Fe 2 O 3 nanoparticles (Fe 2 O 3 NPs) are one of the components of food additives numbered E172 and have been widely used as food pigments to color sweets. Although a large number of studies have reported that Fe 2 O 3 NPs could induce hepatotoxicity, the pathogenesis is still unclear, especially the subacute effects on the metabolic network after oral exposure. Therefore, it is necessary to define a highly sensitive strategy to investigate the potential effects of Fe 2 O 3 NPs and the mechanism. In this study, an animal experiment showed that Fe 2 O 3 NPs had no obvious toxic effects on body weight, histopathology and oxide stress. In order to further investigate the potential effects of Fe 2 O 3 NPs in vivo, a more sensitive LC-MS/MS-based lipidomic study was performed. The results of multivariate statistical analysis and western blot analysis showed that Fe 2 O 3 NP exposure significantly affects the hepatic glycerophospholipid metabolism, decreasing triacylglycerol, diglyceride, lysophosphatidylethanolamine and free fatty acids, and increasing phosphatidylcholine, lysophosphatidylinositol and coenzyme Q9. These data provide further insight into the hepatic subacute effects of Fe 2 O 3 NPs obtained by conventional toxicology methods.