With the aim of developing an electrochemical sensor for the detection of glutamate, in the present study, Ni@NC nanoparticles were prepared by embedding Ni derived from Ni-based metal–organic frameworks in N-enriched carbon via a high-temperature calcination process. Further, the resulting material was investigated as an electrode modifier for sensor applications. Cyclic voltammetry measurements revealed that the Ni@NC nanoparticles prepared at 700 °C exhibited the most prominent activity for the electrocatalytic oxidation of glutamate, affording an amperometric response in a linear range of glutamate concentration of 5 × 10−3–500 μM with a low detection limit of 1.67 × 10−3 μM (S/N = 3), which was comparable with previously reported data. Furthermore, this electrochemical sensor demonstrated a high rate of recovery in real samples, providing a valuable method for the rapid detection of glutamate. As a potential candidate for glutamate sensing, the Ni@NC system can be expected to find application in many biomedical fields.