高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (06): 1188.doi: 10.3969/j.issn.0251-0790.2012.06.013

• 分析化学 • 上一篇    下一篇

小鼠血清中内源性代谢物的GC/TOF-MS分析

吴胜明1, 封波2, 程建华1, 李海静1, 方均建1, 颜贤忠1, 魏来2, 董方霆1   

  1. 1. 国家生物医学分析中心, 北京 100850;
    2. 北京大学人民医院肝病研究所, 北京 100044
  • 收稿日期:2011-06-29 出版日期:2012-06-10 发布日期:2012-06-10
  • 通讯作者: 吴胜明, 男, 实验师, 主要从事代谢组学研究; 董方霆, 女, 研究员, 主要从事药物分析与代谢组学研究. E-mail:wsmwyk@163.com
  • 基金资助:

    国家科技部创新方法工作专项(批准号: 2009IM031700)、国家自然科学基金(批准号: 30973676)、重大传染病专项(批准号: 2008ZX10002-016)和国家自然科学基金(批准号: 81001419)资助.

GC/TOF-MS-based Analytical Method of Endogenous Metabolites for Mouse Serum

WU Sheng-Ming1, FENG Bo2, CHENG Jian-Hua1, LI Hai-Jing1, FANG Jun-Jian1, YAN Xian-Zhong1, WEI Lai2, DONG Fang-Ting1   

  1. 1. National Center of Biomedical Analysis, Beijing 100850, China;
    2. Hepatology Institute, Peking University People’s Hospital, Beijing 100044, China
  • Received:2011-06-29 Online:2012-06-10 Published:2012-06-10

摘要: 采用气相色谱/飞行时间质谱(GC/TOF-MS)联用仪建立了小鼠血清的代谢组学分析方法. 通过对硅烷化试剂的优化和去卷积分析, 共检测到269个峰, 其中相似度在800以上的代谢产物有46个; 以核糖醇为内标, 任意选取18种内源性代谢产物考察此方法的精密度和稳定性, 并通过14种标准氨基酸的混标溶液进行定量分析考察本方法的线性关系. 结果表明, 14种氨基酸在2.78~113.20 ng/ SymbolmA@ L浓度范围内线性关系良好; 18种内源性代谢产物的变异系数均在15%以内, 具有较好的稳定性; 并利用8种标准单糖, 通过肟化反应解决了糖在多个位置出峰的难点. 该方法可用于代谢组学研究, 并通过相关数据处理找出生物标志物, 为疾病诊断提供了新的思路.

关键词: 气相色谱/飞行时间质谱, 肟化, 硅烷化, 代谢组学

Abstract: A gas chromatography/time of flight-mass spectrometer(GC/TOF-MS) method of metabonomic analysis for small mouse serum was established. With optimizing derivatization, 269 peaks of GC/TOF-MS chromatogram were detected. Among them, 46 peaks with the similarity over 800 were identified for endogenous metabolites. 18 endogenous metabolites were selected randomly to evaluate the precision and stability of the method, and 14 mixture standard of amino acids were selected for quantitative analysis. The results showed that linearities of the 14 mixture standards were good over the range of 2.78—113.20 ng/μL, precision for intra-day and inter-day tests were satisfying, and the precision values were all below 15% for 18 endogenous metabolites. Since trimethyl-silyl(TMS) derivatization of monosaccharides often results in the formation of multiple peaks, oximation can resolve the problem smoothly. The results suggest that the method could be usefully applied to metabolomic studies, by data processing, biological markers can be identified and a new way can be presented for disease diagnosis.

Key words: Gas chromatography/time of flight-mass spectrometer(GC/TOF-MS), Oximation, Silylation, Metabolomics

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