Rapid detection of Escherichia coli by flow injection analysis coupled with amperometric method using an IrO2–Pd chemically modified electrode

An amperometric method for the rapid detection of Escherichia coli (E. coli) by flow injection analysis (FIA) using an IrO₂–Pd chemically modified electrode (CME) was developed in this paper. The method is based on a good marker β-d-galactosidase which was found in E. coli strains. β-d-galactosidase was produced by the induction of isopropyl β-d-thiogalactopyranoside (IPTG) and released from E. coli cells through the permeabilization of both polymyxin B nonapeptide and lysozyme to E. coli cells wall. The released β-d-galactosidase could catalyze the hydrolysis of the substrate p-aminophenyl β-d-galactopyranoside (PAPG) in the culture medium to produce 4-aminophenol which was proportional to the concentration of E. coli. Hence, E. coli could be detected by the determination of 4-aminophenol. An IrO₂–Pd CME, which showed high sensitivity in determination of 4-aminophenol, was prepared as the electro-detector in FIA. The amplified response current of 4-aminophenol obtained at the IrO₂–Pd CME was linear with the concentration of E. coli ranging from 2.0 × 10² to 1.0 × 10⁶ cfu/mL, the detection limit of this method to E. coli was 150 cfu/mL and the complete assay could be performed in 3 h.     

Amperometric method for rapid detection of Escherichia coli by flow injection analysis using a bismuth nano-film modified glassy carbon electrode

A new method for rapid detection of Escherichia coli (E. coli) was developed by flow injection analysis (FIA) using bismuth nano-film modified glassy carbon electrode (BiNFE) in this paper. The method depended on a good marker β-d-glucuronidase which is found in E. coli strains. β-d-Glucuronidase was produced by the induction of methyl-β-d-glucuronide sodium (MetGlu), then released from E. coli cells through the permeabilization of cell membrane caused by polymyxin B nonapeptide and lysozyme. The released β-d-glucuronidase could catalyze the hydrolysis of the substrate 4-nitrophenyl β-d-glucuronide (PNPG) in the culture medium to produce 4-nitrophenol. Since 4-nitrophenol is electroactive and its quantity is proportional to the concentration of E. coli, E. coli could be determined by electroanalysis of 4-nitrophenol. The BiNFE was fabricated by an electrodeposition of metallic bismuth onto a glassy carbon electrode, which showed a high sensitivity in determination of 4-nitrophenol when used in conjunction with FIA system. Experimental results showed that the amplified response current of 4-nitrophenol obtained at the BiNFE was linear with the concentration of E. coli ranging from 1.5 × 10² to 1.0 × 10⁶ cfu/ml, the detection limit of this method for E. coli is 100 cfu/ml, and the complete assay was performed in 3 h.     

A disposable amperometric enzyme immunosensor for rapid detection of Vibrio parahaemolyticus in food based on agarose/Nano-Au membrane and screen-printed electrode

Recent outbreaks of foodborne illnesses continue to support the need for rapid and sensitive methods for detection of foodborne pathogens. A disposable electrochemical immunosensor for detection of Vibrio parahaemolyticus (VP) based on the screen-printed electrode (SPE) coated with agarose/Nano-Au membrane and horseradish peroxidase (HRP) labeled VP antibody (HRP-anti-VP) has been developed in this paper. Then, the immunosensor was characterized by cyclic voltammetry (CV) and laser scanning confocal microscope (LSCM). The immunosensor was incubated with the one-step immunoassay format involving VP for 30 min at room temperature (25 ± 0.5 °C). The access of the active center of HRP catalyzing the oxidation reaction of thionine by H₂O₂ was partly inhibited by VP, which connected on the surface of the immunosensor by immunoreaction. VP could be quantificationally detected according to the shift of reduction current while CV was used as electrochemical means to detect the products of the enzymatic reaction. Under the optimum conditions of immunoreaction and electrochemical detection, VP was rapidly detectable by sigmoidal curve method and form a linear calibration between 10⁵ and 10⁹ cfu/ml with an associated detection limit of 7.374 × 10⁴ cfu/ml (S/N = 3). The immunosensor had acceptable specificity, reproducibility, stability and accuracy, indicating that the immunosensor could satisfy the need of practical sample detection.     

