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

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Author

  • Lars Barregard
  • Peter Moller
  • Trine Henriksen
  • Vilas Mistry
  • Gudrun Koppen
  • Pavel Rossner Jr.
  • Radim J. Sram
  • Allan Weimann
  • Henrik E. Poulsen
  • Robert Nataf
  • Roberta Andreoli
  • Paola Manini
  • Tim Marczylo
  • Patricia Lam
  • Mark D. Evans
  • Hiroshi Kasai
  • Kazuaki Kawai
  • Yun-Shan Li
  • Kazuo Sakai
  • Rajinder Singh
  • Friederike Teichert
  • Peter B. Farmer
  • Rafal Rozalski
  • Daniel Gackowski
  • Agnieszka Siomek
  • Guillermo T. Saez
  • Concha Cerda
  • Karin Broberg Palmgren
  • Christian Lindh
  • Bakhtiar Hossain
  • Siamak Haghdoost
  • Chiung-Wen Hu
  • Mu-Rong Chao
  • Kuen-Yuh Wu
  • Hilmi Orhan
  • Nilufer Senduran
  • Raymond J. Smith
  • Regina M. Santella
  • Yali Su
  • Czarina Cortez
  • Susan Yeh
  • Ryszard Olinski
  • Steffen Loft
  • Marcus S. Cooke
Show all

Summary, in English

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.

Publishing year

2013

Language

English

Pages

2377-2391

Publication/Series

Antioxidants & Redox Signaling

Volume

18

Issue

18

Document type

Journal article

Publisher

Mary Ann Liebert, Inc.

Topic

  • Cell and Molecular Biology

Status

Published

Research group

  • Genetic Occupational and Environmental Medicine

ISBN/ISSN/Other

  • ISSN: 1557-7716