Dengue fever is a worldwide public health problem, and efforts to eradicate it have focused on controlling the dengue vector, Aedes aegypti. This study aims to assess the toxicity and effect of commercial eugenol and piperine on Ae. aegypti larvae through enzyme detoxification and histopathological changes in the midgut. Laboratory-reared Ae. aegypti larvae were treated with various concentrations of commercial eugenol and piperine and observed after 24, 48, and 72 h. Biochemical methods were used to assess detoxification enzyme activity for acetylcholinesterase, glutathione S-transferase, and oxidase, and changes in the midgut were examined using routine histological examination. In terms of larvicidal activity, piperine exceeded eugenol. Piperine and eugenol had LC₅₀ and LC₉₀ values of 3.057 and 5.543 μM, respectively, and 6.421 and 44.722 μM at 24 h. Piperine and eugenol reduced oxidase activity significantly (p < 0.05), but increased acetylcholinesterase and glutathione S-transferase activity significantly (p < 0.05). After being exposed to piperine and eugenol, the food bolus and peritrophic membrane ruptured, the epithelial layer was interrupted and irregular, the epithelial cells shrank and formed irregularly, and the microvilli became irregular in shape. Commercial piperine and eugenol behave as potential larvicides, with processes involving altered detoxifying enzymes, specifically decreased oxidase function and increased GST activity, as well as midgut histological abnormalities.