磷酸甲基化DNA和RNA二核苷酸的Rp和Sp非对映异构体分离的优化。

Optimization of the separation of the Rp and Sp diastereomers of phosphate-methylated DNA and RNA dinucleotides.

作者信息

Coenen A J, Henckens L H, Mengerink Y, van der Wal S, Quaedflieg P J, Koole L H, Meijer E M

机构信息

DSM Research, Department FA-CO, Geleen Netherlands.

出版信息

J Chromatogr. 1992 Apr 3;596(1):59-66. doi: 10.1016/0021-9673(92)80202-6.

DOI:10.1016/0021-9673(92)80202-6

PMID:1381717

Abstract

The separation by reversed-phase high-performance liquid chromatography of Rp and Sp diastereomers of phosphate-methylated DNA and RNA dinucleotides was studied with respect to pH, organic modifier type and concentration and reversed-phase packing material. Drylab G was used to deduce optimum conditions. On the basis of the observed discrepancies between the computer predictions and experimental results, the gradient operation procedure with volatile buffers was improved. By repetitive chromatography on a 250 x 22 mm I.D. reversed-phase column, fourteen diastereomeric pairs were obtained in at least 97% purity and 60% yield, in amounts of 10-100 mg.

摘要

研究了反相高效液相色谱法对磷酸甲基化的DNA和RNA二核苷酸的Rp和Sp非对映异构体进行分离时，pH、有机改性剂类型和浓度以及反相填料的影响。使用Drylab G软件推导最佳条件。基于计算机预测和实验结果之间观察到的差异，改进了使用挥发性缓冲液的梯度操作程序。通过在一根内径250×22 mm的反相柱上进行重复色谱分离，获得了14对非对映异构体，纯度至少为97%，产率为60%，产量为10 - 100 mg。