Separation of 5-alkyluracils and purine bases in hydrolysates of enzymatically synthesized nucleic acids by high-performance ion-pair liquid chromatography.

Reversed-phase and reversed-phase ion-pair chromatographic methods were used to determine 5-alkyluracils in the presence of purine bases (mainly adenine) in the hydrolysates of enzymatically synthesized DNA. The effect of ionic strength, pH, concentration of the ion-pairing agent and methanol on the selectivity between alkyluracils and purine bases was examined in order to simplify the routine work and to reduce the time necessary for analysis. Optimal conditions could be developed for the isocratic separation of the various mixtures obtained by hydrolysis of the products of enzymatic synthesis.