Electrochemical biosensor for detection of formaldehyde in rain water

BACKGROUD: This study describes the construction of an electrochemical formaldehyde biosensor based on poly(glycidyl methacrylate‐co‐3‐methylthienyl methacrylate)/formaldehyde dehydrogenase/polypyrrole [poly(GMA‐co‐MTM)/FDH/PPy] composite film electrode. Formaldehyde dehydrogenase (FDH) was chemically immobilized via the epoxy groups of the glycidyl methacrylate (GMA) side chain of the polymer. Formaldehyde measurements were conducted in 0.1 mol L^?1, pH 8 phosphate buffer solution (PBS) including 0.1 mol L^?1 KCl, 0.5 mmol L^?1 of NAD⁺ (cofactor of the enzyme) and 1 mmol L^?1 of 1,2‐napthoquinone‐4‐sulfonic acid sodium salt (NQS) as mediator with an applied potential of ? 0.23 V (vs. Ag/AgCl, 3 mol L^?1 NaCl). Analytical parameters of the biosensor were calculated and discussed. The biosensor was tested in rain water samples. RESULTS: Sensitivity was found to be 15 000 per mmol L^?1 (500 nA ppm^?1) in a linear range between 0.1 ppm and 3 ppm (3.3–100 µmol L^?1). A minimum detectable concentration of 4.5 ppb (0.15 µmol L^?1) (S/N = 3) with a relative standard deviation (RSD) of 0.73% (n = 5) was obtained from the biosensor. Response time of the biosensor was very short, reaching 99% of its maximum response in about 4 s. The biosensor was also tested for formaldehyde measurements in rain water samples. Formaldehyde concentrations in samples were calculated using the proposed biosensor with recovery values ranged between 92.2 and 97.7% in comparison with the colorimetric Nash method. CONCLUSION: The poly(GMA‐co‐MTM)/FDH/PPy) electrode showed excellent measurement sensitivity in comparison with other formaldehyde biosensor studies. Strong chemical bonding between the enzyme and the copolymer was created via the epoxy groups of the composite film. The proposed biosensor could be used successfully in rain waters without a pretreatment step. © 2012 Society of Chemical Industry     

Development of a new two‐enzyme biosensor based on poly(pyrrole‐co‐3,4‐ethylenedioxythiophene) for lactose determination in milk

A new lactose biosensor was developed by preparing a suitable copolymer of polypyrrole and poly(3,4‐ethylenedioxythiophene) synthesized using the electropolymerization method. Pyrrole and 3,4‐ethylenedioxythiophene monomers were deposited in the presence of sodium dodecylbenzene sulphonic acid on a platinum disc electrode, which was used as the working electrode. The sensor is based on the serial reactions of β‐galactosidase and galactose oxidase immobilized on a copolymer‐modified platinum disc electrode. Successful synthesis of the enzyme‐immobilized copolymer was confirmed by FT‐IR spectrometry, SEM, and electrochemical analysis. The response of the enzyme electrode to lactose was determined by cyclic voltammetry at + 0.40 V. The response time of the biosensor was found to be from 8 to 10 s, and the upper limit of the linear working portion was found to be at a lactose concentration of 2.30 mM with a detection limit of 1.4 × 10^?5 M. The apparent Michaelis–Menten constant was found to be 0.65 mM of lactose. The effects of interferents were also investigated. Lactose concentrations determined by the biosensor were in good agreement with those measured by the reference methods. Our results show that the developed biosensor has a significant potential to the determination of lactose concentration in milk. POLYM. ENG. SCI., 58:839–848, 2018. © 2017 Society of Plastics Engineers     

Construction of a choline biosensor through enzyme immobilization on a poly(2‐hydroxyethyl methacrylate)‐grafted Teflon film

An amperometric choline biosensor was constructed by immobilizing choline oxidase (ChO) on poly(2‐hydroxyethyl methacrylate) (PHEMA)‐grafted Teflon (polytetrafluoroethylene, PTFE) film. Grafting was achieved by γ irradiation. PHEMA‐grafted Teflon films were activated with epichlorohydrin or glutaraldehyde to achieve covalent immobilization of enzyme onto the film. To decrease the diffusional barrier caused by the enzyme‐immobilized film, the film was stretched directly on the electrode. The PHEMA‐grafted Teflon film, therefore, had to have appropriate mechanical properties. Glucose oxidase (GOD) was used in the determination of optimum immobilization conditions, then these were applied to ChO. With GOD, the effect of activation type and film position in electrode on enzyme activity was studied and the highest catalytic activity was obtained when the enzyme was immobilized using glutaraldehyde and the film was stretched over the electrode surface. Further studies revealed that the films activated with glutaraldehyde, immobilized in 2 mg/mL ChO concentration, and stretched directly on the electrode were suitable (specific activity, 0.427 ± 0.068 U mg^?1) for use in the choline biosensor. The linear working range of this biosensor was found to be 52–348 μM, with a 40 ± 5 μM minimum detection limit. The response of the sensor, however, decreased linearly upon repeated use. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2007     

