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Zucker Diabtic Fatty rats is a common model used for the studies related to type 2 diabetes. Metabolomics, proven by previous studies, can be used for the prediction of phenotype. This investigation held untargeted metabolomics analysis on the serum metabolites of the Zucker Diabetic Fatty rats, intended to gain insights in the study of the pathology of type 2 diabetes. LCMS method (liquid chromatography coupled with mass spectrometry) is an analytical chemistry technique used to separate and purify complex mixtures. After identifying metabolites using MS2 library spectrum matching, this investigation used Metabo Analyst to compare fatty rats and lean rats. Lastly, by applying over representation analysis, we identified seven metabolic pathways that are important for humans, which assists in discovery of therapeutic targets and the developments of diabetes biomarkers.