Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2'-deoxyguanosine.

2.50
Hdl Handle:
http://hdl.handle.net/10146/269552
Title:
Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2'-deoxyguanosine.
Authors:
Barregard, Lars; Moller, Peter; Henriksen, Trine; Mistry, Vilas; Koppen, Gudrun; Rossner, Pavel; Sram, Radim J.; Weimann, Allan; Poulsen, Henrik E.; Nataf, Robert; Andreoli, Roberta; Manini, Paola; Marczylo, Tim; Lam, Patricia; Evans, Mark D.; Kasai, Hiroshi; Kawai, Kazuaki; Li, Yun-Shan; Sakai, Kazuo; Singh, Rajinder; Teichert, Friederike; Farmer, Peter B.; Rozalski, Rafal; Gackowski, Daniel; Siomek, Agnieszka; Saez, Guillermo T.; Cerda, Concha; Broberg, Karin; Lindh, Christian; Hossain, Mohammad Bakhtiar; Haghdoost, Siamak; Hu, Chiung-Wen; Chao, Mu-Rong; Wu, Kuen-Yuh; Orhan, Hilmi; Senduran, Nilufer; Smith, Raymond J.; Santella, Regina M.; Su, Yali; Cortez, Czarina; Yeh, Susan; Olinski, Ryszard; Loft, Steffen; Cooke, Marcus S.
Abstract:
Abstract Aims: Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed 'spot' samples showed strong correlations with 24 h excretion (the 'gold' standard) of urinary 8-oxodG (r(p) 0.67-0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine- or SG-adjusted first morning samples are recommended.
Citation:
Antioxid. Redox Signal. 2013, 18(18):2377-2391
Journal:
Antioxidants & Redox Signaling
Issue Date:
31-Jan-2013
URI:
http://hdl.handle.net/10146/269552
DOI:
10.1089/ars.2012.4714
PubMed ID:
23198723
Additional Links:
http://online.liebertpub.com/doi/abs/10.1089/ars.2012.4714
Type:
Article
Language:
en
ISSN:
1557-7716
Sponsors:
Some of the authors of this work were partners in, and this work was partly supported by, ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6 th Framework Program, Priority 5:"Food Quality and Safety" (Contract No. FOOD-CT-2005-513943), and also ECNIS 2 , a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologies
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorBarregard, Larsen_GB
dc.contributor.authorMoller, Peteren_GB
dc.contributor.authorHenriksen, Trineen_GB
dc.contributor.authorMistry, Vilasen_GB
dc.contributor.authorKoppen, Gudrunen_GB
dc.contributor.authorRossner, Pavelen_GB
dc.contributor.authorSram, Radim J.en_GB
dc.contributor.authorWeimann, Allanen_GB
dc.contributor.authorPoulsen, Henrik E.en_GB
dc.contributor.authorNataf, Roberten_GB
dc.contributor.authorAndreoli, Robertaen_GB
dc.contributor.authorManini, Paolaen_GB
dc.contributor.authorMarczylo, Timen_GB
dc.contributor.authorLam, Patriciaen_GB
dc.contributor.authorEvans, Mark D.en_GB
dc.contributor.authorKasai, Hiroshien_GB
dc.contributor.authorKawai, Kazuakien_GB
dc.contributor.authorLi, Yun-Shanen_GB
dc.contributor.authorSakai, Kazuoen_GB
dc.contributor.authorSingh, Rajinderen_GB
dc.contributor.authorTeichert, Friederikeen_GB
dc.contributor.authorFarmer, Peter B.en_GB
dc.contributor.authorRozalski, Rafalen_GB
dc.contributor.authorGackowski, Danielen_GB
dc.contributor.authorSiomek, Agnieszkaen_GB
dc.contributor.authorSaez, Guillermo T.en_GB
dc.contributor.authorCerda, Conchaen_GB
dc.contributor.authorBroberg, Karinen_GB
dc.contributor.authorLindh, Christianen_GB
dc.contributor.authorHossain, Mohammad Bakhtiaren_GB
dc.contributor.authorHaghdoost, Siamaken_GB
dc.contributor.authorHu, Chiung-Wenen_GB
dc.contributor.authorChao, Mu-Rongen_GB
dc.contributor.authorWu, Kuen-Yuhen_GB
dc.contributor.authorOrhan, Hilmien_GB
dc.contributor.authorSenduran, Niluferen_GB
dc.contributor.authorSmith, Raymond J.en_GB
dc.contributor.authorSantella, Regina M.en_GB
dc.contributor.authorSu, Yalien_GB
dc.contributor.authorCortez, Czarinaen_GB
dc.contributor.authorYeh, Susanen_GB
dc.contributor.authorOlinski, Ryszarden_GB
dc.contributor.authorLoft, Steffenen_GB
dc.contributor.authorCooke, Marcus S.en_GB
dc.date.accessioned2013-02-14T11:57:30Z-
dc.date.available2013-02-14T11:57:30Z-
dc.date.issued2013-01-31-
dc.identifier.citationAntioxid. Redox Signal. 2013, 18(18):2377-2391en_GB
dc.identifier.issn1557-7716-
dc.identifier.pmid23198723-
dc.identifier.doi10.1089/ars.2012.4714-
dc.identifier.urihttp://hdl.handle.net/10146/269552-
dc.description.abstractAbstract Aims: Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed 'spot' samples showed strong correlations with 24 h excretion (the 'gold' standard) of urinary 8-oxodG (r(p) 0.67-0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine- or SG-adjusted first morning samples are recommended.en_GB
dc.description.sponsorshipSome of the authors of this work were partners in, and this work was partly supported by, ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6 th Framework Program, Priority 5:"Food Quality and Safety" (Contract No. FOOD-CT-2005-513943), and also ECNIS 2 , a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologiesen_GB
dc.languageENG-
dc.language.isoenen
dc.relation.urlhttp://online.liebertpub.com/doi/abs/10.1089/ars.2012.4714en_GB
dc.rightsArchived with thanks to Antioxidants & redox signalingen_GB
dc.subjectOxidative stressen_GB
dc.subjectBiomarkersen_GB
dc.subject8-oxo-7,8-dihydro-2'-deoxyguanosineen_GB
dc.subjectChromatographicen_GB
dc.subjectELISAen_GB
dc.subjectVariabilityen_GB
dc.titleHuman and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2'-deoxyguanosine.en
dc.typeArticleen
dc.identifier.journalAntioxidants & Redox Signalingen_GB

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