An improved DNA biosensor built by layer-by-layer covalent attachment of multi-walled carbon nanotubes and gold nanoparticles

In this work, we synthesized a type of compound, MWCNTs-CONH-(CH₂)₂-SH, via carboxylation, and investigated a thickness-tunable multilayer films DNA biosensor built by layer-by-layer (LBL) covalent attachment of gold nanoparticles (GNPs) and multi-walled carbon nanotubes (MWCNTs) on an Au electrode. Fourier transform infrared (FT-IR) spectra were used to identify the products formed in the step of carboxylation; scanning electron microscopy (SEM) and cyclic voltammetry (CV) were used to study the film assembly process. The hybridization events were monitored through measurement the signal of intercalated doxorubicin by differential pulse voltammetry (DPV), and the oxidation peak currents show a good linear relationship with the logarithm of the concentration of target DNA from 5.0 × 10⁻¹⁰ to 1.0 × 10⁻¹¹ M with a detection limit of 6.2 pM. The improved DNA biosensor has a good stability and reproducibility.     

Voltammetric determination of bisoprolol fumarate in pharmaceutical formulations and urine using single-wall carbon nanotubes modified glassy carbon electrode

The electrochemistry of bisoprolol fumarate (BF) has been investigated by differential pulse voltammetry at a single-wall carbon nanotubes (SWNTs) modified glassy carbon electrode (GCE). The prepared electrode showed an excellent electrocatalytic activity towards the oxidation of BF leading to a marked improvement in sensitivity as compared to bare glassy carbon electrode where electrochemical activity for the analyte cannot be observed. The SWNTs-modified GCE exhibited a sharp anodic peak at a potential of ∼950 mV for the oxidation of BF. Under optimum conditions linear calibration curve was obtained over the BF concentration range 0.01-0.1 mM in 0.5 M phosphate buffer solution (pH 7.2) with a correlation coefficient of 0.9789 and detection limit of 8.27 × 10⁻⁷ M. The modified electrode has been applied for the drug determination in human urine with no prior extraction and in commercial tablets. The proposed method has also been validated.     

Direct voltammetric determination of gold nanoparticles using graphite-epoxy composite electrode

A new voltammetric method for a direct determination of gold nanoparticles, based on adsorption and electrochemical detection of colloidal gold, is described. In this protocol, the absorption of gold nanoparticles onto the rough surface of graphite-epoxy composite electrode is followed by their electrochemical oxidation in 0.1 M HCl medium at a potential of +1.25 V. The resulting tetrachloroaurate ions generated near the electrode surface are detected by differential pulse voltammetry (DPV). The DPV response is linear in the range from 4.7 × 10⁸ to 4.7 × 10¹¹ nanoparticles cm⁻³ with a limit of detection of 1.8 × 10⁸ gold nanoparticles cm⁻³. The surface characteristics of the composite electrode are investigated and the parameters that affect the complete analytical detection process of gold nanoparticles are optimized.     

Electrocatalytic oxidation and simultaneous determination of uric acid and ascorbic acid on the gold nanoparticles-modified glassy carbon electrode

A new gold nanoparticles-modified electrode (GNP/LC/GCE) was fabricated by self-assembling gold nanoparticles to the surface of the l-cysteine-modified glassy carbon electrode. The modified electrode showed an excellent character for electrocatalytic oxidization of uric acid (UA) and ascorbic acid (AA) with a 0.306 V separation of both peaks, while the bare GC electrode only gave an overlapped and broad oxidation peak. The anodic currents of UA and AA on the modified electrode were 6- and 2.5-fold to that of the bare GCE, respectively. Using differential pulse voltammetry (DPV), a highly selective and simultaneous determination of UA and AA has been explored at the modified electrode. DPV peak currents of UA and AA increased linearly with their concentration at the range of 6.0 × 10⁻⁷ to 8.5 × 10⁻⁴ mol L⁻¹ and 8.0 × 10⁻⁶ to 5.5 × 10⁻³ mol L⁻¹, respectively. The proposed method was applied for the detection of UA and AA in human urine with satisfactory result.     

