Stella Girousi

Study of interactions between DNA-ethidium bromide (EB) and dna-acridine orange (AO), in solution, using hanging mercury drop electrode (HMDE)

The interaction of ethidium bromide (EB) and acridine orange (AO) with double stranded (ds), thermally denatured (ss) and supercoiled (sc) DNA, in solution, was studied by alternating current voltammetry (AC voltammetry) at the hanging mercury drop electrode (HMDE) in 0.3 M NaCl+50 mM sodium phosphate buffer (pH 8.5). Their interaction with DNA is shown to be time dependent and completely different. The changes at peak 2 (peak at -1.20 V) of dsDNA form and the appearance of peak 3 (peak at -1.42 V) in scDNA form are presented as criteria declaring the different mechanism of interaction of EB and AO with DNA. Additionally, the appearance of a new peak at around -0.44 V as a result of DNA and AO interaction, differentiates the studied behaviors. The comparison of the electrochemical behaviors of these compounds highlights the differences in the mechanism of interaction.

Study of interactions between actinomycin D and DNA on carbon paste electrode (CPE) and on the hanging mercury drop (HMDE) surface

The interaction of actinomycin (ACTD) with double stranded (ds) calf thymus DNA and single stranded (ss) DNA was studied at the carbon paste electrode surface by means of transfer voltammetry in 0.2 M phosphate buffer solution (pH 7.4). Accordingly the interaction of actinomycin (ACTD) with ds calf thymus DNA, ss DNA and supercoiled (sc) DNA was studied using hanging mercury drop electrode in 0.3 M NaCl, and 50 mM sodium phosphate buffer (pH 8.5). The different electrochemical behaviours are presented and compared in the article.

Sensitive detection of cyclophosphamide using DNA-modified carbon paste, pencil graphite and hanging mercury drop electrodes

The interaction of cyclophosphamide (CP) with calf thymus double-stranded DNA (dsDNA) and thermally denatured single-stranded DNA (ssDNA) immobilized at the carbon paste (CPE) and pencil graphite electrodes (PGE), was studied electrochemically based on oxidation signals of guanine and adenine using differential pulse voltammetry (DPV). As a result of the interaction of CP with DNA, the voltammetric signals of guanine and adenine increased in the case of dsDNA while a slight increase was observed in ssDNA. The effect of experimental parameters such as the interaction time between CP and DNA forms and the concentration of CP, were studied using DPV with CPE and PGE. Additionally, reproducibility and detection limits were determined using both electrodes. A comparison of the analytical performance between CPE and PGE was done. Our results showed that these two different DNA biosensors could be used for the sensitive, rapid and cost effective detection of CP itself as well as of CP-DNA interaction. Furthermore, the interaction of CP with dsDNA and ssDNA was studied in solution and at the electrode surface by means of alternating current voltammetry (ACV) in 0.3M NaCl and 50mM sodium phosphate buffer (pH 8.5) supporting electrolyte, using a hanging mercury drop electrode (HMDE) as working electrode. The conclusions of this study were mainly based on tensammetric peaks I (at -1.183V) and II (-1.419V) of DNA. This study involved the interaction of CP with surface-confined and solution phase DNA where experimental parameters, such as the concentration of CP and the interaction time, were studied. By increasing the concentration of CP, an increase of peak II was observed in both ds and ssDNA, while an increase of peak I was observed only in the case of dsDNA. An overall conclusion of the study using HMDE was that the interaction of CP with surface-confined DNA significantly differed from that with solution phase DNA. The increase of peaks I and II was lower in the case of interaction of CP with surface-confined DNA, probably due to steric positioning of DNA at the electrode surface.

Synthesis, characterization, DNA binding properties and antioxidant activity of a manganese(II) complex with NO6 chromophore

The mononuclear complex [Mn(thiophenyl-2-carboxylate)(2)(H(3)tea)] (1), where H(3)tea=triethanolamine has been synthesized, characterized, while the DNA binding properties and the antioxidant activity were studied. The crystal structure of 1 is also reported. The interaction between 1 and calf thymus double stranded (ds) DNA was electrochemically at a calf thymus DNA (CT-DNA) modified carbon paste electrode (CPE) and spectrophotometrically investigated, respectively. Adsorptive transfer stripping voltammetry showed that the interaction mode between 1 and CT-DNA depends on manganese(II) complex concentration. UV studies between 1 and CT-DNA revealed that 1 can bind to CT-DNA by the intercalative binding mode and the binding constant has been calculated. A competitive study with acridine orange (AO) showed that 1 exhibits the ability to displace the DNA-bound AO, since 1 binds to the DNA in competition with AO. In vitro antioxidant activity of free ligands and 1 was evaluated using five different antioxidant assays: a) interaction with 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) stable free radical, b) the ΗΟ mediated oxidation of DMSO, c) scavenging of superoxide anion radicals, d) inhibition of lipid peroxidation and e) soybean lipoxygenase (LOX) inhibition. The results revealed the selectivity of the manganese complex to different free radicals as a consequence of its physicochemical feature. In particular, 1 presents significant inhibitory activity on LOX, with IC(50)=30 μM and selectivity to the inhibition of superoxide anion radicals and thus a promising candidate as a superoxide dismutase (SOD) biomimetic.

