Discovering oxidized polar lipids in microalgae lipidome using liquid chromatography mass spectrometry approaches

Microalgae are rich in polar lipids, such as glycolipids, phospholipids and betaine lipids esterified with omega-3 polyunsaturated fatty acids (PUFA), which are prone to oxidation. Under stress conditions, the redox balance in microalgae is altered, leading to an abnormal production of reactive oxygen species (ROS) that can affect surrounding lipids. While few oxidized polar lipids have been described to play important roles in mammals and possess interesting bioactive properties, their occurrence in microalgae is poorly described, hindering their exploitation as novel bioactive compounds. To enhance our understanding of these lipids, we conducted a targeted analysis of oxidized polar lipids in lipid extracts obtained from five microalgae species: Chlorella vulgaris, Chlorococcum amblystomatis, Scendesmus obliquus, Nanochloropsis oceanica and Phaeodactylum ticornutum, using reverse-phase liquid chromatography mass spectrometry (RP-LC-MS). A detailed analysis of the LC-MS data identified 150 oxidized polar lipid species across different classes of phospholipids, glycolipids and betaine lipids in the five microalgae species. These modified oxidized lipids featured oxygenated species with 1–3 additional oxygen atoms in the fatty acyl chains. The predominant oxidized fatty acids esterified to these lipids were omega-3 eicosapentaenoic acid (EPA, 20:5 n-3) and α-linonelic acid (ALA, 18:3 n-3). Notably, most oxidized lipid species were identified in diacylglyceryl-N,N,N-trimethylhomoserine (DGTS) in C. amblystomatis, S. obliquus and N. oceanica, monogalactosyldiacylglycerol (MGDG) in P. tricornutum, and phosphatidylethanolamine (PE) in C. vulgaris. Furthermore, distinct fragmentation patterns across lipid classes allowed the unequivocal identification of oxidized polar lipids. These findings reveal a diverse array of oxidized polar lipids in microalgae, predominantly enriched with oxidized omega-3 PUFA, highlighting microalgae as a natural source of oxidized polar lipids that may serve as a natural reservoir of bioactive omega-3 oxylipins.