resource Discovery, Dalian 116023, Peoples R China
2.Shanghai Univ Tradit Chinese Med, Shanghai, Peoples R China
3.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China

Recommended Citation:
Liu, Hui-Xin,He, Yu-Qi,Hu, Ying,et al. Determination of UDP-glucuronosyltransferase UGT2B7 activity in human liver microsomes by ultra-performance liquid chromatography with MS detection[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2008,870(1):84-90.
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