Current research project was conducted to study the urinary excretion of deferoxamine, ferrioxamine, iron and zinc in β-thalassemia population. Oxidative stress and antioxidant status was also measured in β-thalassemic patients under indigenous conditions. Blood specimens were collected from both healthy and thalassemic subjects of age less than 15 years without drug administration. Hematological (red blood cells, hemoglobin, MCH, MCV, MCHC, plt, hct, RDW, eiosinophils, monocytes, neutrophils, lymphocytes) and biochemical parameters (triglycerides, cholesterol, LDL, ALT, AST, urea, total protein, albumin, globulin and creatinine) were determined in both healthy and thalassemic populations to check the health status. Oxidant and antioxidant status was studied with the help of spectrophotometer and HPLC. Thalassemic patients were observed in state at higher level of oxidative stress because free iron in body acts as free radical to enhance the oxidative stress. Antioxidants are (either enzymatic or non enzymatic) substances in body which reacts with these free radicals to stop chain reaction. So, the levels of antioxidant vitamins (E, C and β-carotene) were significantly decreased in β- thalassemic patients. Urine samples were collected at 0, 6, 8, 12, 24 h from β-thalassemic patient after 500 mg subcutaneous infusion of deferoxamine. Concentration of deferoxamine and ferrioxamine was determined with the help of HPLC highest concentration of DFO and FO was recorded in urine samples collectes at 6 th hour. % dose of DFO and FO excreted (3 and 2.97 percent) in β-thalassemic patients within twenty four hours. While the concentration of iron and zinc was determined with the help of AAS and cumulative amount excreted of zinc and iron after 24 hours were 1.57 ± 0.06 mg and 30.47 ± 1.47 respectively in thalassemic patients indicating enhanced excretion of zinc. This study would help in clinical investigations and to design better therapeutic protocols under indigenous conditions.