A sensitivity-controlled hydrogen peroxide sensor based on self-assembled Prussian Blue modified electrode

In this communication, a hydrogen peroxide (H₂O₂) sensor based on self-assembled Prussian Blue (PB) modified electrode was reported. Thin film of PB was deposited on the electrode by self-assembly process including multiple sequential adsorption of ferric ions and hexacyanoferrate ions. The as-prepared PB modified electrode displayed sufficient stability for practical sensing application. At an applied potential of ?0.05 V vs. Ag/AgCl (sat. KCl), PB modified electrode with 30 layers exhibited a linear dependence on H₂O₂ concentration in the range of 1 × 10^?6–4 × 10^?4 M (r = 0.9998) with a sensitivity of 625 mA M^?1 cm^?2. It was found that the sensitivity of H₂O₂ sensors could be well controlled by adjusting the number of deposition cycles for PB preparation. This work demonstrates the feasibility of self-assembled PB modified electrode in sensing application, and provides an effective approach to control the sensitivity of PB-based amperometric biosensors.     

A 3D-impedimetric immunosensor based on foam Ni for detection of sulfate-reducing bacteria

A 3D-immunosensor based on simple and efficient trapping platform (foam Ni) combining with adsorption of gold nanoparticles and specific recognition of biological/chemical molecular has been reported for detection of sulfate-reducing bacteria (SRB) using electrochemical impedance spectroscopy (EIS). The impedance spectra were also used to characterize the successful construct and stepwise modification of the impedimetric immunosensors. This results show that a linear relationship between electron-transfer resistance (R_ct) values and the logarithm of the SRB concentrations was obtained for the SRB concentration range of 2.1 × 10¹–2.1 × 10⁷ cfu/ml. Additionally, the fabricated immunosensor shows a high selectivity against other bacteria.     

Amperometric sensing of dopamine using a single-walled carbon nanotube covalently attached to a conical glass micropore electrode

A single-walled carbon nanotube (SWNT) is covalently attached to the interior surface of a conical glass micropore electrode (GME) to create a novel amperometric dopamine sensor (SWNT/NH-GME). The SWNT/NH-GME combines the advantages of excellent transport properties of the cone-shaped micropore with the characteristics of a SWNT, exhibiting a dramatic electrocatalytic effect on the oxidation of dopamine (DA). Cyclic voltammetry and amperometric methods were employed to study the electrochemical behavior of the SWNT/NH-GME. The results showed that the SWNT/NH-GME sensor exhibited an excellent immunity from ascorbic acid interference and was able to measure DA concentrations with a detection limit of 4.2 × 10^?7 mol/L (S/N = 3).     

Ferrocene-modified Fe3O4@SiO2 magnetic nanoparticles as building blocks for construction of reagentless enzyme-based biosensors

A novel amperometric glucose biosensor was developed by entrapping glucose oxidase (GOD) in chitosan (CS) composite doped with ferrocene monocarboxylic acid-modified magnetic core-shell Fe₃O₄@SiO₂ nanoparticles (FMC-AFSNPs). It is shown that the obtained magnetic bio-nanoparticles attached to the surface of a carbon paste electrode (CPE) with the employment of a permanent magnet showed excellent electrochemical characteristics and at the same time acted as mediator to transfer electrons between the enzyme and the electrode. Under optimal conditions, this biosensor was able to detect glucose in the linear range from 1.0 × 10⁻⁵ to 4.0 × 10⁻³ M with a detection limit of 3.2 μM (S/N = 3). This immobilization approach effectively improved the stability of the electron transfer mediator and is promising for construction of biosensor and bioelectronic devices.     

Selective determination of uric acid with DNA doped polymers modified carbon fiber microelectrode

New biocomposite materials, based on the incorporation of DNA doped p-aminobenzensulfonic acid, was fabricated by electrochemical method. A carbon fiber microelectrode modified by this thin film was fabricated for selective determination of uric acid (UA) in the presence of a larger amount of ascorbic acid (AA). It was found that the voltammetric oxidation peak separation between UA and AA is about 260 mV at the modified electrode. A linear response of the peak current versus the concentration was found in the range of 8 × 10⁻⁷–6 × 10⁻⁴ M with correlation coefficient of 0.9991 and the detection limit was 5 × 10⁻⁷ M (s/n = 3) at the 5 × 10⁻⁴ M AA. The presence of high concentration AA did not interference the determination. The electropolymerized film was characterized by SEM techniques. The modified electrode shows good sensitivity, selectivity and stability.     

