Objectives: The aim of this research was to determine the knowledge of pharmacists about drug and food interactions. The research was conducted throughout Croatia, in the form of an online survey. In addition to the pharmacist knowledge test, the goal was a self-assessment of the knowledge of each pharmacist who filled out this survey, because this is one of the ways for them to test their knowledge and thus perhaps try to work on the topic of questions to which they did not know the answer. Materials and methods: In order to examine the knowledge of pharmacists, a cross-sectional survey was conducted using an anonymous survey questionnaire during June 2022. The processed data was collected online through a Google Forms form, the link of which was forwarded to the respondents. Respondents included in this research are pharmacists in Croatia who wanted to participate anonymously in an online survey. Results: A total of 208 pharmacists participated in the research, of which only 51 respondents declared that they believed they had sufficient knowledge about drug-food interactions, but as many as 187 respondents recognized that older patients are the age group most prone to possible drug-food interactions. Other results vary. The largest share of all pharmacists recognized the interaction of theophylline and coffee (94.4%), and tetracycline and milk (92.21%), only half of the respondents knew about the influence of a diet rich in proteins on the effect of levodopa, as well as the influence of wheat bran on the effect of digoxin. Conclusion: The results of this research showed that the majority of respondents are not sure of their knowledge or think that they do not know enough about drug-food interactions, and they use the Summary of Properties Description as the most common source of information. Also, the results showed no differences in knowledge about interactions between younger and older pharmacists (<30y and ≥30y). Moreover, they did not show high levels of knowledge of the respondents, however most of them knew about more familiar interactions, such as interactions with grapefruit juice. Although this research had certain limitations, this is one of the first studies on this topic in Croatia.