Electrooxidation of glucose in alkaline media is a critical issue as a clean source for Fuel cell application. The using of noble metals as anodes obstructs the economic cost of the glucose fuel cells. That is why the trend is towards the recent doped metal oxides designs as low-cost anodes electrocatalysts. Here, Nickel ferrite (NiFe2O4) nanoparticles (NFNPs) were simply fabricated by the formation of citrate framework followed by combustion and calcination as a promising anode for glucose fuel cells. The as-designed NFNPs were investigated in terms of FE-SEM, TEM, EDX and XRD analysis. The surface morphology has a sponge-like structure with an irregular shape of pores. The successful formation of a porous network can be related to the releasing of considerable amounts of gaseous products during the combustion process at 180 oC and calcination process at 400 oC. The synthesized Nickel ferrite nanoparticles showed enhanced electrocatalytic characteristics at different glucose contents up to 60.0 mM. Besides the high produced current, the enhanced stability and charge transfer were observed for the application of NFNPs in glucose electrooxidation.