Background: Oxidative stress in patients with beta thalassemia major is mainly caused by tissue injury due to over production of free radical by secondary iron overload and alteration in serum antioxidant level. Present study is conducted to evaluate the iron overload in beta thalassemia major patients attending a tertiary care hospital in Miraj, Maharashtra. Materials and Methods: A comparative cross sectional study is done to evaluate the iron overload in beta thalassemia major patients of the age less than or equal to 12 years attending the pediatric OPD who were currently being transfused and managed for the clinical symptoms and manifestations of the disease during January 2016 to December 2016. There were 47 beta thalassemia patients and 50 non beta thalassemia patients hence used as controls. Mean levels of the parameters were compared in both the groups. P value of less than 0.05 was taken as significant. Results: Cases and controls were comparable in their demographic characteristics (for age P–0.888 ; for gender proportion P=0.902). Serum level of iron and ferritin was significantly higher in the patients as compared to controls (P<0.05) whereas total iron binding capacity was significantly lower in patients (P<0.05). Melandialdehyde (MDA) levels were also significantly higher in patients (P<0.05) whereas vitamin C levels were significantly lower in patients (P<0.05). Conclusion: Iron overload is seen in beta thalassemia major patients who were currently being transfused. Iron overload leads to tissue injury by free radicals and hence oxidative stress in patients. This leads to increased lipid peroxidation (high MDA) and decreased antioxidant capacity (lower vitamin C levels). Recommendation: Iron chelation therapy with monitoring of the parameters can help to reduce the iron overload and its complications in beta thalassemia patients.