In this study, magnetic coupled with Magnetic dispersive micro-solid phase extraction was applied for simultaneous separation and preconcentration of sulfadiazine, sulfamethoxazole, and sulfamethazine and determination by HPLC-UV. The effect of important parameters including amount of magnetic sorbent, type and volume of elution solvent, extraction time, sample volume, and desorption time were assessed. At the optimum conditions, the linear ranges of 5-250 (µg L-1) were for three analytes. Moreover, the LODs (S/N = 3) of 1.8, 1.6, and 1.7 (µg L-1), and the LOQ (S/N = 10) of 6.1, 5.5, and 5.9 (µg L-1) were acquired for sulfadiazine, sulfamethoxazole, and sulfamethazine respectively. The efficiencies of ≤ 97.3 % and the relative standard deviations of less than 3.5 % were also obtained for application of the method in complex matrixes.