Benzenthonium chloride (BEC, Hyamine 1622) is a quaternary ammonium surfactant with cationic properties widely used in cleaning, sanitation, and medical products that can become harmful to humans and also to the environment. This study aimed to evaluate its acute effects on Cyprinus carpio fish in terms of oxidative stress and morphological changes on hepatic tissue in order to show the sub-lethal toxicity of BEC. Fish were exposed to 1 mg/L BEC for 24, 48, and 96 h, and the liver samples were collected. The most significant changes were noticed after 96 h of exposure when the entire antioxidant enzyme system was affected. The activities of catalase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase decreased by 44%, 31%, 30%, and 45%, respectively, compared to control. Glucose-6-phosphate dehydrogenase activity decreased by 29% after 96 h of control, inducing a reduction of NADPH formation which decreased by half the level of reduced glutathione, the main non-enzymatic antioxidant. These effects correlated with the raised value of lipid peroxidation after 96 h and the morphology changes on hepatic tissue, such as cytoplasmic vacuolization and nuclear hypertrophy that could affect the normal function of the liver. All of these results showed acute toxicity of BEC on C. carpio after 96 h of exposure, causing oxidative stress response at the hepatic level.