The paper uses fluorescence spectroscopy and ultraviolet-visible absorption spectroscopy to study the effect of ultrasonic treatment (40 kHz / 30 min， 50 kHz / 30 min) on the binding of chlorogenic acid (CGA) and bovine serum albumin(BSA). Analyze the fluorescence quenching effect of CGA on BSA by fluorescence emission spectroscopy； analyze the microenvironmental changes of BSA tyrosine (Tyr) and tryptophan (Trp) residues after CGA binding by synchronous fluorescence spectroscopy to determine the effect of CGA on the conformation of BSA； use mathematical equations such as Stern-Volmer equation and Acharya equation to calculate its quenching constant， binding constant， and number of binding sites； then use warfarin (WARF) and ibuprofen (ibuprofen， IBU) two protein markers to determine the binding site； and use ultraviolet spectroscopy to measure the changes in BSA conformation to compare the effects of different ultrasonic frequencies. The results showed that CGA had the most significant fluorescence quenching effect on BSA at the ultrasonic frequency of 53 kHz， and the conformation of CGA-BSA complex could be changed by ultrasonic treatment to some extent， and the combination of CGA and BSA could be enhanced to some extent， but the reaction force type would not be changed. The results of this study will provide a useful reference for the effect of sonication on the binding of protein and ligand.