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

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.