Background: Atalantia monophylla Correa. a small tree belongs to the family Rutaceae. It is distributed throughout India and in Tamil Nadu the species is commonly seen in foothills of dry vegetation. Objective: The aim was to hydrodistillate and analyze the chemical composition of essential oil from the fresh leaves of A. monophylla Correa. collected in two different seasons (December, 2013 and May, 2014) and to evaluate antioxidant and antibacterial activities of isolated essential oil. Materials and Methods: Chemical composition of isolated essential oil was analyzed by gas chromatography, gas chromatography coupled with mass spectrometry. Antioxidant activity of oil was assessed using five different antioxidant test systems. Antibacterial activity of oil was tested against six pathogenic bacteria by broth dilution method. Results: Essential oil obtained from the leaves collected during May, 2014 had shown more compounds. Antioxidant activity of oil was moderate when compared with positive control. Minimum inhibitory concentration value of oil was ranges between 139.32 ± 0.001 and 541.11 ± 0.003 µg/mL against all the tested bacteria. Conclusion: Result clearly indicates essential oil collected during May, 2014 showed more bioactive compounds.