In this work, a simple, facile, and sensitive magnetic solid-phase extraction method was developed for the extraction and enrichment of three representative steroid hormones before high-performance liquid chromatography analysis. Gold-modified Fe O nanoparticles, as novel magnetic adsorbents, were prepared by a rapid and environmentally friendly procedure in which polydopamine served as the reductant as well as the stabilizer for the gold nanoparticles, thus successfully avoiding the use of some toxic reagents. To obtain maximum extraction efficiency, several significant factors affecting the preconcentration steps, including the amount of adsorbent, extraction time, pH of the sample solution, and the desorption conditions, were optimized, and the enrichment factors for three steroids were all higher than 90. The validity of the established method was evaluated and good analytical characteristics were obtained. A wide linearity range (0.8-500 μg/L for all the analytes) was attained with good correlation (R ≥ 0.991). The low limits of detection were 0.20-0.25 μg/L, and the relative standard deviations ranged from 0.83 to 4.63%, demonstrating a good precision. The proposed method was also successfully applied to the extraction and analysis of steroids in urine, milk, and water samples with satisfactory results, which showed its reliability and feasibility in real sample analysis.