Objective: An ultrafast liquid chromatography-hybrid triple quadrupole/linear ion trap mass spectrometry (UFLC-MS/MS) combined with statistical analysis method was used to investigate the changes of endogenous small molecule compounds and nephrotoxic mechanism in the rat serum induced by Radix et Rhizoma Tripterygii and to explore biomarkers associated with renal toxicity as well as metabolism pathways. Method: After intragastric administration for 14 days

serum biochemical indexes was detected by kit method and nephridial tissue section was observed.The serum samples were added acetonitrile for protein precipitation and analyzed by UFLC-MS/MS with full scan mode under positive ion scan mode

then principle component analysis (PCA) and principle component analysis discriminant analysis (PCA-DA) were used for data processing. Result: Compared with the blank group

serum biochemical indexes[creatinine (Cr) and blood urea nitrogen (BUN)] of Radix et Rhizoma Tripterygii ethanol extract group increased significantly and histopathological section damaged obviously which showed that the animal module prepared well.Nine significant different metabolites

such as dimethylglycine

lysine

phenylalanine

lysolecithin

phosphatidyl choline

phosphatidylserine and diglyceride

were identified as biomarkers of nephrotoxicity induced by Radix et Rhizoma Tripterygii. Conclusion: Nephrotoxic mechanism induced by Radix et Rhizoma Tripterygii may be related to the metabolism of amino acid and phospholipids

and it provides reference relating to the early clinical toxicity prevention of Radix et Rhizoma Tripterygii.