Glucose level surveillance in mankind is considered the greatest need of recent times to achieve healthy lifestyle goals due to its health-related concern for humans. To fulfill the need, Nickel-based compounds emerge with excellent bio-mimic properties. However, due to the poor electrical properties of oxides and hydroxides of Ni, developing a supporting base that can enhance electron transportation is mandatory. The present work is dedicated to non-enzymatic glucose detection by electrochemical means on the Ni3N thin film on an Indium-doped Tin Oxide (ITO) substrate as a working electrode in an alkaline medium. The Ni3N films use HiPIMS as a deposition technique due to its high ionization rate of sputtered species and dense plasma which helps the Ni ions to react with nitrogen in the formation of a perfect Ni3N phase, and it is a green process that can contribute to sustainable manufacturing. Ni3N thin film deposited at a 60 % N2 flow ratio shows the largest crystallinity and catalytic properties, which helps this modified electrode to have significant potential for glucose detection. The Ni3N modified electrode acquires a lower linear range of 0.001–1.25 mM with sensitivity and Limit of Detection (LOD) of 337.46 µAmM–1cm−2, and 0.78 µM, respectively, and a higher linear range of 1.25–7.3161 mM with a sensitivity of 158.58 µAmM–1cm−2. The amperometric measurements reveal the fast response of the electrode towards glucose detection at 2.46 secs. The key measures of the electrode according to the corresponding measurements are satisfactory selectivity, reproducibility, repeatability, and operational stability. The modified electrode shows a significant response for real samples such as commercially available honey, apple juice, and biological fluids.