The essential oil (EO) from the leaves of Onychopetalum periquino, obtained by hydrodistillation, was analyzed by gas chromatography coupled with mass spectrometry (GC-MS), and also was investigated for its larvicidal activity against Aedes aegypti larvae. Thirteen compounds, representing 91.31% of the crude oil, were identified.Major compounds were sesquiterpenes, including β-elemene (53.16%), spathulenol (11.94%) and β-selinene (9.25%). The EO showed high larvicidal activity with a lethal concentration (LC 50 ) of 63.75 μg/mL and 100% mortality at 200 μg/mL. These results represent the first report about the chemical composition of O. periquino and the first larvicidal evaluation with Onychopetalum species.