A Novel Potentiometric Glucose Biosensor Based on Polyaniline Semiconductor Films

Abstract

Application of polyaniline semiconductor films to potentiometric biosensor development provides certain advantages comparing with the known systems. Using self-doped polyaniline instead of common polymer as pH transducer the stable potentiometric response of 70 mV/pH was obtained. Taking as an example glucose biosensor we showed that polyaniline based electrode possessed three-four fold increased potential shift than glucose-sensitive field-effect transistor did. One can increase the sensitivity of potentiometric biosensor using thick ion-exchange membranes (in our case Nafion) in order to concentrate product near electrode surface. Such sensor possessed higher response time.     

Potentiometric detection of organic acids in liquid chromatography using conducting oligomer electrodes

A conducting oligomer electrode was used for the potentiometric detection of organic acids in reversed phase liquid chromatography (LC). The conducting material consisted of a mixture of a phenylene vinylene trimer with a polycarbonate host polymer and iodine. A glassy carbon electrode was coated with this material by evaporation from a chloroform solution. A theoretical model was given to describe the observed potentiometric responses. The analysis conditions were optimized to obtain both efficient separations, and sensitive potentiometric responses. Detection limits in the nanogram level were attained when a 1 mM phosphoric acid solution was used as the eluent, which were comparable to the values obtained with low-wavelength UV detection. Calibration curves showed a logarithmic dependence on an injected amount for amounts higher than 5 nmol, and a linear dependence for injected amounts below this value. The response times of the electrode were smaller than 1 s at typical LC flow-rates. The reproducibility for consecutive injections was 5%.     

Fabrication of Redox‐Mediator Supported Potentiometric Nitrate Biosensor with Nitrate Reductase

Fabrication of a more superior nitrate potentiometric biosensor than previously achieved with NaR and NADH has been accomplished by co‐entrapment of redox mediators and NaR into polypyrrole (PPy) film during galvanostatic polymerization of pyrrole. The replacement of NADH with redox mediators such as thionin acetate (ThAc), safranin (Saf), and azure A (AzA) gave more sensitive potentiometric responses, better minimum detectable concentration, linear concentration range and response time for nitrate than possible with NADH. The co‐entrapment of ThAc, Saf, AzA and methyl viologen (MV) with NaR into PPy films also improved the Nernstian behavior of the electrode process beyond the capability of the PPy‐NaR‐NADH biosensor. Substantial reduction in volume and quantity of cofactor/mediator and, hence cost, was achieved by the replacement of NADH with a redox mediator. Only 50 μM of AzA was required to form a PPy‐NaR‐AzA biosensor which gave the most sensitive potentiometric response for nitrate, achieving a minimum detectable concentration of 10 μM, a linear concentration range of 50–5000 μM and a response time of 2–4 s.     

Potentiometric Biosensor System Based on Recombinant Urease and Creatinine Deiminase for Urea and Creatinine Determination in Blood Dialysate and Serum

A biosensor system for simultaneous determination of creatinine and urea in blood serum and dialysate samples was developed. It consisted of creatinine and urea biosensors based on a potentiometric transducers with two identical pH‐sensitive field‐effect transistors. In creatinine biosensor, creatinine deiminase immobilized via photopolymerization in PVA/SbQ polymer on one transistor served as a biorecognition element, while bovine serum albumin in PVA/SbQ polymer placed on the second transistor was used for reference. The urea biosensor was created in the same way but recombinant urease was used instead of creatinine deiminase. The linear ranges of creatinine and urea measurement were 0.02–2 mM and 0.5–15 mM, correspondingly, which allowed simultaneous determination of the metabolites. Response time of the biosensor system was 2–3 min; RSD of responses did not exceeded 5 %. The biosensors demonstrated absence of non‐selective response towards components of blood dialysate and serum. Urea and creatinine concentrations were determined in 20 samples of blood dialysate and serum. The results correlated well with traditional methods of analysis. Creatinine and urea biosensors were stable during five months of storage (during this time the responses decreased by about 10 %). The proposed biosensor system can be effectively used for analysis of serum samples and for hemodialysis control.     

