Abstract: 【Objective】 This study aimed to investigate the effects of long-term intragastric gavage of rock tea solubles on serum metabolites of mice， and regulatory functions of lipid， glucose， and amino acid in metabolism using untargeted metabolomics， so as to provide theoretical references for rock tea development and utilization.【Method】 Male SPF C57BL/6 mice were randomly assigned into a control group （n=5） and a rock tea intervention group （n=5）. The rock tea solubles were prepared using the water extraction method. The intragastric gavage of rock tea solubles in mice was performed in the rock tea intervention group， while the solubles were replaced by purified water； the mice received intragastric gavage once daily at a volume of 50 μL and a dose of 450 mg/（kg·d）. At the end of the 8-week intervention， blood samples were collected from mice （fasted for 12 h before sampling）anesthetized with isoflurane. Serum was then subjected to untargeted metabolomic analysis using liquid chromatography-tandem mass spectrometry （LC-MS/MS）. Differential metabolites and key signaling pathways were identified through cluster analysis and KEGG signaling pathway enrichment analysis.【Result】 The 8-week intervention with rock tea solubles significantly altered the serum metabolites of mice. A total of 287 metabolites were significantly up-regulated （P<0.05， the same below） and 150 were significantly down-regulated. Significant up-regulations were observed in three glycerophospholipids， two fatty acyls， as well as amino acids and their metabolites such as phenylalanine-aspartate， N-acetylornithine， and Ile-Gly-Ser； down-regulations were observed in seven amino acids and their metabolites， four organic acids and their metabolites， two heterocyclic compounds， one bile acid， and one terpenoid. KEGG signaling pathway enrichment analysis revealed significant enrichment in signaling pathways associated with neural excitation， fat metabolism， and inflammation regulation including caffeine metabolism， glycerophospholipid metabolism， fatty acid metabolism， and modulation of TRP channels by inflammatory mediators.【Conclusion】 Long-term intervention of intragastric gavage of rock tea solubles may exert beneficial health effects by influencing serum metabolites and involving in regulations of signaling pathways associated with lipid and glucose metabolism， amino acid metabolism， anti-inflammation， immunity，and anti-fatigue.