Hydrodistillation of Brazilian basil (Ocimum basilicum L.) yielded 0.4% of essential oil (EO), with activity against eight important postharvest deteriorating fungi. The crude EO completely inhibited the radial growth of the fungi at a concentration of 0.1%. Thirty-two components were identified in crude EO by Kováts retention index, mass spectrometry and standards. Preparative TLC-bioautography of the crude EO presented only one band with an antifungal activity greater than that of crude EO with the following chemical composition (%): 1,8-cineole (6.2), linalool (36.3), camphor (9.7), Alpha-terpineol (3.8), methyl chavicol (9.1) and eugenol (34.9). Based on these results, these compounds were considered to be the major fungitoxic components. This is the first study in the literature that presents data obtained simultaneously on the activity of crude basil EO against eight important post-harvest deteriorating fungi, its unequivocal chemical identification and quantification (area %).