West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, China.
School of Chemical Engineering, Sichuan University, Chengdu, 610065, China.
Anal Biochem. 2023 Mar 1;664:115045. doi: 10.1016/j.ab.2023.115045. Epub 2023 Jan 16.
Cascade reactions catalyzed by natural uricase and mimic peroxidase (MPOD) have been applied for uric acid (UA) detection. However, the optimal catalytic activity of MPOD is mostly in acidic conditions (pH 2-5), mismatching the optimal catalytic alkaline environment of uricase. In this paper, using CuSO and urea as raw materials, a MPOD with high catalytic activity in alkaline environment was synthesized by hydrothermal method. Then, based on coupling reaction of uricase/UA/MPOD/guaiacol (GA) system, a novel spectrophotometric method was established to detect 5-60 μmol/L UA (limit of detection = 3.14 μmol/L (S/N = 3)) and accurately quantified serum UA (275.6 ± 39.9 μmol/L, n = 5) with 95-105% of standard addition recovery. The results were consistent with commercial UA kit (p > 0.05). The MPOD could replace natural POD to reduce the cost of UA detection due to simple preparation and cheap raw materials, and is expected to achieve the specific detection of some substances, like glucose and cholesterol, combined with glucose oxidase and cholesterol oxidase.
级联反应由天然尿酸酶和模拟过氧化物酶(MPOD)催化,已被用于尿酸(UA)检测。然而,MPOD 的最佳催化活性大多在酸性条件下(pH 2-5),与尿酸酶的最佳催化碱性环境不匹配。在本文中,使用 CuSO 和尿素作为原料,通过水热法合成了一种在碱性环境中具有高催化活性的 MPOD。然后,基于尿酸酶/UA/MPOD/愈创木酚(GA)体系的偶联反应,建立了一种新的分光光度法来检测 5-60 μmol/L UA(检测限=3.14 μmol/L(S/N=3)),并准确地定量了血清 UA(275.6±39.9 μmol/L,n=5),标准添加回收率为 95-105%。结果与商用 UA 试剂盒一致(p>0.05)。由于制备简单、原料便宜,MPOD 可以替代天然过氧化物酶,降低 UA 检测成本,有望与葡萄糖氧化酶和胆固醇氧化酶结合,实现对某些物质(如葡萄糖和胆固醇)的特异性检测。