Comparison of different buffer systems for separation of 15 nucleosides by capillary electrophoresis

Abstract

The most suitable background electrolytes (BGEs) for simultaneous separation of 15 nucleosides by different modes of capillary electrophoresis (CE) were obtained. Various modes of CE were performed including capillary zone electrophoresis (CZE), capillary electrophoresis-electrospray ionization-time of flight mass spectrometry (CE-ESI-TOF/MS) and micellar electrokinetic chromatography (MEKC). The electrolyte buffers using sodium tetraborate decahydrate, disodium hydrogen phosphate, sodium acetate, sodium bicarbonate, ammonium acetate or 1, 2-diamino-ethane (DEA) were tested, and the best of them were systematically optimized. In CZE mode, the nucleosides could not be separated completely with sodium tetraborate decahydrate or disodium hydrogen phosphate as BGEs, demonstrating the limited applicability of the two buffer systems for complex samples. However, with 300 mmol/L DEA (containing 2% acetone) as BGE, 15 nucleosides could be separated with good resolution and peak shape, which proved that the DEA buffer was most suitable in CZE. The best buffer system in MEKC mode was 25 mmol/L disodium hydrogen phosphate with 70 mmol/L sodium dodecyl sulfate (SDS), and it was successfully applied for the separation of the nucleosides in Chinese Anthopleura lanthogrammica Berkly. The optimum buffer system for CE-ESI-TOF/MS analysis was 20 mmol/L ammonium acetate (pH 10.0). In the positive ion mode, the MS signals of each compound were better than those in the literature using DEA as BGE. The results of this study demonstrated the applicability of different buffer systems for the simultaneous separation of 15 nucleosides, and were helpful for the development of CE method in complex sample separation.