PDF(Pubmed)
Abstract：
Nicotinamide adenine dinucleotide (NAD) is a cofactor in redox reactions and an essential mediator of energy metabolism. The redox balance between NAD+ and NADH affects various diseases, cell differentiation, and aging, and in recent years there has been a growing need for measurement techniques with improved accuracy. However, NAD(H) measurements, representing both NAD+ and NADH, have been limited by the compound\'s properties. We achieved highly sensitive simultaneous measurement of NAD+ and NADH under non-ion pairing, mobile phase conditions of water, or methanol containing 5 mM ammonium acetate. These were achieved using a simple pre-treatment and 7-min analysis time. Use of the stable isotope 13C5-NAD+ as an internal standard enabled validation close to BMV criteria and demonstrated the robustness of NAD(H) determination. Measurements using this method showed that brain NAD(H) levels correlate strongly with plasma NAD(H) levels in the same mouse, indicating that NAD(H) concentrations in brain tissue are reflected in plasma. As NAD(H) is involved in various neurodegenerative diseases and cerebral ischemia, as well as brain diseases such as mitochondrial myopathies, monitoring changes in NADH levels in plasma after drug administration will be useful for development of future diagnostics and therapeutics.
摘要：
烟酰胺腺嘌呤二核苷酸（NAD）是氧化还原反应中的辅因子，是能量代谢的重要介质。NAD+和NADH之间的氧化还原平衡影响各种疾病，细胞分化，和衰老，近年来，人们越来越需要提高测量精度的技术。然而,NAD(H)测量,代表NAD+和NADH，受到化合物属性的限制。我们实现了非离子配对下NAD+和NADH的高灵敏度同时测量，水的流动相条件，或含有5mM乙酸铵的甲醇。这些是使用简单的预处理和7分钟的分析时间来实现的。使用稳定同位素13C5-NAD+作为内标使得验证接近BMV标准,并证明NAD(H)测定的稳健性。使用这种方法的测量表明，在同一只小鼠中，大脑NAD（H）水平与血浆NAD（H）水平密切相关。表明脑组织中的NAD(H)浓度反映在血浆中。由于NAD(H)参与多种神经退行性疾病和脑缺血,以及脑疾病如线粒体肌病，监测给药后血浆中NADH水平的变化将有助于开发未来的诊断和治疗方法.
