High-performance liquid-chromatographic analysis of bases and nucleoside in mouse testes following in vivo thymidine...

In order to measure accurately bases and nucleosides in vivo, a method has been developed using high-performance liquid chromatography. This method is reproducible under conditions of constant pressure, temperature and pH and is rapid, requiring approximately 1 h for a complete chromatographic separation of bases and nucleosides. The method has been used to measure quantitatively the bases and nucleosides in mouse testes following in vivo administration of thymidine. Groups of male mice were given thymidine by intra-peritoneal injection and were killed at various time intervals and the testes removed for extraction procedures. There was a significant increase in the thymidine concentration in the testes 1 h after injection, which fairly quickly returned to the control value thereafter. There was a subsequent reduction in uridine concentration. This analytical method allows deoxyribonucleotide (dNTP) precursor pool sizes to be determined in germinal cells. It may also be useful for other tissues in vivo.