As COVID-19 restrictions are removed, instructors are faced with two questions: should the course be taught online, via mixed-mode, or through traditional methods, and what effect did online instruction have on students in the course in regards regarding enrolled across eight semesters of metabolism courses using traditional, mixed-mode, and online methods. Post-hoc analysis of a repeated-measures ANOVA determined that while mixed-mode outperformed traditional methods on metabolism exams involving introductory concepts on the first exam (4.47 ±4.24, p= 0.012) and on the cumulative final exam (8.15 ±4.24, p= <0.001), traditional methods were superior to both mixed-mode (9.21 ±4.24, p= <0.001) and online methods (6.37 ±5.76, p= 0.006) at teaching alternative pathways of metabolism and lipid, nucleotide, and amino acid synthesis pathways. All methods showed poor student scores on glycolysis, TCA cycle, oxidative phosphorylation topics. Online methods performed nearly equally to mixed-mode, though there was not a significant difference on the first exam's content as was found with mixed-mode. It is evident that, based on this study, a different approach to teaching basic central metabolic cycles such as glycolysis, the TCA cycle, and the electron transport chain is necessary to improve student understanding of these topics.