Nutraceutical Group, Healthcare New Business Division, Teijin Limited, Hino, Tokyo, Japan.
Discovery DMPK Research Group, Toxicology & DMPK Research Department, Teijin Institute for Bio-Medical Research, Teijin Pharma Limited, Hino, Tokyo, Japan.
Anal Bioanal Chem. 2023 Feb;415(5):775-785. doi: 10.1007/s00216-022-04469-7. Epub 2022 Dec 11.
Nicotinamide adenine dinucleotide (NAD) is a coenzyme essential for energy production. Recently, associations between NAD and aging-related diseases have been reported, and NAD precursors that increase NAD concentration in the body have been acknowledged as anti-aging supplements. However, there have been only a few studies on the link between aging or aging-related diseases and human blood NAD concentration because NAD and its precursors are unstable in blood and difficult to measure. Therefore, we aimed to construct a quantitative NAD measurement method that is simpler than the existing methods. The calibration standards of NAD showed good linearity (0.9936 to 0.9990) in the range of 0.25 to 200 μM, and the lower limit of quantification was 0.5 to 2 μM. We found that QIAcard FTA DMPK-B maintained NAD stability of 85% or more for at least 2 weeks at 4 °C and 1 week at room temperature using the dried blood spot method. Additionally, NAD stability in the blood extraction solution was more than 90% for 2 months. To our knowledge, there has been no report on a quantitative NAD measurement method in human whole blood that can be performed with as little as 5 μL of blood and can be easily implemented at both medical clinics and private homes. Our simple and convenient method has the potential to become the gold standard for NAD measurement in blood. It is expected to contribute to the acceleration of research on the correlation between aging or aging-related diseases and NAD concentration in human blood.
烟酰胺腺嘌呤二核苷酸（NAD）是一种辅酶，对能量产生至关重要。最近，有报道称 NAD 与衰老相关疾病有关，并且已经承认增加体内 NAD 浓度的 NAD 前体是抗衰老补充剂。然而，由于 NAD 和其前体在血液中不稳定且难以测量，因此关于衰老或衰老相关疾病与人体血液 NAD 浓度之间的联系的研究很少。因此，我们旨在构建一种比现有方法更简单的定量 NAD 测量方法。NAD 的校准标准在 0.25 至 200 μM 的范围内表现出良好的线性（0.9936 至 0.9990），定量下限为 0.5 至 2 μM。我们发现，使用干血斑法，QIAcard FTA DMPK-B 在 4°C 下至少 2 周和室温下 1 周可将 NAD 的稳定性保持在 85%或更高水平。此外，血液提取溶液中的 NAD 稳定性在 2 个月内超过 90%。据我们所知，目前尚无关于可使用少至 5 μL 血液且可在医疗诊所和私人住宅中轻松实施的人类全血中定量 NAD 测量方法的报道。我们简单方便的方法有可能成为血液中 NAD 测量的金标准。它有望促进研究衰老或衰老相关疾病与人类血液中 NAD 浓度之间的相关性。
