Fatty acids have been widely used as trophic biomarkers in marine mammals. However, for the South American sea lion, the most abundant otariid in the eastern South Pacific, there is no information about blubber fatty acids and their link to diet. Here, we compare fatty acid profiles of sea lions from two distinct oceanographic regions in northern and southern chile. their fatty acids vary greatly between regions, suggesting dietary differences at a spatial scale. The fatty acid C22:6ω3 was more abundant in sea lions from the northern region, likely associated with consumption of anchovy, cephalopods, and crustaceans, which are rich in that fatty acid, and have been reported as their main prey items. Sea lions from the southern region were richer in C22:1 and C20:1, characteristic of teleost fish, suggesting a piscivorous diet. Males displayed a more diverse fatty acid composition than females, suggesting a wider trophic niche. few individual sea lions within the southern region had unusually high levels of C18:2ω6, commonly found in terrestrial environments. This suggests consumption of farmed salmon, whose diet is usually based on terrestrial sources. this demonstrates how human intervention is being reflected in the tissues of a top predator in a natural environment. Deciphering trophic interactions of top predators is important to understand the structure and functioning of whole ecosystems. The South American sea lion (Otaria byronia) is a top predator with a wide distribution range along the eastern Pacific as well as the western Atlantic coasts of South America 1,2. They are an abundant species, with population estimates of up to 250,000 individuals, and some populations increasing steadily 1,3. South American sea lions are considered to be opportunistic predators, therefore, their diet is expected to vary in space and time depending on food availability 4. In fact, decreases in their population in some areas have been linked to declines in the availability of certain prey species 5. Unsurprisingly Muñoz et al. 4 reported a great variation in diet composition between northern and southern populations of South American sea lions in Chile. Similarly, Sepúlveda et al. 6 demonstrated temporal differences in their diet composition in a population from southern Chile. Most of our knowledge on the trophic ecology of South American sea lions relies on traditional methods such as stomach and scat content analyses 7,8 , or on observations of feeding events, which are relatively rare and limited in aquatic environments 9-11. Through these methods, a broad-spectrum diet has been described 12,13. These methods provide valuable information about recently consumed diets but are not ideal to determine long-term dietary trends, since this would require of repeated sampling throughout long time periods 14,15. Furthermore, identifying inter-individual variation in diet preferences is very challenging due to, for example, the difficulty of assigning scats to particular individuals in crowded rookeries 16 .