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

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。

Metabolic Fingerprint Analysis of Endogenous Metabolites From Blood Serum and Urine of Normal Rats Based on Derivation Reaction Combined with Gas Chromatography-Mass Spectrometry

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.