Research has demonstrated the presence of a variety of chemical substances in the essential oils of species belonging to the Croton genus and its potential insecticide. The objective of this research was to evaluate the larvicidal activity of the essential oil obtained from the aerial part of Croton tetradenius on Aedes aegypti, as well as its yield and chemical composition, in relation to different periods of collection of the botanical material. Eight essential oil concentrations were evaluated, using 4 replicates per treatment, using 30 larvae per replicate, totaling 120 larvae per treatment. The chemical composition analysis was carried out using Gas Chromatography coupled to Mass Spectrometry. The yield of the essential oil in the extraction carried out in February was lower. The most toxic essential oil for A. aegypti larvae was extracted in August. The analysis of the chemical composition revealed the presence of 60, 48 and 62 compounds in the essential oils for the months of February, May and August, respectively. The essential oil obtained from the aerial part of C. tetradenius has shown to be promising for use in A. aegypti integrated control programs, however, in order to maximize this toxic effect, the collection period should be taken into account.