Fabricating a highly active and stable nanocatalyst material displays a great concern when constructing a commercially viable ethylene glycol (EG) fuel cell. Herein, nickel oxide nanostructures were grown onto graphite (NO/T) using coprecipitation and calcination protocol at different temperatures. Techniques for XRD, TEM, SEM, and EDX investigations were used to look into the produced crystal planes, shape, and elemental mapping of synthesized nickel oxide nanoparticles. Different NO/T nanocatalyst electroactivities were examined in order to oxidize EG molecules in basic electrolyte. The surface area values of calcined nanostructures at 200°C and 300°C were much higher than those measured at NO/T‐400 by 7.62 and 3.71 folds to explain their outstanding performance. Some kinetic information for varied NO/T nanocatalysts was derived including the electron transfer coefficient, rate constant, and surface coverage values. Electron transfer rate constants of 0.1946, 0.3734, 0.0113, and 0.0303 s−1 were calculated at NO/T‐200, NO/T‐300, NO/T‐400, and NO/T‐500, respectively. Increased oxidation current densities could be achieved when NO/T nanomaterials were subjected to lowered calcination temperatures. NO/T‐200 and NO/T‐300 nanocatalysts also displayed decreased Eonset for alcohol oxidation process by 20 and 41 mV in relation to that at NO/T‐400. Moreover, chronoamperometric experiments revealed the prevalence of the stable behavior during EG oxidation at these nanostructures, especially for calcined ones at 200°C and 300°C. Much reduced poisoning rates were measured at NO/T‐200 (0.171 s−1) and NO/T‐300 (0.067 s−1) when contrasted to that at NO/T‐500 (0.727 s−1). Increasing the alcohol and supporting electrolyte concentrations was beneficial in improving the charge transfer characteristics of NO/T nanocatalysts as demonstrated by EIS measurements. This study supports the promising activity of NiO nanoparticles onto graphite as anode materials for direct alcohol fuel cells.