A label-free fluorimetric detection of biothiols based on the oxidase-like activity of Ag+ ions

作者:Li, Ru; Lei, Cuihua; Zhao, Xian-En; Gao, Yue; Gao, Han; Zhu, Shuyun; Wang, Hua*
来源:Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy , 2018, 188: 20-25.
DOI:10.1016/j.saa.2017.06.056

摘要

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 0.50-30.0 mu M for Cys, 1.0-45.0 mu M for Hcy and 0.50-40.0 mu M for GSH. The detection limits (S/N = 3) of Cys, Hcy and GSH were 110 nM, 200 nM and 150 nM, respectively. Subsequently, the developed fluorimetric method was successfully applied for the detection of biothiols in human serum.