整合NMR和基于多个LC-MS的非向代谢学,对血清样本进行全面分析
Tereza Kacerova1, Elisabete Pires1, John Walsby-Tickle1
1Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, OX1 3TA, UK.
Analytica chimica acta
|April 27, 2025
概括
这项研究引入了一种新的血清制备方法,用于核磁共振 (NMR) 和液态染色体质谱 (LC-MS) 代谢学. 这种高效的方法增强了从单个样本中检测代谢物和生物标志物的发现.
科学领域:
- 代谢学 代谢学 代谢学
- 分析化学 分析化学
- 生物标志物发现发现
背景情况:
- 质谱 (MS) 和核磁共振 (NMR) 是生物流体代谢学中疾病机制和生物标志物发现的关键工具.
- 由于样品制备的需求不同,这些技术往往没有得到充分利用,例如NMR的化溶剂和MS的蛋白质去除.
- 基于MS的方法中的兼容性问题和蛋白质生物标记物的潜在损失限制了综合分析.
研究的目的:
- 开发一种统一的血清制备协议,用于使用NMR和多LC-MS的顺序非向代谢学分析.
- 通过这两种技术,研究化溶剂和缓冲剂对代谢物检测的影响.
- 为了优化样本准备,提高代谢物丰富度和更广泛的代谢组覆盖范围.
主要方法:
- 开发并验证了一种用于组合NMR和多LC-MS分析的新型血清样本准备协议.
- 使用非向性NMR和多个LC-MS平台分析了人类血清样本.
- 评估了脱溶剂,缓冲剂和蛋白质去除方法 (溶剂沉,MWCO过) 对代谢物概况的影响.
主要成果:
- 在LC-MS分析之前,在NMR样品制备中使用化溶剂时,没有观察到显著的融入代谢物中.
- 与NMR兼容的缓冲器没有对LC-MS性能产生负面影响.
- 蛋白质去除策略显著影响了代谢物丰富度,突出了其在样本准备中的关键作用.
结论:
- 单个血清分量可以有效地用于NMR和多LC-MS的同时非定向分析,从而提供更有效的工作流程.
- 这种综合方法减少了样本量要求,并扩大了代谢组覆盖范围.
- 该研究提供了对样品制备对生物流体代谢学影响的关键见解,证明了NMR和LC-MS的兼容性和互补性.
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