An ultrasensitive platform is accessible for the selective detection of chloramphenicol (CLP) based on NiO/ZnO inclusion of higher electrical conductivity and surface area has promoted voltammetry detection. The NiO/ZnO material has been prepared through an eco-friendly co-precipitation (CPT) method at room temperature. Here, we have designed and synthesized an effective electrocatalyst of NiO/ZnO with an average size of approximately 55.04 nm through a one-step synthetic protocol. The potential utility of the developed electrode was proved by applying them to the analytical determination of CLP in real samples. The NiO/ZnO modified SPCE demonstrated a significantly higher electrocatalytic effect for CLP reduction, at the potential of – 0.62 V in pH 7.0. The calibration curve for CLP linear range is 0.05 μM – 418.5 μM, and the detection limit is low as 0.07 μM. It is concluded the NiO/ZnO electrode is an effective electrode mediator for the selective detection of CLP. The homogenous hexagonal plate-like NiO/ZnO modified SPCE showed exceptional performance for the CLP determination in real food samples (milk and honey) and biotic samples (human urine).