School of Life and Health Sciences, Aston University, Birmingham, UK.
J Lipid Res. 2013 Jul;54(7):1812-24. doi: 10.1194/jlr.M034330. Epub 2013 May 13.
Lipidome profile of fluids and tissues is a growing field as the role of lipids as signaling molecules is increasingly understood, relying on an effective and representative extraction of the lipids present. A number of solvent systems suitable for lipid extraction are commonly in use, though no comprehensive investigation of their effectiveness across multiple lipid classes has been carried out. To address this, human LDL from normolipidemic volunteers was used to evaluate five different solvent extraction protocols [Folch, Bligh and Dyer, acidified Bligh and Dyer, methanol (MeOH)-tert-butyl methyl ether (TBME), and hexane-isopropanol] and the extracted lipids were analyzed by LC-MS in a high-resolution instrument equipped with polarity switching. Overall, more than 350 different lipid species from 19 lipid subclasses were identified. Solvent composition had a small effect on the extraction of predominant lipid classes (triacylglycerides, cholesterol esters, and phosphatidylcholines). In contrast, extraction of less abundant lipids (phosphatidylinositols, lyso-lipids, ceramides, and cholesterol sulfates) was greatly influenced by the solvent system used. Overall, the Folch method was most effective for the extraction of a broad range of lipid classes in LDL, although the hexane-isopropanol method was best for apolar lipids and the MeOH-TBME method was suitable for lactosyl ceramides.
脂质组学是一个不断发展的领域，因为人们越来越了解脂质作为信号分子的作用，这依赖于对存在的脂质进行有效且有代表性的提取。目前有许多适用于脂质提取的溶剂系统在广泛使用，但尚未对它们在多种脂质类别中的有效性进行全面调查。为了解决这个问题，使用来自正常脂质志愿者的人 LDL 来评估五种不同的溶剂提取方案[Folch、Bligh 和 Dyer、酸化的 Bligh 和 Dyer、甲醇（MeOH）-叔丁基甲基醚（TBME）和正己烷-异丙醇]，并通过配备极性切换的高分辨率仪器对提取的脂质进行 LC-MS 分析。总的来说，从 19 个脂质亚类中鉴定出了 350 多种不同的脂质种类。溶剂组成对主要脂质类别的提取（三酰甘油、胆固醇酯和磷脂酰胆碱）影响较小。相比之下，较少的脂质（磷脂酰肌醇、溶血磷脂、神经酰胺和胆固醇硫酸盐）的提取受到所用溶剂系统的极大影响。总的来说，Folch 方法最适合提取 LDL 中广泛的脂质类别，尽管正己烷-异丙醇方法最适合提取非极性脂质，而 MeOH-TBME 方法适合提取乳糖基神经酰胺。
