Methylene Blue Dye Test for Rapid Qualitative Detection of Hydroxyl Radicals Formed in a Fenton's Reaction Aqueous Solution

A new procedure, the methylene blue dye test, qualitatively indicates the presence of hydroxyl radicals through the immediate, distinct bleaching of methylene blue dye on a paper test strip. This method employs a simple procedure requiring inexpensive materials, without the addition of competitive probe chemicals that potentially can interfere with the reaction. A Fenton's reaction with an Fe2+:H2O2 molar ratio of 1:20 generated hydroxyl radicals in Milli-Q water. The presence and absence of hydroxyl radicals were determined prior to and following quenching of the Fenton's reaction with 10% sodium sulfite, respectively. Bleaching of methylene blue dye, due to the presence of hydroxyl radicals in a sample, was indicated by a discoloration from a dark blue color to an almost white color, concentrated at the point of application, with a dark blue outline. A lack of bleaching indicated the absence of hydroxyl radicals in the sample. The presence of hydroxyl radicals was verified by benzoic acid chemical probe experiments with thin-layer chromatography (TLC) and spectrophotometric wavelength scans. The presence of hydroxyl radicals was indirectly determined by detection of hydroxylated benzoic acids on TLC plates and a violet solution color with a peak absorbance at a wavelength close to 520 nm.