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基于衍生化反应和GC-MS技术测定大鼠血液和尿液中内源性代谢物轮廓谱(1)
http://www.100md.com 2012年1月1日 夏鑫华 刘梅 李欢
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     摘 要:目的:建立基于硅烷基衍生化反应的气相色谱-质谱联用技术(GC-MS)的代谢组学测定方法,初步研究大鼠血液和尿液中内源性代谢物轮廓谱。方法:通过硅烷基化反应,将生物样品中的氨基酸、脂肪酸、有机酸、糖类和固醇类物质制成热稳定性强、易挥发的硅烷酯或醚,通过优化色谱条件对大鼠血液和尿液中的内源性代谢物进行分析测定。结果:通过优化衍生化条件和色谱条件,建立了最佳的GC-MS测定方法,并进行了方法学考察。获得了大鼠尿液和血液中的内源性代谢物轮廓谱,并鉴定了其中23个主要色谱共有峰。结论:优化的衍生化方法结合GC-MS技术可以从代谢水平对内源性代谢物进行分析,为后期的病理模型中内源性生物标志物的发现提供最佳的测定技术和方法。

    关键词:代谢指纹图谱;气相色谱-质谱联用(GC-MS);血液;尿液

    中图分类号:R-332 文献标识码:A 文章编号:1673-7717(2012)01-0041-03

    Metabolic Fingerprint Analysis of Endogenous Metabolites

    From Blood Serum and Urine of Normal Rats Based on Derivation Reaction

    Combined with Gas ChromatographyMass Spectrometry

    XIA Xinhua1, LIU Mei2,LI Huan1

    (1.The First Affiliated Hospital of Guangzhou Medical College, Guangzhou 510120,Guangdong,China;

    2.School of Chinese Material Medica of Guangzhou University of Chinese Medicine, Guangzhou 510006,Guangdong,China)

    Abstract:Objective: A metabolomic method based on derivation reaction combined with gas chromatographymass spectrometry was established to investigate metabolic fingerprints of the endogenous metabolites from serum and urine of normal rats. Method: Amino acids, fatty acids, organic acids, glucose, and sterols were transferred into ethers or esters which were heat stable and evaporated easily. Then, the chromatographic conditions were optimized to analyze the endogenous metabolites from serum and urine of the rats. Results: The optimum method was established and validated. The metabolic fingerprints of the endogenous metabolites were obtained by using the method described above. Twenty three chromatographic peaks in the metabolic fingerprints were identified by mass data and standard references. Conclusion: The metabolic fingerprints based on the derivation reaction combined with GC-MS could analyze the endogenous metabolites at metabolic level, which could supply the best method and tool for the biomarkers of pathological animal model in further research.

    Key words:Metabolic fingerprint;GC-MS;serum;urine

    随着系统生物学的提出和迅速发展,代谢组学也成为了继基因组学、蛋白质组学之后,又一个是受到广泛关注的新兴研究领域[1]。代谢组学研究的是生物体各种细胞整合在一起的代谢物变化,以生物体的体液或组织为研究对象,着重强调生物体的代谢物谱在疾病或环境刺激下的整体应激和变化[2]。体内的代谢物会因机体的病理变化而产生某种相应的变化,对正常机体的代谢产物进行测定并建立代谢物指纹轮廓谱;当疾病产生时,再测定相应的代谢物轮廓谱 ......

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