Introduction: Long-term inhalation anesthetics exposure may provide health risks that cannot be completely ruled out. Aim of the study: The target of this research was to see how waste anesthetic gases influenced DNA damage and oxidative damage. Subjects and Methods: Two groups of healthcare workers from the department of anesthesiology joined in the study: Group I comprised of 15 staff who had operated for at least ten years, while Group II consisted of 15 healthcare members who had worked in operating rooms for more than ten years. In addition to control group consisted of 15 employees who had never been exposed to anesthetic agents. The comet assay was used to detect the damage of DNA in whole blood samples. Serum malondialdehyde (MDA) values were used to assess oxidant stress, and superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities were used to assess antioxidant response (SOD). Results: The result indicates that the mean comet variables have increased (tail DNA percentage, olive tail moment, and tail moment). In Group I, the most DNA damage was observed. There were important positive associations between years of anesthetic gas exposure and damage to DNA. In addition, in comparison to the control group, there was a significant decrease in serum GPX activity and rises in serum levels of both MDA and SOD activity in Groups I and II. Conclusions: According to the results, years of working as an anesthetic can damage DNA and lipid peroxidation. As a result, safe waste anesthetic gas concentration limits in operating rooms, as well as precautions for professionals who might be exposed, are important.