Group-selective prefractionation and analysis of nucleosides and catecholamines by high-performance liquid chromatographic techniques

Abstract

The use of borate as ligand in conventional affinity chromatography has found numerous applications in biochemical research [6], as for example for the clean-up of ribonucleosides and catecholamines in physiological fluids, the separation of DNA and RNA, the isolation of glycosidated hemoglobins, separation of aminoacylated RNA from free RNA, ligand mediated (‘piggyback’) chromatographic enrichment of enzymes or the separation of base Q containing tRNA from base Q free tRNA. With the development of borate functionalized silica borate affinity chromatography has also been turned out to work under HPLAC conditions. By use of a column switching technique we could introduce a combined HPLAC/HPLC method particularly suitable for the on-line clean-up and analysis of ribonucleosides in complex matrices. We now have expanded the application of our on-line system for the clean-up and analysis of the adrenergic amines from spiked physiological matrix which means a powerful improvement compared to the system introduced by [7] for the on-line analysis just of one of the dopamine catabolites. The method described should be the method of choice for the majority of applications mentioned above as it greatly decreases the analysis time, is suitable for automation and in conjunction with a data-processing system, is applicable to routine clinical analysis.