Development of environmental low-load-type analytical system of marine lipids

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The aim of this study was to develop an environmental low-load-type analytical system of marine lipids using high-performance liquid chromatography (HPLC), which reduces a consumption of organic solvents as much as possible. For this purpose, HPLC in conjunction with electrospray ionization mass spectrometry (ESI-MS) and HPLC with fluorescent detection were done using conventional ODS columns (250 x 4.6mm x i.d.). Chiral-phase HPLC and reversed-phase HPLC with UV detections were also applied using slurry-packed fused-silica capillaries (40cm x 0.1-0.3mm i.d.) with a various kind of chiral stationary phases and ODS, respectively. The chiral-phase HPLC analysis was optimized by using a ternary mobile phase, n-hexane-1,2-dichloroethane-ethanol. Using these techniques, we obtained successful separations of the molecular species of the glycerolipids (mono-, di-, triacylglycerols, alkylglycerols, phosphatidylcholines, phosphatidyl-ethanolamines, phosphatidylglycerols, phosphatidylinositols) including enantiomers and diastereomers. The glycerolipids were chromatographed as intact forms, 3,5-dinitrophenylurethanes and 1-anthrylurethanes. In the present study, we also achieved the separation and identification of the reverse isomers of glycerolipids, such as 1,2-diacylglycerols of 20:5-16:0 and 22:6-16:0, using reversed-phase HPLC/ESI-MS. The present study provides environmental low-load-type analytical methods of marine lipids, which can reduce a consumption of organic solvents compared to conventional HPLC methods.