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基于碳点的应用和酪氨酸酶活性抑制的酪氨酸酶和阿特拉津的荧光测定法。

Fluorometric Assay of Tyrosinase and Atrazine Based on the Use of Carbon Dots and the Inhibition of Tyrosinase Activity.

机构信息

School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu, 211166, PR China.

Department of Pharmacy, Jiuting hospital of Songjiang District, Shanghai, 201651, PR China.

出版信息

J Fluoresc. 2024 Mar;34(2):765-774. doi: 10.1007/s10895-023-03308-x. Epub 2023 Jun 26.

Abstract

Sensitive and convenient strategy of tyrosinase (TYR) and its inhibitor atrazine is in pressing demand for essential research as well as pragmatic application. In this work, an exquisite label-free fluorometric assay with high sensitivity, convenience and efficiency was described for detecting TYR and the herbicide atrazine on the basis of fluorescent nitrogen-doped carbon dots (CDs). The CDs were prepared via one-pot hydrothermal reaction starting from citric acid and diethylenetriamine. TYR catalyzed the oxidation of dopamine to dopaquinone derivative which could quench the fluorescence of CDs through a fluorescence resonance energy transfer (FRET) process. Thus, a sensitive and selective quantitative evaluation of TYR can be constructed on the basis of the relationship between the fluorescence of CDs and TYR activity. Atrazine, a typical inhibitor of TYR, inhibited the catalytic activity of TYR, leading to the reduced dopaquinone and the fluorescence was retained. The strategy covered a broad linear range of 0.1-150 U/mL and 4.0-80.0 nM for TYR and atrazine respectively with a low detection limit of 0.02 U/mL and 2.4 nM/mL. It is also demonstrated that the assay can be applied to detect TYR and atrazine in spiked complex real samples, which provides infinite potential in application of disease monitoring along with environmental analysis.

摘要

由于基础研究和实际应用的迫切需要,开发一种对酪氨酸酶(TYR)及其抑制剂莠去津具有高灵敏度、便捷性和高效性的灵敏检测策略至关重要。本工作基于荧光氮掺杂碳点(CDs),描述了一种灵敏、便捷的无标记荧光分析方法,用于检测 TYR 和除草剂莠去津。CDs 通过柠檬酸和二乙烯三胺一锅水热反应制备。TYR 催化多巴胺氧化生成多巴醌衍生物,通过荧光共振能量转移(FRET)过程猝灭 CDs 的荧光。因此,基于 CDs 荧光与 TYR 活性之间的关系,可以构建一种灵敏、选择性定量评价 TYR 的方法。莠去津是 TYR 的典型抑制剂,它抑制 TYR 的催化活性,导致多巴醌减少,荧光保留。该策略对 TYR 和莠去津的线性范围分别为 0.1-150 U/mL 和 4.0-80.0 nM,检测限分别为 0.02 U/mL 和 2.4 nM/mL。实验还证明,该测定法可用于检测加标复杂实际样品中的 TYR 和莠去津,为疾病监测和环境分析的应用提供了无限潜力。

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