Determination of β-glucosidase activity in soils with a bioanalytical sensor modified with multiwalled carbon nanotubes

Soil microorganisms and enzymes are the primary mediators of soil biological processes, including organic matter degradation, mineralization, and nutrient recycling. They play an important role in maintaining soil ecosystem quality and functional diversity. Moreover, enzyme activities can provide an indication of quantitative changes in soil organic matter. β-Glucosidase (β-Glu) activity has been found to be sensitive to soil management and has been proposed as a soil quality indicator because it provides an early indication of changes in organic matter status and its turnover. The aims of the present study were to test and use a simple and convenient procedure for the assay of β-Glu activity in agricultural soil. The method described here is based on the enzymatic degradation of cellobiose by β-Glu present in the soil sample and the subsequent determination of glucose produced by the enzymatic reaction using screen-printed carbon electrodes modified with multiwalled carbon nanotubes (SPCE-CNT) equipped with coimmobilized glucose oxidase and horseradish peroxidase enzymes. The potential applied to the SPCE-CNT detection was −0.15 V versus a Ag/AgCl pseudo-reference electrode. A linear calibration curve was obtained in the range 2.7–11.3 mM with a correlation coefficient. In the present study, an easy and effective SPCE-CNT-modified electrode allowed an improved amperometric response to be achieved and this is attributed to the increased surface area upon electrode modification.     

Electrochemical detection of DNA methylation using a glassy carbon electrode modified with a composite made from carbon nanotubes and β-cyclodextrin

We describe a method for detecting DNA methylation. It is based on direct oxidation of DNA bases at a glassy carbon electrode (GCE) modified with film of a multiwalled carbon nanotube-β-cyclodextrin composite. This nano-structured film causes a strong enhancement on the oxidation current of DNA bases due to its large effective surface area and extraordinary electronic properties. Well-defined peaks were obtained as a result of electro-oxidation of guanine (at 0.67 V), adenine (at 0.92 V), thymine (at 1.11 V), cytosine (at 1.26 V), and 5-methylcytosine (at 1.13 V; all data vs. saturated calomel electrode (SCE)). The potential difference between 5-methylcytosine and cytosine (130 mV) is large enough to enable reliable simultaneous determination and analysis. The interference by thymine can be eliminated by following the principle of complementary pairing between purine and pyrimidine bases in DNA. The modified electrode was successfully applied to the evaluation of 5-methylcytosine in a fish sperm DNA, the methylation level of cytosine was found to be 7.47 %, and the analysis process took less than 1 h.     

Sensitive and selective stripping voltammetric determination of copper(II) using a glassy carbon electrode modified with amino-reduced graphene oxide and β-cyclodextrin

We describe a square wave anodic stripping voltammetric (SWASV) platform for the determination of Cu(II). It is based on the use of amino-reduced graphene oxide (NH₂-rGO) and β-cyclodextrin (β-CD) that were self-assembled on the surface of a glassy carbon electrode (GCE). The hydrophilicity and electrochemical performance of the resulting modified GCE were investigated by measurement of static contact angles, cyclic voltammetry and electrochemical impedance spectroscopy. Cu(II) was reduced at −1.1 V and then reoxidized at −0.012 V. Under optimum experimental conditions, the modified GCE exhibited excellent SWASV response in that the stripping peak currents (when sweeping between −0.3 and +0.25 V) depends on the concentration of Cu(II) in the 30 nM to 100 μM range. The limit of detection is 2.8 nM (at 3σ/slope). The modified GCE displaying good reproducibility, is stable, highly sensitive and selective. It was successfully applied to the determination of Cu(II) in synthetic and real water samples. The fast electron transfer rate and simple preparation of the NH₂-rGO/β-CD composite makes it a promising electrode material for applications in sensing of heavy metal ions.

Amino-modified rGO and β-cyclodextrin form an attractive material for use in an electrochemical platform for highly sensitive and selective determination of Cu(II).

     

Electrochemical chiral recognition of tryptophan using a glassy carbon electrode modified with β-cyclodextrin and graphene

We report on a method for electrochemical enantioselective recognition of tryptophan (Trp) enantiomers. It is based on competitive host-guest interaction between a deoxy-(2-aminoethylamino)-β-cyclodextrin (CD) bound to graphene nanosheets and the Cu(II) complexes of the Trp enantiomers via a ligand exchange mechanism. Chiral recognition was investigated via cyclic voltammetry and electrochemical impedance spectroscopy. The results reveal that the CD bound to graphene displays a stronger interaction with the Cu(II) complex of L-Trp than to that of D-Trp. The method was applied to the determination of the ratio of Trp enantiomers in mixtures.

