Intraspecific competition for food can be especially high in colonial breeding seabirds. To minimize colony-induced or annual foraging challenges, diet may vary among individuals, but few studies have simultaneously investigated the effects of both extrinsic conditions (e.g. colony or year effects) and parameters of an individual (e.g. sex, age or individual quality) on diet in seabirds. Using stable isotope analyses, we studied the diet of 214 Adélie penguins Pygoscelis adeliae of known sex, age and breeding quality, nesting in 2 colonies on Ross Island, Antarctica, over 3 breeding seasons. During the study, δ15N and δ13C isotope values were lower in penguins breeding at Cape Crozier compared to those at Cape Bird, revealing a difference in prey proportions. Cape Bird penguins were estimated to consistently consume more energy-rich silverfish Pleuragramma antarctica, while birds at Cape Crozier ate more crystal krill Euphausia crystallorophias. We also found inter-annual differences in diet, with a higher dietary fish proportion in both colonies during 2011. Males had significantly higher δ15N values, indicating a higher fish consumption than females. This sexual segregation in diet was particularly pronounced at Cape Bird, where the overall isotopic niche was wider than at Cape Crozier. Differences in diet among adults of varying ages only existed at Cape Bird, where middle-aged penguins consumed more fish than old and young penguins. This study provides evidence that Adélie penguin diet is largely driven by annual, seasonal and local abundances of prey, with only some individuals selectively foraging for more nutritional prey if prey choices are present.