Multiwalled Carbon Nanotubes‐Chitosan Modified Single‐Use Biosensors for Electrochemical Monitoring of Drug‐DNA Interactions

A multiwalled carbon nanotubes (CNT)‐chitosan (CHIT) modified pencil graphite electrode (CNT‐CHIT/PGE) was developed for the first time herein for electrochemical monitoring of the interaction of an anticancer drug, mitomycin C (MC) and DNA. The characterization of unmodified PGE, CHIT/PGE, CNT/PGE and CHIT‐CNT/PGE were performed by scanning electron microscopy and cyclic voltammetry techniques. The oxidation signals of MC and guanine were measured before and after interaction at the surface of CNT‐CHIT/PGEs using differential pulse voltammetry. Electrochemical impedance spectroscopy technique was also successfully utilized for monitoring of the interaction process at the surface of CNT‐CHIT/PGEs in different interaction times.     

Electrochemical Biosensing of DNA Immobilized Poly(Vinylferrocenium) Modified Electrode

The polymer, poly(vinylferrocenium) (PVF⁺) modified electrode was developed as the first time herein for the improved electrochemical sensing of DNA based on the oxidation signals of polymer and guanine. The morphologies of polymer film and DNA immobilized polymer film were examined using scanning electron microscope (SEM). The electrochemical behavior of polymer modified electrode was investigated by using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in the absence/presence of DNA. Experimental parameters, such as the polymeric film thickness, the DNA immobilization time, the concentration of buffer solution, pH and DNA concentration were examined in order to obtain more sensitive and selective electrochemical signals. After optimization studies, DNA hybridization was also investigated.     

Carbon Nanotubes Modified Graphite Electrodes for Monitoring of Biointeraction Between 6‐Thioguanine and DNA

In this present study, single‐walled carbon nanotubes (SWCNT) modified disposable pencil graphite electrodes (SWCNT‐PGEs) were developed for the electrochemical monitoring of anticancer drug, and its interaction with double stranded DNA (dsDNA). Under this aim, SWCNT‐PGEs were applied for the first time in the literature to analyse of 6‐Thioguanine (6‐TG), and also to investigate its interaction with DNA by voltammetric and impedimetric methods. The surface morphologies of PGE and SWCNT‐PGE were explored using scanning electron microscopy (SEM) and electrochemical characterization of unmodified/modified electrodes was performed by cyclic voltammetry (CV). Experimental parameters; such as, the concentration of 6‐TG and its interaction time with dsDNA were optimized by using differential pulse voltammetry (DPV). Additionally, the interaction of 6‐TG with dsDNA was studied in case of different interaction times by electrochemical impedance spectroscopy (EIS) in contrast to voltammetric results. The detection limit of 6‐TG was found to be 0.25 μM by SWCNT‐PGE.     

Electrochemical Sensor Based on Oxidation of 2,8‐Dihydroxyadenine to Monitor DNA Damage in Calf Thymus DNA

A novel and reliable direct electrochemical method has been established to monitor DNA damage in acid hydrolyzed calf thymus DNA, based on the determination of 2,8‐dihydroxyadenine (2,8‐DHA). A single‐wall carbon nanotubes (SWCNT) modified edge plane pyrolytic graphite electrode (EPPGE) has been used as a sensor to monitor the DNA damage. 2,8‐DHA the main in vivo adenine oxidation product undergoes oxidation at ～395 mV at SWCNT modified EPPGE using square wave voltammetry (SWV). The sensor exhibits potent and persistent electron‐mediating behavior. A well‐defined oxidation peak for the oxidation of 2,8‐DHA was observed at modified electrode with lowering of peak potential and increase in peak current as compared to bare EPPGE. At optimal experimental conditions, the catalytic oxidative peak current was responsive with the 2,8‐DHA concentrations ranging from 0.05 nM to 100 nM. The detection limit was 3.8×10^?11 M and limit of quantification was 1.27×10^?10 M. The modified electrode exhibited high stability and reproducibility.     

Chitosan/Ionic Liquid Composite Electrode for Electrochemical Monitoring of the Surface‐Confined Interaction Between Mitomycin C and DNA

In the present study a chitosan/ionic liquid modified pencil graphite electrode (CHIT‐IL‐PGEs) was developed for the first time for enhanced electrochemical monitoring of nucleic acid, and the interaction of the anticancer drug Mitomycin C (MC) and calf thymus double stranded DNA (dsDNA) by measuring the oxidation signals of MC and guanine in the same voltammetric scale. Differential pulse voltammetry, cyclic voltammetry and electrochemical impedance spectroscopy techniques were used to evaluate the performance of the CHIT‐IL based biosensor on electrochemical monitoring of DNA, and drug‐DNA interaction. The experimental parameters, IL, dsDNA and MC concentration and the interaction time were then optimized.     

