A selective and simple biosensor was prepared by immobilizing chitosan/nickelnanoparticles/multi‐walled carbon nanotubes biocomposite on the glassy carbon electrode surface for voltammetric quantification of neotame. The properties and morphology of the modified electrode surfaces were characterized by scanning electron microscope (SEM), energy dispersive X‐ray analysis (EDX). Electro oxidation of neotame on this modified surface was examined through cyclic voltammetry (CV) and square wave voltammetry (SWV) techniques. The biocomposite modified surface (Chi/NiNPs/MWCNTs/GCE) proposed in this study showed good electrocatalytic activity for neotame with an improved voltammetric peak current at 1.004 V, unlike the bare glassy carbon electrode (GCE) surface and several other modified surfaces. Under optimum conditions, Chi/NiNPs/MWCNTs/GCE gave linear SWV responses at the range of 2 μM ∼50 μM for neotame with 0.84 μM determination limit. This voltammetric sensor was successfully employed for the quantification of neotame on food samples and showed long‐term stability, advanced voltammetric behavior, and good repeatability. Selective, accurate, and precise determination of neotame highlight the importance of this electrode in monitoring the control of food additives and ensures attract a great deal of attention.