In this study platform, electrocatalytic detection of the antibiotic chloramphenicol (CAP) in phosphate buffer (pH 7) was easily achieved using a carbon paste electrode modified with NiO nanoparticles (note NiO-CPE). The peak reduction potential of chloramphenicol on the modified electrode is at (− 0.60 V/NiO-CPE vs. Ag/AgCl), its electrochemical behavior is completely irreversible, and controlled by adsorption phenomena. An excellent electrocatalytic activity has been demonstrated by the modified elaborated electrode towards the NO2 attracting group on the side chain of chloramphenicol. The structure and chemical composition of the NiO-CPE sensor were analyzed by SEM, and the X-ray diffraction results indicated that nickel oxide microcrystals were formed on the carbon sheets. The electrochemical characteristics of the NiO-CPE sensor were examined by cyclic voltammetry and electrochemical impedance spectroscopy in comparison with the unmodified carbon. Since the DPV technique allows plotting the linearity curve between the electrocatalytic current intensity of the Chloramphenicol peak and their concentration, the proposed sensor showed a remarkable detection limit of 1.08 × 10– 8 mol/L M (S/N = 3) and a wide determination range from 100 to 0.1 μM for Chloramphenicol. The developed sensor was successfully applied in the detection of Chloramphenicol in real samples.