Electrochemical Biosensor Based on surfactant doped polypyrrole (PPy) matrix for lactose determination

Lactose biosensor based on surfactant doped polypyrrole (PPy) was developed. Galactose oxidase and β‐galactosidase was coimmobilized in PPy matrix during electropolymerization process with the presence of sodium dodecylbenzene sulphonic acid as surfactant. Bi‐enzyme entrapped PPy was characterized with Fourier transform infrared spectroscopy (FTIR), cyclic voltammetry (CV), and scanning electron microscopy (SEM). The response of the enzyme electrode was measured by CV in the range of ?0.1 to 1.0 V versus Ag/AgCl which was due to the electrooxidation of enzymatically produced H₂O₂. The effect of lactose concentration was investigated. Response time of biosensor was found to be 8–10 s (the time required to obtain the maximum peak current) and upper limit of the linear working portions was found to be 1.22 mM lactose concentration with a detection limit of (2.6 × 10^?6 M). The apparent Michaelis–Menten constant was found to be 0.117 mM lactose. The effects of interferents (ascorbic acid and uric acid) were determined. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40200.     

Polypyrrole–multiwalled carbon nanotubes composites as immobilizing matrices of ascorbate oxidase for the facile fabrication of an amperometric vitamin C biosensor

An amperometric vitamin C biosensor was facilely fabricated by the immobilization of ascorbate oxidase (AO) on polypyrrole (PPy)–multiwalled carbon nanotubes (MWCNTs) composites with a one‐step electrodeposition technique in a 0.05M phosphate buffer solution (pH 6.5). The cyclic voltammetry, IR spectral analysis, electrochemical impedance spectroscopy, and scanning electron microscopy measurements indicated that AO was successfully immobilized on the PPy–MWCNT composites. The optimization of the biosensor parameters, including the working potential, pH, and temperature, was investigated in detail. The proposed biosensor showed a linear range of 5 × 10^?5 to 2 × 10^?2 M with a detection limit of 0.3 μM, a sensitivity of 25.9 mA mM^?1 cm^?2, and a current response time less than 20 s under the optimized conditions. The apparent Michaelis–Menten constant together with the apparent activation energy indicated that the proposed biosensor exhibited a high bioaffinity and a good enzyme activity. In addition, the biosensor also showed good operational and storage stabilities. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

A novel multienzyme electrode for the determination of citrate

A novel amperometric biosensor for the determination of citric acid in food samples and fermentation broths has been developed. The sensor is composed of citrate lyase (CL, EC 4.1.3.6), oxaloacetate decarboxylase (OAC, EC 4.1.1.3) and pyruvate oxidase (POP, EC 1.2.3.3), co-immobilized in gelatin, and an amperometric transducer. A Clark-type O₂-electrode and a modified Clark-type H₂O₂-electrode were alternatively used as a transducer. The biosensor covers a linear detection range from 1 μmol dm^?3 to 1 mmol dm^?3 citrate, with a response time of 2·5 min for the steady state response. The lower detection limit for citrate is 0·5 μmol dm^?3. The response of the sensor remained constant for 8 days and decreased to 25% after 18 days at 20–23°C. The results obtained from citrate determinations in food samples and fermentation broths agree well with those determined by enzymatic sample anlaysis. The relative standard deviation for citrate determinations with the new biosensor was 2·2% (n = 7).     