Alcohol dehydrogenase amperometric biosensor based on a colloidal gold-carbon nanotubes composite electrode

A novel ethanol biosensor based on the bulk incorporation of alcohol dehydrogenase (ADH) into a colloidal gold (Au_coll)-multiwalled carbon nanotubes (MWCNTs) composite electrode using Teflon as binding material is reported. The composite Au_coll-MWCNTs-Teflon electrode exhibited significantly improved electrooxidation of NADH when compared with other carbon composite electrodes, including those based on carbon nanotubes. Amperometric measurements for NADH at +0.3 V showed significant differences in sensitivity between Au_coll-MWCNTs-Teflon and MWCNTs-Teflon composite electrodes. Incorporation of ADH into the bulk electrode material allowed the construction of a mediatorless ethanol biosensor. Both the enzyme loading and the NAD⁺ concentration in solution were optimized. The ADH-Au_coll-MWCNTs-Teflon biosensor allowed a limit of detection for ethanol of 4.7 μmol l⁻¹, which is remarkably better than those reported for other CNTs-based ADH biosensors. The apparent Michaelis-Menten constant was 4.95 mmol l⁻¹, which is much lower than that reported by immobilization of ADH onto a gold electrode. Both repeatability of the ethanol amperometric measurements, reproducibility with different biosensors, lifetime and storage ability can be, in general, advantageously compared with other ADH-CNTs biosensors. The biosensor was applied for the rapid determination of ethanol in commercial and certified beer samples.     

Layer-by-layer construction of multi-walled carbon nanotubes, zinc oxide, and gold nanoparticles integrated composite electrode for nitrite detection

A novel composite electrode of Au/ZnO/MWCNTs/GC has been constructed for the electrochemical detection of nitrite, where ZnO thin film and Au nanoparticles are electrodeposited through layer-by-layer onto MWCNTs/GC substrate. The resulting electrode is characterized by scanning electron microscopy and energy-dispersed X-ray spectroscopy. Its electrocatalytic activity toward the electro-oxidation of nitrite has been examined and compared to various modified electrodes, including MWCNTs/GC, Au/ZnO/GC, Au/MWCNTs/GC, and ZnO/MWCNTs/GC via cyclic voltammetry. The electrodeposition time for ZnO and the Au loading amount together with the solution pH are investigated to achieve optimal conditions for the electrode fabrication and nitrite detection. Linear relationship between current response and nitrite concentration is observed in the range from 7.8 × 10⁻⁷ to 4.0 × 10⁻⁴ M and the limit of the detection is 4.0 × 10⁻⁷ M (S/N = 3). The influence of various anions and cations on the nitrite detection has been studied. The proposed method is also employed for the determination of nitrite in real samples.     

A novel electrochemical DNA biosensor based on graphene and polyaniline nanowires

A novel DNA biosensor based on oxidized graphene and polyaniline nanowires (PANIws) modified glassy carbon electrode was developed. The resulting graphene/PANIw layers exhibited good DPV current response for the complementary DNA sequences. The good electron transfer activity might be attributed to the effect of graphene and PANIw. Graphene and PANIw nanolayers film with highly conductive and biocompatible nanostructure were characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The immobilization of the probe DNA on the surface of electrode was largely improved due to the unique synergetic effect of graphene and PANIw. Under optimum conditions, the biosensor exhibited a fast amperometric response, high sensitivity and good storage stability for monitoring DNA. The current response of the sensor increases linearly with the concentration of target from 2.12 × 10⁻⁶ to 2.12 × 10⁻¹² mol l⁻¹ with a relative coefficient of 0.9938. The detection limit (3σ) is 3.25 × 10⁻¹³ mol l⁻¹. The results indicate that this modified electrode has potential application in sensitive and selective DNA detection.     

A sensitive biosensor for cyanide detection using modified glassy carbon electrode with reduced graphene oxide and immobilization of horseradish peroxidase

Cyanide is a highly poisonous and hazardous substance which may release into the environment from natural sources or industrial effluent; therefore, cyanide detection is a fundamental step to prevent environmental pollution and secure health and safety. In this study, we prepared a sensitive amperometric inhibition biosensor for cyanide detection by immobilization of horseradish peroxidase (HRP) enzyme and reduced graphene oxide (rGO) on the surface of glassy carbon electrode (GCE). To do so, we performed the amperometric measurement by modified GCE to test its efficiency in detecting cyanide. The optimum conditions of pH equal to 7.5, −100 mV applied potential, 0.7 μM mediator concentration, and 0.5 mM substrate concentration were found. Then, experiments were performed at different boundary conditions in a range of 0.1 to 10 μM cyanide concentration at optimal conditions and a low detection limit of 0.01 μM was obtained. Also, the possible mechanism of inhibition was analyzed based on the Michalis–Menten equation and non-competitive inhibition was observed. Due to high sensitivity, low detection limit, and low cost, this biosensor is proposed as a useful method for cyanide determination in real samples.     

