Objective: To establish a method for the determination of bisphenol S in urine using dispersive liquid-liquid microextraction (DLLME) coupled with high performance liquid chromatography (HPLC) . Methods: The acetonitrile, octanol were used as extraction solvent, dispersive solvent respectively, for the preconcentration of bisphenol S. The optimal extraction conditions were optimized by single factor rotations, and methodological performance index were tested. Results: The linear correlation coefficient of bisphenol S in the range of 0.0-160 μg/L is greater than 0.999. The detection limit of this method was 0.76 μg/L, and the recovery rates were 88.06%-103.81%. The intra-and inter-day precisions were 1.78%-2.85% and 2.65%-4.25%, respectively. Conclusion: The method is reliable and sensitive. It is suitable for the determination of bisphenol S in urine samples for occupational exposure populations and non-professional.