[Objective] To explore the antibacterial effect of 17 Chinese herbs and their compounds on Staphylococcus aureus in dairy cow mastitis in vitro. [Methods] Staphylococcus aureus isolated and preserved from milk samples of cows with mastitis in a dairy farm in Shandong Province was used as the study object. Drug sensitivity test and tube double dilution method were used to determine the drug sensitivity of Staphylococcus aureus to 17 traditional Chinese medicines, and the minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC) were determined. In this study, a combination of two Chinese herbs highly sensitive to Staphylococcus aureus was selected to make a compound compound. The MIC of the compound against Staphylococcus aureus was determined by microbroth dilution checkerboard method, and the graded inhibitory concentration (FIC) of the compound was calculated. [Results] Scutellaria baicalensis, forsythia and gallnut were highly sensitive to Staphylococcus aureus. The results of MIC and MBC detection for Staphylococcus aureus were 7.813 mg/mL and 15.625 mg/mL, respectively. The results of MIC and MBC detected by forsythia and gallnut were 15.625 mg/mL and 31.25 mg/mL, respectively. The results of the detection of MIC for Staphylococcus aureus were 15.625 mg/mL for scutellaria baicalensis + forsythia, scutellaria baicalensis + gallnut and forsythia + gallnut, and there was no correlation between the two compounds. [Conclusion] The antibacterial effect of staphylococcus aureus on dairy cow mastitis may not be enhanced after the composition of single Chinese herbs.