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Publication Detail

Title: 8-Oxo-2'-deoxyguanosine as a biomarker of tobacco-smoking-induced oxidative stress.

Authors: Mesaros, Clementina; Arora, Jasbir S; Wholer, Ashley; Vachani, Anil; Blair, Ian A

Published In Free Radic Biol Med, (2012 Aug 01)

Abstract: 7,8-Dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dGuo) is a useful biomarker of oxidative stress. However, its analysis can be challenging because 8-oxo-dGuo must be quantified in the presence of dGuo, without artifactual conversion to 8-oxo-dGuo. Urine is the ideal biological fluid for population studies, because it can be obtained noninvasively and it is less likely that artifactual oxidation of dGuo can occur because of the relatively low amounts that are present compared with hydrolyzed DNA. Stable isotope dilution liquid chromatography-selected reaction monitoring/mass spectrometry (LC-SRM/MS) with 8-oxo-[(15)N(5)]dGuo as internal standard provided the highest possible specificity for 8-oxo-dGuo analysis. Furthermore, artifact formation was determined by addition of [(13)C(10)(15)N(5)]dGuo and monitoring of its conversion to 8-oxo-[(13)C(10)(15)N(5)]dGuo during the analytical procedure. 8-Oxo-dGuo concentrations were normalized for interindividual differences in urine flow by analysis of creatinine using stable isotope dilution LC-SRM/MS. A significant increase in urinary 8-oxo-dGuo was observed in tobacco smokers compared with nonsmokers either using simple urinary concentrations or after normalization for creatinine excretion. The mean levels of 8-oxo-dGuo were 1.65ng/ml and the levels normalized to creatinine were 1.72μg/g creatinine. Therefore, stable isotope dilution LC-SRM/MS analysis of urinary 8-oxo-dGuo complements urinary isoprostane (isoP) analysis for assessing tobacco-smoking-induced oxidative stress. This method will be particularly useful for studies that employ polyunsaturated fatty acids, in which a reduction in arachidonic acid precursor could confound isoP measurements.

PubMed ID: 22613262 Exiting the NIEHS site

MeSH Terms: Biomarkers/urine; Calibration; Case-Control Studies; Chromatography, High Pressure Liquid/standards; DNA Damage; Deoxyguanosine/analogs & derivatives*; Deoxyguanosine/urine; Humans; Limit of Detection; Mass Spectrometry/standards; Oxidative Stress*; Reference Standards; Smoking/urine*

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