ANALYSIS OF URINARY 8-HYDROXYDEOXYGUANOSINE BY CAPILLARY ELECTROPHORESIS AND SOLID-PHASE EXTRACTION

Abstract

Urinary 8-hydroxydeoxyguanosine (8OHdG) has been considered as an excellent marker of individuals at high risk of developing cancer. Until now, urinary 8OHdG has largely been measured by high-performance liquid chromatography with electrochemical detection. A new method for the analysis of urinary 8OHdG by high-performance capillary electrophoresis has been developed and optimized in our laboratory. A single step solid-phase extraction procedure was optimized and used for extracting 8OHdG from human urine. Separations were performed in an uncoated silica capillary (50 cm × 50 μm i.d.) using a P/ACE MDQ system with UV detection. The separation of 8OHdG from interfering urinary matrix components is optimized with regard to pH, applied voltage, pressure injection time and concentration of SDS in running buffer. The detection limit of this method is 0.4 μg/ml, the linear range is 0.8–500 μg/ml, the correlation coefficients levels is better than 0.999. The developed method is simple, fast and good reproducibility, furthermore, it requires a very small injection volumes and low costs of analysis, which makes it possible to provide a new noninvasive assay for an indirect measurement of oxidative DNA damage.

Keywords:

• 8-Hydroxydeoxyguanosine
• Capillary electrophoresis
• Solid-phase extraction
• Urine sample

Acknowledgments