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. 2016 May 5;291(28):14540–14554. doi: 10.1074/jbc.M115.711267

FIGURE 2.

FIGURE 2.

LC-MS analysis of human epidermal extracts for linoleate and linoleate triol-containing ceramides. A, total ion current profile (m/z 950–1350) of LC-MS analysis of a MeOH/CHCl3 extract of human epidermis using an Advantage 5-μm silica column (250 × 4.6 mm) run with gradient elution of hexane/IPA/HAc (95:5:0.1) to hexane/IPA/HAc (75:25:0.1) over 30 min using a Waters Alliance 2690 HPLC system coupled to a TSQ Vantage mass spectrometer (Thermo Scientific). The arrow at 20 min marks the retention time of putative linoleate triol-containing ceramides. B, extracted ion chromatogram of the molecular species eluting at 20 min. C, selected ion monitoring of C32 ceramide species to the respective daughter ions of m/z 279 (linoleate) and m/z 329 (linoleate triols), confirming identification of EOS (m/z 1075–279 [M + Cl]-), EOS-triol (m/z 1149–329 [M − H + CH3CO2H]-), and Glu-EOS (m/z 1237–279 [M + Cl]-).