Tin oxide nanoparticles-polymer modified single-use sensors for electrochemical monitoring of label-free DNA hybridization

In this study, SnO₂ nanoparticles (SNPs)-poly(vinylferrocenium) (PVF⁺) modified single-use graphite electrodes were developed for electrochemical monitoring of DNA hybridization. The surfaces of polymer modified and polymer-SNP modified pencil graphite electrodes (PGEs) were firstly characterized by using SEM analysis. The electrochemical behaviours of these electrodes were also investigated using the differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) techniques. The polymer-SNP modified PGEs were then tested for the electrochemical sensing of DNA based on the changes at the guanine oxidation signals. Experimental parameters, such as; different modifications in DNA oligonucleotides, DNA probe concentrations were examined to obtain more sensitive and selective electrochemical signals for nucleic acid hybridization. After optimization studies, DNA hybridization was investigated in the case of complementary of hepatitis B virus (HBV) probe, mismatch (MM), and noncomplementary (NC) sequences.     

Enhanced Electrocatalytic Oxidation of Paracetamol at DNA Modified Gold Electrode

Cysteine monolayer has been assembled onto bare gold electrode (SAM/Au), and subsequently deoxyribonucleic acid (DNA) has been successfully immobilized at the SAM/Au electrode. The thus modified electrode is assigned DNA/SAM/Au. Modification steps of the electrode were followed electrochemically using K₄[Fe(CN₆)] electrochemical marker. Also, the build‐up of the modified electrode composition is followed using EDX and the crystallographic orientation is inspected using XRD. The electrochemical behavior of paracetamol (PC) at DNA/SAM/Au electrode is investigated. Interestingly, the sluggish irreversible behavior of PC at the bare gold electrode is converted to a quasi‐reversible one at DNA/SAM/Au electrode pointing to some interaction between the immobilized DNA and PC. The enhanced electrochemical behavior of PC at modified DNA/SAM/Au electrode is successfully used for a sensitive electrochemical determination of PC. Square wave voltammetry (SWV) was used for this purpose. The concentration of PC was in linear relation with the peak current at the optimum conditions within the range 10.0–110.0 μg mL^?1 with correlation coefficient (R²) of 0.998. Also, the standard deviation (SD) and relative standard deviation (RSD) were calculated and found to be 0.817 and 1.52, respectively, indicating the significance of the present method.     

Electrochemical Biosensor Based on Carbon Ionic Liquid Electrode Modified with Nafion/Hemoglobin/DNA Composite Film

A new electrochemical biosensor was constructed by immobilization of hemoglobin (Hb) on a DNA modified carbon ionic liquid electrode (CILE), which was prepared by using 1‐ethyl‐3‐methylimidazolium tetrafluoroborate (EMIMBF₄) as the modifier. UV‐vis absorption spectroscopic result indicated that Hb remained its native conformation in the composite film. The fabricated Nafion/Hb/DNA/CILE was characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). A pair of well‐defined redox peaks was obtained on the modified electrode, indicated that the Nafion and DNA composite film provided an excellent biocompatible microenvironment for keeping the native structure of Hb and promoting the direct electron transfer rate of Hb with the basal electrode. The electrochemical parameters of Hb in the composite film were further calculated with the results of the charge transfer coefficient (α) and the apparent heterogeneous electron transfer rate constant (k_s) as 0.41 and 0.31 s^?1. The proposed electrochemical biosensor showed good electrocatalytic response to the reduction of trichloroacetic acid (TCA), H₂O₂, NO and the apparent Michaelis–Menten constant (K_M^app) for the electrocatalytic reaction was calculated, respectively.     

Poly(vinylferrocenium) coated disposable pencil graphite electrode for DNA hybridization

Poly(vinylferrocenium) (PVF⁺) modified electrode was developed in this study for the electrochemical sensing of DNA based on the oxidation signals of polymer, adenine and guanine. Experimental parameters, such as; polymeric film thickness, DNA immobilization time and DNA concentration were examined in order to obtain more sensitive and selective electrochemical signals. After optimization studies, DNA hybridization was investigated.     

