The hydrodistilled essential oil from flowering aerial parts of Ocimum gratissimum L. (Lamiaceae) growing desolately in South India was examined to determine its composition. The oil was analyzed by gas chromatography equipped with flame ionization detector (GC–FID) and gas chromatography coupled with mass spectrometry (GC– MS). Forty-one constituents were identified, representing 99.4% of the total oil. The main components were identified as eugenol (57.1%), α-bulnesene (15.6%), and β-caryophyllene (14.2%). Phenylpropanoids (57.3%) and sesquiterpene hydrocarbons (31.6%) were the prominent groups of compounds, followed by oxygenated sesquiterpenes (6.5%), oxygenated monoterpenes (3.0%), and monoterpene hydrocarbons (1.0%). The compound α-bulnesene was identified for the first time in this report. The essential oil was found to be eugenol–α-bulnesene–β-caryophyllene chemotypes.