Development Of A GC-MS Approach For The Metabonomic Study On The Liver Injury In Mice

Metabonomics, defined as "the quantitative measurement of the multiparametric time-related metabolic responses of a complex system to a pathophysiological intervention or genetic modification", has been widely used in the research of pharmacology and biological sciences. Mass spectrometry combined with chromatographic separations, has become an important technique for the metabonomic study, due to the excellent separating and qualitative abilities with a high sensitivity. Gas chromatography-mass spectrometry (GC-MS) is a mature technique with searchable standard database, which facilitates the identification of metabolites. Thus GC-MS is considered as an ideal tool for the metabonomic study. However, the applications of GC-MS to metabonomics are limited by its lack of ability to analyze non-volatile or semi-volatile components.In this thesis, the chemical derivatization was carried out, and after that, a GC-MS approach was established for characterizing the metabolite profiles of the liver tissue homogenate and plasma from the liver injured mice. The main content of this study is presented as follows:(1) A stable and reliable GC-MS analytical method was established to obtain the metabolite profiles of the liver tissue homogenate and plasma of mice. NIST 2002, a. commercial mass spectral database, was used for rapid identification of the detected metabolites. The procedures of sample preparation, derivatizative conditions and chromatographic conditions were optimized, and validation of the method was carried out. The results showed that GC-MS analytical method was stable and reliable. It was capable to describe the biochemical composition of biological samples.(2) The established GC-MS analytical method was applied to the metabonomic study on the carbon tetrachloride-induced acute liver injury in mice. Principal components analysis (PCA) was used for recognizing the different metabolic patterns between the dosed and control groups. The time- and dose-dependent metabolic changes were discovered from the score plots of the metabonomic data. These changes were highly related to the damage of liver, which was confirmed by liver histological examinations.