It has been demonstrated that a positive correlation exists between clinical knowledge and retained concepts in basic sciences. Studies have demonstrated a modest attrition of anatomy knowledge over time, which may be influenced by students' perceived importance of the basic sciences and the learning styles adopted. The aims of this study were to: (1) conduct a cross-sectional evaluation of the retention of anatomical knowledge in preclinical (second-year) and clinical (fifth-year) chiropractic students at Murdoch University; and (2) examine students' perceptions of factors that may influence their anatomy knowledge retention. Second- and fifth-year chiropractic students at Murdoch University were invited to participate in the study. Ninety-one students voluntarily participated. The Carpal Bone Test, previously utilized to determine the retention of anatomical knowledge, was utilized to determine the extent to which participants retained gross anatomy knowledge. Participants also completed a questionnaire specifically designed to identify the factors that may have influenced their retention of gross anatomy knowledge. A two-sided Pearson chi-square test of association was used to ascertain statistically significant differences in carpal bone retention and students' responses between the two cohorts. Seventy percent of the fifth-year (clinical) chiropractic students correctly identified all eight carpal bones compared to only six percent of second-year chiropractic students. The majority of participants in both cohorts believed that gross anatomy knowledge is of clinical importance. The use of mnemonics and the clinical application of anatomy knowledge were identified as factors that significantly influenced participants' gross anatomy knowledge retention within this study.