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Trachinotus ovatus (TO) is an important cultured marine fish, which is popular with customers for its high muscle quality and favorable price. In this study, lipids in viscera, bone and muscle of TO were extracted and analyzed to identify unique applications. Total lipid content based on dry mass was 34.50% in muscle, 46.25% in bone, and 64.35% in viscera. In the lipid composition of TO oil (TOO), the contents of free fatty acids, diglycerides and phospholipids ranged from 3.54% to 7.21%, 1.88%–4.66% and 4.91%–6.35%, respectively. Phospholipids were mainly composed of phosphatidylcholine and phosphatidylethanolamine. Triglycerides were the major component in TOO for all tissues, the content range was 83.16%–89.39%. The main fatty acids of triglyceride fraction in TOO were palmitic acid (~24%), oleic acid (~26%), and linoleic acid (~26%). It is interesting to observe that palmitic acid was preferentially distributed at the sn-2 position. Ultra-high performance combined-phase chromatography combined with quadrupole time-of-flight mass spectrometry analysis showed that 1-oleoyl-2-palmitoyl-3-linoleoylglycerol was the highest content of triglyceride molecule in TOO, accounting for about 17%, followed by 1,3-dioleoyl-2-palmitoylglycerol. These unique composition and positional distribution in TOO are very similar to those of human milk fat. Therefore, TOO has a great potential in the preparation of human milk fat substitutes.

1-Oleoyl-2-palmitoyl-3-linoleoylglycerol (OPL) is a critical structured lipid in human milk fat and it plays an important role in infants' health. The development of an efficient method for such compound preparation is necessary. In this study, one- and two-step methods for OPL production by enzymatic acidolysis of fractionated palm stearin with mixed acids were compared in solvent-free and solvent systems. The results demonstrated that in the solvent system, the OPL contents by the one- and two-step methods were 45.8% and 56.7%, respectively. In the solvent-free system, the one- and two-step methods resulted in the formation of 43.7% and 54.8% OPL, respectively. Subsequently, acidolysis conditions of the two-step method were optimized. Under the optimum conditions, the content of OPL was 57.7%, and 83.2% of palmitic acid was at the sn-2 position. The prepared OPL has potential to be applied in infant formulas.