Objective: Enhanced knowledge retention and a preference towards a deep learning approach are desirable pedagogical outcomes of case-based learning (CBL). The CBL literature is sparse with respect to these outcomes, and this is especially so in the area of biochemistry. The present study determined the effect of CBL vs. non CBL on knowledge retention in an undergraduate biochemistry course; it also investigated associations of learning approach, age and gender. Methods: We used the Revised Two-Factor Study Process Questionnaire, a retention test, final exam grades and other demographic information to statistically compare academic outcomes of students subjected to either CBL or non-CBL active learning techniques. Results: We showed that students exposed to CBL in a second year course performed significantly better on a retention test conducted nine months after the final exam, and that there was a positive correlation between a deep learning approach and higher retention scores. We did not find an association between gender and age with the retention of biochemistry concepts. Conclusions: Our findings suggest that use of CBL in undergraduate biochemistry education may confer benefits in terms of retention of knowledge of key concepts.