Development of an amperometric biosensor based on a redox‐mediator‐doped polypyrrole film

An amperometric biosensor was developed for the quantitative estimation of phenolic compounds in aqueous media. The enzyme tyrosinase [poly(phenol oxidase) (PPO)] was adsorbed onto a hexacyanoferrate(II)‐ion‐doped conducting polypyrrole (PPY) film deposited on an indium tin oxide (ITO) coated glass‐plate support. The PPO activity in the PPO/Fe²⁺‐PPY/ITO film was assayed as a function of the concentration of phenolic compounds. Cyclic voltammetric studies were carried out on this enzyme electrode, and the surface morphology of the enzyme‐immobilized polymer film was studied with scanning electron microscopy. The results of the amperometric response of the PPO/Fe²⁺‐PPY/ITO film showed sensitivities of 0.14, 0.21, and 0.36 A M^?1cm^?2 and linear response ranges of 9.9–84.7, 6.7–72.6, and 3.9–48.8 μM for phenol, catechol, and p‐chlorophenol, respectively. The PPO/Fe²⁺‐PPY/ITO electrode exhibited a response time of about 50 s and was stable for about 12 weeks at 4°C. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 93: 927–933, 2004     

A novel microbial BOD biosensor developed by the immobilization of P. Syringae in micro‐cellular polymers

BACKGROUND: A biochemical oxygen demand (BOD) sensor, based on an immobilized Pseudomonas syringae in highly porous micro‐cellular polymer (MCP) in combination with a dissolved oxygen electrode, has been developed for the analysis of biodegradable organic compounds in aqueous samples. Microorganisms were immobilized in a molded MCP disk and a wastewater sample was injected into the biocomposite disk by a flow injection system. Dissolved oxygen (DO) changes as a measure of soluble BOD was read with a DO probe placed into a flow cell carrying biocatalytically activated disk. RESULTS: Optimal response of the MCP BOD sensor was obtained at pH 6.8 and 25 °C with a typical response time of 3–5 min for a 2 mm thick molded polymeric disk. The sensor showed detection linearity over the range 5–100 mg L^?1 BOD₅ (r² > 0.99) at a flow rate of 0.6 mL min^?1. The repeatability and reproducibility of the sensor response were found to be 3.08% and 7.77%, respectively. BOD values produced with this biosensor for various municipal and industrial wastewaters correlated well with those determined by the conventional 5‐day BOD test. CONCLUSION: This new biosensor was different from present amperometric BOD biosensor configurations in which the biocatalyst (microbial/enzymatic) is placed between cellulose and Teflon membranes installed on a DO probe. The use of a molded MCP disk coniainng microbial activity offers better stability and lifetime for commercial use in environmental monitoring. Copyright © 2008 Society of Chemical Industry     

Electrochemical sensing of NADH on NiO nanoparticles-modified carbon paste electrode and fabrication of ethanol dehydrogenase-based biosensor

In this work, an electrochemical β-nicotinamide adenine dinucleotide (NADH) sensor based on a carbon paste electrode modified with nickel oxide nanoparticles (NiONPs) was developed. The key highlights of this work are ease of preparation of the NiONPs-modified carbon paste electrode (NiONPs/MCPE), and its high sensitivity to NADH. The electrochemical characterization of NiONPs/MCPEs was performed via cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The electrochemical oxidation response of NADH was investigated by differential pulse voltammetry and chronoamperometry. The results indicated that the electrocatalytic effects of NiONPs on the response current of NADH significantly facilitated the electron transfer and improved the performance of the biosensor. Compared to bare carbon paste electrode (BCPE), the oxidation potential was shifted toward more negative potentials and the oxidation current was increased remarkably. Under optimum conditions, NADH could be detected in the range from 1.0 × 10^?4 to 1.0 mmol L^?1 with lower detection limit (0.05 μmol L^?1). The proposed NADH sensor demonstrated fast and reproducible response. Furthermore, an ethanol biosensor was prepared using NiONPs and NAD⁺-dependent alcohol dehydrogenase enzyme giving linear responses over the concentration range of 1.6 and 38 mmol L^?1 of ethanol.     

A highly selective electrochemical biosensor for Hg using hemin as a redox indicator

A highly selective electrochemical biosensor for the detection of Hg²⁺ in aqueous solution has been developed. This sensor is based on the strong and specific binding of Hg²⁺ by two DNA thymine bases (T–Hg²⁺–T). The hemin worked as a redox indicator to generate a readable electrochemical signal. Short oligonucleotide strands containing 5 thymine (T₅) were used as probe. Thiolated T₅ strands were self-assembled through Au–S bonding on gold electrode. In the presence of Hg²⁺, the specific coordination between Hg²⁺ and thymine bases resulted in more stable and porous arrangement of oligonucleotide strands, so hemin could be adsorbed on the surface of gold electrode and produced an electrochemical signal, which was monitored by differential pulse voltammetry (DPV). The DPV showed a linear correlation between the signal and the concentration of Hg²⁺ over the range 0–2 μM (R² = 0.9983) with a detection limit of 50 nM. The length of probe DNA had no significant impact on the sensor performance. This electrochemical biosensor could be widely used for selective detection of Hg²⁺.     

