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Understanding the metabolic determinants of post-exercise appetite-regulation would facilitate the development of adjunctive-therapeutics to supress compensatory eating behaviours and improve the efficacy of exercise as a weight loss treatment. Metabolic responses to acute exercise are however dependent on pre-exercise nutritional practices, including carbohydrate intake. We therefore aimed to determine the interactive effects of dietary carbohydrate and exercise on plasma hormonal and metabolite responses and explore mediators of exercise-induced changes in appetite-regulation across nutritional states. In this randomised crossover study, participants completed four 120 min visits: (i) control (water) followed by rest; (ii) control followed by exercise (30 min at ∼75% V̇O2 max); (iii) carbohydrate (75 g maltodextrin) followed by rest; and (iv) carbohydrate followed by exercise. An ad libitum meal was provided at the end of each 120 min visit, with blood sample collection and appetite assessment performed at pre-defined intervals. We found that dietary carbohydrate and exercise exerted independent effects on the hormones GLP-1 (Carbohydrate: 16.8 pmol/L, Exercise: 7.4 pmol/L), ghrelin (Carbohydrate: -48.8 pmol/L, Exercise: -22.7 pmol/L) and glucagon (Carbohydrate: 9.8 ng/L, Exercise: 8.2 ng/L) that were linked to the generation of distinct plasma 1H-NMR metabolic phenotypes. These metabolic responses were associated with changes in appetite and energy intake, and plasma acetate and succinate were subsequently identified as potential novel mediators of exercise-induced appetite and energy intake responses. In summary, dietary carbohydrate and exercise independently influence gastrointestinal hormones associated with appetite regulation. Future work is warranted to probe the mechanistic importance of plasma acetate and succinate in post-exercise appetite-regulation.