Mass spectrometry in combination with a chiral column and multichannel-MRM allows comprehensive analysis of glycosphingolipid molecular species from mouse brain
Mass spectrometry in combination with a chiral column and multichannel-MRM allows comprehensive analysis of glycosphingolipid molecular species from mouse brain
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DOI：
10.1016/j.carres.2020.107959
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发表时间：
2020-04-01
影响因子：
3.1
通讯作者：
Yokoyama, Kazuaki
中科院分区：
文献类型：
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作者：
Fujiwara, Yuko;Hama, Kotaro;Yokoyama, Kazuaki
鞘糖脂（Glycosphingolipids, GSLs）仅存在于哺乳动物细胞的质膜外小叶，具有多种结构，包括不同种类的糖和不同分子种类的神经酰胺部分。建立测量GSL类中每个分子种的方法有助于GSL的功能表征和揭示鞘糖脂病发病机制的细节。使用从未用于脂质分析的IF-3手性柱，我们开发了一种液相色谱-质谱（LC-MS）方法来分离基于糖和神经酰胺部分的各种GSLs。为了详细检查GSLs，使用了多通道-多反应监测（multichannel-MRM）模式，覆盖范围为500-2000 Da。在正离子模式下检测到的碰撞能量较高的常见碎片离子为m/z 264和292，分别来源于d18:1和d20:1离子。在多通道mrm中，这两个物种被用作产物离子，用于同时测量中性GSLs、神经节苷脂和硫脂。利用该方法对小鼠脑内GSLs进行综合分析发现，神经节苷类和LacCer的主要分子种为d18:1-C18:0和d20:1-C18:0，而硫脂类的主要分子种为d18:1-C24:0和d18:1-C24:1。结果揭示了多样化的GSL脂肪酸谱。综上所述，通过结合IF-3手性柱和多通道mrm方法成功检测到GSLs的多种分子种类，基于该LC-MS方法的代谢组学方法将有助于GSLs的功能分析和在分子水平上发现鞘糖脂病的早期生物标志物。
Glycosphingolipids (GSLs) exist exclusively in the outer leaflet of plasma membrane in mammalian cells and have diverse structures including different classes of sugars and various molecular species of ceramide moieties. Establishing methods that measure each molecular species in GSL classes should aid functional characterization of GSLs and reveal details about the mechanism of pathogenesis in glycosphingolipidoses. Using an IF-3 chiral column that has never been used for lipid analyses, we developed a liquid chromatography-mass spectrometry (LC-MS) method to separate various GSLs based on sugar and ceramide moieties. To examine GSLs in detail a multichannel-multiple reaction monitoring (multichannel-MRM) mode was used and covered a range of 500-2000 Da. Common fragment ions detected with higher collision energy in the positive ion mode were m/z 264 and 292, and are derived from d18:1 and d20:1 ions, respectively. Both species were used as product ions in the multichannel-MRM for the simultaneous measurement of neutral GSLs, gangliosides and sulfatides. Comprehensive analysis of GSLs in mouse brain using this method revealed that for gangliosides and LacCer, d18:1-C18:0 and d20:1-C18:0 were the major molecular species, whereas d18:1-C24:0 and d18:1-C24:1 were the major molecular species of sulfatides. The results revealed a diverse GSL fatty acid profile. In conclusion, by combining IF-3 chiral column and the multichannel-MRM method various molecular species of GSLs were detected successfully, and a metabolomics approach based on this LC-MS method should facilitate functional analysis of GSLs and the discovery of early biomarkers of glycosphingolipidoses at the molecular level.