This review is focused on genotoxic 2-nitrofluorene, which is a marker for the presence of nitrated polycyclic aromatic hydrocarbons, and summarizes the current knowledge about the negative effects of 2-nitrofluorene and its metabolites on living organisms, especially on their DNA. These findings obtained via in vivo investigations are compared with information obtained using electrochemical DNA biosensors, which represent very promising in vitro alternative to the study of processes proceeding in living organisms during the interaction of their DNA with various xenobiotic compounds.
- Klíčová slova
- DNA biosenzory,
- MeSH
- elektrochemické techniky metody využití MeSH
- lidé MeSH
- polycyklické aromatické uhlovodíky * škodlivé účinky MeSH
- poškození DNA * MeSH
- xenobiotika metabolismus MeSH
- znečištění životního prostředí MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
A simple electrochemical DNA biosensor based on a glassy carbon electrode (GCE) was prepared by adsorbing double-stranded DNA (dsDNA) onto the GCE surface and subsequently used for the detection of dsDNA damage induced by hydroxyl radicals. Investigation of the mutual interaction between hydroxyl radicals and dsDNA was conducted using a combination of several electrochemical detection techniques: square-wave voltammetry for direct monitoring the oxidation of dsDNA bases, and cyclic voltammetry and electrochemical impedance spectroscopy as indirect electrochemical methods making use of the redox-active indicator [Fe(CN)6](4-/3-). Hydroxyl radicals were generated electrochemically on the surface of a boron-doped diamond electrode and chemically (via the Fenton's reaction or the auto-oxidation of Fe(II)). The extent of dsDNA damage by electrochemically generated hydroxyl radicals depended on the current density applied to the generating electrode: by applying 5, 10, and 50mAcm(-2), selected relative biosensor responses decreased after 3min incubation from 100% to 38%, 27%, and 3%, respectively. Chemically generated hydroxyl radicals caused less pronounced dsDNA damage, and their damaging activity depended on the form of Fe(II) ions: decreases to 49% (Fenton's reaction; Fe(II) complexed with EDTA) and 33% (auto-oxidation of Fe(II); Fe(II) complexed with dsDNA) were observed after 10min incubation.
- MeSH
- adsorpce MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- DNA chemie genetika MeSH
- elektrochemie MeSH
- elektrody MeSH
- hydroxylový radikál farmakologie MeSH
- peroxid vodíku chemie MeSH
- poškození DNA * MeSH
- sklo chemie MeSH
- uhlík chemie MeSH
- železo chemie MeSH
- Publikační typ
- časopisecké články MeSH
Interactions of various xenobiotic compounds with deoxyribonucleic acid (DNA) represent the most important aspects of biological studies in clinical and toxicological analyses, drug discovery, and pharmaceutical development processes. In recent years, a growing interest in electrochemical investigation of such supramolecular interactions has arisen. Monitoring the changes in electrochemical signals of DNA and/or observing the mutual interactions of DNA with examined analytes provide good evidence for the interaction mechanism to be elucidated. Moreover, the DNAanalyte interactions can also be used for sensitive determination of the analyte itself. This short review summarizes our results obtained during the last five years in the field of novel electrochemical DNA biosensors utilizing carbon-based transducers as substrates for immobilization of DNA. Its aim is to provide evidence that the electrochemical approach (employing simple, fast, sensitive, and inexpensive DNA biosensors as tools for investigation and detection of DNA damage) brings a new insight into human health protection and leads to better understanding of the interaction mechanism between xenobiotic compounds and DNA.
- MeSH
- cytotoxicita imunologická MeSH
- DNA * analýza chemie ultrastruktura MeSH
- elektrochemie * MeSH
- elektrody MeSH
- léčivé přípravky MeSH
- lidé MeSH
- noxy * analýza toxicita MeSH
- oxidační stres MeSH
- poškození DNA * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Polycyclic aromatic hydrocarbons are an important class of compounds ubiquitous in both the living and working environment. Since they are proven carcinogens and/or mutagens, the development of highly sensitive analytical methods for their determination is among the most demanding tasks in environmental analysis nowadays. A simple electrochemical biosensor (DNA/GCE), based on a glassy carbon electrode (GCE) and low molecular weight double-stranded DNA from salmon sperm immobilized onto the electrode surface, was used in this study for the development of a differential pulse voltammetric method for sensitive determination of a representative of polycyclic aromatic hydrocarbons – anthracene. The spontaneous accumulation of anthracene into the DNA structure (intercalation between the double-stranded DNA base pairs) was used to increase the sensitivity of the determination. The influence of pH on electrochemical behavior of anthracene was investigated at the bare GCE in buffered ethanolic–aqueous solutions. The optimal medium found was ethanol – 0.04 mol l–1 Britton-Robinson buffer pH 5.0 (1:1, v/v), which was also used for the subsequent determination of anthracene at the DNA/GCE, with an analyte accumulation time of 5 min. The limit of quantification (LQ) of anthracene at the bare GCE was 2.2 mol l–1, while the LQ of 0.15 mol l–1 was reached at the DNA/GCE. Moreover, the applicability of the newly developed sensitive voltammetric method was successfully verified on model samples of gravel and sand, with the added/found recoveries of 98 % and 96 %, respectively.
- MeSH
- anthraceny * analýza MeSH
- biosenzitivní techniky MeSH
- DNA MeSH
- elektrochemické techniky * metody přístrojové vybavení využití MeSH
- karcinogeny životního prostředí analýza škodlivé účinky MeSH
- kontaminace potravin analýza prevence a kontrola MeSH
- molekulární sondy - techniky MeSH
- polycyklické aromatické uhlovodíky * analýza škodlivé účinky MeSH
- Publikační typ
- práce podpořená grantem MeSH
Optimum conditions were found for the determination of nanomolar and subnanomolar concentrations of genotoxic 2-aminofluoren-9-one (2-AFN) by adsorptive stripping differential pulse voltammetry (AdSDPV) at a hanging mercury drop minielectrode (HMDmE) after preliminary separation and preconcentration by solidphase extraction (SPE) in LiChrolut RP-18 E (500 mg) columns. The adsorbed analyte was eluted with acetone, the solution then evaporated to dryness, and the residue dissolved in acetate buffer (pH 4.0) (AcB). A sample was then measured by AdSDPV at the HMDmE, with the accumulation potential 100 mV (vs. Ag|AgCl in 1 mol l–1 KCl) and the accumulation time 120 s. Linear calibration curves were obtained for 2·10–11 – 1·10–9 mol l–1 2-AFN, with the limit of quantification (LQ) ≈ 4·10–11 mol l–1. The method was verified by the determination of 2-AFN in model samples of drinking and river water at concentrations 4·10–11 – 1·10–9 and 2·10–10 – 1·10–9 mol l–1 (LQ ≈ 6·10–11 and 4·10–10 mol l–1, respectively).
- MeSH
- chemické znečištění vody * MeSH
- elektrochemické techniky * metody přístrojové vybavení využití MeSH
- fluoreny analýza MeSH
- indikátory a reagencie * MeSH
- pitná voda MeSH
- polarografie * metody přístrojové vybavení využití MeSH
- polycyklické aromatické uhlovodíky * škodlivé účinky toxicita MeSH
- vodní hospodářství MeSH
- Publikační typ
- práce podpořená grantem MeSH