Electrostatic assembly of calf thymus DNA on multi-walled carbon nanotube modified gold electrode and its interaction with chlorpromazine hydrochloride

The electrostatic assembly of calf thymus DNA on multi-walled carbon nanotubes (MWNTs)-modified gold electrode via poly(diallyldimethylammonium chloride) (PDDA), a cationic polyelectrolyte, has been studied. Piezoelectric quartz crystal impedance (PQCI) technique was employed to monitor the assembly of DNA in real time. Electrochemical impedance spectroscopy was also used to characterize the modification process. It has been found that modification of the electrode with MWNTs has significantly enhanced the effective electrode surface area in addition to providing negatively charged groups for the electrostatic assembly of cationic polyelectrolyte. PDDA plays a key role in the attachment of DNA to MWNTs and acts as a bridge to connect DNA with MWNTs, though the direct adsorption of DNA on MWNTs has been observed. Moreover, the interaction between DNA and chlorpromazine hydrochloride was observed with PQCI technique. This shows that DNA assembled on MWNTs still has the ability to interact with drug molecules, which has great importance in the construction of new types of miniature DNA biosensors.     

Electrochemical fabrication of self assembled monolayer using ferrocene-functionalized gold nanoparticles on glassy carbon electrode

Electrochemical (EC) quantification of 11-Mercaptoundecylferrocene (thiolated-ferrocene substrate, Fc-SH) onto gold nanoparticles (AuNPs) was carried out through cyclic voltammetric (CV) investigations in different base electrolyte aqueous media on glassy carbon (GC) electrode. The electrochemical data including peak current, peak potential values revealed the Fc-SH–AuNPs redox system to be adsorption controlled in terms of self assembled monolayers (SAMs). The electrochemically generated SAMs were found quite stable as these maintained the redox activity upto 3 months after repeated CV scans at 298 K. The overall output of this research can be utilized in two ways: application of electroactivity and stability of these SAMs in detection of biologically important molecules and also towards the development of EC biosensors.     

Second-order data obtained from differential pulse voltammetry: Determination of tryptophan at a gold nanoparticles decorated multiwalled carbon nanotube modified glassy carbon electrode

Three-way data obtained from different pulse heights of differential pulse voltammetry (DPV) was analyzed using multivariate curve resolution by alternating least squares (MCR-ALS) algorithm. Differential pulse voltammograms of tryptophan were recorded at a gold nanoparticles decorated multiwalled carbon nanotube modified glassy carbon electrode (GCE/MWCNTs-nanoAu). The determination of tryptophan was performed even in the presence of unexpected electroactive interference(s). Both the simulated and experimental data were non-bilinear. Therefore a potential shift algorithm was used to correct the observed shift in the data. After correction, the data was augmented and MCR-ALS was applied to the augmented data. A relative error of prediction of less than 8% for the determination of the simulated analyte of interest and tryptophan in synthetic samples indicated that the methodology employing voltammetry and second-order calibration could be applied to complex analytical systems.     

A simple method for the synthesis of PtRu nanoparticles on the multi-walled carbon nanotube for the anode of a DMFC

We report the synthesis of PtRu nanoparticles on the multi-walled carbon nanotubes (MWCNTs) by a simple sodium borohydride reduction method. Transmission electron microscopy (TEM) analysis indicated that well-dispersed small (2-3 nm) PtRu particles were formed on the MWCNTs. X-ray diffraction (XRD) analysis confirmed the formation of the PtRu alloy on the MWCNTs. X-ray photoelectron spectroscopy (XPS) measurements revealed that 70.4% Pt and 61.0% Ru are present in their metallic states. Cyclic voltammetry (CV) and chronoamperometry results demonstrated that the PtRu/MWCNT synthesized by this method exhibited a higher methanol oxidation current than did the PtRu/MWCNT synthesized by the more complex method using sodium borohydride as the reducing agent and tetraoctyl ammonium bromide as the stabilizer. Finally, the direct methanol fuel cell (DMFC) performance test showed that the PtRu/MWCNT nanocatalyst used at the anode of the fuel cell yielded higher performance than did the commercial E-TEK PtRu/C catalyst.     