Sensitively Electrochemical Sensing for Sequence‐Specific Detection of Phosphinothricin Acetyltransferase Gene: Layer‐by‐Layer Films of Poly‐L‐Lysine and Au‐Carbon Nanotube Hybrid

An electrochemical DNA sensing film was constructed based on the multilayers comprising of poly‐L ‐lysine (pLys) and Au‐carbon nanotube (Au‐CNT) hybrid. A precursor film of mercaptopropionic acid (MPA) was firstly self‐assembled on the Au electrode surface. pLys and Au‐CNT hybrid layer‐by‐layer assembly films were fabricated by alternately immersing the MPA‐modified electrode into the pLys solution and Au‐CNT hybrid solution. Cyclic voltammetry was used to monitor the consecutive growth of the multilayer films by utilizing [Fe(CN)₆]^3?/4? and [Co(phen)₃]^3+/2+ as the redox indicators. The outer layer of the multilayer film was the positively charged pLys, on which the DNA probe was easily linked due to the strong electrostatic affinity. The hybridization detection of DNA was accomplished by using methylene blue (MB) as the indicator, which possesses different affinities to dsDNA and ssDNA. Differential pulse voltammetry was employed to record the signal response of MB and determine the amount of the target DNA sequence. The established biosensor has high sensitivity, a relatively wide linear range from 1.0×10^?10 mol/L to 1.0×10^?6 mol/L and the ability to discriminate the fully complementary target DNA from single or double base‐mismatched DNA. The sequence‐specific DNA related to phosphinothricin acetyltransferase gene from the transgenically modified plants was successfully detected.     

Selective Detection of Dopamine in Presence of Ascorbic Acid by Use of Glassy‐Carbon Electrode Modified with Amino‐β‐Cyclodextrin and Carbon Nanotubes

A glassy carbon electrode modified with per‐6‐amino‐β‐cyclodextrin (β‐CDNH₂) and functionalized single‐walled carbon nanotubes (SWCNT‐COOH) was elaborated. This structure was investigated for the detection of dopamine acid (DA) in presence of ascorbic acid (AA). The sensor behavior was studied by cyclic voltammetry, square wave voltammetry and electrochemical impedance spectroscopy. The analysis results show that the electrode modification with CD derivative improves the sensitivity and selectivity of the DA recognition; the electrochemical response was further improved by introduction of SWCNT‐COOH. The sensor shows good and reversible linear response toward DA within the concentration range of 7×10^?7–10^?4 M with a detection limit of 5×10^?7 M.     

基于聚硫堇膜对DNA杂交的直接电化学检测

In this work, the application of a conducting polymer, poly（thionine）, modified electrode as matrix to DNA immobilization as well as transducer to label-free DNA hybridization detection was introduced. The electropolymerization of thionine onto electrode surface was carried out by a simple two-step method, which involved a preanodization of glassy carbon electrode at a constant positive potential in thionine solution following cyclic voltammetry scans in the solution. Electrochemical detection was performed by differential pulse voltammetry in the electroactivity potential domain of poly（thionine）. The resulting poly（thionine） modified electrode showed a good stability and electroactivity in aqueous media during a near neutral pH range. Additionally, the pendant amino groups on the poly（thionine） chains enabled poly（thionine） modified electrode to immobilize phosphate group terminated DNA probe via covalent linkage. Hybridization process induced a clear decrease in poly（thionine） redox current, which was corresponding to the decrease in poly（thionine） electroactivity after double stranded DNA was formed on the polymer film. The detection limit of this electrochemical DNA hybridization sensor was 1.0 × 10^-10mol/L. Compared with complementary sequence, the hybridization signal values of 1-base mismatched and 3-base mismatched samples were 63.9% and 9.2%, respectively.     

Multi‐Walled Carbon Nanotubes (MWCNT)‐Ionic Liquid‐Modified Carbon Paste Electrode: Probing FurazolidoneDNA Interactions and DNA Determination

The interactions of furazolidone (Fu) with double‐stranded calf thymus DNA (dsDNA) on the multi‐walled carbon nanotubes‐ionic liquid‐modified carbon paste electrode (MWCNT‐IL‐CPE) have been studied by cyclic voltammetry. In the presence of DNA, the cathodic peak current of Fu decreased and the peak potential shifted to a positive potential, indicating the intercalative interaction of Fu with DNA. The binding constant of Fu with DNA and stoichiometric coefficient has been determined according to the Hill's model. This electrochemical method was further applied to the determination of DNA. Two linear calibration curves were obtained for DNA detection in the concentration ranges of 0.03–0.10 and 0.10–4.0 μg l^?1 with a detection limit of 0.027 μg l^?1. The method was successfully applied to analyze Fu in serum samples.     