Novel potentiometric sensors for the selective determination of domperidone

The fabrication and electrochemical response characteristics of two novel potentiometric sensors for the selective determination of domperidone (DOM) are described. The two fabricated sensors incorporate DOM–PTA (phosphotungstic acid) ion pair as the electroactive material. The sensors include a PVC membrane sensor and a carbon paste sensor. The sensors showed a linear, stable, and near Nernstian slope of 56.5 and 57.8 mV/decade for PVC membrane and carbon paste sensors, respectively over a relatively wide range of DOM concentration (1.0 × 10^?1–1.0 × 10^?5 and 1.0 × 10^?1–3.55 × 10^?6 M). The response time of DOM–PTA membrane sensor was less than 25 s and that in the case of carbon paste sensor was less than 20 s. A useful pH range of 4–6 was obtained for both types of sensors. A detection limit of 7.36 × 10^?5 M was obtained for PVC membrane sensor and 1.0 × 10^?6 M was obtained for carbon paste sensor. The proposed sensors showed very good selectivity to DOM in the presence of a large number of other interfering ions. The analytical application of the developed sensors in the determination of the drug in pharmaceutical formulations such as tablets was investigated. The results obtained are in good agreement with the values obtained by the standard method. The sensors were also applied for the determination of DOM in real samples such as urine by the standard addition method.     

A New Sensor for Determination of Anionic Surfactants in Detergent Products with Carbon Nanotubes as Solid Contact

A new solid‐state sensor for potentiometric determination of surfactants with a layer of multi‐walled carbon nanotubes was prepared. As a sensing material, 1,3‐didecyl‐2‐methylimidazolium–tetraphenylborate ion‐pair was used. The investigated sensor showed a Nernstian response for both dodecylbenzenesulphonate (DBS, 57.6 mV/decade of activity between 5 × 10^?7 to 1 × 10^?3 M) and sodium lauryl sulfate (LS, 58.4 mV/decade of activity between 2 × 10^?7 to 2 × 10^?3 M). It responded in 8–10 s for each ten‐fold concentration change in the range of 1 × 10^?6 to 3 × 10^?3 M. The detection limits for DS and DBS were 2 × 10^?7 and 3 × 10^?7 M, respectively. The sensor revealed a stable response (signal drift 2.6 mV/h) and exhibited satisfactory selectivity performances for LS over most of the anions generally used in surfactant‐based commercial detergents. The main application of this sensor was the end‐point determination in potentiometric titrations of anionic surfactants. The (diisobutyl phenoxy ethoxy ethyl)dimethyl benzyl ammonium chloride (Hyamine), cetyltrimethylammonium bromide, hexadecylpyridinium chloride monohydrate (HDPC) and 1,3‐didecyl‐2‐methylimidazolium chloride were tested as potential cationic titrants, and all exhibited analytically usable titration curves with well‐defined equivalence points. The standard solution of HDPC was used as a cationic titrant by all potentiometric titrations. The operational life‐time of the sensor described was prolonged to more than 3 months.     

Comparison of Two Enzyme Sequences for a Novel L-Malate Biosensor

Two novel amperometric biosensors for the determination of L -malic acid in food samples have been compared. Both sensors make use of a Clark-type O₂-electrode but differ in the enzymes used. The first sensor is composed of malate dehydrogenase (decarboxylating), also known as ‘malic enzyme’ (MDH(dec.), EC 1.1.1.40) and pyruvate oxidase (POP, EC 1.2.3.3). It covers a linear detection range from 1 μmol dm⁻³ to 0·9 mmol dm⁻³ L -malate, with a response time of 1·5 min (t₉₀) and a relative standard deviation of 3·5%. Measurements with real samples offered a good correlation with the standard enzymatic assay (difference ±7%). Stored at room temperature, the response of the sensor is constant for 8 days. The second biosensor is based on the three enzyme sequence malate dehydrogenase (MDH, EC 1.1.1.37), oxaloacetate decarboxylase (OAC, EC 4.1.1.3) and pyruvate oxidase (POP, EC 1.2.3.3). It has a non-linear calibration curve. Concentrations from 5 μmol dm⁻³ to 1 mmol dm⁻³ L -malate can be detected, within a response time of 1·5 min and with a relative standard deviation of 20%. The lower detection limit for L -malate is 2 μmol dm⁻³. The response is constant for 10 days when the sensor is stored at room temperature.     

