利用双同位素介导液相色谱-串联质谱法(dimeLC-MS/MS)绝对定量生物样本中的烟酰胺单核苷酸

IF 5.4 Q1 GERIATRICS & GERONTOLOGY
Junya Unno, Kathryn F. Mills, Tairo Ogura, Masayuki Nishimura, Shin-ichiro Imai
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引用次数: 0

摘要

烟酰胺腺嘌呤二核苷酸(NAD+)是包括衰老在内的基本生物现象所必需的代谢物。烟酰胺单核苷酸(NMN)是一种关键的 NAD+ 中间体,作为一种有效的 NAD+ 增强化合物,已在小鼠和人体中进行了广泛测试。然而,准确测量生物样本中的 NMN 长期以来一直是该领域的一项挑战。在此,我们利用液相色谱-三重四极杆质谱法(LC-MS/MS)和双同位素 NMN 标准,建立了一种精确、定量测量 NMN 的方法。在这一新方法中,生物样本的基质效应得到了适当的调整,并且在样本处理过程中可追溯 NMN 的去向。我们已经证明,这种方法可以准确量化小鼠血浆中的 NMN 含量,并证实 NMN 可以快速、直接地被血液循环和细胞吸收。这种双同位素介导的 LC-MS/MS(dimeLC-MS/MS)可以很容易地扩展到其他与 NAD+ 相关的代谢物,成为 NAD+ 生物学的可靠标准方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Absolute quantification of nicotinamide mononucleotide in biological samples by double isotope-mediated liquid chromatography-tandem mass spectrometry (dimeLC-MS/MS)

Absolute quantification of nicotinamide mononucleotide in biological samples by double isotope-mediated liquid chromatography-tandem mass spectrometry (dimeLC-MS/MS)

Nicotinamide adenine dinucleotide (NAD+) is an essential metabolite for fundamental biological phenomena, including aging. Nicotinamide mononucleotide (NMN) is a key NAD+ intermediate that has been extensively tested as an effective NAD+-boosting compound in mice and humans. However, the accurate measurement of NMN in biological samples has long been a challenge in the field. Here, we have established an accurate, quantitative methodology for measuring NMN by using liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) with double isotopic NMN standards. In this new methodology, the matrix effects of biological samples were properly adjusted, and the fate of NMN could be traced during sample processing. We have demonstrated that this methodology can accurately quantitate NMN levels in mouse plasma and confirmed quick, direct NMN uptake into blood circulation and cells. This double isotope-mediated LC-MS/MS (dimeLC-MS/MS) can easily be expanded to other NAD+-related metabolites as a reliable standard methodology for NAD+ biology.

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来源期刊
NPJ Aging and Mechanisms of Disease
NPJ Aging and Mechanisms of Disease Medicine-Geriatrics and Gerontology
自引率
0.00%
发文量
0
审稿时长
8 weeks
期刊介绍: npj Aging and Mechanisms of Disease is an online open access journal that provides a forum for the world’s most important research in the fields of aging and aging-related disease. The journal publishes papers from all relevant disciplines, encouraging those that shed light on the mechanisms behind aging and the associated diseases. The journal’s scope includes, but is not restricted to, the following areas (not listed in order of preference): • cellular and molecular mechanisms of aging and aging-related diseases • interventions to affect the process of aging and longevity • homeostatic regulation and aging • age-associated complications • translational research into prevention and treatment of aging-related diseases • mechanistic bases for epidemiological aspects of aging-related disease.
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