Measuring NAD+ levels in mouse blood and tissue samples via a surrogate matrix approach using LC-MS/MS
Measuring NAD+ levels in mouse blood and tissue samples via a surrogate matrix approach using LC-MS/MS
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DOI：
10.4155/bio.14.8
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发表时间：
2014-01-01
期刊：
影响因子：
1.8
通讯作者：
Deng, Yuzhong
中科院分区：
文献类型：
--
作者：
Liang, Xiaorong;Yang, Lulu;Deng, Yuzhong
背景：NAD(+)是一种内源性分析物，在血液样本采集过程中不稳定，这两者都给定量带来了障碍。此外，目前的NAD(+)样本采集程序需要现场使用强酸处理，以稳定小鼠血细胞中的NAD(+)。结果：冰冻小鼠血液标本解冻前加入酸可使NAD(+)稳定。为了便于分析小鼠血液和组织样本中的NAD(+)，提出了一种简单的样本收集程序。建立了LC-MS/MS法测定小鼠血液和各种组织样品中NAD(+)的方法。所描述的方法用于测量口服烟酰胺磷酸核糖转移酶抑制剂GNE-617后小鼠血液中内源性NAD(+)的水平。结论：本研究为临床前发现研究中NAD(+)样本的高通量筛选提供了一种合适的检测和样本采集程序。
Background: NAD(+) is an endogenous analyte and is unstable during blood sample collection, both of which present obstacles for quantitation. Moreover, current procedures for NAD(+) sample collection require onsite treatment with strong acid to stabilize the NAD(+) in mouse blood cells. Results: NAD(+) can be stabilized by addition of acid before the frozen mouse blood sample was thawed. A simple sample collection procedure was proposed to facilitate the analysis of NAD(+) in mouse blood and tissue samples. A LC-MS/MS method was developed for quantifying NAD(+) in mouse blood and various tissue samples. The described method was used to measure endogenous NAD(+) levels in mouse blood following oral administration of the nicotinamide phosphoribosyltransferase inhibitor GNE-617. Conclusion: This study presents a suitable assay and sample collection procedure for high throughput screening of NAD(+) samples in preclinical discovery studies.