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A comparison of five lipid extraction solvent systems for lipidomic studies of human LDL

机译:五个脂质提取溶剂系统用于人LDL脂质组学研究的比较

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摘要

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.
机译:流体和组织的脂质组概况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的有效和代表性的提取。尽管尚未对跨多种脂质类别的有效性进行全面研究,但通常使用许多适用于脂质提取的溶剂系统。为了解决这个问题,使用了来自高血脂志愿者的人类低密度脂蛋白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)和己烷-异丙醇]和提取的脂质在配备极性切换的高分辨率仪器中通过LC-MS分析。总体上,从19种脂质亚类中鉴定出350多种脂质。溶剂组成对主要脂质类别(三酰基甘油酯,胆固醇酯和磷脂酰胆碱)的提取影响很小。相反,较不丰富的脂质(磷脂酰肌醇,溶血脂质,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,尽管己烷-异丙醇方法最适合非极性脂质,而MeOH-TBME方法最适合于乳糖基神经酰胺,但Folch方法对LDL中多种脂质类别的提取最为有效。

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