开发和验证使用液体染色学-并联质谱法 (LC-MS/MS) 来定量235种血代谢物的有针对性的代谢学方法
Kangkang Xu1,2,3, Franz Berthiller1,2, Barbara U Metzler-Zebeli2,4
1Institute of Bioanalytics and Agro-Metabolomics, Department of Agricultural Sciences, BOKU University, 3430 Tulln, Austria.
Molecules (Basel, Switzerland)
|February 13, 2025
概括
这项研究开发了向液体染色学-并联质谱法 (LC-MS/MS) 方法,用于量化猪血中的235种代谢物. 这些经过验证的方法可以为生物医学研究提供全面的血代谢组分析.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 生物医学科学 生物医学科学
背景情况:
- 血代谢组分析对于健康监测和医学评估至关重要.
- 血代谢物的复杂性和多样性带来了重大的分析挑战.
- 现有的方法往往需要衍生,限制吞吐量和范围.
研究的目的:
- 开发和验证向液体染色学-并联质谱法 (LC-MS/MS) 方法,以量化猪血中的235种哺乳动物代谢物.
- 建立适用于大规模代谢学研究的可靠分析技术.
- 为了实现全面的分析,而不需要先前的衍生.
主要方法:
- 使用逆相和水友相互作用液态染色学 (RPLC和HILIC) 与并联质谱学 (MS/MS) 相结合.
- 采用了两种选择的反应监测 (SRM) 过渡来识别和量化分析物.
- 实施了快速极性切换和计划的SRM,以提高覆盖率和吞吐量.
- 使用"稀释和射击"样品制备协议,具有双重稀释水平.
主要成果:
- 在猪血中成功量化了来自17个化合物类的235种代谢物.
- 每个样品的分析时间约为40分钟.
- 证明方法验证,包括线性,检测/定量限制,可重复性和恢复.
- 已证实适用于包括人类在内的各种动物物种的血样本.
结论:
- 建立了两种可靠的LC-MS/MS方法,用于全面量化猪血代谢组.
- 开发的方法在生物医学研究中推进了大规模的向代谢学.
- 该方法提供了一个多功能工具,用于分析跨物种的多样化代谢物概况.
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