Sphingolipidomics: High-throughput, structure-specific, and quantitative analysis of sphingolipids by liquid chromatography tandem mass spectrometry
Sphingolipidomics: High-throughput, structure-specific, and quantitative analysis of sphingolipids by liquid chromatography tandem mass spectrometry
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DOI：
10.1016/j.ymeth.2005.01.009
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发表时间：
2005-06-01
期刊：
影响因子：
4.8
通讯作者：
Wang, E
中科院分区：
文献类型：
--
作者：
Merrill, AH;Sullards, MC;Wang, E
鞘脂是一种高度多样化的化合物，不仅作为生物结构的组分，而且作为许多细胞功能的调节剂。由于生物系统中的许多鞘脂是生物活性的，并且通常在结构和代谢上密切相关（例如，复杂鞘脂神经酰胺鞘氨醇鞘氨醇1-磷酸），为了理解鞘脂在给定背景下的作用，必须进行“鞘脂组学”分析，即，所有这些化合物或至少一个关键子集的所有成员的结构特异性和定量测量。液相色谱串联质谱法（LC MS/MS）是目前唯一一种具有必要的结构特异性、灵敏度、定量精度和相对高通量能力的技术，可用于小样本（类似于106个细胞）的此类分析。本文介绍了一系列的协议，已开发的相对快速的分析所有的分子物种，从3-酮鞘氨醇，通过鞘磷脂和一些鞘糖脂（包括目前被认为是鞘脂“第二信使”的所有化合物）使用正相和反相LC分离等轴和同量异位物质（如葡糖神经酰胺和半乳糖神经酰胺）与三重四极杆（用于MS/MS）和混合四极杆-离子阱（用于MS 3）质谱的组合。还讨论了一些剩余的问题，以解决在分析的全部鞘脂类。(c)2005年爱思唯尔公司All rights reserved.
Sphingolipids are a highly diverse category of compounds that serve not only as components of biologic structures but also as regulators of numerous cell functions. Because so many of the sphingolipids in a biological system are bioactive and are often closely related structurally and metabolically (for example, complex sphingolipids ceramide sphingosine sphingosine 1-phosphate), to understand the role(s) of sphingolipids in a given context one must conduct a "sphingolipidomic" analysis-i.e., a structure-specific and quantitative measurement of all of these compounds, or at least all members of a critical subset. Liquid chromatography tandem mass spectrometry (LC MS/MS) is currently the only technology with the requisite structural specificity, sensitivity, quantitative precision, and relatively high-throughput capabilities for such analyses in small samples (similar to 106 cells). This review describes a series of protocols that have been developed for the relatively rapid analysis of all of the molecular species from 3-ketosphinganines through sphingomyelins and some glycosphingolipids (including all the compounds that are presently regarded as sphingolipid "second messengers") using normal- and reverse-phase LC to separate isometric and isobaric species (such as glucosylceramides and gatactosylceramides) in combination with triple quadrupole (for MS/MS) and hybrid quadrupole-ion trap (for MS3) mass spectrometiy. Also discussed are some of the issues remaining to be resolved in the analysis of the full sphingolipidome. (c) 2005 Elsevier Inc. All rights reserved.