Many oxidative stress related diseases like cancer, diabetes, neurodegenerative disorders and cardiovascular diseases are occurring as a result of accumulation of free radicals in the body. Tinospora cordifolia (Guduchi) is a medicinal herb used in the Indian system of medicine due to its health benefits. In the present study, the antioxidant activity of Guduchi extract was investigated using various in vitro methods such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), nitric oxide (NO), hydroxyl (OH), hydrogen peroxide radical scavenging and anti-lipid peroxidation assays. The results indicated that IC50 values of Guduchi extract were 0.997mg/ml in DPPH, 0.72mg/ml in ABTS, 10.7mg/ml in NO, 3.1mg/ml in H2O2, 0.86mg/ml in hydroxyl radical scavenging and 109mg/ml in antilipid peroxidation. The results revealed that Guduchi extract has promising antioxidant activity and could serve as potential source of natural antioxidants.