Evaluation of separation properties of stationary phases in supercritical fluid chromatography; deazapurine nucleosides case study
Abstract
A series of biologically important 7-deazapurine nucleosides was used for evaluation of selectivity of three stationary phases designed for supercritical fluid chromatography. The sorbents of stationary phases tested were diethylamine, diol and 2-picolylamine bonded on ethylene bridged hybrid particles.
The highest selectivity for the tested nucleosides showed neutral diol column. All twelve tested nucleosides were baseline resolved in simple mobile phase composed of carbon dioxide/ethanol 79/21 (v/v).
The effect of basic isopropylamine and mixed isopropylamine and trifluoroacetic acid mobile phase additives on retention and selectivity was also assessed. The interactions participating in the separation systems were revealed by linear free energy relationship.
The obtained results demonstrated the significant contribution of hydrogen bond acidity and basicity on chromatographic behavior of the diol- and 2-picolylamine-based stationary phases.