Studies of the effect of varying dietary carbohydrate compositions on exercise metabolism of horses are scarce. In the present study, the starch and sugar porportions were altered in the diet to 4 Standardbred horses. In a crossover experiment, the horses were offered a hay and oat diet, where the oats were substituted for barley syrup (BS) at 4 levels (BS 0, 0.5, 1.0 and 1.5 kg). Each diet was consumed for 21 days and both an incremental treadmill exercise test (IE) and a 40 min submaximal exercise test (SE) were performed. During the IE there was a significant (P<0.05) increase in mean V̇O2 on BS 1.5 and at the highest speed respiratory exchange ratio (RER) was significantly lower on BS 1.0 and 1.5. During the SE, RER and HR were significantly higher on BS 1.0 and 1.5. There were no significant effects of diets on plasma levels of glucose, lactate and insulin. Following the SE there was a significant decrease in the muscle glycogen content on BS 0, which was not observed in the other diets. In addition, there was a numerical decrease in the glycogen utilisation with increasing proportions of sugar in the diet. In conclusion, this study indicates that horses on diets with a high sugar inclusion respond with an increased heart rate during exercise of lower intensities; and that the response in V̇O2 and RER may differ depending on the exercise intensity. In addition, glycogen utilisation during submaximal exercise was lowered when daily sugar intake was increased. However, further investigation is needed to evaluate whether this strategy of feeding could be beneficial for exercising horses competing over endurance distances.