methods Targeted analysis of ganglioside and sulfatide molecular species by LC/ESI-MS/MS with theoretically expanded multiple reaction monitoring
methods Targeted analysis of ganglioside and sulfatide molecular species by LC/ESI-MS/MS with theoretically expanded multiple reaction monitoring
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发表时间：
2008
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通讯作者：
Kazutaka Ikeda;T. Shimizu;R. Taguchi
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作者：
Kazutaka Ikeda;T. Shimizu;R. Taguchi
基质辅助激光解吸/电离质谱（MALDI-MS）主要应用于通过TLC分析从其他复杂混合物中分离出来的鞘糖脂，但通过MALDI-MS很难获得TLC上不同点之间每种鞘糖脂的定量分析。因此，开发一种利用液相色谱/电喷雾电离 (LC/ESI)-MS 的便捷方法引起了人们的兴趣。然而，先前报道的方法不足以分离和区分每种神经节苷脂类别。在此，我们报告了一种通过 LC/ESI 串联质谱 (LC/ESI-MS/MS) 结合多反应监测 (MRM) 对理论上预期的神经节苷脂分子种类进行靶向分析的有效方法。针对唾液酸的 MRM 检测使我们能够分析皮摩尔至飞摩尔水平的神经节苷脂标准品，例如 GM1、GM2、GM3、GD1 和 GT1。此外，通过理论上扩展的 MRM，还可以在猪脑的鞘糖脂标准混合物和小鼠脑的酸性糖脂提取物中检测到其他神经节苷脂，如 GD2、GD3、GT2、GT3 和 GQ1。我们发现这种方法也适用于鞘糖脂混合物中含有的硫苷脂。此外，我们建立了一种用糖链MRM分离和区分区域异构神经节苷脂的方法，例如GM1a和-1b，GD1a，-1b和-1c，以及GT1a，-1b和-1c。资源。 2008. 49: 2689. ( 2 55 V)、GT1、GT2、GT3 和 GQ1 ( 2 60 V) 和硫苷脂 ( 2 117.5 V)。通过这些 MS/MS 谱图选择用于 MRM 分析的母离子和产物离子对。监测以下 MRM m/z 转变：神经节苷脂的母离子到唾液酸离子 (SA 2 ) 或含唾液酸的糖链，以及脑硫苷脂的母离子到硫酸根离子 (HSO 4 2 )。为了分析区域异构体神经节苷脂，选择源自这些异构体糖链末端区域的特定片段，并在与每种神经节苷脂的 MRM 分析相同的条件下进行操作。
Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) has been applied primarily to the analysis of glycosphingolipids separated from other complex mixtures by TLC, but it is difficult to obtain quantitative profiling of each glycosphingolipid among the different spots on TLC by MALDI-MS. Thus, the development of a convenient approach that utilizes liquid chromatography/ electrospray ionization (LC/ESI)-MS has received interest. However, previously reported methods have been insuffi-cient to separate and distinguish each ganglioside class. Here we report an effective method for the targeted analysis of theoretically expected ganglioside molecular species by LC/ESI tandem mass spectrometry (LC/ESI-MS/MS) in combination with multiple reaction monitoring (MRM). MRM detection specific for sialic acid enabled us to analyze ganglioside standards such as GM1, GM2, GM3, GD1, and GT1 at picomolar to femtomolar levels. Furthermore, other gangliosides, such as GD2, GD3, GT2, GT3, and GQ1, were also detected in glycosphingolipid standard mixtures from porcine brain and acidic glycolipid extract from mouse brain by theoretically expanded MRM. We found that this approach was also applicable to sulfatides contained in the glycosphingolipid mixtures. In addition, we established a method to separate and distinguish regioisomeric gangliosides, such as GM1a and -1b, GD1a, -1b, and -1c, and GT1a, -1b, and -1c with sugar chains the MRM. Res. 2008. 49: 2689. ( 2 55 V), GT1, GT2, GT3, and GQ1 ( 2 60 V), and sulfatides ( 2 117.5 V). The precursor and product ion pairs for the MRM analysis were selected by these MS/MS spectra. The following MRM m/z transitions were monitored: parent ions to sialic acid ions (SA 2 ) or sialic acid-containing sugar chains for gangliosides and parent ions to sulfate ions (HSO 4 2 ) for sulfatides. For the analysis of regioisomeric gangliosides, specific fragments derived from terminal regions of the sugar chain in these isomers were selected and operated under the same conditions of the MRM analysis for each ganglioside.