Vitex gardneriana Schauer (Lamiaceae) popularly known as “jaramataia”, is a shrub commonly found in caatinga biome located in Northeast Brazil. In folk medicine, its leaves have been used as analgesic and anti-inflammatory agents. The chemical composition of the essential oil from leaves obtained by hydrodistillation was analyzed and identified by GC-MS and GC-FID and showing a total of 26 constituents (95.9%) being 2 monoterpenes (0.4%) and 24 sesquiterpenes (95.4%). The main constituents identified were cis-calamenene (29.7%), 6,9-guaiadiene (14.5%) and caryophyllene oxide (14.0%). The essential oil has been demonstrated high larvicidal activity againstAedes aegypti (LC50 = 28.0 μg/mL). In the evaluation of the bioassay with Artemia salina the essential oil showed LC50 = 98.11 μg/mL. Inaddition, the essential oil did not show cytotoxicity (IC50 > 2.50 mg/mL) by the hemolysis assay.