TY - JOUR
T1 - 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
AU - Henriksen, Trine
AU - Hillestrøm, Peter R
AU - Poulsen, Henrik E
AU - Weimann, Allan
AU - Henriksen, Trine
AU - Hillestrøm, Peter René
AU - Poulsen, Henrik E
AU - Weimann, Allan
N1 - Keywords: Automation; Chromatography, Liquid; Deoxyguanosine; Guanosine; Humans; Reproducibility of Results; Sensitivity and Specificity; Specimen Handling; Tandem Mass Spectrometry; Urinalysis
PY - 2009/9/1
Y1 - 2009/9/1
N2 - 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.
AB - 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.
U2 - 10.1016/j.freeradbiomed.2009.06.002
DO - 10.1016/j.freeradbiomed.2009.06.002
M3 - Journal article
C2 - 19501155
SN - 0891-5849
VL - 47
SP - 629
EP - 635
JO - Free Radical Biology & Medicine
JF - Free Radical Biology & Medicine
IS - 5
ER -