Nature remains to be an attractive source of bioactive compounds and natural compounds are extensively being explored for their therapeutic applications. Steroidal glycosides are a major group of secondary metabolites that exhibit broad spectrum of biological activities. Several steroidal glycosides have been reported to possess promising anti-oxidant activities. In the present study we investigate the anti-oxidant potentialities of cholestanol glucoside (CG), a steroidal glycoside produced by Lasiodiplodia theobromae, an endophytic fungus isolated from Saraca asoca, using in vitro assays such as DPPH radical scavenging assay, nitric oxide radical scavenging assay, hydroxyl radical scavenging assay, super oxide radical scavenging assay and hydrogen peroxide scavenging assay. The compound showed significant free radical scavenging capacity with an IC50 value of 16.2 μM. The compound could also effectively scavenge hydrogen peroxide (IC50 value of 7.2 μM) and hydroxyl radicals (IC50 value of 3.6 μM).