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Healthcare workers’ exposure to antineoplastic drugs may occur through handling of the drugs and/or via contact with drug-contaminated surfaces. However, studies have been limited to select departments and/or certain job titles. This may lead to an underestimate of the risk as the drugs circulate within a facility known as the hospital medication system (process flow of drugs). This study aimed to answer the following questions related to antineoplastic drugs and the hospital medication system: 1) is contamination found on surfaces located throughout, 2) are workers throughout occupationally exposed (dermal and urinary contamination), and 3) what factors are associated with surface contamination and occupational exposure? Site observations were conducted to identify which surfaces may be contaminated and the job categories that may contact these surfaces. Wipe samples were collected from potentially-contaminated surfaces and the hands of at-risk healthcare workers. Urine samples were collected from these same workers. Participants were asked to complete a questionnaire regarding their knowledge and usual protective habits regarding antineoplastic drugs and surveyed about contact with these agents on their work shift. Drug residual was measurable on surfaces located throughout the hospital medication system. Determinants associated with increased surface contamination were the drug preparation and drug administration stages of the medication system as well as having more job categories responsible for drug transport. Up to 11 job categories per facility may have an exposure risk and the maximum dermal contamination levels for every job category exceeded the limit of detection. Factors associated with increased dermal contamination were working in acute care hospitals, female personnel, working as a porter, nurse, transport, unit clerk or other roles in the drug administration unit and having a duty to handle antineoplastic drugs. Urinary drug contamination of participants was higher than in non-hospital controls confirming that exposure is occurring in the workplace. Being a pharmacy receiver, pharmacy technician, porter, nurse, or unit clerk and a facility having more job categories responsible for drug transport were associated with increased urinary contamination. This is believed to be the first study examining environmental contamination and occupational exposure to antineoplastic drugs across the entire hospital medication system.