The number of patients with end-stage renal disease is growing rapidly in Asia. Patients with end stage renal disease failure (ESRD) undergoing long term hemodialysis (HD) are at risk for high oxidative stress. There is paucity of information with regard to oxidative stress during HD. Therefore, the aim of the present study was to evaluate catalase enzyme activity in erythrocytes and the total antioxidant capacity (TAC) in plasma before, during and after dialysis in hemodialysis patients. The cross-sectional study carried out in thirty four patients (16 males and 18 females) having mean age of 55 ± 16 years undergoing HD three times per week were included. The enzyme activity of catalase was measured by UV Spectrophotometric method and total antioxidant capacity in plasma were observed by ferric reducing ability of plasma (FRAP) assay method. All tests were performed before, during and after the process of dialysis. The prevalence of erythrocyte catalase enzyme activity was observed to be significantly decreased (P < 0.001) during (107.01 ± 36.2) and after (105.30 ± 36.75) dialysis as compared to its activity before (159.75 ± 30.98) dialysis. On the other hand, a total antioxidant capacity in plasma was noticed to be significantly elevated (P < 0.001) during (648.52 ± 14.66) and after (573.08 ± 15.16) dialysis as compared to before (486.61 ± 14.86) dialysis. The study concluded that dialysis process affects erythrocyte catalase enzyme activity as well as total antioxidant capacity in plasma of patients. HD exacerbates oxidative stress and disturbances in antioxidant enzymes in patients.