Development of a new biosensor for mediatorless voltammetric determination of hydrogen peroxide and its application in milk samples

A new biosensor for the voltammetric detection of hydrogen peroxide was developed based on immobilization of catalase on a clinoptilolite modified carbon paste electrode using bovine serum albumin and glutaraldehyde. The biosensor response was evaluated according to electrode composition, reaction time, solution pH and temperature. The voltammetric signals were linearly in proportion to H₂O₂ concentration in the range 5.0 × 10⁻⁶–1.0 × 10⁻³ M with a correlation coefficient of 0.9975. The detection limit is 8.0 × 10⁻⁷ M and the relative standard deviation for 4.0 × 10⁻⁴ M hydrogen peroxide was 1.83% (n = 6). The biosensor exhibited high sensitivity, and it was determined that it could be used for more than 2 months. In addition, the biosensor was successfully applied for the determination of hydrogen peroxide in milk samples.     

Electropreparation of (±)‐10‐camphorsulfonate‐doped poly(o‐phenylenediamine) in acetonitrile and its use in determination of glucose

Poly(o‐phenylenediamine) (PoPD) film has been electrochemically prepared on Pt electrode in an acetonitrile–water medium containing o‐phenylenediamine (oPD) monomer and (±)‐10‐camphorsulfonic acid (HCSA) by using the cyclic voltammetry (CV). The PoPD film (PoPD–CSA) has been characterized by FTIR, CV, EIS, FESEM, and conductivity measurement. The glucose biosensor (Pt/PoPD–CSA/GOx) has been prepared from the PoPD coated electrode by immobilizing glucose oxidase (GOx) enzyme using glutaraldehyde. The biosensor shows a low detection limit and wide linear working range, a good reusability, long‐term stability, and anti‐interference ability. The Pt/PoPD–CSA/GOx has possesses higher sensitivity (2.05 μA/mmol L^?1) and affinity to glucose due to the use of CSA ion as dopant. The linear concentration ranges of Pt/PoPD–CSA/GOx have been found to be 9.6 × 10^?3 to 8.2 mmol L^?1 from calibration curve and 4.6 × 10^?2 to 100 mmol L^?1 from the relationship between the (1/glucose concentration) and (1/current difference). © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39864.     

Covalent immobilization of glucose oxidase to poly(O‐amino benzoic acid) for application to glucose biosensor

A biosensor for glucose utilizing glucose oxidase (GOX) covalently coupled to poly(o‐amino benzoic acid) (PAB; a carboxy‐group‐functionalized polyaniline) is described. Amperometric response measurements conducted via unmediated and mediated (with ferrocene carboxylic acid and tetrathiafulvalene) reoxidation of GOX show that glucose can be detected over a wide range of concentrations. An enzyme‐conducting polymer‐mediator model provides for better charge transport in a biosensor. The optimal response, obtained at pH 5.5 and 300 K, lies in the 1–40 mM range. A kinetic plot yields the value of the apparent Michaelis–Menten constant, K_m^app. The operational stability of the PAB‐based glucose biosensor was experimentally determined to be about 6 days. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 78: 662–667, 2000     

Molecularly imprinted electrochemical sensor for the determination of ampicillin based on a gold nanoparticle and multiwalled carbon nanotube‐coated pt electrode

A novel molecularly imprinted electrochemical sensor was developed for the sensitive and selective determination of ampicillin (AMP). The sensor was prepared on a platinum electrode modified with multiwalled carbon nanotubes (MWCNTs), gold nanoparticles (AuNPs), and a thin film of molecularly imprinted polymers (MIPs). MWCNTs and AuNPs were introduced to enhance the sensor's electronic transmission and sensitivity. The molecularly imprinted polymer (MIP) was synthesized using AMP as the template molecule, methacrylic acid as functional monomer, and ethylene glycol maleic rosinate acrylate (EGMRA) as cross‐linker. The performance of the proposed imprinted sensor was investigated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). The results showed that the imprinted film displayed a fast and sensitive response to AMP. Under optimal conditions, response peak current had a linear relationship with the concentration of AMP in the range of 1.0 × 10^?8 mol/L to 5.0 × 10^?6 mol/L and a detection limit of 1.0 × 10^?9 mol/L (S/N = 3). This imprinted sensor was used to detect AMP in food samples with recoveries of 91.4–105%. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40613.     

