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Electrochemical Determination of Oxidative Damaged DNA with High Sensitivity and Stability Using a Nanocarbon Film

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Abstract

We describe the electrochemical determination of oxidative damaged DNA by using a nanocarbon film electrode combined with a high performance liquid chromatography (HPLC) system. The nanocarbon film was formed by employing the electron cyclotron resonance sputtering method, and has a nano-crystalline sp2 and sp3 mixed bond structure with an atomically flat surface. This film electrode provided the high electrode activity and stability needed to quantitatively detect oxidative damaged DNA, 8-hydroxy-2'-deoxyguanosine (8-OHdG), by direct electrochemical oxidation. The coefficient of variation (C.V.) value of 1 µM 8-OHdG at our film electrode was 0.75% (n = 12), which constitutes superior reproducibility to that of a conventional glassy carbon (GC) electrode (9.28%, n = 12) in flow-injection analysis. This was because the nanocarbon film suppressed fouling for the oxidized product of 8-OHdG owing to its hydrophilically ultraflat and chemically stable surface. We also investigated the performance of HPLC with an electrochemical detection (HPLC-ECD) system using our nanocarbon film electrode. The detection limit for 8-OHdG at the nanocarbon film electrode was 3 nM, which was superior to the detection limit of the GC electrode (7.2 nM). Furthermore, this electrode was more suitable for use in a urinary 8-OHdG experiment than the GC electrode. The concentration of urinary 8-OHdG in the urinary sample was 8.30 nM. These results indicate that this HPLC-ECD system with our nanocarbon film electrode enables us to realize an accurate, sensitive, reproducible and easy to use analysis technique.

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Authors and Affiliations

  1. National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, 305-8566, Tsukuba, Ibaraki, Japan

    Dai Kato & Ryoji Kurita

  2. University of Tsukuba, 1-1-1 Tenno-dai, 305-8571, Tsukuba, Ibaraki, Japan

    Mayuri Komoriya & Kohei Nakamoto

  3. MES-Afty Corporation, 2-35-2 Hyoe, 192-0918, Hachioji, Tokyo, Japan

    Shigeru Hirono & Osamu Niwa

Authors
  1. Dai Kato
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  2. Mayuri Komoriya
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  3. Kohei Nakamoto
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  4. Ryoji Kurita
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  5. Shigeru Hirono
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  6. Osamu Niwa
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Correspondence to Dai Kato.

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Kato, D., Komoriya, M., Nakamoto, K. et al. Electrochemical Determination of Oxidative Damaged DNA with High Sensitivity and Stability Using a Nanocarbon Film. ANAL. SCI. 27, 703–707 (2011). https://doi.org/10.2116/analsci.27.703

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  • DOI: https://doi.org/10.2116/analsci.27.703

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