Selective detection of dopamine in the presence of uric acid using a gold nanoparticles-poly(luminol) hybrid film and multi-walled carbon nanotubes with incorporated β-cyclodextrin modified glassy carbon electrode

Gold nanoparticles-poly(luminol) (Plu-AuNPs) hybrid film and multi-walled carbon nanotubes with incorporated β-cyclodextrin modified glassy carbon electrode (β-CD-MWCNTs/Plu-AuNPs/GCE) was successfully prepared for simultaneous determination of dopamine (DA) and uric acid (UA). The surface of the modified electrode has been characterized by X-ray photo-electron spectroscopy (XPS), energy dispersive X-ray spectroscopy (EDS), field-emission scanning electron microscope (SEM) and transmission electron microscope (TEM). Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) have been used to investigate the β-CD-MWCNTs/Plu-AuNPs composite film. Gold nanoparticles anchored into poly(luminol) film exhibited catalytic activity for DA. MWCNTs with incorporated β-CD can greatly promote the direct electron transfer. In 0.10 M phosphate buffer solution (PBS, pH 7.0), the DPV response of the β-CD-MWCNTs/Plu-AuNPs/GCE sensor to DA is about 8-fold as compared with the Plu-AuNPs/GCE sensor, and the detection limit for DA is about one order of magnitude lower than the Plu-AuNPs/GCE sensor. The steady-state current response increases linearly with DA concentration from 1.0 × 10⁻⁶ to 5.6 × 10⁻⁵ M with a low detection limit (S/N = 3) of 1.9 × 10⁻⁷ M. Moreover, the interferences of ascorbic acid (AA) and uric acid (UA) are effectively diminished. The applicability of the prepared electrode has been demonstrated by measuring DA contents in dopamine hydrochloride injection.     

An electrochemical immunosensor for the tumor marker α-fetoprotein using a glassy carbon electrode modified with a poly(5-formylindole), single-wall carbon nanotubes,and coated with gold nanoparticles and antibody

We report on a label-free electrochemical immunosensor for α-fetoprotein (α-FP). It is based on the use of a glassy carbon electrode that was first modified with conducting poly(5-formylindole) and single-walled carbon nanotubes (P5FIn/SWNTs), and then coated with gold nanoparticles and the respective antibody. The presence of aldehyde groups warrants direct immobilization of the antibody and results in a convenient method for fabricating of the immunosensor. Gold nanoparticles (GNPs) were deposited on the P5FIn/SWNTs composite material, and the modified electrode was applied to the detection of α-FP. The analytical signal is obtained by measuring the change of amperometric response at a typical working voltage of 100 mV before and after the immunoreaction. The detection limit is 200 fg mL^?1. The immunosensor is simple, sensitive, specific and reproducible. It has the potential for reliable point-of-care diagnosis of tumor or other diseases. Figure

A simple electrochemical immunosensor based on conducting poly(5-formylindole) and single-walled carbon nanotubes composite was fabricated to detect alpha-fetoprotein. The detection limit is 200 fg mL^?1. This immunosensor is simple, sensitive, specific and reproducible.     

Covalent modification of multiwalled carbon nanotubes with a low molecular weight chitosan

Covalent modification of shortened multiwalled carbon nanotubes (MWNTs) with a natural low molecular weight chitosan (LMCS) was accomplished by the nucleophilic substitution reaction. The LMCS modified MWNTs (MWNT-LMCS) were characterized by FTIR, solid-state 13C NMR, and XPS spectroscopies, thermogravimetric analysis, and transmission electron microscopy. The results revealed that amino and primary hydroxyl groups of the LMCS participated mainly in the formation of the MWNT-LMCS conjugates. The MWNT-LMCS consists of 58 wt.% LMCS, and about four molecular chains of the LMCS were attached to 1000 carbon atoms of the nanotube sidewalls. As a novel derivative of the MWNTs, the MWNT-LMCS not only solved in DMF, DMAc and DMSO, but also in aqueous acetic acid solution.     

Electrochemical oxidation behavior of guanosine-5��-monophosphate on a glassy carbon electrode modified with a composite film of graphene and multi-walled carbon nanotubes, and its amperometric determination

The electrochemical oxidation of guanosine-5??-monophosphate (GMP) was studied with a glassy carbon electrode modified with a composite made from graphene and multi-walled carbon nanotubes. GMP undergoes an irreversible oxidation process at an oxidation peak potential of 987?mV in phosphate buffer solution. Compared to other electrodes, the oxidation peak current of GMP with this electrode was significantly increased, and the corresponding oxidation peak potential negatively shifted, thereby indicating that the modified material exhibited electrochemical catalytic activity towards GMP. Chronocoulometry demonstrates that the material also effectively increases the surface area of the electrode and increases the amount of GMP adsorbed. Under the optimum conditions, the oxidation current is proportional to the GMP concentration in the range from 0.1 to 59.7???M with a correlation coefficient of 0.9991. The detection limit is 0.025???M (at S/N?=?3).

Investigation of complexes of zinc(II) and cadmium(II)DI-n-butyldithiocarbamates with 2,2′-bipyridyl

A new mixed-ligand complex, Cd(S₂CN(C₄H₉)_{2 2})₂(2,2′-Bipy), was synthesized. A polycrystal X-ray diffraction analysis was performed (DRON-3M and DRON-UM1 diffractometers, CuK_α radiation, Ni filter) and the crystal structure was determined [Enraf-Nonius CAD-4 automatic diffractometer, MoK_α radiation, 2440 nonzero independent reflections, 153 refined structural parameters, R is 0.11 for I>2σ(I)]. Crystal data for C₂₈H₄₄CdN₄S₄ : a = 28.716(4), b = 6.848(6), c = 17.188(2) Å, space group Pcca, V-3380.2(7) ų, Z = 4, M = 679.42, dcaU.= 1.335 g/cm3. The structure consists of monomeric molecules in which the cadmium atom has a distorted octahedral environment. The polycrystal diffraction analysis revealed that the complex is isostructural with the defined complex Zn(S₂CN(C₄Hg)₂)₂(2,2′-Bipy). A crystal-chemical search on metal dialkyldithiocarbamates in the Cambridge Structural Database was accomplished and isostructural pairs of Zn and Cd metal complexes were found.     

Sensitive immunoassay for the β-agonist ractopamine based on glassy carbon electrode modified with gold nanoparticles and multi-walled carbon nanotubes in a film of poly-arginine

We are presenting an electrochemical immunosensor for the determination of the β-agonist and food additive ractopamine. A glassy carbon electrode (GCE) was modified with gold nanoparticles and a film of a composite made from poly(arginine) and multi-walled carbon nanotubes. Antibody against ractopamine was immobilized on the surface of the modified GCE which then was blocked with bovine serum albumin. The assembly of the immunosensor was followed by electrochemical impedance spectroscopy. Results demonstrated that the semicircle diameter increases, indicating that the film formed on the surface hinders electron transfer due to formation of the antibody-antigen complex on the modified electrode. Under optimal conditions, the peak current obtained by differential pulse voltammetry decreases linearly with increasing ractopamine concentrations in the 0.1 nmol?L^?1 to 1 μmol?L^?1 concentration range. The lower detection limit is 0.1 nmol?L^?1. The sensor displays good stability and reproducibility. The method was applied to the analysis of spiked swine feed samples and gave satisfactory results. Figure

Immunoassay for ractopamine based on glassy carbon electrode modified with gold nanoparticles and a film of a composite made from poly (arginine) and multi-walled carbon nanotubes was proposed. Under optimal conditions, the peak currents obtained by differential pulse voltammetry decreases linearly with increasing ractopamine concentrations in the 0.1 nmol?L^?1 to 1 μmol?L^?1 concentration range. The detection limit is 0.1 nmol?L^?1.     

