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In the present study, restricted-access material based on surfactant-coated dodecyl-functionalized magnetite nanoparticles (C12-Fe3O4) were prepared and developed to extract steroid hormones from environmental and biological samples.The magnetite nanoparticles were synthesized by a solvothermal reduction method, and functionalized with dodecyltriethoxysilane.Furthermore, C12-Fe3O4 nanoparticles were modified with different surfactants by self-assembly adsorption.Several types of surfactants, Tween 20, Tween 40, Tween 60 and Tween 80, Span 40, Span 60 and Span 85 were investigated.Tween 20-C12-Fe3O4 and Tween 40-C12-Fe3O4 showed good anti-interference ability.Besides, surfactant modified C12-Fe3O4 exhibited good dispersibiility in aqueous solution, which could be appreciated in extraction.The factors that may influence the extraction, including the amount of magnetite nanoparticles, extraction/desorption time, the amount of salt addition, the type and volume of desorption solvent, the volume of methanol addition and pH of sample solution, were investigated.Tween 20-Fe3O4-C12 nanoparticles were found to give the satisfactory recovery, reproducbilities and low LODs.This novel material was promising as an anti-interference adsorbent for complex sample.