Simple and Efficient One-Pot Extraction Method for Phospholipidomic Profiling of Total Oil and Lecithin by Phosphorus-31 Nuclear Magnetic Resonance Measurements

The present study reports an efficient method using ethanol and hexane for lipid extraction (ETHEX) that is simpler and faster than the FOLCH (methanol/chloroform) and PALC (ethanol/hexane, a multi-step and time-consuming method) methods for determination of the phospholipid (PL) and fatty acid contents, using hoki roe as a model system. Substantial differences were found with the PALC and ETHEX methods, resulting in higher total lipid (14.6 ± 0.35 and 14.3 ± 0.08%, respectively) and lecithin (4.95 ± 0.08 and 4.89 ± 0.35%, respectively) yields compared to the FOLCH method (total lipid, 12.9 ± 0.35%; lecithin, 3.15 ± 0.35%). Phospholipids (LDPG, CL, LPS, SM, PE, LPC, PI, and PC) were found to partition in the methanol aqueous layer with the FOLCH method. Better phosphorus-31 nuclear magnetic resonance resolution and detection of PL, including lyso-PL, was obtained using D₂O. The best extraction and detection of PL was achieved with the novel ETHEX method using D₂O.