BACKGROUNDEugenia calycina is an endemic species in the Brazilian savannah (the Cerrado) and it is threatened with extinction. Several species of Eugenia are used as insecticides or insect repellents. No data are available on the larvicidal activity of E. calycina. The chemical composition of the essential oil (EO) from leaves of Eugenia calycina was analyzed by gas chromatography coupled to mass spectrometry (GC–MS) and the larvicidal activity against Aedes aegypti larvae in the third stage of development was studied.RESULTSOxygenated and non‐oxygenated sesquiterpenes were identified, and the main compounds were bicyclogermacrene, spathulenol, and β‐caryophyllene. The EO was fractionated in a chromatographic column and three compounds were isolated and identified: spathulenol, aromadendrane‐4β,10α‐diol, and 1β‐11‐dihydroxy‐5‐eudesmene. It is the first time that the last two compounds have been identified in E. calycina. The exposure times in the larvicidal test were 24 h and 48 h and the LC50 values obtained were 199.3 and 166.4 μg mL−1. The cytotoxicity of the EO in mammalian cells (HeLa and Vero) was evaluated for 24 and 48 h of incubation. The cytotoxic concentrations of the EO for HeLa and Vero cells (266.8 ± 46.5 and 312.1 ± 42.5 μg mL−1, respectively) in 48 h of exposure were higher than the LC50, showing low cytotoxicity at the concentration exhibiting larvicidal activity, resulting in a positive selectivity index.CONCLUSIONThese results indicate that the EO of E. calycina showed high activity against the A. aegypti larvae but lower cytotoxicity to mammalian cells. The leaves of E. calycina are therefore a very promising source of natural larvicidal products. © 2020 Society of Chemical Industry