Assay of urinary 8-hydroxy-2'-deoxyguanosine by capillary electrophoresis with spectrophotometric detection using a high-sensitivity detection cell and solid-phase extraction.
A sensitive capillary electrophoretic method featuring spectrophotometric detection using a commercial Z-cell was devised for the assay of 8-hydroxy-2'-deoxyguanosine (8OHdG) in human urine. Solid-phase extraction (SPE) based on hydrophilic-lipophilic-balanced RP sorbent was utilized for urine sample pretreatment and analyte preconcentration. The separation was carried out in conventional fused-silica capillaries employing a Z-cell with hydrodynamic sample injection (at 50 mbar for 12 s). The BGE (pH* 9.2, adjusted with 1 M NaOH) contained 0.15 M boric acid and 10% v/v ACN. The detection wavelength was 282 nm. The calibration curve for 8OHdG (measured in spiked urine) was linear in the range 10-1000 ng/mL; R(2) = 0.9993. The LOD was 3 ng/mL (11 nmol/L) of 8OHdG. Determination of the 8OHdG urinary levels was possible even in healthy individuals.
Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského, Hradec Králové, Czech Republic.
Klára
K
Petrů
Jitka
J
Siroká
Lucie
L
Bydžovská
Lenka
L
Krčmová
Miroslav
M
Polášek
D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*
D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine
8-Hydroxy-2′-deoxyguanosine; CE; Oxidative stress; Urine