Tinospora cordifolia (Willd) is a medicinal plant, commonly known as Guduchi or Giloy. It has multifaceted implications as an immunomodulatory, anti-inflammatory and antioxidant drug. The present study was carried out for the Comparative analysis of free radical scavenging activity of various extracts of Tinospora cordifolia stem and their Protective Effect on Oxidation of biomolecules. In the present study, the antioxidant potential of different solvent extracts of stem were evaluated by both enzymatic and non-enzymatic methods. The stem of Tinospora cordifolia was extracted with ethanol and distilled water through both hot and cold extraction methods. The total phenolic and flavonoid content of the extracts were determined by Folin-Coicalteu method and aluminium chloride colorimetric method respectively. The In-vitro bioassays were done to evaluate the antioxidant activity through ABTS radical scavenging assay, Reducing ability (FRAP) assay, Lipid peroxidation assay (LPO) and Peroxidase assay. The phenolic contents of the extracts as gallic acid equivalents were found to be highest in ethanolic extract of test drug. In ATBS assay and FRAP assay, the free radical scavenging activity was found to be highest with ethanolic extract followed by aqueous extract of test drug. Protective effect on biomolecule oxidation can be inferred by Peroxidase assay and Lipid peroxidation assay which is also found to be highest with ethanolic extract. The data obtained from this study clearly establish the higher antioxidant potency of the ethanolic extract of T. cordifolia stem compared to its aqueous extract.