Electrochemical methods for simultaneous determination of trace amounts of dopamine and uric acid using a carbon paste electrode incorporated with multi-wall carbon nanotubes and modified with α-cyclodextrine

In this work, we investigate the electrochemical activity of dopamine (DA) and uric acid (UA) using both a bare and a modified carbon paste electrode as the working electrode, with a platinum wire as the counter electrode and a silver/silver chloride (Ag/AgCl) as the reference electrode. The modified carbon paste electrode consists of multi-walled carbon nanotubes (>95%) treated with α-cyclodextrine, resulting in an electrode that exhibits a significant catalytic effect toward the electro-chemical oxidation of DA in a 0.2-M Britton–Robinson buffer solution (pH 5.0). The peak current increases linearly with the DA concentration within the molar concentration ranges of 2.0 × 10⁻⁶ to 5.0 × 10⁻⁵ M and 5.0 × 10⁻⁵ to 1.9 × 10⁻⁴ M. The detection limit (signal to noise >3) for DA was found to be 1.34 × 10⁻⁷ M, respectively. In this work, voltammetric methods such as cyclic voltammetry, chronoamperometry, chronocuolometry, differential pulse and square wave voltammetry, and linear sweep and hydrodynamic voltammetry were used. Cyclic voltammetry was used to investigate the redox properties of the modified electrode at various scan rates. The diffusion coefficient (D, cm² s⁻¹ = 3.05 × 10⁻⁵) and the kinetic parameters such as the electron transfer coefficient (α = 0.51) and the rate constant (k, cm³ mol⁻¹ s⁻¹ = 1.8 × 10³) for DA were determined using electrochemical approaches. By using differential pulse voltammetry for simultaneous measurements, we obtained two peaks for DA and UA in the same solution, with the peak separation approximately 136 mV. The average recovery was measured at 102.45% for DA injection.     

Sensitive immunoassay for the β-agonist ractopamine based on glassy carbon electrode modified with gold nanoparticles and multi-walled carbon nanotubes in a film of poly-arginine

We are presenting an electrochemical immunosensor for the determination of the β-agonist and food additive ractopamine. A glassy carbon electrode (GCE) was modified with gold nanoparticles and a film of a composite made from poly(arginine) and multi-walled carbon nanotubes. Antibody against ractopamine was immobilized on the surface of the modified GCE which then was blocked with bovine serum albumin. The assembly of the immunosensor was followed by electrochemical impedance spectroscopy. Results demonstrated that the semicircle diameter increases, indicating that the film formed on the surface hinders electron transfer due to formation of the antibody-antigen complex on the modified electrode. Under optimal conditions, the peak current obtained by differential pulse voltammetry decreases linearly with increasing ractopamine concentrations in the 0.1 nmol•L⁻¹ to 1 μmol•L⁻¹ concentration range. The lower detection limit is 0.1 nmol•L⁻¹. The sensor displays good stability and reproducibility. The method was applied to the analysis of spiked swine feed samples and gave satisfactory results.

Immunoassay for ractopamine based on glassy carbon electrode modified with gold nanoparticles and a film of a composite made from poly (arginine) and multi-walled carbon nanotubes was proposed. Under optimal conditions, the peak currents obtained by differential pulse voltammetry decreases linearly with increasing ractopamine concentrations in the 0.1 nmol•L⁻¹ to 1 μmol•L⁻¹ concentration range. The detection limit is 0.1 nmol•L⁻¹.

     

An amperometric penicillin biosensor with enhanced sensitivity based on co-immobilization of carbon nanotubes, hematein, and β-lactamase on glassy carbon electrode

An amperometric penicillin biosensor with enhanced sensitivity was successfully developed by co-immobilization of multi-walled carbon nanotubes (MWCNTs), hematein, and β-lactamase on glassy carbon electrode using a layer-by-layer assembly technique. Under catalysis of the immobilized enzyme, penicillin was hydrolyzed, decreasing the local pH. The pH change was monitored amperometrically with hematein as a pH-sensitive redox probe. MWCNTs were used as an electron transfer enhancer as well as an efficient immobilization matrix for the sensitivity enhancement. The effects of immobilization procedure, working potential, enzyme quantity, buffer concentration, and sample matrix were investigated. The biosensor offered a minimum detection limit of 50 nM (19 μg L⁻¹) for penicillin V, lower than those of the conventional pH change-based biosensors by more than two orders of magnitude. The electrode-to-electrode variation of the response sensitivity was 7.0% RSD.     

