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

环境低负荷型海洋脂质分析系统的研制

基本信息

批准号：
16580164
负责人：
ITABASHI Yutaka
金额：
$ 2.37万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

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.

本研究的目的是开发一种环境低负荷型海洋脂质分析系统，使用高效液相色谱（HPLC），这将减少有机溶剂的消耗尽可能多。为此目的，使用常规ODS柱（250 x 4.6mm x i.d.）进行HPLC结合电喷雾电离质谱（ESI-MS）和HPLC荧光检测。手性相HPLC和反相HPLC与UV检测也使用浆料填充的熔融石英毛细管（40 cm X 0.1-0.3mm i. d.）分别用不同的手性固定相和ODS固定。采用正己烷-1，2-二氯乙烷-乙醇三元移动的流动相优化了手性HPLC分析。使用这些技术，我们获得了成功的分离的分子种类的甘油脂（单，二，三酰基甘油，烷基甘油，磷脂酰胆碱，磷脂酰乙醇胺，磷脂酰甘油，磷脂酰肌醇），包括对映异构体和非对映异构体。对完整形式、3，5-二硝基苯氨基甲酸乙酯和1-蒽基氨基甲酸乙酯的甘油脂进行色谱分析。本研究还利用反相HPLC/ESI-MS技术对1，2-二酰基甘油（20：5-16：0和22：6-16：0）的反相异构体进行了分离和鉴定。