An amperometric biosensor based on covalent immobilization of ascorbate oxidase on biocompatiable and low-toxic poly(thiophene-3-acetic acid) matrix

The biocompatiable and low-toxic poly（thiophene-3-acetic acid）（PTAA） matrix was successfully electrosynthesized in ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate（BmimPF₆） in comparison with the electrosynthesis of PTAA matrix in acetonitrile（ACN）.Ascorbate oxidase（AO） was used as a model for the development and application of biosensor.Vitamin C（VC） biosensors were facilely fabricated by the covalent immobilization of AO molecules on PTAA matrices electrosynthesized in ACN containing tetrabutylammonium tetrafluoroborate and BmimPF₆, respectively.Electrochemical impedance spectroscopy,scanning electron microscopy and FTIR spectroscopy indicated that AO molecules were covalently immobilized on PTAA matrices.Parameters of the as-obtained biosensors such as working potential,pH and temperature have been optimized.The amperometric biosensor based on PTAA matrix electrosynthesized in BmimPF₆ exhibited wider linear range,lower detection limit,higher sensitivity and bioaffinity,and better operational and storage stability than that electrosynthesized in ACN under optimal conditions.The as-obtained biosensor based on PTAA matrix electrosynthesized in BmimPF₆ was employed for the detection of VC content in commercial juices,and the result was close to the data given by manufacturers.Excellent results indicate that the PTAA matrix electrosynthesized in ionic liquid is a promising platform for the covalent immobilization of biologically-active species and the development of biosensors.     

Potentiometric Cholesterol Biosensor Based on ZnO Nanowalls and Stabilized Polymerized Lipid Film

A novel potentiometric cholesterol biosensor was fabricated by immobilization of cholesterol oxidase into stabilized lipid films using zinc oxide (ZnO) nanowalls as measuring electrode. Cholesterol oxidase was incorporated into the lipid film prior polymerization on the surface of ZnO nanowalls resulting in a sensitive, selective, stable and reproducible cholesterol biosensor. The potentiometric response was 57 mV/ decade concentration. The sensor response had no interferences by normal concentrations of ascorbic acid, glucose, and urea, proteins and lipids. The present biosensor could be implanted in the human body because of the biocompatibility of the lipid film.     

Chemically modified electrode based on poly[tetra(4-aminophenyl)porphyrin] as a pH sensor

A chemically modified platinum electrode with coated poly[tetra(4-aminophenyl)porphyrin] has been used as a potentiometric pH sensor. It gives a linear response over the pH range 1.5-13.7 with a slope of 55 mV/pH (at 20 degrees ). The sensor has fair resistance to erosion of hydrofluoric acid and to interference of a coexisting redox couple. The sensor can be used for pH determination and end-point indication for potentiometric titration of hydrofluoric acid with sodium hydroxide. The a.c. impedance of the polymer membrane has also been studied.     

Flow Potentiometric Injection Analysis of Uric Acid Using Lipid Stabilized Films with Incorporated Uricase on ZnO Nanowires

A novel potentiometric uric acid biosensor was fabricated by immobilization of uricase into stabilized lipid films using zinc oxide (ZnO) nanowires as measuring electrode. Uricase was incorporated into the lipid film prior polymerization on the surface of well aligned ZnO nanowires resulting in a sensitive, selective, stable and reproducible uric acid biosensor. The potentiometric response was twice as large from previously reported values due to the presence of a cationic lipid in the lipid film. The sensor response had no interferences by normal concentrations of ascorbic acid, glucose, urea, proteins and lipids.     

Coated wire silver-ion selective electrode based on a N,N'-bis(2-thienylmethylene)-1,2-diaminobenzene.

A novel membrane coated platinum-wire electrode (MCPWE) based on N,N'-bis(2-thienylmethylene)-1,2-diaminobenzene (BTMD) for highly selective determination of Ag+ ion has been developed. The influences of membrane composition and pH on the potentiometric responses of electrode were investigated. The potentiometric responses are independent of the pH of the test solution in the range of 5.0 - 9.0. The electrode shows a linear response for Ag+ ion over the concentration range of 1.0 x 10(-60 to 1.0 x 10(-1) M with a lower detection limit of 6.0 x 10(-7) M. The electrode possesses a Nernstian slope of 59.7 mV decade(-1) and a fast response time of < or = 17 s and can be used for at least 2 months without any observable deviation. The proposed electrode displayed very good selectivity for Ag+ ion with respect to NH4+ and alkali, alkaline earth and some common transition metal ions. The practical utility of the electrode has been demonstrated by its use as the indicator electrode in the potentiometric titration of an AgNO3 solution with a NaI solution and in determination of the silver content of a developed radiological film.     

