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Ultra-performance liquid chromatographic-electrospray mass spectrometric determination (UPLC-ESI-MS) of O-demethylated metabolite of paeonol in vitro: Assay development, human liver microsome activities and species differences
Liu, Hui-Xin1,2; Hu, Ying1; He, Yu-Qi3; Liu, Yong1; Li, Wei1,2; Yang, Ling1
KeywordUltra-performance Liquid Chromatography Paeonol Human Liver Microsomes O-demethylation Enzyme Kinetics Species Differences
Source PublicationTALANTA
2009-10-15
DOI10.1016/j.talanta.2009.06.018
Volume79Issue:5Pages:1433-1440
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordRISK-ASSESSMENT ; CYTOCHROME-P450 ENZYMES ; INHIBITION ; DRUGS ; RAT ; PHARMACOKINETICS ; GLUCURONIDATION ; OXIDATION ; ADDUCT
AbstractA simple and sensitive method for determination of the O-demethylation activity of rat. dog, minipig. and human liver micrsomes toward paeonol using ultra-performance liquid chromatography with mass detection (UPLC-MS) has been developed. The method uses chemically synthesized O-demethylated metabolite of paeonol (2,4-dihydroxyacetophenone, DHA) as a standard for method validation. Validation was done 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 30:70). Selective ion reaction (SIR) monitor was specific for paeonol, DHA and I.S. The method was specific since there were no interference peaks from the reaction matrix. The calibration curve for DHA was linear from 0.5-100 mu M with r(2) = 0.9999. The newly developed method has good precision and accuracy. The method was successfully used to determine the kinetics of DHA activities toward paeonol in liver microsomes from different species. Dog liver microsomes (DLMs) were the most active in paeonol O-demethylation (709.7 pmol/min/mg protein) followed by rat liver microsomes (RLMs) (579.6 pmol/min/mg protein), HLMs (569.3 pmol/min/mg protein), and then minipig liver microsomes (PLMs) (417.3 pmol/min/mg protein). The developed method was appropriated for rapid screening paeonol O-demethylation activity in liver microsomes from different species. (C) 2009 Elsevier B.V. All rights reserved.
Language英语
WOS IDWOS:000269288400037
Citation statistics
Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/141303
Collection中国科学院大连化学物理研究所
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Lab Pharmaceut Resource Discovery, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China
3.Shanghai Univ Tradit Chinese Med, Shanghai, Peoples R China
Recommended Citation
GB/T 7714
Liu, Hui-Xin,Hu, Ying,He, Yu-Qi,et al. Ultra-performance liquid chromatographic-electrospray mass spectrometric determination (UPLC-ESI-MS) of O-demethylated metabolite of paeonol in vitro: Assay development, human liver microsome activities and species differences[J]. TALANTA,2009,79(5):1433-1440.
APA Liu, Hui-Xin,Hu, Ying,He, Yu-Qi,Liu, Yong,Li, Wei,&Yang, Ling.(2009).Ultra-performance liquid chromatographic-electrospray mass spectrometric determination (UPLC-ESI-MS) of O-demethylated metabolite of paeonol in vitro: Assay development, human liver microsome activities and species differences.TALANTA,79(5),1433-1440.
MLA Liu, Hui-Xin,et al."Ultra-performance liquid chromatographic-electrospray mass spectrometric determination (UPLC-ESI-MS) of O-demethylated metabolite of paeonol in vitro: Assay development, human liver microsome activities and species differences".TALANTA 79.5(2009):1433-1440.
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