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题名: Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach
作者: Liu Xi-Dong1;  Zhu Jun2;  Cong Yu-Ting1;  Hu Liang-Hai1;  Ye Ming-Liang2;  Gu Jing-Kai1;  Zou Han-Fa2
关键词: Uridinediphosphate glucuronosyl transferase ;  Multiple reaction monitoring ;  Liver microsome ;  Cytochrome P450 ;  Liquid chromatography-tandem mass spectrometry
刊名: CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
发表日期: 2014
DOI: 10.3724/SP.J.1096.2014.30741
卷: 42, 期:1, 页:10-15
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Strategy for absolute quantification of metabolic enzymes in rat liver microsomes was developed by using shotgun-based proteomic approach. Rat liver microsome was digested by trypsin and the specific peptides of drug metabolic enzymes were determined by multiple reaction monitoring (MRM)-based LC-MS/MS. Firstly, standard curves were employed for the quantification of P450 and uridinediphosphate glucuronosyl transferase (UGT), which showed good linearity with the r values were all larger than 0.995 and the low limit of quantification was <= 10 nmol/L. Secondly, the synthetic isotope labeled specific peptide was served as internal standard for the quantification of UGT1A1. The stable isotope labeled specific peptide showed the same behavior with unlabeled peptide by LC-MS/MS, and the labeled peptide showed good linearity in matrix solution. The content of UGT1A1 was 17.30 nmol/g protein and 18.23 nmol/g protein obtained by standard curve and stable isotope dilution method, respectively. In summary, the results obtained by two methods were consistent, however, the stable isotope dilution method was more convenient than by standard curve, and it was suitable for high throughput determination in complex system.
关键词[WOS]: QUANTITATIVE RT-PCR ;  ENZYME REACTOR ;  PROTEINS ;  PEPTIDE ;  SAFETY
语种: 英语
WOS记录号: WOS:000333673500002
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/145619
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.Jilin Univ, Coll Life Sci, Changchun 130012, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China

Recommended Citation:
Liu Xi-Dong,Zhu Jun,Cong Yu-Ting,et al. Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach[J]. CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,2014,42(1):10-15.
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