To comprehensively elucidate the possible mechanism of short-term administration of emodin on the metabolic disturbance in vivo, our study utilized both LC/MS and NMR platforms for untargeted metabolomics studies. These analyses aimed to identify potential biomarkers associated with emodin-induced metabolic disturbances in mice. Subsequently, we delved into lipidomics to explore lipid metabolism systematically and determined the position of C=C double bonds in unsaturated lipids.