Background: Students who receive e-learning in nursing education state that they cannot learn the lessons effectively. This study aims to evaluate the effect of multimedia teaching materials on students' academic achievement and motivation in e-learning. Methods: The quasi-experimental study was performed on 70 students. The experimental group students were presented with multimedia materials, and the control group students were taught the e-learning course. Results: In this study, regarding the effect of multimedia-based education on students’ Instructional Materials Motivation Survey, the mean score of the control group was 85.25 ± 11.95, and the mean score of the experimental group was 94.44 ± 11.82 after the intervention. A moderately significant positive correlation was found between the experimental group's average academic achievement and total scale scores. Conclusion: The study results show that the participants accepted the important role of multimedia as practical learning compared to traditional learning methods. Given the impact of multimedia-based education on academic achievement, this approach can be used to teach other core subjects to nursing students.