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European Standards Committee on oxidative DNA Damage

Contract number : QLK1-1999-00568
Contract type : Concerted Action
Total cost : € 501.850
EC contribution : € 501.850
Starting date : 1/02/2000
Duration : 36 Months
Scientific Officer : Rosanna d'Amario
Project website : not yet available
Dr Andrew Collins
The Rowett Research Institute
Greenburn Road
AB21 9SB Bucksburn, Aberdeen
United Kingdom
Tel.: +44-1224 716634
Fax: +44-1224 716629
E-mail: a.collins @

Oxidative damage to DNA may contribute to cancer aetiology and is thought to be a factor in ageing. Measurement of 8-oxo-guanine as a marker of DNA oxidation is problematic, and there is an urgent need for standardisation of methods. Spurious oxidation of guanine during sample preparation must be eliminated. This Concerted Action will develop standard protocols and validate methods by distributing standard samples to participants. The final objective will be to reach a consensus on the background level of DNA oxidation in normal human lymphocytes.


The principal objectives of this Concerted Action are:

  • to validate HPLC, GC-MS and LC-MS-MS methods used for measurement of 8-oxo-7,8-dihydroguanine (8-oxo-gua) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) by the use of reference standard DNA samples analysed in parallel in different laboratories, including heavy labelled standards for mass spectrometry;
  • to increase the sensitivity and reliability of these 'conventional' methods;
  • to measure DNA oxidation in parallel on identical samples using 'conventional' methods and repair endonuclease-based methods;
  • to reach a consensus on the average level of oxidation in normal human DNA.

The role of oxidative stress in the aetiology of human disease has recently been recognised. Oxidative changes to DNA are chemically well defined; over 100 modifications have been described, the most abundant being 8-oxo-guanine. This oxidised base is potentially mutagenic; in in vitro replication systems, it leads to misincorporation of adenine in the opposite strand. There is some evidence that, at least in animals, the frequency of 8-oxo-guanine in the DNA increases with age. Advanced analytical methods such as GC-MS, HPLC with electrochemical detection, and recently LC-MS-MS have been employed to measure the background level of 8-oxo-gua (8-oxo-dG) in normal human cells. Estimates range over two or three orders of magnitude. An alternative approach, based on the use of a lesion-specific repair endonuclease that introduces strand breaks into DNA at sites of 8-oxo-gua, suggests that the level of damage may be even lower than the lowest of the direct estimates. It is clear that oxidation of gua during sample preparation for GC-MS and HPLC is a serious artefact. Elimination of this problem, standardisation of protocols and reduction of variability and errors in the different assays are essential if we are to achieve a sound judgment of the amount of damage present - a prerequisite for assessing the importance of oxidative DNA damage in the aetiology of diseases such as cancer, Down's syndrome, cystic fibrosis and premature ageing syndromes such as Werner's syndrome as well as its role in normal ageing. This Concerted Action will provide for the systematic exchange of standards and samples between participants, with regular meetings to discuss the results and to optimise methodology. In addition, scientists will take part in exchanges between laboratories to facilitate the adoption of improvements in technique. At each stage, rigorous analysis of the reported analytical results will be carried out, and progress towards the objectives will be very easy to assess.


University of Florence
Department of Preclinical and Clinical Pharmacology
Viale Pieraccini 6
Florence 50139
Département de Recherche Fondamentale sur la Matière Condensée
17 avenue des Martyrs
38054 Grenoble
University Libre de Bruxelles
Institut de Pharmacie
Laboratoire de Chimie Bioanalytique de Toxicologie et de Chimie et Physique Appliqués
CP 205/1
Bd du Triomphe
1050 Bruxelles
Medical Faculty of Comenius University
Department of Medical Chemistry, Biochemistry and Clinical Biochemistry
Sasinkova 2
Bratislava 81372
Slovak Republic
Department of Molecular and Genetic Toxicology
Institute of Preventive and Clinical Medicine
Limbova 14
Bratislava 83301
Health & Consumer Science Institute
Food Research Norwich Research
Park Colney
Norwich NR4 7UA
United Kingdom
Institut für Pharmazie
Johannes Gutenberg-Universität
Staudinger Weg 5
55099 Mainz
Université Bordeaux 1
351 Cours de la Libération
Talence 33405
Institut für Lebensmittelchemie
Universität Karlsruhe
Kaiserstraße 12
Karlsruhe 76128
Centre for Mechanisms of Human Toxicity
University of Leicester
138 Lancaster Road
Leicester LE1 9HN
United Kingdom
University of Occupational & Environmental Health
Institute of Industrial Ecological Sciences
Department of Environmental Oncology
1-1 Iseigaoka
Kitakyushu 807-8555
Rayne Institute
Cardiovascular Research
King's College London
St Thomas' Hospital
London SE1 7EH
United Kingdom
Department of Environment and Occupational Medicine
Institute of Public Health
Panum, Blegdamsvej 3
Copenhagen 2200
CNT Karolinska Institutet
Stockholm S-14157
Department of Clinical Biochemistry
Ludwik Rydygier Medical University Bydgoszcz
Karlowicza 24
Bydgoszcz 85-092
Université Catholique de Louvain
Laboratoire de Biologie Cellulaire
Unité de Biologie Animale
Croix du Sud. 5
1348 Louvain-La-Neuve
Quality and Safety Assurance Department
Metabolism and Biomarkers Group
Nestec S.A.
Nestlé Research Centre
PO Box 44
Lausanne 26, 1000
Department of Pharmacy & Pharmacology
University of Bath
Claverton Down
Bath BA2 7AY
United Kingdom
Molecular Epidemiology Unit
University of Leeds
Algernon Firth Building
Leeds LS2 9JT
United Kingdom
Departamento de Fisiología
Facultad de Medicina
Universitat de Valencia
Avda. Blasco Ibañez
17, Valencia 46071
Faculté de Pharmacie
Université de Rennes 1
Avenue du Professeur Léon Bernard
Rennes 35043
Division of Chemical Pathology
Centre for Mechanisms of Human Toxicity
University of Leicester
Lancaster Road
Leicester LE1 9HN
United Kingdom
Deutsches Krebsforschungszentrum
H0600 Molekulare Genomanalyse
PO Box 101949
Im Neuenheimer Feld 280
69120 Heidelberg
Laboratoire de Biologie du Stress Oxydatif
Faculté de Médecine-Pharmacie
Université Joseph Fourier Grenoble 1
Avenue du Maquis du Grésivaudan
38700 La Tronche
Division of Chemical Pathology
Centre for Mechanisms of Human Toxicity
University of Leicester
PO Box 138
Lancaster Road
Leicester LE1 9HN
United Kingdom
Safety of Medicines
Astra Zeneca Pharmaceutical
Mereside, Alderley Park
Macclesfield SK10 4TG
United Kingdom

Fifth Framework Programme

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