Glycerol ester (GE) is a kind of important lipid in milk, which varies greatly depending on many factors. In this study, a novel pseudotargeted lipidomics strategy, named SWATH-MS2&1, was developed for the detection of GEs in milk and the Folch method was selected for the sample preparation. The developed method exhibited a competitive alternative to the acknowledged pseudotargeted strategy, including wider coverage (12 more GEs detected), higher repeatability (12 more GEs, whose coefficient of variation < 0.3), better linearity (5 more GEs, whose R2 > 0.8), and similar sensitivity (only 2 GEs less than P-MRM after dilution). SWATH-MS2&1 was applied in the investigation of GEs from different milk samples. The orthogonal partial least-squares difference analysis of 219 GEs identified from SWATH-MS2&1 showed satisfying differentiation of different milk samples, and 76 GEs were screened out as potential markers. Our findings demonstrated that SWATH-MS2&1 could offer an accurate method to measure a wide spectrum of GEs in milk.
Keywords: DIA; HPLC-Q-TOF-MS; SWATH; SWATH-MS2&1; glycerol ester; lipid extraction; lipidomics; milk; pseudotargeted omics.