Analytical procedures for the preparation, isolation, analysis and preservation of reduced nicotinamides
Analytical procedures for the preparation, isolation, analysis and preservation of reduced nicotinamides
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DOI：
10.2174/157341106778520508
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发表时间：
2006-10-01
影响因子：
1.8
通讯作者：
Kohen, Amnon
中科院分区：
文献类型：
--
作者：
Markham, Kelli A.;Kohen, Amnon
含有辅因子的烟酰胺在生物系统中普遍存在。因此，当在体外或体内研究烟酰胺依赖性系统时，同位素标记的烟酰胺辅因子通常用于揭示感兴趣的生物系统的机制。这些辅因子的还原形式是不稳定的，对光、氧、pH、温度和其他因素敏感。因此，它们的合成、纯化、分析和保存一直是一个持续的挑战。由于这些稳定性问题，使用烟酰胺辅因子的研究，特别是使用还原形式的那些，已经被降级为在实验使用之前合成所需材料的重复循环。这种做法在利用复杂同位素标记模式的研究中是不切实际的。已经报道的用于各种烟酰胺辅因子的合成、分离和长期保存的许多努力导致程序不再实用或具有成本效益，而其他程序则提供了当今可用的最佳解决方案。在这篇小型评论中介绍并比较了其中的一些努力。在此，我们描述了我们用于不稳定烟酰胺辅因子的合成、分离、纯化、分析和长期保存的分析和合成方法，其实现了优异的通用性、产率、纯度、HPLC分辨率和长期稳定性。
Nicotinamide containing cofactors are ubiquitous in biological systems. As a result, when studying nicotinamide dependent systems in vitro or in vivo, isotopically labeled nicotinamide cofactors are often used to reveal the mechanism of the biological system of interest. The reduced form of these cofactors is labile and is sensitive to light, oxygen, pH, temperature and other factors. Consequently, their synthesis, purification, analysis, and preservation have been an ongoing challenge. Due to these stability issues, studies using nicotinamide cofactors, especially those utilizing the reduced forms, have been relegated to repetitive cycles of synthesizing the needed material just prior to experimental use. Such practice is not practical in studies utilizing complex isotopic labeling patterns. Many of the efforts that have been reported for synthesis, separation and long-term preservation of the various nicotinamide cofactors, led to procedures that are no longer practical or cost effective while others present the best solutions available today. Some of these efforts are presented and compared in this mini-review. Herein we describe our analytical and synthetic methodologies for the synthesis, separation, purification, analysis and long term preservation of labile nicotinamide cofactors, which achieve excellent versatility, yields, purity, HPLC resolution, and long term stability.