Retention behavior of lipids in reversed-phase ultrahigh-performance liquid chromatography-electrospray ionization mass spectrometry.

•RP-UHPLC/MS two sub–2μm C18 columns used for study of retention behavior of lipids.•HR-MS/MS in both polarity modes with high mass accuracy used for identification.•Method is applied for human plasma, human erythrocytes, human urine and porcine brain.•Over 400 lipids from 14 classes and 5 lipid categories are unambiguously identified.•Retention times vs. fatty acyl chain length or DB number fitted by polynomial regressions.