Objective: Taking into account the significant role of sports in Croatian society, the aim of this study was to assess the knowledge and skills of pharmacists in Croatia in assisting professional athletes to prevent unintentional use of prohibited substances. Methods: The research was conducted during July 2024 through telecommunication interaction with 80 randomly selected public pharmacies from Zagreb, Rijeka, Osijek and Zadar. Using the simulated patient method, the professional water polo player, following the established interview protocol, requested the advice from the Master of Pharmacy on taking a salbutamol inhaler (a WADA-prohibited substance with conditional requirements). During and immediately after each interview, additional information requested from the patient as well as advice given to the patient were recorded on the data collection form, after which the data were coded and analyzed. Results: The majority of pharmacists (70.0%) did not request any additional information about the patient, his health status or therapy. A total of 34 respondents (42.5%) gave incorrect recommendations to the patient, whilst two pharmacists (2.5%) pointed out that they did not have the required information nor did they know how to find it. Only six pharmacists (7.5%) completely, correctly and exclusively pointed out the conditions for using inhaled salbutamol in sport. Conclusion: Most of the respondents lacked required knowledge and skills to enable the delivery of comprehensive information to protect athletes from anti-doping violations. The observed incompetence indicates the need for additional education in sport-related pharmacy in order to optimize care for athlete-patients.