Patthy M, Balla T, Arányi P
Institute for Drug Research, Budapest, Hungary.
J Chromatogr. 1990 Dec 7;523:201-16. doi: 10.1016/0021-9673(90)85023-o.
A detailed study of all the major chromatographic variables affecting the retention behaviour and separation of myo-inositol phosphates in reversed-phase ion-pair chromatographic systems was carried out. The parameters studied included the eluent concentration of the pairing ion, the eluent concentration of the organic modifier and the buffer salt, the pH of the eluent, the minimum column plate count necessary for the separation of the inositol trisphosphate isomers and isocratic and gradient modes of separation. The retention behaviour of some common nucleotides and sugar phosphates was also investigated as these phosphates present chromatographic interference problems in biochemical studies based on the cellular incorporation of [32P]Pi. The separation methods developed appear to be superior to established anion-exchange separation techniques in terms of separation speed and "mildness" of the chromatographic conditions.
对反相离子对色谱系统中影响肌醇磷酸酯保留行为和分离的所有主要色谱变量进行了详细研究。研究的参数包括配对离子的洗脱液浓度、有机改性剂和缓冲盐的洗脱液浓度、洗脱液的pH值、分离肌醇三磷酸异构体所需的最小柱板数以及等度和梯度分离模式。还研究了一些常见核苷酸和糖磷酸酯的保留行为，因为这些磷酸盐在基于[32P]Pi细胞掺入的生化研究中存在色谱干扰问题。就分离速度和色谱条件的“温和性”而言，所开发的分离方法似乎优于既定的阴离子交换分离技术。
