基于银离子模拟酶活性的生物硫醇无标记荧光检测法。
A label-free fluorimetric detection of biothiols based on the oxidase-like activity of Ag ions.
机构信息
Shandong Provincial Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong Province 273165, China.
Shandong Provincial Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong Province 273165, China; Institute of Medicine and Materials Applied Technologies, Qufu Normal University, Qufu City, Shandong Province 273165, China.
出版信息
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Jan 5;188:20-25. doi: 10.1016/j.saa.2017.06.056. Epub 2017 Jul 2.
In this work, a label-free and sensitive fluorimetric method has been developed for the detections of biothiols including cysteine (Cys), homocysteine (Hcy), and glutathione (GSH), based on the specific biothiol-induced inhibition of the oxidase-like activity of silver ions (Ag). It is well established that o-phenylenediamine (OPD) can be oxidized by Ag ions to generate fluorescent 2,3-diaminophenazine (OPDox). The introduction of biothiols would inhibit the oxidation of OPD by Ag due to the strong coordination between biothiols and Ag. The changes of fluorescence intensities obtained in the Ag-OPD system exhibited good linear correlations in the ranges of 0.50-30.0μM for Cys, 1.0-45.0μM for Hcy and 0.50-40.0μM for GSH. The detection limits (S/N=3) of Cys, Hcy and GSH were 110nM, 200nM and 150nM, respectively. Subsequently, the developed fluorimetric method was successfully applied for the detection of biothiols in human serum.
在这项工作中，开发了一种基于银离子（Ag）的氧化酶样活性被生物硫醇特异性抑制的无标记和灵敏的荧光法，用于检测生物硫醇，包括半胱氨酸（Cys）、同型半胱氨酸（Hcy）和谷胱甘肽（GSH）。众所周知，邻苯二胺（OPD）可以被 Ag 离子氧化生成荧光 2,3-二氨基吩嗪（OPDox）。由于生物硫醇与 Ag 之间的强配位作用，生物硫醇的引入会抑制 Ag 对 OPD 的氧化。在 Ag-OPD 体系中，获得的荧光强度变化在 Cys 的 0.50-30.0μM、Hcy 的 1.0-45.0μM 和 GSH 的 0.50-40.0μM 范围内表现出良好的线性相关性。Cys、Hcy 和 GSH 的检测限（S/N=3）分别为 110nM、200nM 和 150nM。随后，该荧光法成功应用于人血清中生物硫醇的检测。
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