The fabrication of enzyme‐free glucose sensors is highly demanded for the biological, clinical, and food applications. In this study, we have developed a green method for tuning the surface properties of nickel‐cobalt bimetallic oxide (NiCo2O4) by adding mustard (Brassica Campestris) leaves extract during hydrothermal growth. The mustard (Brassica Campestris) leaves extract is rich with a variety of phytochemicals, which can easily tune the surface properties of NiCo2O4 nanostructures, thereby paving the way toward the development of sensitive and selective non‐enzymatic glucose sensors. The effect of various amounts of mustard (Brassica Campestris) leaves extract (0–20 ml) was also studied to find out the optimal conditions for growing surface‐modified NiCo2O4 nanostructures. The morphology and crystalline structure of the nanomaterials were studied by scanning electron microscopy (SEM) and powder X‐ray diffraction (XRD) techniques, respectively. The presence of an increasing quantity of mustard (Brassica Campestris) extract keeps the crystalline structure and the morphology of the NiCo2O4 nanostructures unaltered but changes their dimensions. All nanostructures show the same cubic spinel structure of NiCo2O4 and a morphology of spherical urchins composed of nanorods, but the diameter of the urchins decreases from ~10 μm to several nanometers, thus increasing the surface area of the nanomaterial. Furthermore, NiCo2O4 nanostructures were deposited onto glassy carbon electrodes (CGE), showing excellent catalytic properties toward the enzyme‐free detection of glucose using cyclic voltammetry. Importantly, the intensity of the oxidation current peak was linear over a wide range of glucose concentrations (from 0.1 to 10 mM) and the limit of detection (LOD) was estimated around 0.001 mM. Additionally, NiCo2O4 nanostructures grown in the presence of 20 ml of mustard leaf extract demonstrated good repeatability and excellent selectivity for glucose, without interference by other components such as urea, lactic acid, uric acid, ascorbic acid, as well as potassium and sodium ions. The combined results attest that mustard leaf extract has high potential as a green approach to improve the electrochemical properties of nanostructured materials, and could be useful for a wide range of materials for future electrochemical applications.