We measured concentrations of essential fatty acids (EFAs) in four size categories of planktonic organismsseston (10-64 m), microzooplankton (100-200 m), mesozooplankton (200-500 m), and macrozooplankton (Ͼ500 m)-and in rainbow trout (Oncorhynchus mykiss) in coastal lakes. Size-dependent patterns in concentrations of specific fatty acids (FAs) are important for ecosystem function, because planktivorous fish and some invertebrates are size-selective predators. We demonstrate that the retention of individual FAs differs among the four size categories of planktonic organisms in our study systems. Changes in individual EFA concentrations within the planktonic food web were similar in all coastal lakes sampled, which indicates the generality of our findings. Although concentrations of arachidonic acid, eicosapentaenoic acid (EPA), and linoleic acid increased steadily with plankton size, the concentration of ␣-linolenic acid decreased slightly in larger size fractions of zooplankton. Concentrations of another EFA, docosahexaenoic acid (DHA), declined sharply from mesozooplankton to the cladoceran-dominated macrozooplankton size class. Our results indicate that the retention of EFAs, as a function of plankton size, is related, in part, to the taxonomic composition of planktonic food webs. We suggest that, in general, zooplankton exhibit an EPA-retentive metabolism with increasing body size, whereas different taxonomic groups within the planktonic food web retain DHA differently. Finally, we conclude that EPA is highly retained in zooplankton, whereas in rainbow trout DHA is highly retained.