Electro-oxidation nitrite based on copper calcined layered double hydroxide and gold nanoparticles modified glassy carbon electrode

In this paper, a novel nitrite sensor was constructed based on electrodeposition of gold nanoparticles (AuNPs) on a copper calcined layered double hydroxide (Cu-CLDH) modified glassy carbon electrode. Electrochemical experiments showed that AuNPs/CLDH composite film exhibited excellent electrocatalytic oxidation activity with nitrite due to the synergistic effect of the Cu-CLDH with AuNPs. The fabricated sensor exhibited excellent performance for nitrite detection within a wide concentration interval of 1–191 μM and with a detection limit of 0.5 μM. The superior electrocatalytic response to nitrite was mainly attributed to the large surface area, minimized diffusion resistance, and enhanced electron transfer of the Cu-CLDH and AuNPs composition film. This platform offers a novel route for nitrite sensing with wide analytical applications and will supply the practical applications for a variety of simple, robust, and easy-to-manufacture analytical approaches in the future.     

Highly sensitive and simultaneous determination of hydroquinone and catechol at poly(thionine) modified glassy carbon electrode

A simple and highly sensitive electrochemical method for the simultaneous and quantitative detection of hydroquinone (HQ) and catechol (CT) was developed, based on a poly(thionine)-modified glassy carbon electrode (GCE). The modified electrode showed excellent electrocatalytic activity and reversibility towards the oxidation of both HQ and CT in 0.1 M phosphate buffer solution (PBS, pH 7.0). The peak-to-peak separations (ΔE_p) between oxidation and reduction waves in CV were decreased significantly from 262 and 204 mV at the bare GCE, to 63 and 56 mV, respectively for HQ and CT at the poly(thionine) modified GCE. Furthermore, the redox responses from the mixture of HQ and CT were easily resolved in both CV and DPV due to a difference in the catalytic activity of the modified GCE to each component. The peak potential separation of ca. 0.1 V was large enough for the simultaneous determination of HQ and CT electrochemically. The oxidation peak currents of HQ and CT were linear over the range from 1 to 120 μM in the presence of 100 and 200 μM of HQ and CT, respectively. The modified electrode showed very high sensitivity of 1.8 and 1.2 μA μM⁻¹ cm⁻² for HQ and CT, respectively. The detection limits (S/N = 3) for HQ and CT were 30 and 25 nM, respectively. The developed sensor was successfully examined for real sample analysis with tap water and revealed stable and reliable recovery data.     

Poly(2-amino-4-thiazoleacetic acid)/multiwalled carbon nanotubes modified glassy carbon electrodes for the electrochemical detection of copper(II)

Poly(2-amino-4-thiazoleacetic acid)/multiwalled carbon nanotubes modified glassy carbon electrodes obtained by electropolymerization of 2-amino-4-thiazoleacetic acid, were used for the voltammetric determination of copper ions. The voltammetric response of copper ions at poly(2-amino-4-thiazoleacetic acid)/multiwalled carbon nanotubes modified glassy carbon electrodes was evaluated by differential pulse stripping voltammetry. The peak currents were linearly dependent on the concentrations of the copper ions in the range from 7.0 × 10⁻⁷ M to 5.0 × 10⁻⁵ M, with a coefficiency of 0.9987. The detection limit is 5.0 × 10⁻¹⁰ M calculated for a signal-to-noise ratio of 3 (S/N = 3). And it could be used for the simultaneous determination of copper and cadmium ions. The proposed method was successfully applied to the determination of copper ions in natural water. The concentration of Cu²⁺ was calculated to be 2.0 × 10⁻⁵ M by standard addition method. The recovery rate was 94%.     

Amperometric biosensor for nitrite and hydrogen peroxide based on hemoglobin immobilized on gold nanoparticles/polythionine/platinum nanoparticles modified glassy carbon electrode

BACKGROUND: This paper describes a convenient and effective strategy to construct a highly sensitive amperometric biosensor for nitrite (NO₂^?) and hydrogen peroxide (H₂O₂). First, Pt nanoparticles (PtNPs) were electrodeposited on a glassy carbon electrode (GCE) surface, which promoted electron transfer and enhanced the loading of poly‐thionine (PTH). Subsequently, thionine (TH) was electropolymerized on the PtNPs/GCE, and gold nanoparticles (AuNPs) were assembled onto the PTH film to improve the absorption capacity of hemoglobin (Hb) and further facilitate electron transfer. Finally, Hb was immobilized onto the electrode through the AuNPs. RESULTS: Cyclic voltammetry (CV) and scanning electron microscopy (SEM) were used to characterize the fabrication process of the sensing surface. Under optimum conditions, the biosensors can be used for the determination of NO₂^? in the concentration range 70 nmol L^?1 to 1.2 mmo L^?1 and of H₂O₂ in the range 4.9 µmol L^?1 to 6.8 mmol L^?1. The detection limits (S/N = 3) were 20 nmol L^?1 and 1.4 µmol L^?1, respectively. CONCLUSION: The biosensor exhibits good analytical performance, acceptable stability and good selectivity. Copyright © 2011 Society of Chemical Industry     

