The copper-coated iron oxide nanoparticles with core-shell were produced by deposition of a Cu shell on Fe3O4 NPs through reduction of Cu2+ ions in solution using NaBH4. Subsequently, the organosulfur compound, bis-(2,4,4-trimethylpentyl)-dithiophosphinic acid (b-TMP-DTPA), was used to form self-assembled monolayer in order to modify sorbent's surface via covalent bonding between Cu and thiol (-SH) terminal groups. The prepared magnetic nanoparticles were characterized by using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), transmission electron microscope (TEM), vibrating sample magnetometer (VSM) and thermo gravimetric analysis (TGA). Then, the application of this new sorbent was investigated to extract the steroid drugs in ointment samples with the aid of ultrasound.An external magnetic field was applied to collect the magnetic nanoparticles (MNPs). The extracted analytes were desorbed using acetonitrile. The obtained extraction solution was analyzed by HPLC-UV. The main affecting factors on the extraction efficiency including pH, sonication time, amount of sorbent, salt concentration, and desorption conditions were optimized in detail. Under the optimum conditions, good linearity was obtained in the range of 2.5-250.0 μg/ L with reasonable linearity (R2 > 0.99) and the limits of detection (LODs) ranged between 0.5 and 1.0 μg/L (based on S/N = 3). Repeatability (intra-day precision) based on five replicates and preconcentration factors were calculated to be 3.6％-4.7％ and 87116,respectively.Relative recoveries in ointment samples at two spiked levels of the target analytes were obtained in the range of 90.0％-103.2％. The results illustrated that the Fe3O4@Cu@ b-TMP-DTPA NPs have the capability of extraction of steroid drugs from ointment samples.