Abstract—Prospects for the use of imidazolium ionic liquids (ILs) as extractants of sex steroid hormones (estrogens and androgens) in microextraction methods (dispersive liquid–liquid microextraction, DLLME, and magnetic solid-phase microextraction, mSPME) are identified. The key parameters of DLLME using C6MImNTf2 IL that affect extraction efficiency are optimized using the design of experiments technology. High recoveries (88–99%) are achieved. An approach of dynamic IL immobilization on the surface of magnetic nanoparticles (MNPs) for the extraction of steroids under mSPME conditions is proposed. Two types of MNP pre-coating are studied: hydrophilic based on silica and hydrophobic with oleic acid. The capabilities of C8MImBF4 IL as a MNP surface modifier for the efficient extraction of steroids are revealed. Optimum conditions provided high degrees of recovery (83–97%), with an exception of estriol (60%). The limits of detection are 0.26–1.29 ng/mL. Limitations of the method related to the partial removal of IL from the surface of NPs are revealed; they reduce the reproducibility of the results for estriol.