Use of a fundamental elution protocol for the development of reversed-phase high-performance liquid chromatography enabling rapid simultaneous determination of purines, pyrimidines and allied compounds commonly found in human biological fluids.

Many high-performance liquid chromatographic methods for the analysis of nucleosides and bases have been developed for a specific requirement. Most are not directly applicable to the separation of all the purines and pyrimidines found in biological fluids. A method has been developed to separate a wide range of compounds found in human urine and plasma in a single run without unduly long run times. Special attention has been paid to sample preparation. Isocratic systems derived from this method for the analysis of specific compounds and enzyme assay mixtures over even shorter periods are described.