Separation of nucleosides and nucleotides by reversed-phase high-performance liquid chromatography with volatile buffers allowing sample recovery

✍ Scribed by Charlotte Yu Ip; Daniel Ha; Paul W. Morris; Mark L. Puttemans; Duane L. Venton

• Publisher: Elsevier Science
• Year: 1985
• Tongue: English
• Weight: 479 KB
• Volume: 147
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(85)90025-9

✦ Synopsis

Reversed-phase high-performance liquid chromatography using a C,8 column with volatile buffers as the eluant was applied to the separation of a number of nucleosides and nucleotides. Groups of seven nucleosides and five nucleoside monophosphates were separated isocratically employing 0.1 M trimethylammonium acetate and 2% acetonitrile at pH 7.0. Groups of seven nucleoside diphosphates and seven nucleoside triphosphates were separated with 0.1 M triethylammonium bicarbonate and 2% acetonitrile titrated to a pH of 7.1 with acetic acid. The techniques described give resolution and separations comparable to nonvolatile buffers. Moreover, the eluant trimethylammonium acetate or triethylammonium bicarbonate buffer can easily be removed in vacua from the column effluent, making the technique useful for preparative separations of these compounds. The observed elution pattern of nucleoside phosphates suggests that "paired-ion" chromatography is involved in the separation. 0 1985 Academic PESS, IIIC.