Ip C Y, Ha D, Morris P W, Puttemans M L, Venton D L
Anal Biochem. 1985 May 15;147(1):180-5. doi: 10.1016/0003-2697(85)90025-9.
Reversed-phase high-performance liquid chromatography using a C18 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 vacuo 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.
使用C18柱和挥发性缓冲液作为洗脱剂的反相高效液相色谱法被用于多种核苷和核苷酸的分离。在pH 7.0条件下，采用0.1 M乙酸三甲基铵和2%乙腈等度洗脱，分离出七组核苷和五组核苷单磷酸。在pH 7.1条件下，用0.1 M碳酸氢三乙铵和2%乙腈（用乙酸滴定）分离出七组核苷二磷酸和七组核苷三磷酸。所述技术提供的分离度和分离效果与非挥发性缓冲液相当。此外，洗脱剂乙酸三甲基铵或碳酸氢三乙铵缓冲液可轻松地在真空中从柱流出物中除去，使得该技术可用于这些化合物的制备分离。观察到的核苷磷酸洗脱模式表明，“离子对”色谱法参与了分离过程。