Determination of nitrite with the electrocatalytic property to the oxidation of nitrite on thionine modified aligned carbon nanotubes

A thionine modified aligned carbon nanotubes (ACNTs) electrode was fabricated and was used to electrochemically determine nitrite. The thionine modified ACNTs electrode exhibited enhanced electrocatalytic behavior to the oxidation of nitrite. The electrochemical mechanism of the thionine/ACNTs electrode towards the oxidation of nitrite was discussed. The thionine modified ACNTs electrode exhibited fast response towards nitrite with a detection limit of 1.12 × 10⁻⁶ mol  L⁻¹ and a linear range of 3 × 10⁻⁶ – 5 × 10⁻⁴ mol  L⁻¹. The proposed method was successfully applied in the detection of nitrite in real samples.     

Anodic electrochemiluminescent behavior of lucigenin on MWNT/GCE

The electrochemiluminescent (ECL) behavior of lucigenin on a multiwall carbon nanotubes modified glassy carbon electrode (MWNT/GCE) during anodic scanning was studied. A strong and stable anodic ECL signal was found on MWNT modified electrode, which results from the oxidation reaction between lucigenin and the oxidation production of OH^-. The effects of electrode materials, pH and scan rate on the ECL intensity were studied, and the possible ECL mechanism was also proposed. Under the optimized conditions, the ECL intensity was found to be linear with concentration of lucigenin in the range of 5.0 × 10^?7–5.0 × 10^?6 mol/L with a detection limit of 2.0 × 10^?7 mol/L. Superoxide dimutase (SOD) was found to be able to inhibit this ECL system, based on which a sensitive ECL methods for detection of SOD had been established.     

Porous cuprous oxide microcubes for non-enzymatic amperometric hydrogen peroxide and glucose sensing

Using porous cuprous oxide (Cu₂O) microcubes, a simple non-enzymatic amperometric sensor for the detection of H₂O₂ and glucose has been fabricated. Cyclic voltammetry (CV) revealed that porous Cu₂O microcubes exhibited a direct electrocatalytic activity for the reduction of H₂O₂ in phosphate buffer solution and the oxidation of glucose in an alkaline medium. The non-enzymatic amperometric sensor used in the detection of H₂O₂ with detection limit of 1.5 × 10^?6 M over wide linear detection ranges up to 1.5 mM and with a high sensitivity of 50.6 μA/mM. This non-enzymatic voltammetric sensor was further utilized in detection of glucose with a detection limit of 8.0 × 10^?7 M, a linear detection range up to 500 μM and with a sensitivity of ?70.8 μA/mM.     

A self-assembled monolayer based electrochemical immunosensor for detection of leukemia K562A cells

A novel strategy to quantify the cell number of leukemia K562A cells using electrochemical immunosensor was developed by effective surface immunoreaction between P-glycoprotein (P-gp) on cell membrane and P-gp mouse monoclonal antibody bound on an epoxysilane monolayer modified glassy carbon electrode. The surface morphologies of the epoxysilane monolayer and the bound antibodies were studied with atomic force microscopy. The binding of target K562A cells onto the immobilized antibodies increased the electron-transfer impedance of electrochemical probe, which depended linearly on the cell concentration in the range of 5.0 × 10⁴–1.0 × 10⁷ cells mL⁻¹. The detection limit of the immunosensor was 7.1 × 10³ cells mL⁻¹. The proposed strategy showed acceptable reproducibility with an RSD of 3.4% for the linear slope and good precision with the RSD of 3.7% and 3.0% examined at the cell concentrations of 2.0 × 10⁶ and 1.0 × 10⁷ cells mL⁻¹.     

An ionophore–Nafion modified bismuth electrode for the analysis of cadmium(II)

A novel ionophore–Nafion modified bismuth electrode is described for sensitive and selective anodic stripping analysis of cadmium(II). The electrode is prepared by coating the glassy carbon electrode with the cadmium ionophore N,N,N′,N′-tetrabutyl-3,6-dioxaoctanedi(thioamide) and Nafion composite. Bismuth is deposited in situ on the electrode surface by plating simultaneously with cadmium in sample solution. Numerous key variables affecting the current response of cadmium have been optimized. The electrode has a linear concentration range of 0.5–10 nM with a deposition time of 180 s. The detection limit is 1.3 × 10^?10 M and the relative standard deviations for 0.5 and 7 nM cadmium are 6.5% and 4.5%, respectively. The proposed electrode shows excellent selectivity over other heavy metals, such as copper, lead and indium. The attractive performance of such electrode offers a feasible way to monitor trace cadmium(II) rapidly and precisely in complex matrixes.     

