The hydrolysis of cyclic 2′-3′ Nucleotides by ammonia during paper chromatography and its influence on quantitative determinations

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Abstract
It was found that the presence of NH3 or (NH4)2CO3 in isopropanol-water solvent systems used for the chromatography of cyclic nucleotides results into appreciable losses of the cyclic compounds which are hydrolysed to nucleotides. This happens almost exclusively in the preliminary phase of the chromatography i. e. before the compounds are covered with the solvent. In order to use such solvents for quantitative purposes, it is necessary to add the NH3 or (NH4)2CO3 to the system only after the compounds to be analysed are covered with isopropanolwater. The hydrolysis presumably takes place by a preferential adsorption of water to the paper during the equilibration period with the local formation of a too alkaline NH3 solution. It is probable that the same mechanism operates in solvent systems containing essentially an organic solvent, water and ammonia or compounds giving rise to ammonia.