Forage is the primary feed source for livestock in tropical regions and energy is one of the most important nutrients for ruminant nutrition. The effects of harvest management of Marandu palisade grass (Brachiaria brizantha cv. Marandu Syn. Urochloa brizantha cv. Marandu) on non-structural carbohydrate (NSC) concentrations were evaluated. A plot (Experiment 1) and a greenhouse study (Experiment 2) were conducted in 2013–14. In Experiment 1, treatments were the factorial arrangement of two harvest times and two vertical canopy layers (upper and intermediate), distributed in a completely randomized design with five replicates. In Experiment 2, treatments were the factorial arrangement of six harvest times and two morphological fractions (leaf blade and pseudostem). In both experiments, NSC concentration increased during the day. Upper and intermediate canopy layers had greater NSC concentration at 15.00 than 06.00 h during spring and summer. In addition, the magnitude of NSC increase was greater in the upper than intermediate canopy layer and in spring than summer. Marandu palisade grass shows greater digestibility in the afternoon than morning, representing an opportunity to optimize energy concentration through harvest management.