The present study investigated the effects of mixed high-carbohydrate (CHO) meals (breakfast and lunch) with different glycaemic indices (GI) on substrate metabolism during rest throughout the postprandial periods and during subsequent exercise. Nine recreationally active males completed two trials, high glycaemic index (HGI) and low glycaemic index (LGI), separated by 7 d in a randomised crossover design. In each trial, participants consumed breakfast and lunch, both of which were followed by a 3 h resting postprandial period. Following this, participants completed a 60 min run at 70 % of V O2max. The plasma glucose and serum insulin concentrations following both meals were significantly higher in the HGI trial than in the LGI trial (P<0.05). Serum insulin concentrations remained higher throughout the postprandial period following lunch in the HGI trial compared with the LGI trial (P<0.05). The total amount of fat oxidised was higher during the 3 h rest following lunch in the LGI trial than in the HGI trial (P<0.01) and subsequently CHO oxidation was lower (P<0.005). No significant differences in substrate utilisation were observed throughout the subsequent run. At 45 and 60 min, plasma glucose concentrations were higher in the LGI trial v. the HGI trial (P<0.05). The results of the present study provide further support that the GI concept can be successfully applied to mixed meals. The results also suggest that meals composed of LGI CHO may be more beneficial for maintaining a favourable metabolic milieu during the postprandial periods. Furthermore, during subsequent exercise, plasma glucose concentrations were better maintained following the LGI CHO meals.