Huber C G, Oefner P J, Bonn G K
Institute of Radiochemistry, Leopold-Franzens University, Innsbruck, Austria.
J Chromatogr. 1992 May 22;599(1-2):113-8. doi: 10.1016/0021-9673(92)85463-4.
Detritylated oligonucleotides were separated by reversed-phase high-performance liquid chromatography on highly cross-linked polystyrene-based particles having a mean particle diameter of 2.3 microns. The addition of poly(vinyl alcohol) during polymerization, which resulted in the presence of hydroxyl groups on the surface of the poly(styrene-divinylbenzene) beads, was necessary to obtain baseline resolution of phosphorylated oligodeoxyadenylic acids with a chain length of up to 30 bases. The impact of temperature was investigated and optimum resolution was achieved at 40 degrees C. At pH 7.0, the retention times of oligonucleotides were found to depend on the ratio of bases and to increase in the order of C less than G less than A less than T. Under the same conditions, it was possible to separate phosphorylated from dephosphorylated oligonucleotides, the former being eluted earlier. Recoveries ranged from 92 to 100%.
去三苯甲基化寡核苷酸通过反相高效液相色谱法在平均粒径为2.3微米的高度交联聚苯乙烯基颗粒上进行分离。在聚合过程中添加聚乙烯醇，这导致聚（苯乙烯 - 二乙烯基苯）珠粒表面存在羟基，对于获得链长高达30个碱基的磷酸化寡聚脱氧腺苷酸的基线分离是必要的。研究了温度的影响，在40℃时实现了最佳分离度。在pH 7.0时，发现寡核苷酸的保留时间取决于碱基比例，并按C＜G＜A＜T的顺序增加。在相同条件下，可以将磷酸化寡核苷酸与去磷酸化寡核苷酸分离，前者洗脱较早。回收率在92%至100%之间。
