Trimeresurus stejnegeri is one of the top ten venomous snakes in China, and its bite causes acute and severe diseases. Elucidating the metabolic changes of the body caused by Trimeresurus stejnegeri bite will be beneficial to the diagnosis and treatment of snakebite. Thus, an animal pig model of Trimeresurus stejnegeri bite was established, and then the metabolites of serum and urine were subse-quently screened and identified in both ESI+ and ESI-modes identified by ultra-performance liquid chro-matography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS) methods. There are 9 dif-ferential metabolites in serum, including Oleic acid, Lithocholic acid, Deoxycholic acid, Hypoxanthine, etc. There are 11 differential metabolites in urine, including Dopamine, Thiocysteine, Arginine, Indoleac-etaldehyde, etc. Serum enrichment pathway analysis showed that 5 metabolic pathways, including Tryp-tophanuria, Liver disease due to cystic fibrosis, Hartnup disease, Hyperbaric oxygen exposure and Biliary cirrhosis, the core metabolites in these pathways, including deoxycholic acid, lithocholic acid, trypto-phan and hypoxanthine, changed significantly. Urine enrichment pathway analysis showed that 4 meta-bolic pathways, including Aromatic L-Amino Acid Decarboxylase, Vitiligo, Blue Diaper Syndrome and Hyperargininemia, the core metabolites in these pathways including dopamine, 5-hydroxyindole acetic acid and arginine. Taken together, the current study has successfully established an animal model of Tri-meresurus stejnegeri bite, and identified the metabolic markers and metabolic pathways of Trimeresurus stejnegeri bite. These metabolites and pathways may have potential application value and provide a thera-peutic basis for the treatment of Trimeresurus stejnegeri bite.