Eugenia pitanga (O.Berg) Nied., Myrtaceae, popularly known as pitanga-do-campo, is a species native from Pampa biome, in southern of Brazil and is popularly used as antidiarrheal, anti-rheumatic, stimulant and febrifuge. The aim of this work was to determine the chemical composition of essential oil from fresh leaves of Eugenia pitanga (O.Berg) Nied. Preliminaries data about antileishmanial activity in vitro of essential oil against promastigotes forms of Leishmania amazonensis are presented. The method to determine the chemical composition of the essential oil from fresh leaves of E. pitanga (EoEp) was gas chromatography-mass spectrometry. EoEp was assayed in vitro for antileishmanial activity against promastigotes of Leishmania amazonensis. The EoEp comprised 57 components and was rich in oxygenated sesquiterpenes (72.97 %) with spathulenol (15.34 %), globulol (10.93 %) and (2E,6E) methyl farnesoate (7.40 %) as the major constituents. EoEp was effective against L. amazonensis promastigotes with IC50 value of 6.10 ± 1.80 μg/mL. This work reports for the first time the composition of essential oil from leaves of E. pitanga from Pampa Biome, and it suggests the importance of continuing the studies in order to explain the antileishmanial activity.