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通过使用校准常数,通过质谱测量对NAD+/NADH和NADP/NAPH+/NADP+进行快速量化

Qiuyuan Guo¹, Mike Lingjue Wang¹, Michaela Schwaiger-Haber¹

¹Departments of Chemistry, Genetics, and Medicine, Washington University, St Louis, MO, USA; Center for Mass Spectrometry and Metabolic Tracing, Washington University, St Louis, MO, USA.

|December 13, 2025

概括

使用质谱仪精确测量细胞氧化还原平衡. 一种新方法使用校准常数来快速确定NAD+/NADH和NADP/NADP+的比率,比传统测试提高了准确性.

科学领域:

生物化学生物化学
代谢学代谢学代谢学
分析化学分析化学

背景情况:

细胞的氧化还原状态至关重要,由NAD+/NADH和NADP/NADP+比率表示.
对于氧化还原比的传统质谱法是耗时的,因为需要绝对度的确定.
现有的方法通常依赖于标准,限制了快速分析.

研究的目的:

开发一种基于质谱的更简单,更快速的策略来确定细胞氧化还原比.
建立一种准确量化NAD+/NADH和NADP/NAPH+的方法,没有真正的或同位素标记的标准.
为了验证新方法的准确性,与已建立的商业测试进行对比.

主要方法:

开发了一种使用质谱的代谢学工作流.
引入了从相对信号强度变化的一致速率中得出的校准常数概念.
应用校准常数来将相对质谱信号转换为准确的氧化还原比.

主要成果:

证明氧化和减少辅因子的相对信号强度在给定的质谱仪上以可预测的速度变化.
展示了校准常数允许相对强度快速转化为准确的氧化还原比.
验证了质谱法,与商业色度测试相比,显示出更高的准确性.

结论:

开发的质谱法提供了一种更准确,更有效的方法来确定细胞氧化还原状态.
校准常量简化了氧化还原比的确定,消除了耗时绝对量化或昂贵标准的需要.
这种方法增强了质谱在代谢学和氧化还原生物学研究中的实用性.

关键词:

校准常量是指校准常量.在 LC/MS 中使用.代谢学代谢学代谢学这是NAD+) /NADH.NADPH/NADP ((+) 是一个尼古丁胺胺腺因二核酸是什么尼古丁胺胺腺氨酸二核酸酸盐

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