Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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

Barregard, Lars ; Moller, Peter ; Henriksen, Trine ; Mistry, Vilas ; Koppen, Gudrun ; Rossner Jr., Pavel ; Sram, Radim J. ; Weimann, Allan ; Poulsen, Henrik E. and Nataf, Robert , et al. (2013) In Antioxidants & Redox Signaling 18(18). p.2377-2391
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Antioxidants & Redox Signaling
volume
18
issue
18
pages
2377 - 2391
publisher
Mary Ann Liebert, Inc.
external identifiers
  • wos:000319871200001
  • scopus:84878591374
  • pmid:23198723
ISSN
1557-7716
DOI
10.1089/ars.2012.4714
language
English
LU publication?
yes
id
70b6e36a-8f83-4b01-924f-e925fbaf9634 (old id 3926997)
date added to LUP
2016-04-01 14:13:49
date last changed
2022-04-06 17:28:11
@article{70b6e36a-8f83-4b01-924f-e925fbaf9634,
  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.}},
  author       = {{Barregard, Lars and Moller, Peter and Henriksen, Trine and Mistry, Vilas and Koppen, Gudrun and Rossner Jr., Pavel and Sram, Radim J. and Weimann, Allan and Poulsen, Henrik E. and Nataf, Robert and Andreoli, Roberta and Manini, Paola and Marczylo, Tim and Lam, Patricia and Evans, Mark D. and Kasai, Hiroshi and Kawai, Kazuaki and Li, Yun-Shan and Sakai, Kazuo and Singh, Rajinder and Teichert, Friederike and Farmer, Peter B. and Rozalski, Rafal and Gackowski, Daniel and Siomek, Agnieszka and Saez, Guillermo T. and Cerda, Concha and Broberg Palmgren, Karin and Lindh, Christian and Hossain, Bakhtiar and Haghdoost, Siamak and Hu, Chiung-Wen and Chao, Mu-Rong and Wu, Kuen-Yuh and Orhan, Hilmi and Senduran, Nilufer and Smith, Raymond J. and Santella, Regina M. and Su, Yali and Cortez, Czarina and Yeh, Susan and Olinski, Ryszard and Loft, Steffen and Cooke, Marcus S.}},
  issn         = {{1557-7716}},
  language     = {{eng}},
  number       = {{18}},
  pages        = {{2377--2391}},
  publisher    = {{Mary Ann Liebert, Inc.}},
  series       = {{Antioxidants & Redox Signaling}},
  title        = {{Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2 '-deoxyguanosine}},
  url          = {{http://dx.doi.org/10.1089/ars.2012.4714}},
  doi          = {{10.1089/ars.2012.4714}},
  volume       = {{18}},
  year         = {{2013}},
}