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The hyaluronic acid receptor is typically overexpressed in cancer cells, and hyaluronic acid targeting has gained attention in cancer treatment. In this work, cytotoxic potential of magnetic iron oxide nanoparticles co-conjugated with hyaluronic acid and limonene in the MDA-MB-231 cell line was characterized. Physicochemical characteristics of the nanoparticles were studied by FT-IR, XRD, TEM, SEM-EDS, DLS, zeta potential and VSM analyses. Cytotoxic effects of Fe₃O₄@Glu-HA-LIM NPs in cancer and normal cells were studied by MTT assay. Flow cytometry and AO/PI staining were employed to study cell cycle phases and cell apoptosis levels in the cancer cells. ROS levels in the treated and control cells were also quantified. According to physicochemical characterization, Fe₃O₄@Glu-HA-LIM NPs were correctly synthesized and exhibited spherical shape, particle diameter of 29 to 69 nm, surface charge of -52.5mV, hydrodynamic size of 245.4 nm and magnetic saturation of 60.607emu/g at 9000Oe. The 24 -hour IC₅₀ of Fe₃O₄@Glu-HA-LIM NPs in the MDA-MB-231 cell line was 227 µg/mL and Fe₃O₄@Glu-HA-LIM NP displayed lower cytotoxicity in the normal cell line (24-hour IC₅₀=483 µg/mL), which is potentially attributed to hyaluronic acid receptor targeting and induction of significant oxidative stress in cancer cells. Treatment of cancer cells with Fe₃O₄@Glu-HA-LIM NPs caused cell cycle arrest mainly at the S and G2/M phases. Moreover, exposure to Fe₃O₄@Glu-HA-LIM NPs elevated ROS levels in the cancer cell line by 9.27 folds. This study demonstrates the promising cytotoxic potential of Fe₃O₄@Glu-HA-LIM NPs in the MDA-MB-231 cell line, which can be considered for chemotherapy of triple-negative breast cancer.