Highly sensitive and selective colorimetric detection of glutathione based on Ag [I] ion–3,3′,5,5′-tetramethylbenzidine (TMB)

Glutathione (GSH) plays an important role in the biological system and serves many cellular functions. Since all of the biothiols possess similar functional groups, it is still challenging to selectively detect GSH over cysteine (Cys) and homocysteine (Hcy). In this work, a novel and simple colorimetric method for discriminative detection of glutathione (GSH) over Cys and Hcy is developed. The proposed method is based on the fact that Ag [I] ion could oxidize 3,3',5,5',-tetramethylbenzidine (TMB) to the oxidized TMB to induce a blue color and an absorption peak centered at 652 nm. However, the introduction of GSH could cause the reduction of oxidized TMB and it could also combine with Ag(+), both of which result in a blue color fading and a decrease of the absorbance at 652 nm. Based on this finding, we propose a method to qualitatively and quantitatively detect GSH by naked eyes and UV-vis spectroscopy, respectively. The proposed method shows a low detection limit of 0.1 µM by naked eyes and 0.05 µM with the help of UV-vis spectroscopy. In addition, this method has great potential in discriminatively detecting GSH over other amino acid and biothiols. More importantly, this method is simple and fast without the preparation of nanomaterials and has also been successfully applied to the detection of GSH in biological fluids.