Application of gamma irradiation for inactivation of three pathogenic bacteria inoculated into meatballs

In this research, the effect of gamma irradiation on the inactivation of Escherichia coli O157:H7 (ATCC 33150), Staphylococcus aureus (ATCC 2392) and Salmonella typhimurium (NRRL 4463) inoculated into Tekirdag meatballs was investigated. The meatball samples were inoculated with pathogens and irradiated at the absorbed doses of 1, 2.2, 3.2, 4.5 and 5.2 kGy. E. coli O157:H7 count in 1 kGy irradiated meatballs stored in the refrigerator for 7 days was detected to be 4 log cfu/g lower than the count in nonirradiated samples (p<0.05). S. aureus counts were decreased to 4 log cfu/g after being exposed to irradiation at a dose of 1 kGy. Although it was ineffective on elimination of S. typhimurium, irradiation at a dose of 3.2 kGy reduced E. coli O157:H7 and S. aureus counts under detectable values in the meatballs. However, none of the test organisms were detected in the samples after irradiation with 4.5 kGy doses.     

A study on size effect of carboxymethyl starch nanogel crosslinked by electron beam radiation

The formation of carboxymethyl starch (CMS) nanogel with 50 nm less particle size was carried out through a radiation crosslinked process on the electron beam (EB) linear accelerator. Changes of intrinsic viscosities and weight averaged molecular weight in the CMS concentration, which ranged from 3 to 10 mg ml^?1 in absorbed doses were investigated. There were some new peaks in the ¹H NMR spectra of CMS nanogel compared with those of CMS polymer. These results were anticipated that the predominant intramolecular crosslinking of dilute CMS aqueous solution occurred while being exposed to a short intense pulse of ionizing radiation. Hydrodynamic radius (often called particle size, R_h) and distribution of particle size were measured by a dynamic light scattering technique. The radiation yield of intermolecular crosslinking of CMS solution was calculated from the expression of G_x (Charlesby, 1960, Jung-Chul, 2010). The influence of the “size effect” was demonstrated by testing culture of Lactobacillus bacteria on MRS agar culture medium containing CMS nanogel and polymer. Results showed that the number of Lactobacillus bacteria growing on nanogel containing culture medium is about 170 cfu/ml and on polymer containing culture medium is only 6 cfu/ml.     

Electrospun zirconia-embedded carbon nanofibre for high-sensitive determination of methyl parathion

Carbon nanofibers embedded with ultrafine zirconia nanoparticles (ZrO₂-CNFs) are fabricated via a new methodology. Polyvinylpyrrolidone (PVP) and polymethylmethacrylate (PMMA) binary polymers containing zirconium n-butoxide are first dissolved in dimethylformamide, and the resulting solution is electrospun and heat-treated. The tetragonal zirconia nanoparticles formed, with a size of 5 ± 2 nm in diameter, are uniformly distributed in the carbon nanofibres. Using Nafion as an additive, ZrO₂-CNFs are drop-cast onto the glassy carbon electrode (ZrO₂-CNF/GCE) and the modified electrode is then applied to detect methyl parathion (MP) using differential pulse voltammetry. Two linear relationships are found at the concentration ranges of 1 × 10^− 9–2 × 10^− 8 g/L and 2 × 10^− 8–2 × 10^− 7 g/L, with a detection limit of 3.4 × 10^− 10 g/L (S/N > 3). The electrospun-based ZrO₂-CNF is a very promising coating material for electrochemical sensing of organophosphorus compounds.     

