使用溶剂切换液体色谱学,并联质量谱学进行全面的无目标极性代谢物分析
Jake P Violi1, Connor R Phillips2, David S Gertner2
1School of Chemistry, University of New South Wales, Sydney, NSW, Australia.
Talanta
|January 24, 2025
概括
这项研究开发了一个全面的极性代谢学管道,用于基于细胞的分析. 一种结合酸性和基本色谱的顺序方法显著增加了神经毒性研究中识别的代谢物数量.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 细胞生物学 细胞生物学
背景情况:
- 代谢学分析细胞生化物质,以了解代谢途径和疾病状态.
- 代谢物是多样化的,使全面分析和管道开发复杂化.
- 现有的单流体色谱-质谱方法可能会产生有限的可识别代谢物.
研究的目的:
- 为基于细胞的研究开发一个全面的极性代谢学管道.
- 优化质谱和色谱条件,以提高极性代谢物检测.
- 评估不同的水友互动液体染色学 (HILIC) 方法和顺序分析策略.
主要方法:
- 在各种pH值下优化基于HILIC的代谢学方法.
- 测试了阳性,阴性和极性切换质谱模式.
- 比较单个方法与顺序运行两个优化方法 (酸性和基本性条件).
主要成果:
- 在极性切换模式下结合酸性和基本色谱条件的顺序方法确定了最独特的代谢物.
- 这种方法在单个分析批量内是可行的,因为柱子的pH值可以容忍.
- 一种快速的溶剂切换技术调节了列,并准备了MS源进行顺序分析.
结论:
- 开发的顺序HILIC-MS方法显著提高了基于细胞的样本中极性代谢物的识别.
- 这一全面的管道适用于使用SH-SY5Y神经母细胞瘤细胞进行神经毒性研究.
- 与单个方法相比,优化的序列色谱提供了更完整的代谢概况.
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