The yellowfin tuna is a very abundant tropical tuna species in the western equatorial Atlantic Ocean and an important fishery resource for the Brazilian tuna fleet. In this study we performed stable isotope analysis to better understand the spatial trophodynamics and dietary changes in yellowfin tuna around two insular marine protected areas in Brazil. A total of 65 yellowfin tuna specimens measuring between 47 and 138 cm LT (total length) were sampled around the archipelagos of Fernando de Noronha (FNA; n = 34) and Saint Peter and Saint Paul (SPSPA; n = 31) between July 2018 and September 2019. Bayesian mixing models and generalized additive models were used to investigate the contributions of four different prey items (zooplankton, cephalopods, fish larvae, and flying fish) to yellowfin tuna diet in each area and their potential changes in relation to predator growth. The four prey items were found to have different overall contributions between the two studied areas, with zooplankton being the most important prey in FNA, whereas flying fish was the most relevant prey to the species' diet in SPSPA. Significant changes in the species diet by size were also found, with fish smaller than 90 cm (TL) having a more generalist diet and larger animals relying more on consuming larger and more nutritious prey (i.e., flying fish). Our results suggest that these two marine protected areas play an important role in ocean dynamics, providing important and different foraging grounds for the development of this predator species.