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

• Published in: Free radical biology & medicine Vol. 47; no. 5; pp. 629 - 635
• Main Authors: Henriksen, Trine, Hillestrøm, Peter R., Poulsen, Henrik E., Weimann, Allan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2009
• Subjects: 8-oxodG; 8-oxoGuo; Automation - methods; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Deoxyguanosine - analogs & derivatives; Deoxyguanosine - analysis; Deoxyguanosine - urine; Guanosine - analogs & derivatives; Guanosine - analysis; Guanosine - urine; Human urine; Humans; Mass spectrometry; Oxidative stress; Reproducibility of Results; Sensitivity and Specificity; Specimen Handling - methods; Tandem Mass Spectrometry - instrumentation; Tandem Mass Spectrometry - methods; UPLC; Urinalysis - instrumentation; Urinalysis - methods; Oxidative stress; 8-oxoGuo; UPLC; Human urine; ISTD; Mass spectrometry; 8-oxodG
• ISSN: 0891-5849; 1873-4596; 1873-4596
• DOI: 10.1016/j.freeradbiomed.2009.06.002

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.