Quantitative Biodetection of Anticancer Drug Rituxan with DNA Biosensor Modified PAMAM Dendrimer/Reduced Graphene Oxide Nanocomposite

PAMAM dendrimer/reduced graphene oxide nanocomposite modified pencil graphite electrode (PAMAM/RGO/PGE) was used to fabricate an electrochemical DNA biosensor for determination of Rituxan (RTX) at low concentrations, for the first time. The fabricated biosensor was characterized with FE‐SEM, EIS, and CV techniques. The ds‐DNA/PAMAM/RGO/PGE was used as a working electrode to study the interaction between the RTX and salmon sperm ds‐DNA by DPV technique. Because of the interaction between the drug and DNA leads to a decrease in the guanine oxidation peak current, it was used as an indicator for the determination of the RTX. Under the optimized experimental conditions, a wide linear relationship between RTX concentration and guanine signal was obtained within the range of 7.0 to 60.0 μmol L⁻¹ and 60.0 to 300.0 μmol L⁻¹ with a low detection limit (0.56 μmol L⁻¹). To clarify the interaction mechanism between the RTX and the ds‐DNA, DPV and UV‐Vis measurements were used. The reproducibility, stability, and performance of the constructed biosensor was examined by quantitative measuring RTX in pharmaceutical and human serum samples with good precision (RSD; 2.0–6.0 %) and acceptable recoveries (100.04–101.95 %).     

Oxidation of Sanguinarine and Its Dihydro‐Derivative at a Pyrolytic Graphite Electrode Using Ex Situ Voltammetry. Study of the Interactions of the Alkaloids with DNA

This study describes the oxidation of sanguinarine (SG) and its metabolite dihydrosanguinarine (DHSG) on the surface of a basal‐plane pyrolytic graphite electrode (PGE). Since both alkaloids strongly adsorb onto the surface of pyrolytic graphite, measurements were performed using ex situ voltammetric methods, adsorptive transfer (AdT) cyclic voltammetry (CV) and square‐wave voltammetry (SWV). Oxidation peaks of SG (peak A) and DHSG (peak A*) were observed around the potential of +0.7 V (vs. Ag/AgCl/3 M KCl), depending on the experimental conditions. The voltammetric peaks A and A* are probably related to the oxidation of N‐methylphenanthridine nitrogenous heterocycle of SG and oxidation of DHSG back to SG, respectively. The electrochemical results and optimized AdT SWV were subsequently applied to the study of the interactions of SG and DHSG with DNA in vitro. Analysis of the alkaloid/DNA interactions was based on observing heights of oxidation peaks A and A* after incubation of SG and/or DHSG with supercoiled (sc) DNA [pBSK_(?)]. Electrochemical study of the interactions was supported and complemented with measurements using gel electrophoresis (Topoisomerase I scDNA relaxation assay) and steady‐state and time‐resolved fluorescence spectroscopy. The results suggest that SG intercalates into the double‐stranded structure of scDNA (the SG/base pair ratio is max. 1/4) while increased binding affinity was observed for quaternary cation (SG⁺). DHSG which, unlike SG⁺, does not possess a strictly planar molecular structure, did not show intercalative DNA binding in any of the three methods applied.     

Electrochemical Behavior of 8‐Azaguanine at DNA Langmuir–Blodgett Modified Glassy Carbon Electrode and Its Analytical Application

A highly sensitive electrochemical biosensor for the detection of trace amounts of 8‐azaguanine has been designed. Double stranded (ds)DNA molecules are immobilized onto a glassy carbon electrode surface with Langmuir–Blodgett technique. The adsorptive voltammetric behaviors of 8‐azaguanine at DNA‐modified electrode were explored by means of cyclic voltammetry and square wave voltammetry. Compared with bare glassy carbon electrode (GCE), the Langmuir–Blodgett film modified electrode can greatly improve the measuring sensitivity of 8‐azaguanine. Under the optimum experimental conditions, the Langmuir–Blodgett film modified electrode in pH 3.0 Britton–Robinson buffer solutions shows a linear voltammetric response in the range of 5.0×10^?8 to 1.0×10^?5 mol L^?1 with detection limit 9.0×10^?9 mol L^?1. The method proposed was applied successfully for the determination of 8‐azaguanine in diluted human urine with wonderful satisfactory.     

