Mobile phase and temperature studies in the reversed-phase liquid chromatography of inosine and guanosine compounds - Yi, C.; Fasching, J.L.; Brown, P.R. (Elsevier)
The results are reported of a systematic investigation of the effects of the mobile phase methanol concentration and pH, temperature and flow-rate, separately or in combination, on the reversed-phase liquid chromatography of selected biologically important nucleosides. As expected in a reversed-phase separation, the retention times of each nucleoside decreased proportionally with increasing methanol concentration. The pH effects were dependent on the percentage of methanol present. A methanol selectivity factor is proposed for expressing numerically the effect of methanol concentration on retention.Increasing temperature also decreased retention, but the extent of the effect was dependent on the structure of the nucleoside. Thus, chromatography of a sample at temperatures above ambient could cause peak mergers. A method using experimentally determined retention times of two nucleosides at two temperatures was proposed for determining exact t"0, (void time) values of a system. The t"0, values which were calculated by an equation and determined graphically were in good agreement.
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