The long periods of working hours at the hospital, and exposure to residues of harmful substances such as anticancer drugs and others lead to excessive formation of free radicals in the body. The aim of this study was to evaluate peroxynitrite, folic acid, homocysteine, ceruloplasmin, and copper in hospital cleaning worker staff (janitors) exposed to residue of different types of substances such as patients' blood, urine, and tools, as well as anticancer drugs in the work setting. This study was conducted among 40 subjects. Twenty subjects (from hospital cleaning staff) were considered hospital cleaning worker staff subjects, twenty other subjects were included as a healthy control group. The mean levels of peroxynitrite, folic acid, and ceruloplasmin in the hospital cleaning staff were (90.80± 23.93, 3.305±1.388, and 269.9±71.86) and in a healthy control group were (19.7±6.551, 6.232±1.738, and 129.88±37.98) µmol/L, respectively, which are statistically high significant. In contrast, the mean serum levels of homocysteine in two groups were (10.84±4.695 and 8.21±1.31) µmol/L, respectively, which are statistically significant. The average copper content was also found in both groups, although it was statistically insignificant and measured (129.34±.26.51) µg/dL in the hospital cleaning staff group and (133.91±11.25) µg/dL in the control group. These findings suggest that long-term exposure to residues of various types of substances in the work setting may cause an induced oxidative stress that results in a number of diseases including cancer.