Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics
Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics
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DOI：
10.1194/jlr.d700041-jlr200
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发表时间：
2008-05-01
影响因子：
6.5
通讯作者：
Schwudke, Dominik
中科院分区：
文献类型：
--
作者：
Matyash, Vitali;Liebisch, Gerhard;Schwudke, Dominik
脂质组的准确分析依赖于从所分析的细胞、体液或组织中对脂质种类进行定量且无偏性的提取，这通常通过氯仿两相萃取来实现。我们证明了甲基叔丁基醚（MTBE）萃取能够更快且更纯净地提取脂质，非常适合自动化的鸟枪法脂质组分析。由于MTBE密度低，含脂质的有机相在相分离过程中形成上层，这简化了其收集过程并使滴漏损失最小化。不可萃取的基质在萃取管底部形成致密的沉淀，通过离心很容易去除。严格的测试表明，与“金标准”的Folch法或Bligh和Dyer法相比，MTBE方案对大多数主要脂质类别物质的提取回收率相近或更高。
Accurate profiling of lipidomes relies upon the quantitative and unbiased recovery of lipid species from analyzed cells, fluids, or tissues and is usually achieved by two-phase extraction with chloroform. We demonstrated that methyl-tert-butyl ether (MTBE) extraction allows faster and cleaner lipid recovery and is well suited for automated shotgun profiling. Because of MTBE's low density, lipid-containing organic phase forms the upper layer during phase separation, which simplifies its collection and minimizes dripping losses. Nonextractable matrix forms a dense pellet at the bottom of the extraction tube and is easily removed by centrifugation. Rigorous testing demonstrated that the MTBE protocol delivers similar or better recoveries of species of most all major lipid classes compared with the "gold-standard" Folch or Bligh and Dyer recipes.