Studies have shown that practicing temporally spaced retrieval of previously learned information via formal assessments increases student retention of the information. Our objective was to determine the impact of online homework administered as a first retrieval practice 1 or 5 days after introduction of physiology topics on long-term information retention. Students in two undergraduate courses, Anatomy and Physiology (ASCI 240) and Animal Physiological Systems (ASCI 340), were presented with information on a specific physiological system during each weekly laboratory and then completed an online homework assignment either 1 or 5 days later. Information retention was assessed via an in-class quiz the following week and by a comprehensive final exam at semester’s end (4–13 wk later). Performance on homework assignments was generally similar between groups for both courses. Information retention at 1 wk did not differ due to timing of homework in either course. In both courses, however, students who received homework 5 days after class performed better on final exam questions relevant to that week’s topic compared with their day 1 counterparts. These findings indicate that the longer period between introducing physiology information in class and assigning the first retrieval practice was more beneficial to long-term information retention than the shorter period, despite seemingly equivalent benefits in the shorter term. Since information is typically forgotten over time, we speculate that the longer interval necessitates greater retrieval effort in much the same way as built-in desirable difficulties, thus allowing for stronger conceptual connections and deeper comprehension