Abstract: In order to explore the antimicrobial susceptibility of Staphylococcus epidermidis to florfenicol injection in southern Xinjiang Uygur Autonomous Region, in this experiment, microbroth dilution method was used to determine the minimum inhibitory concentration（MIC） and minimum bactericidal concentration（MBC） of florfenicol injection against Staphylococcus epidermis ATCC 35984 and clinical isolates; the effect on the growth of Staphylococcus epidermidis and the formation of biofilm and surface hydrophobicity were determined; the antibacterial effect of florfenicol injection in vitro and its effect on the integrity of the cell membrane of Staphylococcus epidermidis were determined. The results showed that the MIC values of florfenicol injection for Staphylococcus epidermis ATCC 35984 and clinical isolates were 4 and 2 μg/mL, respectively, and MBC values of them were 16 and 8 μg/mL, respectively. Compared with the bacterial solution without florfenicol injection, 0.5 μg/mL florfenicol injection could significantly inhibit the growth of Staphylococcus epidermidis and 0.25 μg/mL florfenicol injection could significantly inhibit the formation of its biofilm（P＜0.05）. When the concentration was higher than 2 μg/mL, the inhibition effect was very significant（P＜0.01）, which was concentration-dependent. Florfenicol injection could reduce the surface hydrophobicity of Staphylococcus epidermidis and was concentration-dependent. When the concentration of florfenicol injection was 1/2 MIC, it significantly inhibited the growth of Staphylococcus epidermidis ATCC 35984（P＜0.05）, when the concentration was higher than 1 MIC, the growth of Staphylococcus epidermidis ATCC 35984 and clinical isolates could be significantly inhibited in vitro（P＜0.01）. Florfenicol injection could impair the cell membrane integrity of Staphylococcus epidermidis. The results suggested that Staphylococcus epidermidis in southern Xinjiang Uygur Autonomous Region had good drug sensitivity to florfenicol injection, and florfenicol might play bacteriostatic and bactericidal effects by reducing the hydrophobicity on the cell surface of Staphylococcus epidermidis and damaging the cell membrane integrity of Staphylococcus epidermidis.