pH sensor based on polyaniline and aniline–anthranilic acid copolymer films using quartz crystal microbalance and electronic absorption spectroscopy

The pH sensitivity based on conducting polyaniline (PANI) and copolymer of aniline and o‐anthranilic acid (AA) films were studied using quartz crystal microbalance (QCM) technique and UV–Vis spectroscopy. The sensor was constructed from these polymer films coated on the electrode of the QCM. The resonant frequency changes as a function of pH in the range of 2–12 were measured. These changes are quantitative indication of the degree of dedoping or redoping of the polymer films upon the subsequent exposure of the electrode to 0.25 M sulfuric acid and different pH solutions. There are two linear regressions between the frequency change and pH with two different and opposite slopes in the regions from 2 to 9 and 9 to 12. The pH sensitivity of the copolymer film was found to be less than using the PANI film. Thin films of PANI and copolymer, which were chemically polymerized in a sulfuric acid solution, were deposited onto the inner walls of the quartz cuvettes. The UV–Vis absorption spectra of these films were measured in different pH solutions. Relations between the maximum absorption and its wavelength versus pH were constructed. The copolymer film shows some advantages over the PANI film. The difference between the PANI and copolymer films as pH sensors using the QCM and electronic absorption extends from the determination of pK_a for both films. Copyright © 2008 John Wiley & Sons, Ltd.     

Conducting polymer ion sensor electrodes-III. Potentiometric sulfide ion selective electrode

A new potentiometric sensor electrode for sulfide based on conducting polymer films is introduced. The electrode is formed by electrochemically depositing a film of poly(3-methylthiophene) and poly(dibenzo-18-crown-6) onto an alloy substrate. Different methods were used for the electrode preparations. The alloy used has a low melting point, which allowed its use for manufacturing a microsize version of this electrode. The electrode response is stable for 3 days. The working temperature range for this electrode is between 10 and 40 degrees C. The linear dynamic range is 1.0x10(-7)-1.0x10(-2) M and measures total sulfide concentration over a range of pH from 1 to 13. The polymer electrode showed high selectivity for sulfide in the presence of many common interfering anions. The electrode is useful for the measurement of total sulfide in biological environments and can be manufactured in the micron scale. Therefore, it will be useful for the measurement within biofilms.     

One-pot electrosynthesis of polyglycine-like thin film on platinum electrodes as transducer for solid state pH measurements

The anodic oxidation of concentrated glycine based aqueous electrolyte on smooth platinum electrode leads to a strongly grafted polyglycine-like coating on the surface in an irreversible way. Due to the proton affinity towards amino groups of polyglycine (PG), the electrodeposited thin film was used as receptor for solid potentiometric pH sensor. In order to reach local pH measurement, we developed miniaturized microelectrodes on glass substrate thanks to photolithography process. We used silver chloride on silver as the reference electrode. The couple (silver chloride, PG based platinum electrode) of microelectrodes gives linear potentiometric response vs. pH in the range [2-12], reversibly and with a sensitivity of 52.4 mV/pH (for 1 mm electrode size). PG based pH electrode is compared to other organic polymer based pH receptor such as linear polyethylenimine (L-PEI), polyaniline (PANI) and glass membrane.     

Electrochemical Determination of Copper(II) in Water Samples Using a Novel Ion-Selective Electrode Based on a Graphite Oxide–Imprinted Polymer Composite

