Location and analysis of 2-deoxy sugars on chromatograms and pherograms by fluorescence

Abstract

A group of new reagents for the location of 2-deoxy sugars is introduced. A representative reagent is 4'-aminoacetophenone, which gives a green trimethine fluorogen with at least 0.5 microg of 2-deoxyribose, 2-deoxygalactose, or 2-deoxyglucose under neutral or slightly acidic conditions and the orange fluorescent anion under alkaline conditions at a lower limit of approximately 5 microg. Reagents such as 2-thiobarbituric acid, aniline-diphenylamine, and 2,4-pentanedione give vivid fluorescent colours with these sugars. Aniline-diphenylamine reagent was a useful fluorescent agent for locating the purine nucleosides and nucleotides and DNA. A new attachment for the spectropho-tofluorimeter has been evaluated. With this attachment fluorescence excitation and emission spectra are obtained directly from paper, glass fibre, or thin-layer chromatograms and pherograms in sizes up to 20 x 20 cm. Glass plates are handled as readily as paper chromatograms. The fluorescence spectra of the various fluorogens produced after separation were readily obtained directly from the chromatogram or pherogram. 2-Thiobarbituric acid and 4'-aminoacetophenone offered the greatest potential for future application to characterization and assay of 2-deoxy sugars. The aniline-diphenylamine procedure was the most useful for analysis of purine nucleosides and nucleotides and DNA. These various reagents show potential for application to air pollution studies in terms of location, characterization and assay of 2-deoxy sugars and their derivatives.