Introduction: The juice of leaves of Barleria grandiflora has been used by folk across central region of India for the treatment of oral ulcers. The objective of the study is to determine the antioxidant and the antibacterial activity of ethanolic extract of leaves of Barleria grandiflora Dalz (Acanthaceae). Materials and Methods: Phytochemical screening was carried out as per the usual chemical tests. The TLC was carried out to determine the constituents which might be similar to Barleria prionitis. Total phenolic content was determined using Folin-Ciocalteau method. Antioxidant activity was performed by using hydrogen peroxide radical scavenging method and DPPH (1, 1-Diphenyl-2-picrylhydrazyl) radical scavenging activity. Antibacterial activity was carried using agar well diffusion method. Results: Phytochemical screening had shown the presence of carbohydrates, proteins, alkaloids, glycosides, phenolic compounds and flavanoids in ethanolic extracts of leaves of the plant. The total phenolic content was found to be 75.44 mg/10ml. Barleria grandiflora was found to have antioxidant potential comparable to ascorbic acid (vitamin c). It was also found that the ethanolic extract of Barleria grandiflora shows significant antibacterial activity. Zones of inhibition of ethanolic extract of Barleria grandiflora were found comparable to that of standard. Conclusion: Ethanolic extract of leaves of Barleria grandiflora exhibited antioxidant and antibacterial activity which may be the reason for the use of juice of Barleria grandiflora leaves in the treatment of mouth ulcers by some folk.