Department of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan.
Department of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan.
Talanta. 2024 Jun 1;273:125925. doi: 10.1016/j.talanta.2024.125925. Epub 2024 Mar 16.
In the present study, we propose a scheme for detecting HO by using horseradish peroxidase (HRP) adsorbed onto single silica particles and fluorescence microspectroscopy. When the silica particles were immersed in an HRP solution, the HRP concentration in the silica particles increased by a factor of 690 compared to that in the bulk aqueous solution because HRP was adsorbed on the silica surface. When a single particle containing HRP was added to a mixed solution of HO and Amplex Red, fluorescence from resorufin, which was produced by the reaction of HRP, HO, and Amplex Red, was observed. The fluorescence from the resorufin in the particles increased after a single particle was added to the solution, and the release of resorufin was observed. As the concentration of HO (C) decreased, the time it takes for fluorescence intensity to reach its maximum was shorter. The detection limit for HO in the present system was 980 nM. The reaction behavior of a single silica particle was evaluated using a spherical diffusion model, which explains the approximate concentration change of resorufin in the silica particle. The proposed method has the advantages of simple sample preparation and detection, low sample consumption, and a short detection time.
在本研究中，我们提出了一种利用辣根过氧化物酶（HRP）吸附在单个硅胶颗粒上和荧光显微光谱法来检测 HO 的方案。当硅胶颗粒浸入 HRP 溶液中时，由于 HRP 吸附在硅胶表面，硅胶颗粒中的 HRP 浓度比在体相水溶液中的浓度增加了 690 倍。当含有 HRP 的单个颗粒被添加到 HO 和 Amplex Red 的混合溶液中时，观察到由 HRP、HO 和 Amplex Red 反应产生的 Resorufin 的荧光。当将单个颗粒添加到溶液中后，颗粒中的 Resorufin 的荧光强度增加，并观察到 Resorufin 的释放。随着 HO（C）浓度的降低，荧光强度达到最大值所需的时间更短。本系统对 HO 的检测限为 980 nM。使用球形扩散模型评估了单个硅胶颗粒的反应行为，该模型解释了 Resorufin 在硅胶颗粒中的近似浓度变化。该方法具有样品制备和检测简单、样品消耗少、检测时间短的优点。