Impedimetric microbial biosensor based on single wall carbon nanotube modified microelectrodes for trichloroethylene detection

Contamination of soils and groundwaters with persistent organic pollutants is a matter of increasing concern. The most common organic pollutants are chlorinated hydrocarbons such as perchloroethylene and trichloroethylene (TCE). In this study, we developed a bacterial impedimetric biosensor for TCE detection, based on the immobilization of Pseudomonas putida F1 strain on gold microelectrodes functionalized with single wall carbon nanotubes covalently linked to anti-Pseudomonas antibodies. The different steps of microelectrodes functionalization were characterized by electrochemical impedance and atomic force spectroscopies, and analytical performances of the developed microbial biosensor were determined. The impedimetric biosensor response was linear with TCE concentration up to 150 μg L⁻¹ and a low limit of detection (20 μg L⁻¹) was achieved. No significant loss of signal was observed after 4 weeks of storage at 4 °C in phosphate buffer saline pH 7 (three to four measurements a week). After 5 weeks, 90% of the initial value still remained. cis-1,2-Dichloroethylene and vinylchloride, the main TCE degradation products, did not significantly interfere with TCE. The microbial sensor was finally applied to the determination of TCE in natural water samples spiked at the 30, 50 and 75 μg L⁻¹ levels. Recoveries were very good, ranging from 100 to 103%.     

A Single‐Electrode,Dual‐Potential Ferrocene–PNA Biosensor for the Detection of DNA

A Fc–PNA biosensor (Fc: ferrocenyl, C₁₀H₉Fe) was designed by using two electrochemically distinguishable recognition elements with different molecular information at a single electrode. Two Fc–PNA capture probes were therefore synthesized by N‐terminal labeling different dodecamer PNA sequences with different ferrocene derivatives by click chemistry. Each of the two strands was thereby tethered with one specific ferrocene derivative. The two capture probes revealed quasi‐reversible redox processes of the Fc^0/+ redox couple with a significant difference in their electrochemical half‐wave potentials of ΔE_1/2=160 mV. A carefully designed biosensor interface, consisting of a ternary self‐assembled monolayer (SAM) of the two C‐terminal cysteine‐tethered Fc–PNA capture probes and 6‐mercaptohexanol, was electrochemically investigated by square wave (SWV) and cyclic voltammetry (CV). The biosensor properties of this interface were analyzed by studying the interaction with DNA sequences that were complementary to either of the two capture probes by SWV. Based on distinct changes in both peak current and potential, a parallel identification of these two DNA sequences was successful with one interface design. Moreover, the primary electrochemical response could be converted by a simple mathematical analysis into a clear‐cut electrochemical signal about the hybridization event. The discrimination of single‐nucleotide polymorphism (SNP) was proven with a chosen single‐mismatch DNA sequence. Furthermore, experiments with crude bacterial RNA confirm the principal suitability of this dual‐potential sensor under real‐life conditions.     

Hybrid electrochemical biosensor for organophosphorus pesticides quantification

An electrochemical biosensor for organophosphorus (OP) pesticides trace level concentrations determination was developed and characterized. It integrates a hybrid biorecognition element consisting of immobilized Arthrobacter globiformis and free acetylcholinesterase (ACh) with a Clark type oxygen probe transducer. The bacteria convert the ACh-generated choline to betaine with oxygen consumption measured as a Clark probe current change. This change representing the sensor response correlates to the concentration of the OP pesticides inhibiting the Ach-catalyzed acetylcholine hydrolysis to choline. The conditions for maximal sensor response to choline were optimized according to the methodology of design of experiments. The analytical performances of the enzyme substrate determination in a wide concentration range (0.1-20 μmol dm⁻³ of acetylcholine) and different ACh activities were established. It was demonstrated that the biosensor ensures reproducible, accurate and reliable chlorophos quantification reaching a limit of detection (LOD) of 1 nmol dm⁻³ and a sensitivity of 0.0252 μA/p(mol dm⁻³) under optimal experimental conditions. The biosensor response time is 200 s and the storage stability is t_L50 = 49 days for the bacterial membrane at ambient temperature. The device is reusable, the bacterial membrane being not affected by OP. The biosensor was applied to chlorophos determination in contaminated milk.