Endocrine Disruptors(EDs) substances are able to alter the function of the endocrine system and may cause adverse effects. The aim of this work was to optimize the suitable condition of extraction & cleanup in twelve pesticides of Endocrine Disruports from the Sea sand matrix. For the first experiment, three extraction methods were compared to determine better recovery efficiencies using Accelerated Solvent Extractor(ASE), Soxhlet and Sonication extractor from Sea sand sample. The percent recovery of twelve analytes of ASE, Soxhlet and Sonication methods ranged from 47.3% to 111.8%(ASE), 61.4% to 118.9%(soxhlet) and 69.3% to 111.1%(Sonication), respectively. Generally, The experiment results shows that Sonication extraction obtained the highest recovery rates of three extraction methods in the most of analytes. For the second experiments, the extraction efficiencies of various gradient solvents(20% A/H, 50% A/H, 100% Acetone) were compared by sonication solvent extraction method. The mean recovery of twelve analytes using 20%A/H, 50%A/H and 100%(Acetone) obtained 93.1%(20%A/H), 101.3%(50%A/H), 106.4%(100%Acetone), respectively. Generally, the results shows that 50% Acetone/n-Hexane obtained the optimum recovery rates in the most of analytes. For the third experiments, the efficiencies of the cleanup procedure were checked by various gradient solvents(10% A/H, 20% A/H, 50% A/H). The mean recovery of twelve analytes using 10%A/H, 20%A/H and 50% A/H obtained 56.9%(10%A/H), 92.7%(20%A/H), 93.4%(50%A/H), respectively. Generally, the results shows that 20% Acetone/n-Hexane obtained the optimum recovery rates in the most of analytes.