Objective To identify the practice differences among chemotherapy units and the opinions of pharmacists on safety precautions for handling antineoplastic drugs in hospitals. Method The survey was administered to hospital pharmacists in Turkey. The pharmacists who work in chemotherapeutic drug preparation units were asked to participate if they are volunteered. The participating pharmacists were asked questions about the type of chemotherapy units, order and verification process of chemotherapy protocols and potential shortcomings of chemotherapy preparation. The questions were initially assessed in a small sample of pharmacists and subsequently revised and refined. The filled and returned questionnaires were accepted as given consent. Results A total of 97 questionnaires were delivered and 40 (41%) were returned by pharmacists. The median age and practicing years in hospitals of the pharmacists were 38.0 and 6.25 years, respectively. Fully automated chemotherapy units were used by 42.5% of respondents, whereas 30% prepare chemotherapeutics manually; furthermore, 87.5% provide chemotherapeutic drug preparations (admixtures) less than 120 per day. With regard to the drug preparation process, more than half of the respondents 'always' use protective equipment without foot covers (32.3%). Almost 90% of respondents 'always' use a coat, mask and biological safety cabinets; however, 20% of respondents 'often' use closed-system drug transfer sets. More than 50% of pharmacists expressed that they have sufficient knowledge on cytotoxics and the risk of exposure, protective precautions, selection and usage of personal protective equipment and correct storage of cytotoxics. Conclusion Pharmacists acknowledge sufficient understanding of safe preparation and handling of chemotherapeutics, but reported practices by pharmacists in hospitals are not consistent with published recommendations. Providing theoretical and practical educations for pharmacists and developing standard procedures for preparing chemotherapy may be beneficial in improving the preparation and use of antineoplastics in Turkey.