Abstract
Thiochrome, its mono- and diphosphate were synthesized and their physicochemical properties such as paper chromatography, ultraviolet absorption, infrared absorption and fluorescnce spectra were investigated. It was found that the ultraviolet absorption and fluorescence maxima of these compounds were shifted to longer wavelengths with decreasing pH values. In alkaline solutions, the ultraviolet absorption and fluorescence maxima are located at 370 and 450 mμ, respectively. It is of interest to note that the phosphorylation induces no changes in the absorption and the appearance of emission spectra but induces an appreciable decrease in the fluorescence intensity. The ratio of the fluorescence intensity, thiochrome: thiochrome monophosphate: thiochrome diphosphate, is 10:8:9 in the alkaline solution (pH 9 to 12).The concentration of thiamine, thiamine monophosphate, and thiamine diphosphate were determined fluorometrically without dephosphorylation. A reasonable results was obtained when cyanogen bromide was used as an oxidizing agent, whereas an unreasonable result was obtained when potassium ferricyanide was used because of its quenching effect.
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