In this research, polystyrene nanofibers were synthesized by electrospinning method and used as adsorbent in solid-phase extraction of Disulfine blue from aqueous solutions. Some important parameters affecting extraction efficiency including sample flow rate through the adsorbent, ionic strength, pH of the sample solution, weight of adsorbent, volume, and type of eluent were evaluated and optimized. Under the optimized conditions, the calibration graph was linear in the range of 6.4-1000 mu g L-1 with correlation coefficient (r(2)) of 0.9962. The limit of detection and enhancement factor of proposed method, for extraction from 50mL sample solution, were obtained as 2 mu g L-1 and 9.5, respectively. Finally, the proposed method was successfully applied for extraction and measurement of Disulfine blue from real samples, and satisfactory results were obtained.