Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)A novel microwave assisted extraction (MAE) method for fish lipids was optimized by means of a central composite design and validated using a fish tissue standard reference material (SRM, 1946, NIST). Scanning electron microscopy showed that MAE caused total disruption of the fish tissue, thus lipid migration to the extraction solvent was more efficient and faster in comparison to the Folch method. The lipid content of the SRM obtained by MAE (10.10.2g/100g) was similar to that declared on the certificate (10.20.5g/100g). The use of microwave energy did not alter the fatty acid composition of the fish nor formed lipid oxidation derived compounds at higher levels than in the Folch method. The MAE method was applied to fishes with different lipid contents, tilapia, pacu, and hake. MAE is a fast and robust technique with low solvent consumption when compared to the Folch method. Practical applications: MAE presented the following advantages over the Folch method: the lipids can be extracted from up to 12 samples simultaneously, it reduces the extraction time by up to 90%, produces less residues, reduces solvent consumption proportionaly to the mass used by up to 25%, and contributes to a reduction in toxicity in the environment by substituting chloroform with ethyl acetate.1193CAPESCNPqFAPESP [2013/06489-1]Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)