Simultaneous determination of purine metabolites inovine urine and blood plasma by high-performanceliquid chromatography

TL;DRAbstract

High-performance liquid chromatographic methods for directly determining products of nucleic acid and purine catabolism in urine and blood plasma of sheep were developed. Urine and plasma samples were diluted 1:3 with deionized water. Fractionation and quantifi cation of allantoin, uric acid, hypoxanthine and xanthine were performed using two Nova-Pak C 18 columns (300 3.9 mm, Waters). Binary gradient programs and UV detection were used for purine metabolite analysis. Satisfactory fractionation of all analytes in urine and plasma was obtained in less than 19 and 21 min, respectively. The average recoveries of standard compounds added to the assayed samples were ~100%. The low coeffi cient of variation (1-2%) as well as the low detection limits (0.09-0.36 nmol) indicate satisfactory precision, reproducibility and sensitivity of the proposed methods. These chromatographic methods are suitable for routine quantifi cation of purine metabolites in a large number of samples.

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