Fabrication of binary transition metal oxide-based composite over the active surface area of glassy carbon electrode (GCE) stimulates the electrochemical activity due to their morphology, and higher active surface area, etc. The successful formation of (NiO/ZnO) composite has been scrutinized through various microscopic and spectroscopic techniques. Further, NiO/ZnO/GCE tends to exemplify exquisite kinetic performance with a higher electron transfer rate and charge transfer resistance (Rct) of 404 Ω. The electrochemical detection of Rutin (RT) on NiO/ZnO/GCE exemplifies a spacious linear range of 0.049 – 742.5 µM in linear sweep voltammetry (LSV) and differential pulse voltammetry (DPV). Moreover, the proposed NiO/ZnO/GCE sensor symbolizes Nanomolar level detection of RT (11.0 & 13.6 nM) in DPV and LSV with an appraisable sensitivity of 2.52 µA µM−1 cm−2 and 0.62 µA µM−1 cm−2, respectively. The practical feasibility analysis of the proposed NiO/ZnO/GCE sensor features exemplary selectivity, repeatability, reproducibility, and storage stability. On top of that, the real-time monitoring analysis demonstrates exquisite recovery results in blood and orange samples.