Development and validation of urinary nucleosides and creatinine assay by capillary electrophoresis with solid phase extraction

Abstract

For the analysis of metabolite nucleoside profiles, capillary electrophoretic (CE) methods preceded by appropriate solid phase extraction procedures have been developed. The approach has been proposed for the determination of 13 nucleosides and creatinine in human urine. A background solution composed of 100 mM borate–72 mM phosphate–160 mM SDS and a fused silica capillary of 70 cm length to detector and 50 μm i.d. were used. The methods developed were statistically validated for their linearity, trueness, precision and selectivity. Stability of the analyzed nucleoside profiles in urine during storage was checked. Validation parameters of solid phase extraction procedures for urinary nucleosides were evaluated. The developed analytical methods were employed for the analysis of 22 urine samples from healthy patients and cancer patients from the urological ward. Nucleoside profiles were compared among the subjects. It was proved that the methods proposed were suitable for a fast and reliable determination of urinary creatinine and modified nucleoside profiles, which can be further submitted for the metabonomic analysis of cancer patients.