Reversed-Phase Separation of the Major Deoxyribonucleosides and Their Mononucleotides Using Tetrabutylammontum Hetaerons

Publisher Website
Google Scholar
Abstract
Simultaneous separation of the 4 major deoxyribonucleosides and their monophosphate nucleotides was achieved using tetrabutyl ammonium [TBA] phosphate hetaerons with a reversed-phase (C8) packing material. Baseline resolution for all 8 solutes was achieved within 48 min, using 7.5% methanol mobile phase, 2.0 mM in TBA, buffered with 50 mM phosphate at pH 4.8. The effect of methanol and TBA concentrations upon the retention of neutral and anionic solutes was studied in detail. Changes in solute k'' with increasing methanol could be explained by essentially independent phenomena. These are a decrease in the partition coefficient of the TBA cation with increasing organic concentrations, resulting in lower surface charge densities, and a decrease in the hydrophobic interactions of the solutes with the reversed-phase HPLC [high performance liquid chromatography]. The overall effect was a log-linear decrease in k'' with increasing methanol concentration. An empirical equation was derived for the above model which was helpful in determining the optimal separation conditions for the nucleosides and nucleotides.

This publication has 18 references indexed in Scilit: