Comparison of the reversed-phase high-performance liquid chromatographic separations of fluoropyrimidines, pyrimidines, and purines

The reversed-phase, high-performance liquid chromatographic separation of fluoropyrimidines, pyrimidines, and purines was investigated under isocratic conditions at ambient temperature. The performance of nine analytical, commercially available columns with five mobile phases is compared, and capacity and resolution factors are reported. The variables determining resolution are discussed, and the systems accomplishing the desired separation of fluoropyrimidine and pyrimidine bases and nucleosides are described. The best chromatographic results are achieved by using Spherisorb ODS-2 as stationary phase and 0.05 M monobasic ammonium phosphate (pH 3.5) as mobile phase.