Nanozymes (NZs) are nanoparticles that mimic the catalytic properties of natural enzymes. The present work aimed to obtain effective peroxidase mimetics (PO-like NZs), to characterize their morphological properties, estimate the kinetic parameters of NZs and evaluate the prospects of their application in analysis of ethanol. Herein, we have proposed a convenient spectrophotometric method for ethanol assay using reusable alginate beads enriched with alcohol oxidase (AO) and nanoparticles of PtCu (nPtCu) as PO-like NZs, and 3,3′,5,5′-tetramethylbenzidine (TMB) as a chromogen. The linear range for the proposed nPtCu–AO/alginate beads/TMB-based method is from 0.01 mM to 0.15 mM with a limit of detection of 3.3 μM ethanol. The method is used for the quantitative determination of ethanol in alcoholic beverages. The obtained results proved to be in a good correlation with the enzymatic reference method. These results highlight the potential of the nPtCu with PO-like activity in bioanalytical applications. The proposed method, being sensitive, economical and suitable for routine and micro-volume formats, can be used in clinical diagnostics for the detection of ethanol.

中文翻译：

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可重复使用的酒精氧化酶 - nPtCu/藻酸盐珠用于高灵敏度饮料中的乙醇测定

纳米酶（NZ）是模仿天然酶催化特性的纳米颗粒。目前的工作旨在获得有效的过氧化物酶模拟物（PO类NZs），表征其形态特性，估计NZs的动力学参数并评估其在乙醇分析中的应用前景。在此，我们提出了一种方便的分光光度法进行乙醇测定，使用富含乙醇氧化酶（AO）的可重复使用的藻酸盐珠和作为 PO 类 NZ 的 PtCu（nPtCu）纳米粒子，以及 3,3',5,5'-四甲基联苯胺（TMB） ）作为色原。所提出的 nPtCu-AO/藻酸盐珠/TMB 方法的线性范围为 0.01 mM 至 0.15 mM，检测限为 3.3 μM 乙醇。该方法适用于酒精饮料中乙醇的定量测定。所得结果与酶法参考法具有良好的相关性。这些结果凸显了具有 PO 样活性的 nPtCu 在生物分析应用中的潜力。该方法灵敏、经济且适用于常规和微量形式，可用于乙醇检测的临床诊断。