Comparison of the electrocatalytic performance of PtRu nanoparticles supported on multi-walled carbon nanotubes with different lengths and diameters

To investigate the electrocatalytic performance of PtRu nanoparticles supported on multi-walled carbon nanotubes (MWCNTs) with different lengths and diameters, X-ray diffraction, transmission electron microscopy, X-ray photoelectron spectroscopy, and cyclic voltammetry experiments were conducted. It is demonstrated that the length and diameter of MWCNTs play an important role in the electrocatalytic performance of PtRu catalysts. The co-existence of amorphous carbon impurities on the MWCNT10-2 support lowered the accessible surface area of the PtRu nanoparticles, hampered the dispersion of the PtRu nanoparticles, and induced the formation of a low degree of PtRu alloy, thus lowered the electrocatalytic performance of the PtRu/MWCNT10-2 catalyst for methanol oxidation. The highest mass-specific activity of PtRu/MWCNT3050-2 results from a highly accessible PtRu surface and a good dispersion of PtRu particles. Our experimental results also demonstrate that the tube length of MWCNT samples has little effect of the surface area specific activity of the PtRu/MWCNT catalyst, whereas the PtRu nanoparticles supported on the MWCNT samples with large tube diameter tends to exhibit a higher surface area specific activity for methanol oxidation reaction. This result is suggested to be the combined effects of a high degree of PtRu alloying and the high electronic conductivity of these MWCNT samples.     

Direct electrochemical behavior of cytochrome c on DNA modified glassy carbon electrode and its application to nitric oxide biosensor

Cytochrome c/DNA modified electrode was achieved by coating calf thymus DNA onto the surface of glassy carbon electrode firstly, then immobilizing cytochrome c on it by multi-cyclic voltammetric method and characterized by the electrochemical impedance. The electrochemical behavior of cytochrome c on DNA modified electrode was explored and showed a quasi-reversible electrochemical redox behavior with a formal potential of 0.045 ± 0.010 V (versus Ag/AgCl) in 0.10 M, pH 5.0, acetate buffer solution. The peak currents were linearly with the scan rate in the range of 20-200 mV/s. Cytochrome c/DNA modified electrode exhibited elegant catalytic activity for the electrochemical reduction of NO. The catalytic current is linear to the nitric oxide concentration in the range of 6.0 × 10⁻⁷ to 8.0 × 10⁻⁶ M and the detection limit was 1.0 × 10⁻⁷ M (three times the ratio of signal to noise, S/N = 3).     

Gold nanoparticles/water-soluble carbon nanotubes/aromatic diamine polymer composite films for highly sensitive detection of cellobiose dehydrogenase gene

An electrochemical sensor based on gold nanoparticles (GNPs)/multiwalled carbon nanotubes (MWCNTs)/poly (1,5-naphthalenediamine) films modified glassy carbon electrode (GCE) was fabricated. The effectiveness of the sensor was confirmed by sensitive detection of cellobiose dehydrogenase (CDH) gene which was extracted from Phanerochaete chrysosporium using polymerase chain reaction (PCR). The monomer of 1,5-naphthalenediamine was electropolymerized on the GCE surface with abundant free amino groups which enhanced the stability of MWCNTs modified electrode. Congo red (CR)-functionalized MWCNTs possess excellent conductivity as well as high solubility in water which enabled to form the uniform and stable network nanostructures easily and created a large number of binding sites for electrodeposition of GNPs. The continuous GNPs together with MWCNTs greatly increased the surface area, conductivity and electrocatalytic activity. This electrode structure significantly improved the sensitivity of sensor and enhanced the DNA immobilization and hybridization. The thiol modified capture probes were immobilized onto the composite films-modified GCE by a direct formation of thiol–Au bond and horseradish peroxidase–streptavidin (HRP–SA) conjugates were labeled to the biotinylated detection probes through biotin–streptavidin bond. Scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to investigate the film assembly and DNA hybridization processes. The amperometric current response to HRP-catalyzed reaction was linearly related to the common logarithm of the target nucleic acid concentration in the range of 1.0 × 10⁻¹⁵–1.0 × 10⁻¹⁰ M, with the detection limit of 1.2 × 10⁻¹⁶ M. In addition, the electrochemical biosensor exhibited high sensitivity, selectivity, stability and reproducibility.