A reversible competition colorimetric assay for the detection of biothiols based on ruthenium-containing complex

A novel reversible colorimetric sensor, which based on a competitive ligation of Hg(2+) by thiols, cysteine (Cys) or glutathione (GSH), and thiocyanate (SCN) on the N3 dye (bis(4,4'-dicarboxy-2,2'-bipyridine) dithiocyanato ruthenium (II)), was developed for the detection of biothiols. First, Hg(2+) ions coordinate to the sulfur atom of the dyes' SCN groups, and this interaction induces a change in color from red to yellow, owing to the formation of a complex of Hg(2+)-N3. Then, in the presence of biothiols, the red color of N3 is recovered concomitantly with the dissociation of the Hg(2+)-N3 complex, due to the extraction of Hg(2+) by biothiols. Thus the corresponding color variation in the process of the dissociation of the Hg(2+)-N3 complex can be employed for the quantitative detection of thiols using UV-vis spectroscopy. In particular, the transformation can be readily viewed with the naked eye. A good linear relationship between the change in absorbance (ΔAbs) of Hg(2+)-N3 at 461 nm and the thiol concentration was obtained in the range of 0.5-25 μM, and the detection limits are then calculated to be 57 and 52 nM for Cys and GSH, respectively. The proposed colorimetric assay displays a high selectivity for Cys over various other amino acids and GSSG (oxidized glutathione).