The main focus of this study is to prove the therapeutic potential of Traditional formulation (TF) by investigating antioxidant activity in different in vitro models and cytotoxic effect of formulation in normal cell line. In vitro free radical scavenging activity of extracts were studied for DPPH (1,1-diphenyl-2-picryl-hydrazyl), ABTS (ABTS) 2,2’-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid, Superoxide, hydroxyl, lipid peroxidation. Total antioxidant assay, reducing power assay and FRAP (Ferric reducing/antioxidant power) assay were also studied. Cytotoxicity study of the formulation was evaluated on HEK 293 cell line and cell viability was determined using MTT assay. The Traditional formulation showed significant and concentration-dependent antioxidant and free radical scavenging activities. Consequently, the formulation exerted lipid peroxidation inhibitory effect by 79.72% as compared to standard ascorbic acid (89.68%). Moreover,formulation has significant scavenging activities of DPPH, nitric oxide, superoxide, ABTS and hydroxyl free radicals in a concentration - dependent manner by 80%, 81.4%, 72.4%, 78.4% and 80.4%, when compared to the standard compound ascorbic acid by 78.4%,77.5%,70.4%,74.3% and 81.3% respectively. Cytotoxity studies revealed non toxic nature of a traditional formulation on experimental cell line. Our studies suggest that TF contain medicinally important secondary metabolites which have significant antioxidant efficacy and non-toxic in nature. However, the in vivo safety needs to be thoroughly investigated in experimental rodent models prior to its practical application.