title: Electrochemical determination of oxidative damaged DNA with high sensitivity and stability using a nanocarbon film.
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 sp(2) and sp(3) 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.
authors: National Institute of Advanced Industrial Science and Technology, Higashi, Tsukuba, Ibaraki, Japan. dai.kato@aist.go.jp
authors: Dai
authors: D
authors: Kato
authors: Mayuri
authors: M
authors: Komoriya
authors: Kohei
authors: K
authors: Nakamoto
authors: Ryoji
authors: R
authors: Kurita
authors: Shigeru
authors: S
authors: Hirono
authors: Osamu
authors: O
authors: Niwa
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000328:Adult; D002244:Carbon / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D005260:Female; D005898:Glass / Q000737:chemistry; D006801:Humans; D008297:Male; D008875:Middle Aged; D049329:Nanostructures / Q000737:chemistry*; D010084:Oxidation-Reduction
chemical_list: D002244:Carbon; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine