In this paper we describe the synthesis, characterization, and use of a glucose-assisted magnetic aluminum oxyhydroxide@carbon (γ-AlOOH/Fe₃O₄@C) nanocomposite as a novel sorbent for the extraction and pre-concentration of deferasirox (DFX) present in plasma and urine samples using the ultrasonic-assisted dispersive solid-phase microextraction (UA-DSPME) method prior to HPLC-UV measurement. The sorption process was systemically optimized by experimental design and statistical analysis to maximize the extraction efficiency of DFX. The proposed method could be successfully employed for DFX determination in double-distilled water, plasma, and urine samples (real and spiked samples), with recovery range of 96.1–105%. Also, a good linear range from 10 to 3500 ng mL⁻¹ was found with a correlation value of 0.9938, and relative recoveries between 2.09 and 5.22% were achieved adequately. Furthermore, the limit of quantification (LOQ) and the limit of detection (LOD) were found to be 5.421 and 0.163 ng mL⁻¹ (based on a signal-to-noise ratio of 3) which were acceptable values compared to those of other reports.