Striby L, Lafont R, Goutx M
Laboratoire de Microbiologie Marine, CNRS-INSU EP 2032, Marseille, France.
J Chromatogr A. 1999 Jul 23;849(2):371-80. doi: 10.1016/s0021-9673(99)00589-0.
Mono- and diacylglycerols are important intermediates in glycerolipid biodegradation and intracellular signalling pathways. A method for mass determination of these lipid classes in marine particles was developed using the Iatroscan, which combines thin layer chromatography (TLC) and flame ionisation detection (FID) techniques. We improved existing protocols by adding two elution steps: hexane-diethyl-ether-formic acid (70:30:0.2, v/v/v) after triacylglycerol and free fatty acid scan, and acetone 100% followed by chloroform-acetone-formic acid (99:1:0.2, v/v/v) after 1,2 diacylglycerols. Diacylglycerol isomers 1,2 and 1,3 were separated from each other, as well as from free sterols in standards and marine lipids from sediment trap particles. Monoacylglycerols were separated from pigments and galactosyl-lipids in the same trap samples and in a rich pigment phytoplankton extract of Dunaliella viridis. Quantitation of each class in samples was performed after calibration with 0.5 to 2 micrograms of standards. As many as 17 lipid classes can be identified and quantified in samples using this proposed six-step development.
单酰甘油和二酰甘油是甘油脂质生物降解和细胞内信号传导途径中的重要中间体。利用结合了薄层色谱（TLC）和火焰离子化检测（FID）技术的Iatroscan开发了一种测定海洋颗粒中这些脂质类别的质量的方法。我们通过添加两个洗脱步骤改进了现有方案：在三酰甘油和游离脂肪酸扫描后使用己烷 - 二乙醚 - 甲酸（70:30:0.2，v/v/v），在1,2 - 二酰甘油之后依次使用100%丙酮，然后是氯仿 - 丙酮 - 甲酸（99:1:0.2，v/v/v）。二酰甘油异构体1,2和1,3彼此分离，并且与标准品中的游离甾醇以及沉积物捕获颗粒中的海洋脂质分离。在相同的捕获样品以及杜氏盐藻富含色素的浮游植物提取物中，单酰甘油与色素和半乳糖脂分离。在用0.5至2微克标准品校准后对样品中的每一类进行定量。使用此提议的六步展开法，样品中多达17种脂质类别可被鉴定和定量。
