用软X射线,大气压,光电离子化和质谱法进行类分析
Simona Sedláčková1,2, Juha-Pekka Hieta3, Miroslava Blechová1
1Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, CZ-166 10 Prague 6, Czech Republic.
概括
软X射线光电化为蛋白质组学中的类分析提供了一种新的方法. 这种方法增强了电离,特别是在负离子模式下,有助于对具有挑战性的和翻译后修饰的表征.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 质谱测量质量谱测量
背景情况:
- 从底部向上的蛋白质组学通常使用反向相位超高性能液体染色学 (UHPLC) 和电喷离子化 (ESI) 双重质谱法 (MS/MS).
- 传统方法面临的挑战是电离酸性/疏水性和检测后翻译性修改 (PTMs).
- 真空紫外线 (VUV) 大气压光电离 (APPI) 已被探索为一种替代的电离技术.
研究的目的:
- 研究使用一种新型原型APPI源,利用软X射线光子进行分析.
- 为了比较软X射线APPI与VUVAPPI在正离子和负离子模式.
- 评估软X射线APPI在改善酸电离和PTM表征方面的潜力.
主要方法:
- 使用新型软X射线APPI源进行化.
- 软X射线和VUV APPI之间的电离效率和通路的比较.
- 分析软X射线APPI产生的脱质物种 ([M-2H]•-) 的片分裂 (碰撞诱导解离 - CID).
主要成果:
- 与VUV APPI相比,软X射线APPI显示出更高的电离能力.
- 软的X射线光子有效地产生了去化和缺的基离子 ([M - 2H]•-) 在负离子模式下.
- [M-2H]•-的CID产生了独特的结构信息,有利于PTM识别.
结论:
- 软X射线APPI在底部向上蛋白质组学中为类分析提供了一个有希望的进步.
- 使用负离子模式与软X射线APPI提高了和PTMs的检测和表征.
- 这种新的技术为与传统蛋白质组分析方法相关的局限性提供了潜在的解决方案.
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