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Simple enzyme-free detection of uric acid by an in situ fluorescence and colorimetric method based on Co-PBA with high oxidase activity
Analyst ( IF 3.6 ) Pub Date : 2023-12-20 , DOI: 10.1039/d3an01985c Lu Liu 1 , Guang Liu 1 , Xiaomei Mu 1 , Shulin Zhao 1 , Jianniao Tian 1
Analyst ( IF 3.6 ) Pub Date : 2023-12-20 , DOI: 10.1039/d3an01985c Lu Liu 1 , Guang Liu 1 , Xiaomei Mu 1 , Shulin Zhao 1 , Jianniao Tian 1
Affiliation
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In this work, we prepared a simple and low-cost cobalt-doped Prussian blue analog (Co-PBA), which can directly oxidize 10-acetyl-3,7-dihydroxyphenoxazine and 3,3′,5,5′-tetramethylbenzidine (TMB) to produce resorufin (ox-AR) with high fluorescent quantum yield and ox-TMB with blue color, respectively, without the need for unstable H2O2. Using the Michaelis–Menten curve and Lineweaver–Burk equation, the Michaelis–Menten constant of Co-PBA and the substrate TMB was found to be 0.033 mM, which was much lower than horseradish peroxidase and other reported nanozymes, showing satisfactory substrate affinity. Uric acid (UA) can cause erosion of the Co-PBA structure, and it significantly reduces the catalytic activity of Co-PBA, resulting in the decrease of the fluorescence emission signal of ox-AR and the absorption signal of ox-TMB. Based on this, a simple, sensitive, and fast fluorescence/colorimetric dual-mode uric acid detection platform was established. The detection range for UA by fluorescence method is 0.625–40 μM, and the detection limit (LOD, S/N = 3) is as low as 0.389 μM. The detection system was applied to serum samples with good recovery and can be used for field detection of UA in biological samples under different environments to meet different needs.
中文翻译：
基于高氧化酶活性 Co-PBA 的原位荧光比色法简单无酶检测尿酸
在这项工作中，我们制备了一种简单且低成本的钴掺杂普鲁士蓝类似物（Co-PBA），它可以直接氧化10-乙酰基-3,7-二羟基吩恶嗪和3,3',5,5'-四甲基联苯胺（ TMB）分别生产具有高荧光量子产率的试卤灵（ox-AR）和蓝色的ox-TMB，而不需要不稳定的H 2 O 2 。利用米氏曲线和Lineweaver-Burk方程，发现Co-PBA和底物TMB的米氏常数为0.033 mM，远低于辣根过氧化物酶和其他报道的纳米酶，表现出令人满意的底物亲和力。尿酸(UA)会引起Co-PBA结构的侵蚀，显着降低Co-PBA的催化活性，导致ox-AR的荧光发射信号和ox-TMB的吸收信号减弱。在此基础上建立了一个简单、灵敏、快速的荧光/比色双模式尿酸检测平台。荧光法对UA的检测范围为0.625~40 μM，检测限（LOD，S/N = 3）低至0.389 μM。该检测系统应用于血清样本，回收率良好，可用于不同环境下生物样本中UA的现场检测，满足不同需求。
更新日期：2023-12-20
中文翻译：
基于高氧化酶活性 Co-PBA 的原位荧光比色法简单无酶检测尿酸
在这项工作中，我们制备了一种简单且低成本的钴掺杂普鲁士蓝类似物（Co-PBA），它可以直接氧化10-乙酰基-3,7-二羟基吩恶嗪和3,3',5,5'-四甲基联苯胺（ TMB）分别生产具有高荧光量子产率的试卤灵（ox-AR）和蓝色的ox-TMB，而不需要不稳定的H 2 O 2 。利用米氏曲线和Lineweaver-Burk方程，发现Co-PBA和底物TMB的米氏常数为0.033 mM，远低于辣根过氧化物酶和其他报道的纳米酶，表现出令人满意的底物亲和力。尿酸(UA)会引起Co-PBA结构的侵蚀，显着降低Co-PBA的催化活性，导致ox-AR的荧光发射信号和ox-TMB的吸收信号减弱。在此基础上建立了一个简单、灵敏、快速的荧光/比色双模式尿酸检测平台。荧光法对UA的检测范围为0.625~40 μM，检测限（LOD，S/N = 3）低至0.389 μM。该检测系统应用于血清样本，回收率良好，可用于不同环境下生物样本中UA的现场检测，满足不同需求。
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