Electrochemical detection of benzo(a)pyrene and related DNA damage using DNA/hemin/nafion-graphene biosensor.
A novel electrochemical biosensor, DNA/hemin/nafion-graphene/GCE, was constructed for the analysis of the benzo(a)pyrene PAH, which can produce DNA damage induced by a benzo(a)pyrene (BaP) enzyme-catalytic product. This biosensor was assembled layer-by-layer, and was characterized with the use of cyclic voltammetry, electrochemical impedance spectroscopy (EIS) and atomic force microscopy. Ultimately, it was demonstrated that the hemin/nafion-graphene/GCE was a viable platform for the immobilization of DNA. This DNA biosensor was treated separately in benzo(a)pyrene, hydrogen peroxide (H2O2) and in their mixture, respectively, and differential pulse voltammetry (DPV) analysis showed that an oxidation peak was apparent after the electrode was immersed in H2O2. Such experiments indicated that in the presence of H2O2, hemin could mimic cytochrome P450 to metabolize benzo(a)pyrene, and a voltammogram of its metabolite was recorded. The DNA damage induced by this metabolite was also detected by electrochemical impedance and ultraviolet spectroscopy. Finally, a novel, indirect DPV analytical method for BaP in aqueous solution was developed based on the linear metabolite versus BaP concentration plot; this method provided a new, indirect, quantitative estimate of DNA damage.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; Department of Chemistry, Nanchang University, Nanchang 330031, China. Electronic address: ynni@ncu.edu.cn.
Yongnian
Y
Ni
Department of Chemistry, Nanchang University, Nanchang 330031, China.
Pingping
P
Wang
Department of Chemistry, Nanchang University, Nanchang 330031, China.
Haiyan
H
Song
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; Department of Chemistry, Nanchang University, Nanchang 330031, China.
Xiaoyun
X
Lin
School of Chemistry, Physics and Mechanical Engineering, Science and Engineering Faculty, Queensland University of Technology, Brisbane 4001, Australia. Electronic address: s.kokot@qut.edu.au.
Serge
S
Kokot
D001564:Benzo(a)pyrene / Q000032:analysis / Q000633:toxicity*; D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects*; D058266:Dielectric Spectroscopy; D055664:Electrochemical Techniques / Q000379:methods; D004566:Electrodes; D004785:Environmental Pollutants / Q000032:analysis / Q000633:toxicity*; D006108:Graphite / Q000737:chemistry*; D006427:Hemin / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000737:chemistry; D057230:Limit of Detection; D015203:Reproducibility of Results
D004785:Environmental Pollutants; D001564:Benzo(a)pyrene; D006427:Hemin; D006108:Graphite; D004247:DNA; D006861:Hydrogen Peroxide
DNA biosensor; DNA damage; DNA/hemin/nafion–graphene/glassy carbon electrode; Differential pulse voltammetry; Polycyclic aromatic hydrocarbons