The sensitive detection of trace amounts of ofloxacin is very significant in many aspects. Fortunately, creating a new kind of electrode material with a low resistance, wide surface area, and stable microstructures remains a challenging task. In this work, we describe the simple method for fabricating a novel disposable glassy carbon electrode modified by ceramic cerium ferrite CeFeO3 (CFO) for ofloxacin detection. The nanomaterial has been characterized using a series of characterizations such as X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Field emission scanning electron microscopic (FESEM), transmission electron microscopy (TEM) showing the physico-chemical properties of CFO. Under optimum conditions, the CFO based sensor demonstrated a quick and more sensitive voltammetry response towards the ofloxacin oxidation. The calibration curve for ofloxacin is linear in the range from 1 to 55 µM, and the detection limit is as low as 0.06 µM. Furthermore, the repeatability, reproducibility, and stability of the present sensor done with satisfactory results. According to the electrochemical experiment, the synthesized hexagon prism shaped CFO provide strong electron mediator behaviors for the oxidation of ofloxacin. Finally, after basic sample preparation rapidly and reliable determination of ofloxacin in human urine samples was achieved by employing the standard addition process.