This study aimed to identify the active components of Dingshisanyangtoujie Decoction Dry Extract and drug-containing serum by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap/MS/MS) and explore the material basis of pharmacodynamics of Dingshisanyangtoujie Decoction.Methods:Firstly，the main components of Dingshisanyangtoujie Decoction Dry Extract，drug-containing serum，and blank serum were identified by UPLC-Q-Orbitrap/MS/MS，and Xcalibur software was used for data acquisition and analysis.The mass spectral peaks were identified according to the component cleavage database，and the active components of Dingshisanyangtoujie Decoction were determined.The pharmacological action of the active components was clarified by bioinformatics analysis.Results:A total of 18 effective active components of Dingshisanyangtoujie Decoction were identified，including luteolin，4'-O-methoxypuerarin，aloe emodin-8-O-glucoside，naringenin，emodin-8-O-β-D-glucoside，gallic acid，baicalin，and kaempferol.The results of bioinformatics analysis showed that the main active components of Dingshisanyangtoujie Decoction could act on inflammatory diseases，digestive system diseases，and cardiovascular system diseases.Dingshisanyangtoujie Decoction mainly played a role through oxidative phosphorylation，peroxisome proliferator-activated receptor(PPAR) inflammatory signaling pathway，steroid hormone biosynthesis，and fatty acid metabolism pathways.Conclusion:Eighteen effective active components of Dingshisanyangtoujie Decoction are successfully identified，and Dingshisanyangtoujie Decoction has the pharmacological effect of “multiple components，multiple targets，and multiple pathways”.