Fabrication of poly(orthanilic acid)–multiwalled carbon nanotubes composite film-modified glassy carbon electrode and its use for the simultaneous determination of uric acid and dopamine in the presence of ascorbic acid

A multiwalled carbon nanotubes (MWNT) modified glassy carbon electrode (GCE) coated with poly(orthanilic acid) (PABS) film (PABS–MWNT/GCE) has been fabricated and used for simultaneous determination of dopamine (DA) and uric acid (UA) in the presence of ascorbic acid (AA) by differential pulse voltammetry (DPV). Scanning electron microscopy, Fourier transform infrared spectra, and electrochemical techniques have been used to characterize the surface morphology of the PABS–MWNT composite film and the polymerization of ABS on electrode surface. In comparison with the bare GCE and the MWNT-modified GCE, the PABS–MWNT composite film-modified GCE, which combines the advantages of MWNT and the self-doped PABS, exhibits good selectivity and sensitivity for the simultaneous and selective determination of UA and DA in the presence of AA. Due to the different electrochemical responses of AA, DA, and UA, PABS–MWNT/GCE can resolve the overlapped oxidation peak of DA and UA into two well-defined voltammetric peaks with enhanced current responses using both cyclic voltammetry (CV) and DPV. The peak potential separations between DA and UA are 170 mV using CV and 160 mV using DPV, respectively, which are large enough for the selective and simultaneous determination of these species. In the presence of 0.5 mM AA, the DPV peak currents are linearly dependent on the concentration of UA and DA in the range of 6–55 and 9–48 μM with correlation coefficients of 0.997 and 0.993, respectively. The detection limits (S/N = 3) for detecting UA and DA are 0.44 and 0.21 μM, respectively. The PABS–MWNT/GCE shows good reproducibility and stability and has been used for the simultaneous determination of DA and UA in the presence of AA in samples with satisfactory results.     

Fast SPE and HPLC–DAD determination of brucine in human urine using multi-walled carbon nanotubes modified with magnetic nanoparticles

A nanomaterial comprising Fe₃O₄-modified hydroxylated multi-walled carbon nanotubes (Fe₃O₄–MWCNTs–OH) was prepared by a co-precipitation method. Combined with HPLC-photodiode array detector (DAD), Fe₃O₄–MWCNTs–OH was used to determine brucine in human urine. Some important parameters that could influence extraction efficiency of brucine were optimized, including the extraction time, amounts of Fe₃O₄–MWCNTs–OH, pH of sample solutions, desorption solvent and desorption time. Under the optimal conditions, the recoveries of brucine from spiked urine samples were between 93.1 and 104.1%, and the relative standard deviations (RSDs) ranged from 3.1 to 5.7%. The correlation coefficient was 0.9997. The limits of detection and quantification were 6 and 21 ng/mL at a signal-to-noise ratio of 3 and 10, respectively. The results indicated that Fe₃O₄–MWCNTs–OH combined with HPLC–DAD is a promising solid-phase extraction material for the sample pretreatment in the determination of brucine.     

Selective detection of dopamine in the presence of ascorbic acid by use of glassy-carbon electrodes modified with both polyaniline film and multi-walled carbon nanotubes with incorporated β-cyclodextrin

A simple, sensitive, and reliable method based on a combination of multi-walled carbon nanotubes with incorporated β-cyclodextrin (β-CD-MWNTs) and a polyaniline (PANI) film-modified glassy-carbon (GC) electrode has been successfully developed for determination of dopamine (DA) in the presence of ascorbic acid (AA). The PANI film had good anti-interference properties and long-term stability, because of the permselective and protective properties of the conducting redox polymer film. The acid-treated MWNTs with carboxylic acid functional groups promoted the electron-transfer reaction of DA and inhibited the voltammetric response of AA. Sensitive detection of DA was further improved by the preconcentration effect of formation of a supramolecular complex between β-CD and DA. The analytical response of the β-CD-MWNTs/PANI film to the electrochemical behavior of DA was, therefore, better than that of a MWNTs/PANI film, a PANI film, or a bare glassy-carbon (GC) electrode. Under the conditions chosen a linear calibration plot was obtained in the range 1.0 × 10⁻⁷–1.0 × 10⁻³ mol L⁻¹ and the detection limit was 1.2 × 10⁻⁸ mol L⁻¹. Interference from AA was effectively eliminated and the sensitivity, selectivity, stability, and reproducibility of the electrodes was excellent for determination of DA.