© 2019, Turkish Chemical Society. All rights reserved.Electrochemical behavior of naphazoline hydrochloride on a carbon paste electrode that was modified with aluminium oxide nanoparticles (Al2O3NPs) was investigated in a Britton-Robinson (BR) buffer (pH 7.0) using various voltammetric techniques. The results support the presence of an irreversible and diffusion-controlled electrochemical oxidation signal of naphazoline hydrochloride which is approximately at 0.9 V vs. Ag/AgCl. A selective, accurate, and simple square-wave anodic adsorptive stripping voltammetric method was proposed for naphazoline hydrochloride detection. The linear response was within the range of 5.0 × 10-8-3.0 × 10-5 mol L-1 with a detection limit of 2.6 × 10-9 mol L-1 (0.642 μg L-1). In addition, the proposed method was also utilized for naphazoline hydrochloride determination in human serum sample.