Chemotherapy results in increased free radical formation and depletion of tissue antioxidants. Moreover, patients receiving chemotherapy are under emotional stress, which is also accompanied by low antioxidant levels. In the present study, we measured cortisol, the main stress hormone, and total antioxidant capacity (TAC) in serum of 51 cancer patients during chemotherapy. Antioxidant activity was estimated by measuring the influence of serum in oxidation of ABTS (2,2΄-azino-bis (3-ethylbenz-thiazoline-6-sulfonic acid to ABTS+ by methmyoglobin (Antioxidant kit of Cayman). Serum cortisol was measured using an ELISA colorimetric assay. Serum TAC was significantly decreased (75% decrease compared to normal levels, p = 0.001) in all patients during chemotherapy, while blood cortisol concentration was increased by 10%, (p = 0.044). Lower antioxidant levels and higher cortisol concentration were detected in patients receiving chemotherapeutic drugs daily, compared to the ones receiving chemotherapy once a week. A difference between sexes was observed with male patients presenting lower antioxidant status and higher cortisol levels than females. A significant and persistent decrease in antioxidant capacity accompanied by increased cortisol concentration was observed in all patients during chemotherapy. This fact, which is probably generated by biological and emotional stress, increases the probability of harmful side effects and organism weakening and needs to be considered during patients' treatment.