Automated method for the direct analysis of 8-oxo-guanosine and 8-oxo-2'-deoxyguanosine in human urine using ultraperformance liquid chromatography and tandem mass spectrometry.
The potential use of oxidative stress-induced DNA and RNA damage products as biomarkers is an important aspect of biomedical research. There is a need for assays with high specificity and sensitivity that also can be used in molecular epidemiology studies with a large number of subjects. In addition there is a need for assays that can measure more than one product from DNA oxidation. We present a sensitive, precise, and accurate method for quantitative analysis of the oxidized nucleosides 8-oxoGuo and 8-oxodG in human urine. The assay is based on automated sample handling using a BIOMEK 3000 Workstation, and UPLC-ESI(+)-MS/MS analysis. High specificity is evidenced by the use of qualifier ions for both analytes. The quantification limit in urine samples is 1 nM for both analytes. Accuracy and precision were documented, showing average recoveries of 106.2% (8-oxoGuo) and 106.9% (8-oxodG), and overall precision (within-day and between-days) of 6.1 and 4.4%, respectively.
Henriksen
Trine
T
Laboratory of Clinical Pharmacology Q7642, Rigshospitalet, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
Hillestrøm
Peter R
PR
Poulsen
Henrik E
HE
Weimann
Allan
A
D000080242:8-Hydroxy-2'-Deoxyguanosine; D001331:Automation / Q000379:methods; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000652:urine; D006151:Guanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000652:urine; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000295:instrumentation* / Q000379:methods*; D016482:Urinalysis / Q000295:instrumentation / Q000379:methods*
D006151:Guanosine; C046215:8-hydroxyguanosine; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine