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The microbiota-host co-metabolites have proven to be of great importance in physiological studies and their dysregulation has been found to be closely related to various inflammation disorders and beyond.However, simultaneous determination of these endogenous metabolites represents a great challenge.We thus developed an original and feasible method using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) which made it possible to rapidly quantify 11 metabolites spanning succinate, phenylacetylglutamine, hippurate and trimethylamine metabolic pathways within 10 min.