Diazenyl schiff bases: Synthesis,spectral analysis,antimicrobial studies and cytotoxic activity on human colorectal carcinoma cell line (HCT-116)

A series of diazenyl schiff bases have been synthesized by reaction of salicylaldehyde containing azo dyes with various substituted aniline derivatives in the presence of acetic acid as catalyst. The structures of diazenyl derivatives were determined by FTIR, UV–vis, ¹H NMR, ¹³C NMR, CHN analysis, fluorimetric and mass spectroscopic studies. The synthesized derivatives were screened for their in vitro antimicrobial activity against various Gram-positive (S. aureus, B. subtilis, B. cereus), Gram-negative (S. typhi, S. enterica, E. coli, P. aeruginosa) bacterial and fungal (C. albicans, A. niger and A. fumigatus) strains, using cefadroxil (antibacterial) and fluconazole (antifungal) as standard drugs. The diazenyl schiff bases were also screened for their cytotoxicity against human colorectal carcinoma cell line (HCT-116) using 5-fluorouracil as standard drug by Sulforhodamine-B Stain (SRB) assay. The schiff bases exhibited significant activity toward both Gram-positive, Gram-negative bacterial and fungal strains. Most of the synthesized derivatives showed high activity against S. enterica. 4-((2,5-Dichlorophenyl)diazenyl)-2-((3-bromophenylimino)methyl)phenol (SBN-40) was found to be very active against S. aureus, B. cereus and E. coli, with MIC = 0.69 (µM/ml × 10²). The compound 4-((2-bromophenyl)diazenyl)-2-((4-nitrophenylimino)methyl)phenol (SBN-13) possessed comparable activity (IC₅₀ = 7.5 µg/ml) to the standard drug 5-fluorouracil (IC₅₀ = 3.0 µg/ml) against human colorectal carcinoma cell line (HCT-116).     

Amperometric enzyme biosensor for hydrogen peroxide via Ugi multicomponent reaction

A novel strategy based on the Ugi multicomponent reaction was employed for immobilizing horseradish peroxidase on sodium alginate-coated gold electrode. The electrode was employed for constructing an amperometric biosensor device using 1 mM hydroquinone as electrochemical mediator. The electrode showed linear response (poised at −300 mV vs Ag/AgCl) toward H₂O₂ concentration between 70 μM and 8.8 mM at pH 7.0. The biosensor reached 95% of steady-state current in about 12 s and its sensitivity was 33.8 mA/M cm². The electrode retained full initial activity after 30 days of storage at 4 °C in 50 mM sodium phosphate buffer, pH 7.0.     

Zirconium(IV) phosphosulphosalicylate-based ion selective membrane electrode for potentiometric determination of Pb(II) ions

Zirconium(IV) phosphosulphosalicylate, a cation exchanger was synthesized by mixing zirconium oxychloride to a mixture of 5-sulphosalicylic acid and phosphoric acid. The material showed good efficiency for the preparation of an ion-selective membrane electrode. The membrane was characterized affinity for Pb(II) ions. Due to its Pb(II) selective nature, the ion-exchanger was used as an electroactive by XRD and SEM analysis. The electrode responds to Pb(II) ions in a linear range from 1 × 10⁻⁵ to 1 × 10⁻¹ M with a slope of 43.8 mV per decade change in concentration with detection limit of 4.78 × 10⁻⁶ M. The life span of electrode was found to be 90 days. The proposed electrode showed satisfactory performance over a pH range of 4.0–6.5, with a fast response time of 15 s. The sensor has been applied to the determination of Pb(II) ions in water samples of different origins. It has also been used as indicator electrode in potentiometric titration of Pb(II) ion with EDTA.     

Electrochemical determination of rosiglitazone by square-wave adsorptive stripping voltammetry method

Square-wave adsorptive stripping voltammetry technique was used to determine rosiglitazone (ROS) on the hanging mercury dropping electrode (HMDE) surface, in Britton Robinson buffer, pH = 5. The voltammetric cathodic peak was observed at ?1520 mV vs. Ag/AgCl reference electrode. The voltammetric peak response was characterized with respect to pH, supporting electrolyte, accumulation potential, preconcentration time, scan rate, frequency, pulse amplitude, surface area of the working electrode and the convection rate. Under optimal conditions, the voltammetric current is proportional to the concentration of ROS over the concentration range of 5 × 10^?8–8 × 10^?7 mol l^?1 (r = 0.9899) with a detection limit of 3.2 × 10^?11 mol l^?1 using 120 s accumulation time. The developed SW-AdSV procedure showed a good reproducibility, the relative standard deviation RSD% (n = 10) at a concentration level of 5 × 10^?7 mol l^?1 was 0.33%, whereas the accuracy was 101% ± 1.0. The proposed method was successfully applied to assay the drug in the human urine and plasma samples with mean recoveries of 90 ± 0.71% and 86 ± 1.0%, respectively.