Stilbene photochrome-fluorescence-spin molecules: Covalent immobilization on silica plate and applications as redox and viscosity probes

We report herein on the development of a new photochrome-fluorescence-spin method for the quantitative analysis of the redox status and viscosity of a medium. The method of the viscosity measurement is based on the use of double fluorescence-nitroxide molecules. In such hybrid compounds the nitroxide moiety quenches the fluorescence of the fluorophore (stilbene moiety). The reduction of nitroxide by an antioxidant (ascorbic acid) causes a rise of fluorescence of the fluorophore. The rate constant of the stilbene fragment photoisomerization in such systems is dependent upon the viscosity of the media. The synthesized dual stilbene-nitroxide probe was covalently immobilized onto the surface of a quartz plate as an eventual fiber-optic sensor. The immobilization procedure included a cyanogen bromide surface activation followed by smoothing with a protein tether. The rate of fluorescence change was monitored in aqueous-glycerol solutions of different viscosities and content of ascorbic acid. Good correlation was found: (a) between the concentration of ascorbic acid in the sample and the rate of fluorescence increase due to the reduction of the nitroxide moiety, and (b) between the rate constant of photoisomerization and the viscosity of the media. Appropriate calibration would make the determination of the viscosity of a media possible (in a range 1-500 cP), as well as ascorbate content, in a range (1-9)×10^-4 M, with fast single measurement.