Ratiometric fluorescence and Nanozyme colorimetry for dual-modality uric acid detection in complex Biosystems

IF 4.6 2区化学 Q1 SPECTROSCOPY

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy Pub Date : 2025-10-01 DOI:10.1016/j.saa.2025.127008

Wei Li , Yongtao Xu , Wuyuan Pan , Shuqing Tu , Jian Wang , Fang Ke

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Abstract

Humans lack urate oxidase (UOX) due to UOX gene mutations, leading to hyperuricemia-related disorders like gout and kidney disease. Accurate uric acid (UA) quantification is therefore vital for diagnosis and management. Conventional methods are often impractical, while popular enzymatic colorimetric assays, though simpler, suffer from cross-reactivity and matrix interference. Nanozymes, such as graphitic carbon nitride (g-C₃N₄), offer robust, cost-effective alternatives; Mn-doping C₃N₄ (Mn-C₃N₄) enhances its uricase-like activity and stability. However, real-world application of Mn-C₃N₄ is still challenged by selectivity issues and matrix effects. To overcome these limitations, we developed a synergistic dual-detection platform integrating Mn-C₃N₄ nanozymes applied together with the ratiometric fluorescent probe MPNVPI. Under blue-light irradiation (460 nm), Mn-C₃N₄ oxidizing UA to generate H₂O₂ triggers an event: nucleophilic cleavage of MPNVPI releases fluorescent HVMI (emission shift: 500 nm to 568 nm). This label-free, dual-modality assay leverages intrinsic optical properties for ultra-sensitive UA quantification in complex serum matrices. It also enables real-time imaging of UA dynamics in RAW 264.7 cells and zebrafish, demonstrating exceptional sensitivity, selectivity, and monitoring capability for diagnostic applications in metabolic disorders.

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比率荧光和纳米酶比色法在复杂生物系统中检测尿酸的双模态

由于UOX基因突变，人类缺乏尿酸氧化酶（UOX），导致高尿酸血症相关疾病，如痛风和肾脏疾病。因此，准确的尿酸（UA）定量对诊断和治疗至关重要。传统的方法往往是不切实际的，而流行的酶比色测定法，虽然更简单，遭受交叉反应性和基质干扰。纳米酶，如石墨氮化碳（g-C3N4），提供了强大的，具有成本效益的替代品；mn掺杂C3N4 （Mn-C3N4）增强了其类尿酸酶活性和稳定性。然而，Mn-C3N4的实际应用仍然受到选择性问题和矩阵效应的挑战。为了克服这些限制，我们开发了一种协同双检测平台，将Mn-C3N4纳米酶与比例荧光探针MPNVPI结合使用。在蓝光照射（460 nm）下，Mn-C3N4氧化UA生成H2O2触发MPNVPI亲核裂解释放荧光HVMI（发射位移：500 nm ~ 568 nm）。这种无标签，双模分析利用内在光学特性的超敏感UA定量在复杂的血清基质。它还可以实时成像RAW 264.7细胞和斑马鱼的UA动态，显示出卓越的灵敏度、选择性和监测能力，用于代谢紊乱的诊断应用。

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来源期刊

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 物理-光谱学

CiteScore

8.40

自引率

11.40%

发文量

1364

审稿时长

40 days

期刊介绍： Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy (SAA) is an interdisciplinary journal which spans from basic to applied aspects of optical spectroscopy in chemistry, medicine, biology, and materials science. The journal publishes original scientific papers that feature high-quality spectroscopic data and analysis. From the broad range of optical spectroscopies, the emphasis is on electronic, vibrational or rotational spectra of molecules, rather than on spectroscopy based on magnetic moments. Criteria for publication in SAA are novelty, uniqueness, and outstanding quality. Routine applications of spectroscopic techniques and computational methods are not appropriate. Topics of particular interest of Spectrochimica Acta Part A include, but are not limited to: Spectroscopy and dynamics of bioanalytical, biomedical, environmental, and atmospheric sciences, Novel experimental techniques or instrumentation for molecular spectroscopy, Novel theoretical and computational methods, Novel applications in photochemistry and photobiology, Novel interpretational approaches as well as advances in data analysis based on electronic or vibrational spectroscopy.