The maintenance of weight and adipose tissue mass in humans appears to be related to a balance between the rates of oxidation and consumption of macronutrients; yet, little evidence is available on the reproducibility of 24-h macronutrient oxidation rates and how this relates to diet in the days preceding a chamber session. This study examined the reproducibility of 24-h macronutrient oxidation rates, 24-h energy expenditure (EE), and respiratory quotient (RQ) in 30 adults who ate their habitual diets before two 24-h whole body indirect calorimeter sessions. Results showed that the within-subject coefficients of variation (CVws) for 24-h EE and RQ were 2.8 and 2.6%, respectively. CVws for macronutrient oxidations ranged from approximately 15 to 25%. Means comparisons of 24-h EE, RQ, and macronutrient oxidation rates between sessions showed no significant differences, and all variables had significant positive intraclass correlation coefficients (P < 0.05). In conclusion, macronutrient oxidations all showed significant reproducibility for the group and a significant but lower reproducibility for individuals when habitual diet and activity preceded the experimental sessions.