Mitochondria-Based Amperometric Biosensor for the Determination ofL-Glutamic Acid

A mitochondria based amperometric biosensor was developed for the determination of L-glutamic acid. The biosensor was characterized using hexacyanoferrate(III) as the electrochemical mediator. The electrochemical behavior of the redox couple hexacyanoferrate(II)/(III) was improved by incorporating octadecylamine into the enzyme modified carbon paste. L-Glutamic acid was detected at +0.35 V. Linearity was observed in the range 0.4–10 mM of L-glutamic acid (sensitivity 0.189 µA/mM, correlation coefficient 0.997) and in the range 10–100 mM (sensitivity 0.021 µA/mM, correlation coefficient 0.958). The detection limit was 0.1 mM. The electrode maintains 60 % of its activity after 10 days of use. The bioelectrode was used for the determination of L-glutamate in chicken cube samples.

SQUARE-WAVE ANODIC STRIPPING VOLTAMMETRY (SWASV) FOR THE DETERMINATION OF ECOTOXIC METALS, USING A BISMUTH-FILM ELECTRODE

This article reviews the use of square wave anodic stripping voltammetry for the simultaneous determination of ecotoxic metals (Pb, Cd, Cu, and Zn) on a bismuth-film (BiFE) electrode. The BiFE was prepared in situ on a glassy-carbon electrode (GCE) from the 0.1 mol L−1 acetate buffer solution (pH 4.5) containing 200 µg L−1 of bismuth (III). The addition of hydrogen peroxide to the electroanalytical cell proved beneficial for the interference-free determination of Cu (II) together with zinc, lead, and cadmium, using the BiFE. The experimental variables were investigated and optimized with the view to apply this type of voltammetric sensor to real samples containing traces of these metals. The performance characteristics, such as reproducibility, decision limit (CCa), detection capability (CCβ), sensitivity, and accuracy indicated that the method holds promise for trace Cu2+, Pb2+, Cd2+, and Zn2+ levels by employment of Hg-free GCE with SWASV. CCa, and CCβ were calculated according to the Commission Decision of 12 August 2002 (2002/657/EC). Linearity was observed in the range 20–280 µg L−1 for zinc, 10–100 µg L−1 for lead, 10–80 µg L−1 for copper, and 5–50 µg L−1 for cadmium. Using the optimized conditions, the stripping performance of the BiFE was characterized by low limits of detection (LOD). Finally, the method was successfully applied in real as well as in certified reference water samples.

Electrochemical DNA Biosensors Applicable to the Study of Interactions Between DNA and DNA Intercalators

Electrochemical DNA biosensors, based either on carbon paste electrode (CPE) or hanging mercury drop electrode (HMDE) were prepared. These biosensors were used in the study of interaction between double stranded DNA (dsDNA) and single stranded DNA (ssDNA) and acridine orange, a well known DNA intercalator. The different electrochemical behaviors were compared in the article.

Electroanalytical quantification of total dsDNA extracted from human sample using, an ionic liquid modified, carbon nanotubes paste electrode.

Direct electrochemistry of dsDNA has been achieved by using an ionic liquid 1-butyl-4-methylpyridinium hexafluorophosphate modified carbon nanotubes paste electrode (IL-CNTPE). Oxidation peaks appeared at 0.93 and 1.26 V (vs. Ag/AgCl) on the IL- CNTPE after preconcentration of dsDNA in pH 5.0 acetate buffer, which were attributed to the oxidation of guanine and adenine residues on the dsDNA molecule structure. Based on the signal of guanine, under the optimal conditions, very low levels of dsDNA can be detected after 60s accumulation with detection limits of 0.249 mg L(-) 16 pM. Additionally, human DNA from a healthy volunteer is determined by use of the IL-CNTPE and it is found to be 40±2, 14 pM.

Electroanalytical study of the interaction between dsDNA and curcumin in the presence of copper(II).

As a result of the reaction between curcumin (CC) and copper(II) the characteristic peak of curcumin at -1.0V significantly increased, and the peak at -1.6V disappeared. Curcumin forms complex with copper(II). The interaction between double stranded (ds) calf-thymus DNA and curcumin in the presence of Cu(II) was studied in solution, by differential pulse adsorptive transfer voltammetry using carbon paste electrode (CPE) and hanging mercury drop electrode (HMDE). Cu(II)-CC complex generated changes in calf-thymus DNA. The characteristic peak of dsDNA, due to the oxidation of guanine residues, decreased. The increased DNA damage by Cu(II)-CC complex was observed in the presence of various concentrations of the transition metal ions, copper(II).

Square wave anodic stripping voltammetry determination of eco-toxic metals in samples of biological and environmental importance

AbstractSquare wave anodic stripping voltammetry was used in simultaneous determinations of eco-toxic metals (Pb, Cd, Cu and Zn) on bismuth film electrodes. The electrodes were prepared in situ on a glassy-carbon electrode (GCE) from 0.1 M acetate buffer (pH 4.5) containing 200 μg L−1 of bismuth (III), as well ex situ on electrochemically oxidized graphitized polyacrylonitrile carbon fibres from 200 mg L−1 Bi(NO3)3 in 1% HNO3 (aqueous) solution. Preparation of a Bi-modified carbon fibre electrode (CFE) was by cation exchange of Bi+3 ions for H+ of the acidic surface groups of the electro-oxidized carbon fibres, followed by electrochemical reduction to Bi0. For the Bi-GCE the linear range was 20–280 μg L−1 for zinc, 10–100 μg L−1 for lead, 10–80 μg L−1 for copper, and 5–50 μg L−1 for cadmium. For the Bi-CFE it was 20–160 μg L−1 for zinc, 10–100 μg L−1 for lead, 10–100 μg L−1 for copper, and 2–120 μg L−1 for cadmium. For both kinds of bismuth modified carbon electrodes, low limits of detection and satisfactory precision were achieved. The method was successfully applied to certified reference materials of biological (bovine liver) and environmental (mussel tissue) importance.