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小鼠血清中内源性代谢物的GC/TOF-MS分析
引用本文:吴胜明,封波,程建华,李海静,方均建,颜贤忠,魏来,董方霆.小鼠血清中内源性代谢物的GC/TOF-MS分析[J].高等学校化学学报,2012,33(6):1188-1194.
作者姓名:吴胜明  封波  程建华  李海静  方均建  颜贤忠  魏来  董方霆
作者单位: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
基金项目:国家科技部创新方法工作专项,国家自然科学基金,重大传染病专项,国家自然科学基金
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
收稿时间:2011-06-29

GC/TOF-MS-based Analytical Method of Endogenous Metabolites for Mouse Serum
WU Sheng-Ming , FENG Bo , CHENG Jian-Hua , LI Hai-Jing , FANG Jun-Jian , YAN Xian-Zhong , WEI Lai , DONG Fang-Ting.GC/TOF-MS-based Analytical Method of Endogenous Metabolites for Mouse Serum[J].Chemical Research In Chinese Universities,2012,33(6):1188-1194.
Authors:WU Sheng-Ming  FENG Bo  CHENG Jian-Hua  LI Hai-Jing  FANG Jun-Jian  YAN Xian-Zhong  WEI Lai  DONG Fang-Ting
Institution:1 (1.National Center of Biomedical Analysis,Beijing 100850,China; 2.Hepatology Institute,Peking University People’s Hospital,Beijing 100044,China)
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.
Keywords:Gas chromatography/time of flight-mass spectrometer(GC/TOF-MS)  Oximation  Silylation  Metabolomics
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