OBJECTIVES: This study aimed to identify and evaluate instruments used to measure pharmacy students' knowledge of HIV prophylaxis, focusing on both preexposure prophylaxis (PrEP) and postexposure prophylaxis. METHODS: A systematic literature search was conducted using PubMed, Virtual Health Library, Web of Science, Scopus, ScienceDirect, and Embase. Article screening was performed independently by 2 reviewers. Studies assessing pharmacy students' knowledge of HIV prophylaxis were included. FINDINGS: Eight studies involving 1,797 students, mostly from the U.S., were identified. All had a cross-sectional design, with only one assessing knowledge retention. Six assessment tools focused on PrEP, one on PEP, and one on both. While confidence in PrEP was evident, significant gaps were found in PEP awareness, prophylaxis access, and prescribing guidelines. SUMMARY: There is a notable shortage of assessment tools for PEP. Further research is needed to develop validated instruments for measuring knowledge and addressing educational gaps. Longitudinal studies with pre- and post-tests are crucial to evaluate educational interventions and enhance pharmacy students' preparedness for HIV prophylaxis.