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Aiming to valorise the Atlantic Rainforest biodiversity in Santa Catarina, the chemical characterization of the essential oils (EOs) from leaves of Vernonanthura montevidensis (Spreng.) H. Rob. is described for the first time. Fresh leaves collected in the year 2014 and 2015, were submitted to hydrodistillation to give pale blue EOs in yields of 0.21 and 0.19%, respectively. The EOs were characterized by GC-MS and GC-FID quantitative method. The monoterpene β-pinene was the major constituent in both samples reaching a maximum of 26.3%. Besides β-pinene, the monoterpene α-pinene and the sesquiterpene β-caryophyllene, were among the major constituents in both samples. By means of extracted ion chromatogram procedure, it was possible to detect chamazulene, which was associated to the pale blue colour of the essential oils. In the in vitro antimollicute assays, the essential oil was moderately active against Mycoplasma genitalium and M. pneumoniae with MIC values of 250 µg mL -1 .