一种基于LC-MS的混合模式方法,用于从生物样本矩阵中全面分析NAD和相关代谢物
Omprakash Nacham1, Jordan W Brown1, Mohammad Mehdi Maneshi1
1AbbVie, 1 North Waukegan Road, North Chicago, IL, 60044, USA.
Scientific reports
|April 24, 2025
概括
一种新的LC-MS/MS方法准确地分析尼古丁胺氨酸二核酸 (NAD+) 和其代谢物. 这种经过验证的平台证实了补充的NAD+增强效应,增强了我们对NAD+路径生物学的理解.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
背景情况:
- 尼古丁胺胺氨基二核酸 (NAD+) 对于细胞能量至关重要,但其水平随着年龄的增长而下降,与与年龄相关的疾病有关.
- 了解NAD+代谢对于开发有效的补充疗法至关重要.
- 分析极性NAD+代谢物在分析上具有挑战性,因为标准染色学中的保留率较低.
研究的目的:
- 开发和优化一种强大的分析方法来量化NAD+及其代谢物.
- 为了验证这种方法用于生物样本分析和与现有测试的兼容性.
- 为了研究尼古丁胺胺 (NAM) 补充剂对NAD+代谢的影响.
主要方法:
- 开发一种混合模式 (逆相/离子交换) 液态染色学-并联质谱法 (LC-MS/MS) 方法.
- 优化色谱条件,包括移动相离子强度和柱体温度.
- 对线性,准确性和精度的方法资格,以及对NAD-Glo试验的验证.
主要成果:
- LC-MS/MS方法证明了NAD+代谢物分析的高性能.
- 对组织NAD+的LC-MS/MS和NAD-Glo测定结果之间观察到强烈的相关性 (R2 = 0.94).
- 纳米补充剂在小鼠组织中证实了剂量依赖的NAD+增加,在高剂量时观察到特定代谢物水平的变化.
结论:
- 一种优化的LC-MS/MS方法为NAD+代谢组分析提供了可靠的平台.
- 这种方法可以作为基于生物发光的测试的直角验证,如NAD-Glo.
- 这些发现支持使用NAM补充剂来提高NAD+水平,并提供有关NAD+途径动态的见解.
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