A novel copper(II)-selective electrode based on graphite oxide/imprinted polymer composite was developed for the electrochemical monitoring of copper(II) (Cu²⁺) ions. The electrode exhibited highly selective potentiometric response to Cu²⁺ with respect to common alkaline, alkaline earth and heavy metal cations. The composite composition studies indicated that the most suitable composite composition performing the most promising potentiometric properties was 20.0% ionophore (Cu²⁺-ion imprinted polymer), 10.0% paraffin oil, 5.0% multiwalled carbon nanotubes, and 65.0% graphite oxide. The fabricated electrode exhibited a linear response to Cu²⁺ over the concentration range of 1.0?×?10^??6–1.0?×?10^??1?M (correlation coefficient of 0.9998) with a sensitivity of 26.1?±?0.9?mV decade^??1. The detection limit of the fabricated electrode was determined to be 4.0?×?10^??7?M. The electrode worked well in the pH range of 4.0–8.0. The electrode had stable, reversible and fast potentiometric response (3?s). In addition, the electrode had a lifetime of more than 1 year. The analytical applications of the proposed electrode were performed using as an indicator electrode for the potentiometric titration of Cu²⁺ with ethylene diamine tetraacetic acid solution and for the determination of Cu²⁺ of spiked river, dam, and tap water samples. The obtained results for potentiometric titration and water samples were satisfactory.     

Development of electrochemical biosensors based on sol-gel enzyme encapsulation and protective polymer membranes

Protective polymer coatings have been used to enhance the retention of enzymes in sol-gel films as immobilisation phases in electrochemical biosensors. Carbon film electrodes were electrochemically modified with poly(neutral red) (PNR). These electrodes were coated with oxysilane sol-gels incorporating glucose oxidase and an outer coating of carboxylated PVC (CPVC) or polyurethane (PU), with and without Aliquat-336 or isopropyl myristate (IPM) plasticizer, was applied. The biosensors were characterised electrochemically using cyclic voltammetry and amperometry, electrochemical impedance spectroscopy and scanning electron microscopy. Impedance spectra showed that the electrode surface is most active when the sol-gel–GOx layer is not covered with a membrane. However, membranes without plasticizer extend the lifetime of the biosensor to more than 2 months when PU is used as an outer membrane. The linear range of the biosensors was found to be 0.05–0.50 mM of glucose and the biosensor with PU outer membrane exhibited higher sensitivity (ca.117 nA mM⁻¹) in the region of linear response than that with CPVC. The biosensors were applied to glucose measurement in natural samples of commercial orange juice.     

Fabrication of a covalently attached multilayer film electrode containing cobalt phthalocyanine and its application as a potentiometric sensor of iodide ion

Construction of a highly stable covalently attached multilayer film electrode containing cobalt phthalocyanine was achieved by UV irradiation of ionic self-assembled multilayer films of diazo-resins (DAR) and cobalt phthalocyanine tetrasulfonic acid (CoTsPc) tetrasodium salt. The modified electrode had good potentiometric response to iodide ion. The potentiometric response was independent of the pH of the solution between pH 2.5 and 6.0, while it was dependent on the nature of the buffer media. The modified electrode had a linear dynamic range between 4.7×10⁻⁶ M and 0.1 M with a Nernstian slope of 58.8 mV per decade and a detection limit of 3.5×10⁻⁶ M in acetate buffer (0.1 M, pH 4.6). The modified electrode also exhibited a fast response, good stability and repeatability.     

Investigation of a Biosensor Based on Phenylalanine Dehydrogenase Immobilized on a Polymer‐Blend Film for Phenylketonuria Diagnosis

We investigated a L ‐phenylalanine (L ‐phe) biosensor, functionalized through enzyme immobilization on a polymer‐blend film. The electron mediator 3,4‐dihydroxybenzaldehyde (3,4‐DHB) was employed at the electrode surface to improve direct oxidation of NADH to NAD⁺ and no additional reagents is required to be added to the sample solution. The bioactivated electrode was coated with a semi‐permeable cellulose acetate membrane in order to prevent dissolution of biofunctionalized polymer‐blend film. This constructed enzyme electrode is the first selective biosensor for phenylketonuria (PKU) detection. The sensitivity of the enzyme electrode was determined as 12.014 mA/M cm². The Michaelis–Menten and current responses as well as sensitivity of the electrode showed improved values than those of previous works. This selective biosensor presented an excellent electroanalytical response for L ‐phe, with a high steady‐state current being obtained after 20 s. The sensitivity of our biodevice is quite sufficient for the purpose of PKU detection because the reference range of clinical concern for L ‐phenylalanine concentration is C_{L ‐phe}>0.5 mM. This surface‐bioactivated enzyme electrode retained more than 80 % of its electrocatalytic activity after 16 days.     

