Fish are crucial for the fishing industry and essential nutrient provision, including lipids. This study employed a high-throughput lipidomic approach to evaluate and contrast the lipid profiles of three marine fish species (P. crocea, S. fuscens, and C. saira) and one freshwater species (H. molitrix) across head, muscle, and viscera. Over 1000 molecular lipid species across 17 subclasses were identified. Notably, acylated monogalactosyldiacylglycerol (acMGDG) was detected for the first time in these species, with a high prevalence of saturated fatty acids (44.7 %−87.7 %). Glycerolipids (67.7 − 86.3 %) and PLs (10.7 − 31.8 %) were identified as the dominant lipid classes. Marine fish muscles displayed higher PL content than freshwater species, and P. crocea viscera contained over 30 % PLs of total lipids. In particular, ether phosphatidyl ethanolamine incorporated more DHA than ether phosphatidylcholine. The viscera of four fish species also exhibited a significant abundance of diacylglycerol (DG), indicating their potential as functional lipid sources. Multivariate analysis identified triglyceride (TG) (59:13), DG (16:1/22:5), and MGDG (16:0/18:2) as potential biomarkers for differentiating among fish anatomical parts. This study deepens the understanding of the nutritional values of these fish, providing guidance for consumer dietary choices and paving the way for transforming previously underutilized by-products into resources with high-value potential.
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