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Title: Analysis of urinary8-oxo-7,8-dihydro-purine-2’-deoxyribonucleosides by LC-MS/MS and improved ELISA
Authors: Evans, Mark D.
Singh, Rajinder
Mistry, Vilas
Sandhu, Karendeep
Farmer, Peter B.
Cooke, Marcus S.
First Published: Oct-2008
Publisher: Taylor and Francis
Citation: Free Radical Research, 2008, 42 (10), pp. 831-840
Abstract: Non-invasive monitoring of oxidative stress is highly desirable. Urinary 7,8-dihydro- 8-oxo-2’-deoxyguanosine (8-oxodG) is a biologically relevant and convenient analytical target. However, immunoassays can over-estimate levels of urinary 8-oxodG. Measurement of more than one DNA oxidation product in urine would be advantageous in terms of mechanistic information. Urines samples were analysed for 8-oxodG by solid-phase extraction/LC-MS/MS and ELISA. The solid-phase extraction/LC-MS/MS assay was also applied to the analysis of urinary 7,8-dihydro-8-oxo-2’-deoxyadenosine (8-oxodA). Concurring with previous reports, urinary 8- oxodG measured by ELISA was significantly higher than levels measured by LCMS/MS. However, apparent improvement in the specificity of the commercially available Japanese Institute for the Control of Ageing (JaICA) ELISA brought mean LC-MS/MS and ELISA measurements of urinary 8-oxodG into agreement. Urinary 8-oxodA was undetectable in all urines, despite efficient recovery by solid phase extraction. Exploitation of the advantages of ELISA may be enhanced by a simple modification to the assay procedure, although chromatographic techniques still remain the ‘gold standard’ techniques for analysis of urinary 8-oxodG. Urinary 8-oxodA is either not present or below the limit of detection of our instrumentation.
DOI Link: 10.1080/10715760802506323
ISSN: 1029-2470
Type: Article
Rights: This is the author's final draft of the paper published as Free Radical Research, 2008, 42 (10), pp. 831-840. The final published version is available at, Doi: 10.1080/10715760802506323.
Appears in Collections:Published Articles, Dept. of Cancer Studies and Molecular Medicine

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