Electrocatalytic Oxidation of Diethylaminoethanethiol and Hydrazine at Single‐walled Carbon Nanotubes Modified with Prussian Blue Nanoparticles

In this work, edged plane pyrolytic graphite electrode EPPGE was modified with functionalised single‐walled carbon nanotubes and Prussian blue nanoparticles (PB). The modified electrode was characterised by techniques such as TEM, FTIR, XPS, EDX and cyclic voltammetry. The EPPGE‐SWCNT‐PB platform exhibited enhanced electron transport and catalytic efficiency towards the oxidation of Diethylaminoethanethiol (DEAET) and hydrazine compared with the other electrodes studied. The EPPGE‐SWCNT‐PB showed good electrochemical stability in the analytical solution, showing limit of detection in the micromolar range and catalytic rate constant of 3.71×10⁶ and 7.56×10⁶ cm³ mol^?1 s^?1 for DEAET and hydrazine respectively. The adsorption properties of these analytes that impact on their detection at the SWCNT‐PB film modified electrode were evaluated and discussed.     

Electrochemical sensor for ultrasensitive determination of ceftazidime using hollow platinum nanoparticles/reduced graphene oxide/pencil graphite electrode

In this paper, an electrochemical sensor was prepared based on the modification of pencil graphite electrode (PGE) by hollow platinum nanoparticles/reduced graphene oxide (HPtNPs/rGO/PGE) for determination of ceftazidime (CFZ). Initially, rGO was electrodeposited on the electrode surface, and then, hollow platinum nanoparticles were placed on the electrode surface via galvanic displacement reaction of Pt(IV) ions with cobalt nanoparticles (CoNPs) that had electrodeposited on the electrode surface. Several significant parameters controlling the performance of the HPtNPs/rGO/PGE were examined and optimized using central composite design as one optimization methodology. The surface morphology and elemental characterization of the bare PGE, rGO/PGE, CoNPs/rGO/PGE, and HPtNPs/rGO/PGE-modified electrodes was analyzed by field-emission scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, and electrochemical impedance spectroscopy. The electrochemical activity of CFZ on resulting modified electrode was investigated by cyclic voltammetry (CV) and adsorptive differential pulse voltammetry (AdDPV). Adsorptive differential pulse voltammetry indicates that peak current increases linearly with respect to increment in CFZ concentration. CFZ was determined in the linear dynamic range of 5.0 × 10^?13 to 1.0 × 10^?9 M, and the detection limit was determined as 2.2 × 10^?13 M using AdDPV under optimized conditions. The results showed that modified electrode has high selectivity and very high sensitivity. The method was used to determine of CFZ in drug injection and plasma samples.     

Lable‐Free Electrochemical DNA Sensor Based on Gold Nanoparticles/Poly(neutral red) Modified Electrode

We present a new strategy for the label‐free electrochemical detection of DNA hybridization based on gold nanoparticles (AuNPs)/poly(neutral red) (PNR) modified electrode. Probe oligonucledotides with thiol groups at the 5‐end were covalently linked onto the surface of AuNPs/PNR modified electrode via S‐Au binding. The hybridization event was monitored by using differential pulse voltammetry (DPV) upon hybridization generates electrochemical changes at the PNR‐solution interface. A significant decrease in the peak current was observed upon hybridization of probe with complementary target ssDNA, whereas no obvious change was observed with noncomplementary target ssDNA. And the DNA sensor also showed a high selectivity for detecting one‐mismatched and three‐mismatched target ssDNA and a high sensitivity for detecting complementary target ssDNA, the detection limit is 4.2×10^?12 M for complementary target ssDNA. In addition, the DNA biosensor showed an excellent reproducibility and stability under the DNA‐hybridization conditions.     

Electrochemical and Spectroscopic Studies on the Interaction of Gatifloxacin,Moxifloxacin and Sparfloxacin with DNA and Their Analytical Applications

The electrochemical oxidation of the three fluoroquinolone drugs FQs: gatifloxacin GTF, moxifloxacin MXF and sparfloxacin SPF, at the bare and DNA‐modified glassy carbon electrodes has been studied by voltammetric techniques. The three FQs showed one irreversible oxidation peak at potential range 0.85–0.91 V vs. Ag‐AgCl, in phosphate buffer of pH 7.0. Differential pulse voltammetry (DPV) and UV‐absorption spectroscopic techniques were employed to probe the interaction between the FQs and calf thymus double stranded deoxyribonucleic acid (ds CT‐DNA). From electrochemical data, the binding constant between DNA and the gatifloxacin, moxifloxacin and sparfloxacin are calculated to be 3228, 2596 and 2857 M^?1 respectively. Based on electrochemical and spectroscopic results, the mode of binding of fluoroquinolone to DNA through combined effect of intercalation and electrostatic interaction was concluded. A detection scheme based on a preconcentration and differential pulse voltammetric (DPV) determination at dsDNA modified glassy carbon electrode (DNA/GCE) was proposed for the trace determination of the studied analytes. The developed method was successfully applied to the determination of the FQs in pharmaceutical formulations.