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Detection of extracellular 8-oxo-7,8-dihydro-2′-deoxyguanosine as a biomarker of oxidative damage in X-irradiated fibroblast cultures: optimization of analytical procedure

Abstract : We have developed a simple methodology, based on single-step solid-phase extraction followed by isocratic high-performance liquid chromatography coupled with electrochemical detection (HPLC-ECD), to determine extracellular 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) in culture supernatants of normal human dermal fibroblasts. A standard addition method, using externally added 8-oxodG (0.5 and 1 pmol) was employed to eliminate matrix effects arising from the chemically complex, protein-rich medium. Secondly, applying this procedure to X-ray irradiated fibroblasts, we report a significant twofold increase in the levels of 8-oxodG at the radiobiologically relevant dose of 6 Gy. This suggests that extracellular 8-oxodG might be a useful biomarker for oxidative stress following moderate doses of X-irradiation.
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https://hal-normandie-univ.archives-ouvertes.fr/hal-02297398
Contributor : Carine Laurent <>
Submitted on : Thursday, September 26, 2019 - 10:27:46 AM
Last modification on : Wednesday, June 10, 2020 - 12:44:08 PM

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Stéphanie Lagadu, Ivannah Pottier, François Sichel, Carine Laurent, Jean-Louis Lefaix, et al.. Detection of extracellular 8-oxo-7,8-dihydro-2′-deoxyguanosine as a biomarker of oxidative damage in X-irradiated fibroblast cultures: optimization of analytical procedure. Biomarkers, Taylor & Francis, 2010, 15 (8), pp.707-714. ⟨10.3109/1354750X.2010.511269⟩. ⟨hal-02297398⟩

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