A not-stop-flow online normal-/reversed-phase two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry method for comprehensive lipid profiling of human plasma from atherosclerosis patients.

•A not-stop-flow online NP/RP 2D LC-MS method was developed for lipid profiling.•Overall, 540 endogenous lipid species from 17 classes were identified from human plasma.•This novel method showed high sensitivity (ng/mL grade) and good repeatability.•Successful separation of isomers of GalC and GluC was achieved.•The levels of GalC and GluC were different in human plasma from different subjects.