Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low-abundance regime of cellular sphingolipids
Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low-abundance regime of cellular sphingolipids
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DOI：
10.1016/j.ab.2007.08.019
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发表时间：
2007-12-15
影响因子：
2.9
通讯作者：
Han, Xianlin
中科院分区：
文献类型：
--
作者：
Jiang, Xuntian;Cheng, Hua;Han, Xianlin
含有鞘氨醇碱基的鞘脂由数百到数千种不同的化合物组成，其中许多作为生物功能的脂质调节剂。在许多情况下，大量低丰度鞘脂分子种类的全局分析受到阻碍，因为鞘脂分子种类被其他类脂质（例如甘油磷脂）分子种类的数量所掩盖，从而对使用鸟枪法脂质组学测量它们的动态范围施加了严格限制。在此，我们开发了一种简便的方法，通过对脂质提取物进行直接碱性甲醇解，然后提取以去除大部分其他内源性脂质类别，从而使细胞提取物中的鞘脂显著富集。通过直接注入所得的富集溶液，我们使用电喷雾电离质谱法（即鸟枪鞘脂组学）鉴定并定量了多种极低丰度的鞘脂类别（例如鞘氨醇、脑苷脂和溶血鞘磷脂）和分子种类（例如鞘磷脂）。因此，通过利用这些简便的富集技术，对鞘脂组的直接了解已大大扩展，有助于对其在生物系统中的代谢和信号传导功能有新的认识。（C）2007年爱思唯尔公司。保留所有权利。
Sphingolipids that contain a sphingoid base are composed of hundreds to thousands of distinct compounds, many of which serve as lipid regulators of biological functions. The global analysis of the large number of low-abundance sphingolipid molecular species has been hampered in many cases by the sphingolipid molecular species being overwhelmed by the quantity of other classes of lipid (e.g., glycerophospholipid) molecular species present, thereby imposing severe restrictions on the dynamic range of their measurement using shotgun lipidomics. Herein, we developed a facile approach in which the sphingolipids of cellular extracts were dramatically enriched by direct alkaline methanolysis of lipid extracts followed by extraction to remove the large majority of other endogenous lipid classes. Through direct infusion of the resultant enriched solution, we identified and quantitated a variety of very-low-abundance sphingolipid classes (e.g., sphingosine, psychosine, and lysosphingomyelin) and molecular species (e.g., sphingomyelin) using electrospray ionization mass spectrometry (i.e., shotgun sphingolipidomics). Accordingly, through utilization of these facile enrichment techniques, direct penetrance into the sphingolipidomes has been greatly extended, facilitating new insights into their metabolism and signaling functions in biological systems. (C) 2007 Elsevier Inc. All rights reserved.