Increased climate change and human mobility have led to the ecological expansion of highly harmful insect species. Mosquitoes have a major role in transmitting various disease agents that represent significant threats to human and animal health. Phytochemicals offer bio-safe and eco-friendly alternatives for combating disease vectors. This study investigated the effect of margosa oil extract on the proteins' biochemical components, indicating an increase in total protein values of 0.341 and 0.70 for larvae and pupa, respectively. Both groups of acid and alkaline phosphatase increased to 48.40 and 126.53 after 24 h post-treatment of the third larval instar and increased in the pupa stage with values of 11.82 and 46.66, respectively. In addition, acetylcholinesterase raised to 602.40 in the third larval instar and 297.7 in pupa stages after 48 h. of treatment. Up-and-down regulation of these enzymes in different stages and periods of treatment indicate that they disrupt metabolic homeostasis in target mosquitoes. The results confirm that margosa oil and its components have a promising effect as larvicides for mosquito vector control.