Analysis of normal and modified nucleosides in urine samples by high-performance liquid chromatography with different stationary phases.

The main aim of the present work was to study the retention behavior and quantification of nine nucleosides with the use of octadecyl, alkylamide, cholesterol and alkyl-phosphate stationary phases. The influence of organic solvent and buffer concentration on the separation of these compounds was under investigation. The retention factor had the highest values for the octadecyl and cholesterol packing materials. Complete separation of all the studied nucleosides was achieved in case of cholesterol stationary phase. The optimized separation method was applied for the quantification of nucleosides in the urine samples. Calibration plots showed good linearity (R(2) > 0.999) and the limits of detection were in a range of 0.3-0.5 µg/mL, while the limits of quantitation were >0.9 µg/mL. Accuracy was in the range of 5-11%.