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This work reports an enzyme-less glutamate sensor based on nickel oxide modified glassy carbon electrode. The nanoparticles of NiO were synthesized by sol–gel method, and showed high electro-catalytic activity towards glutamate oxidation in 0.1 M NaOH solution. The sensor displays a fast response time of < 5 s and a linear dependency of R
2
0.997 for glutamate concentration of up to 8 mM at an applied potential of + 0.55 V with a sensitivity of 11 µA mM
−1
cm
−2
and a detection limit of 272 μM (S/N = 3). The interference studies of NiO/GC electrode showed a significant current response for glutamate in the presence of uric acid and ascorbic acid. Thereby, this simple NiO nanoparticle based sensor can be promising platform for the development of a robust enzymeless glutamate sensor.