Persistent changes in the fatty acid composition of erythrocyte membranes after moderate intake of n−3 polyunsaturated fatty acids: study design implications

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Abstract
To examine the incorporation of n−3 polyunsaturated fatty acids (PUFAs) into erythrocyte membranes during and after moderate n−3 PUFA intake, 12 healthy men were fed three diets for 6-wk periods in a 3 × 3 crossover design, supplying different amounts of eicosapentaenoic acid (EPA, 20: 5n−3) and docosahexaenoic acid (DHA, 22:6n−3): a control diet, a fish diet (0.15 g EPA/d, 0.41 g DHA/d), and a fish + oil diet (5 g fish oil/d; 0.99 g EPA/d and 0.99 g DHA/d). A 6-wk washout period was allowed between diets. Between 6 and 12 wk after the fish + oil diet, erythrocyte EPA and DHA were still declining and it was only after 18 wk that erythrocyte EPA had returned to baseline whereas DHA had not. Investigators examining variables that are influenced by altered membrane fatty acid composition should be aware of these prolonged effects when designing studies. Protracted washout periods (> 18 wk) make the classic crossover design prohibitive and a parallel design becomes essential.