High-performance liquid chromatographic methods for base and nucleoside analysis in extracellular fluids and in cells

Abstract

High-performance liquid chromatography based methods for the study of the metabolism of purine and pyrimidine bases and nucleosides have been developed. These methods, using 200--50 microliter samples of extracellular fluids and employing isocratic separations, can measure a wide range of compounds. Hypoxanthine, xanthine and uridine concentrations in plasma from normal men are relatively stable. Species differences have been detected: concentrations of cytidine are higher in rat and mouse serum than in man, since the concentrations of uridine are similar; purine/pyrimidine ratios may be different. Fetal calf serum used for tissue culture contains about a 40 times higher concentration of hypoxanthine than the less-effective calf serum. Use of the methods appears to be justified in the assessment of the metabolic damage due to severe hypoxia and/or ischaemia.