Electrochemical synthesis, characterization and some properties of a polymer derived from thioflavin S

Polymeric films derived from thioflavin S were electrosynthesized on mild steel and silver electrodes in sulfuric acid and lithium perchlorate-containing aqueous solutions. The introduction of thioflavin S in an acidic solution protected the surface of steel from corrosion. The electrochemical behavior of the steel coated with a layer of poly(thioflavin) was examined by electrochemical impedance spectroscopy. The films exhibited a capacitive behavior and were semi-conductive in nature. Infra red reflectance measurements of the polymer films at the steel surface showed that the polymer structure retained the aromatic structure of the benzene and thiazole rings with the distinction of a nitrogen quinone vibrational band. Surface morphology of the polymer film was examined with scanning electron microscopy. The films are yellow, compact and dense when electrochemically formed onto steel surfaces when compared to a blue rather porous when formed onto silver electrode. The mechanism of electropolymerization of thioflavin is given and found similar to that of aniline with the possibility of metal chelation with the sulfur and/or nitrogen in the thiazole ring.     

Amperometric l-lactate biosensor based on screen-printed carbon electrode containing cobalt phthalocyanine,coated with lactate oxidase-mesoporous silica conjugate layer

A novel amperometric biosensor for the measurement of l-lactate has been developed. The device comprises a screen-printed carbon electrode containing cobalt phthalocyanine (CoPC-SPCE), coated with lactate oxidase (LOD) that is immobilized in mesoporous silica (FSM8.0) using a polymer matrix of denatured polyvinyl alcohol; a Nafion layer on the electrode surface acts as a barrier to interferents. The sampling unit attached to the SPCE requires only a small sample volume of 100 μL for each measurement. The measurement of l-lactate is based on the signal produced by hydrogen peroxide, the product of the enzymatic reaction. The behavior of the biosensor, LOD-FSM8.0/Naf/CoPC-SPCE, was examined in terms of pH, applied potential, sensitivity and operational range, selectivity, and storage stability. The sensor showed an optimum response at a pH of 7.4 and an applied potential of +450 mV. The determination range and the response time for l-lactate were 18.3 μM to 1.5 mM and approximately 90 s, respectively. In addition, the sensor exhibited high selectivity for l-lactate and was quite stable in storage, showing no noticeable change in its initial response after being stored for over 9 months. These results indicate that our method provides a simple, cost-effective, high-performance biosensor for l-lactate.     

Metallic and non-metallic redox response of conducting polymers

The potentiometric response of electrodes coated with polypyrrole or poly(N-methylpyrrole) films with different doping anions was studied in solutions containing the redox couples: Fe(CN)₆^3−/4−, Ru(NH₃)₆^3+/2+ and Fe(Ill)/Fe(II). The stable potential measured with the electrodes was the potential of the redox couple. The response time was instant for polypyrrole doped with dodecylsulphate ions, PPy(DS) and slow for the polymers doped with mobile anions. On the basis of electrochemical measurements and chemical analysis by EDAX spectroscopy it was found that with the PPy(DS) electrode the potentiometric response was of the ‘metallic’ type, with no change in the oxidation state of the bulk polymer. With the other polymer systems studied reduction or oxidation of the polymer bulk took place when it was in contact with a redox couple in the solution.     

A biosensor prepared by co-entrapment of a glucose oxidase and a carbon nanotube within an electrochemically deposited redox polymer multilayer

A glucose biosensor based on a nanocomposite made by layer-by-layer electrodeposition of the redox polymer into a multilayer containing glucose oxidase (GOx) and single-walled carbon nanotubes (SWCNT) on a screen-printed carbon electrode (SPCE) surface was developed. The objectives of the electrodeposition of redox polymer are to stabilize further the multilayer using a coordinative cross-linked redox polymer and to wire the GOx. The electrochemistry of the layer-by-layer assembly of the GOx/SWCNT/redox polymer nanocomposite was followed by cyclic voltammetry. The resultant biosensor provided stable and reproducible electrocatalytic responses to glucose, and the electrocatalytic current for glucose oxidation was enhanced with an increase in the number of layers. The biosensor displayed a linear range from 0.5 to 6.0mM, a sensitivity of 16.4μA/(mMcm(2)), and a response time of about 5s. It shows no response to 0.05mM of ascorbic acid, 0.32mM of uric acid and 0.20mM of acetaminophen using a Nafion membrane covering the nanocomposite-modified electrode surface.