Phenomenex Luna silica (3 μm, 1.5 mm × 200 mm) was selected as the chromatographic column. Lipids were extracted under A phase (chloroform/methanol/ammonia = 89.5:10:0.5) and B phase (chloroform/methanol/ammonia/water = 55:39:0.5:5.5). Extraction began with a 95% gradient of A phase from 0 to 5 min, then a linear decrease to 60% (in 7 min) for 4 min, a further decline to 30% for 15 min, and return to 95% for the last 5 min. Mass spectrometry multiple reaction monitoring was established for lipid identification and quantitative analysis (Lam et al., 2017 (link), 2018 (link)).
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