Methotrexate (MTX) is one of the most widely used cytostatic drugs belonging to the folic acid antagonists. It is a substance non-classified as a carcinogen in the European Union and by the International Agency for Research on Cancer (IARC) as there is no evidence of its carcinogenicity to humans and animals. Nevertheless, MTX has been placed on the list of dangerous drugs used in chemotherapy, mainly due to genotoxic and teratogenic effects, causing developmental toxicity and reproductive toxicity. Methotrexate was determined in the hospital ward air during the preparation of a medicament at a level of 0.3 mg/m³, as well as on protective gloves and preparatory room surfaces. In most research projects MTX was identified in the urine of health care workers, pharmacists and nursing staff. The highest cumulative concentration of MTX in 112 urine samples was 1416 mg in workers preparing infusions for patients. Studies carried out in pharmacies revealed the presence of MTX in 60% of tests, and the maximum concentration of 15 ng/cm² surface of the tray to count tablets. Legal exposure limit values for MTX in the work environment have not yet been established. Occupational exposure limits have been established by some manufacturers at the level of 0.0003–0.0025 mg/m³. There is an urgent need to establish normative values. It should also be emphasized that MTX is absorbed through the skin, which may significantly increase the exposure and measuring its concentration in the work environment may not be sufficient to estimate the actual exposure. Med Pr 2015;66(2):265–275