2023 Fiscal Year Final Research Report
Development of a direct quantification method for NADP+ using fluorescence derivatization reaction of catechol
| Project/Area Number |
20K05561
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 34020:Analytical chemistry-related
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| Research Institution | Gunma University |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Keywords | NADP+ / 蛍光 / アドレナリン / ペプチド蛍光反応 |
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| Outline of Final Research Achievements |
In this study, we have developed a method for the direct quantification of NADP+ via fluorometric quantification of the fluorescent products by oxidation of adrenaline by NADP+. As a result, we found that this reaction proceeded rapidly in boric acid buffer solution at pH 8.0. We also found that the hydroxy group at the benzyl position of a side chain, which is attached at 4-position of catechol, is essential, and accelerating the elimination of this hydroxy group leads to the rapid formation of the fluorescent product. Under the optimized reaction conditions, NADP+ could be sensitively detected with high selectivity over NADPH.
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| Free Research Field |
分析化学
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| Academic Significance and Societal Importance of the Research Achievements |
生体内酸化還元反応は、細胞の生命活動を司る重要な代謝反応である。中でもNADPHとNADP+の相互変換は多くの反応において補酵素を担っていることから、生体試料に含まれるNADPHとNADP+の分析は生命科学研究や疾病診断において必須の技術である。NADPH検出系は、特徴的な340 nmの最大吸収波長を利用した吸光度法が協力であり、臨床検査にも応用されている。一方、NADP+検出系は、分離技術を使用しない特異的測定法の開発例がない。本研究成果は、高速液体クロマトグラフィー等の分離技術を必要とせず、簡便な吸光度法でNADP+を直接定量できる技術を提供することから、社会的意義が大きいものと考える。
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