Pseudotargeted Lipidomics Strategy Enabling Comprehensive Profiling and Precise Lipid Structural Elucidation of Polyunsaturated Lipid-Rich Echium Oil

Echium oil has great nutritional value as a result of its high content of α-linolenic acid (ALA, 18:3ω-3) and stearidonic acid (SDA, 18:4ω-3). However, the comprehensive lipid profiling and exact structural characterization of bioactive polyunsaturated lipids in echium oil have not yet been obtained. In this study, we developed a novel pseudotargeted lipidomics strategy for comprehensive profiling and lipid structural elucidation of polyunsaturated lipid-rich echium oil. Our approach integrated untargeted lipidomics analysis with a targeted lipidomics strategy based on Paternò–Büchi (PB)–tandem mass spectrometry (MS/MS) using 2-acetylpyridine (2-AP) as the reaction reagent, allowing for high-coverage lipid profiling and simultaneous determination of C═C locations in triacylglycerols (TGs), diacylglycerols (DGs), free fatty acids (FFAs), and sterol esters (SEs) in echium oil. A total of 209 lipid species were profiled, among which 162 unsaturated lipids were identified with C═C location assignment and 42 groups of ω-3 and ω-6 C═C location isomers were discovered. In addition, relative isomer ratios of certain groups of lipid C═C location isomers were revealed. This pseudotargeted lipidomics strategy described in this study is expected to provide new insight into structural characterization of distinctive bioactive lipids in food.

• This study develops a novel pseudotargeted lipidomics approach integrating untargeted UPLC-HRMS with Paterno−Buchi (PB) reaction tandem MS, enabling high-coverage lipid profiling and unambiguous C=C localization in triacylglycerols (TGs), diacylglycerols (DGs), free fatty acids (FFAs), and sterol esters (SEs) from echium oil.
• 162 unsaturated lipids with C=C locations, including 42 groups of ω-3/ω-6 positional isomers were identified in this study. Relative isomer ratios were also determined, providing critical insights into the nutritional value and distinctive bioactivity of echium oil.