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In clinical trials, omega-3 fatty acid supplementation improves symptoms in psychiatric disorders involving dysregulated mood and impulse control, yet it is unclear whether in healthy adults, omega-3 fatty acid supplementation affects mood, impulse control, and the brain systems supporting these processes. Accordingly, this study tested the hypotheses that eciosapentaenoic (EPA) and docosahexaenoic (DHA) acid supplementation reduces negative affect and impulsive behaviors in healthy adults and that these changes correspond to alterations in corticolimbic and corticostriatal brain systems, which support affective and impulsive processes.Healthy volunteers (N = 272) consuming 300 mg/d or less of EPA and DHA were enrolled in a double-blind, randomized, placebo controlled clinical trial. The participants received either capsules providing 1000 mg of EPA and 400 mg of DHA versus identical appearing soybean oil capsules per day for 18 weeks. Negative affect and impulsivity were measured by questionnaire and ecological momentary assessment, as well as functional alterations in corticolimbic and corticostriatal brain systems evoked by standardized functional magnetic resonance imaging tasks.There were no group by time interactions for any questionnaire or ecological momentary assessment measures of mood and impulsivity. Likewise, no group by time interactions were observed for functional magnetic resonance imaging responses evoked within corticolimbic and corticostriatal systems.In healthy adults with low intake of omega-3 fatty acids, moderate-dose supplementation for 18 weeks did not alter affect or impulsive behaviors nor alter corticolimbic and corticostriatal brain functionality.Trial number NCT00663871.