The role of intracontinental migration patterns of European eel (Anguilla anguilla) receives more and more recognition in both ecological studies of the European eel and possible management measures, but small-scale patterns proved to be challenging to study. We experimentally investigated the suitability of fatty acid trophic markers to elucidate the utilization of feeding habitats. Eight groups of juvenile European eels were fed on eight different diets in a freshwater recirculation system at 20°C for 56 days. Three groups were fed on freshwater diets (Rutilus rutilus, Chironomidae larvae, and Gammarus pulex) and four groups were reared on diets of a marine origin (Clupea harengus, Crangon crangon, Mysis spec., and Euphausia superba) and one on commercial pellets used in eel aquaculture. Fatty acid composition (FAC) of diets differed significantly with habitat. FAC of eel muscle tissue seemed to be rather insensitive to fatty acids supplied with diet, but the general pattern of lower n3:n6 and EPA:ARA ratios in freshwater prey organisms could be traced in the respective eels. Multivariate statistics of the fatty acid composition of the eels resulted in two distinct groups representing freshwater and marine treatments. Results further indicate the capability of selectively restraining certain fatty acids in eel, as e.g. the n3:n6 ratio in all treatments was <4, regardless of dietary n3:n6. In future studies on wild eel, these measures can be used to elucidate the utilization of feeding habitats of individual European eel.