Fluorescent Method for the Detection of Biothiols Using an Ag⁺-Mediated Conformational Switch.

In this work, a novel, simple, and time-saving fluorescence approach for the detection of biothiols (glutathione and cysteine) was developed by employing a DNA probe labeled with 2-aminopurine. As an adenine analogue, 2-aminopurine exhibits high fluorescence intensity that can be rapidly quenched in the presence of DNA. In the presence of Ag⁺, the fluorescence increased significantly, which was a result of the formation of cytosine⁻Ag⁺⁻cytosine base pairs and the release of 2-aminopurine. Upon addition of either glutathione or cysteine, the structure of cytosine⁻Ag⁺⁻cytosine was disrupted, a product of the stronger affinity between biothiols and Ag⁺. As a result, the 2-aminopurine-labeled DNA probe returned to its former structure, and the fluorescence signal was quenched accordingly. The detection limit for glutathione and cysteine was 3 nM and 5 nM, respectively. Furthermore, the determination of biothiols in human blood serum provided a potential application for the probe as a diagnostic tool in clinical practice.