A new polyaniline-catalase-glutaraldehyde-modified biosensor for hydrogen peroxide detection.

A new biosensor based on catalase enzyme immobilized on electrochemically constructed polyaniline (PANI) film modified with glutaraldehyde has been developed for the determination of hydrogen peroxide (H2O2) in milk samples. Assembly processes of polyaniline and immobilization of the enzyme were monitored with the help of electrochemical impedance spectroscopy. Amperometric measurements have been performed at cathodic peak (-0.3 V vs. Ag/AgCI) which was attributed to reduction of PANI. Hydrogen peroxide was determined by using amperometric method at -0.3 V. The biosensor responses were correlated linearly with the hydrogen peroxide concentrations between 5.0 × 10-6 and 1.0 × 10-4 M by amperometric method. Detection limit of the biosensor is 2.18 × 10-6 M for H2O2. In the optimization studies of the biosensor, some parameters such as optimum pH, temperature, concentration of aniline, amount of enzyme, and number of scans during electropolymerization were investigated.

Design of a multiwalled carbon nanotube-Nafion-cysteamine modified tyrosinase biosensor and its adaptation of dopamine determination.

In this work, a multiwalled carbon nanotube (MWCNT)-Nafion-cysteamine (CA) modified tyrosinase biosensor brings a new and original perspective to biosensor technology intended for the development of dopamine determination. Dopamine measurements were done at 0.2V with the amperometric method by the developed biosensor system. In addition, in this study dopamine determination was carried out by using the differential pulse voltammetry method between potentials of 0.4 and -0.15 V. In the optimization studies of the biosensor, some parameters such as optimal pH, optimal temperature, optimal enzyme amount, and effect of MWCNT concentration were investigated. Afterward, in the characterization studies, some parameters such as linearity and reproducibility were determined. In the reproducibility experiment, an average value of 1.026 µM, a standard deviation of ±0.03975, and a coefficient of variation of 3.8% were determined for a 1-µM dopamine concentration (n=15). Determination of dopamine was carried out in drug samples by the developed biosensor.

A new non-enzymatic biosensor for the determination of bisphenol-A.

In this study, a new non-enzymatic carbon paste biosensor was developed for the determination of Bisphenol-A (BPA) based on Multiwalled Carbon Nanotube (MWCNT) modified Myoglobin (Mb). The measurement principle of the biosensor was developed based on the inhibition effect of BPA on the heme group of myoglobin in the presence of hydrogen peroxide. With the designed biosensor, measurements were taken in the potential range of (-0.15 V & +0.65 V) using the differential pulse voltammetry (DPV) method in the medium containing K4[Fe(CN)6]. The linear range for BPA was determined to be 100-1000 µM. Response time was calculated as 16 s. The limit of detection was set at 89 µM. As a result, it has been proven that MWCNT modified myoglobin based biosensor is an alternative method that can be used for BPA determination, giving very sensitive and fast results.

Alkaline phosphatase based amperometric biosensor immobilized by cysteamine-glutaraldehyde modified self-assembled monolayer.

Alkaline phosphatase (ALP) was immobilized with cross-linking agents glutaraldehyde and cysteamine by forming a self-assembled monolayer on a screen printed gold electrode. ALP converts p-nitrophenyl phosphate to p-nitrophenol and phosphate. p-Nitrophenol loses H(+) ion and turns into the negatively charged compound p-nitrophenolate at medium pH. As a result, the unstable product formed is measured chronoamperometrically at an application potential of + 0.95 V. The biosensor response depends linearly on p-nitrophenyl phosphate concentration between 0.05 - 0.6 mM with a response time of 40 seconds. Detection limit of the biosensor is 0.033 mM.

Sensitive nitrate determination in water and meat samples by amperometric biosensor.

This study describes a novel biosensor method for specific determination of nitrate in food and water samples by using nitrate reductase (NR) (EC 1.9.6.1) biosensor based on the detection of oxidation peak current of redox mediator, methyl viologen, related to nitrate concentration. The method was shown to be selective and sensitive to determine the nitrate levels of water samples and processed meat samples. Immobilization procedure and also working conditions of the biosensor were optimized. Dynamic range attained with this method was established as (5.0-90.0 x 10(-9) M) for nitrate concentration with a 10 s response time. Limit of detection (LOD) and quantification (LOQ) of the biosensor were calculated as 2.2 x 10(-9) M and 5.79 x 10(-9) M, respectively. Reproducibility experiments was established on repetitive measurements by using a freshly prepared biosensor for avoiding the memory effect. The RSD was calculated as 1.22% at a nitrate concentration of 4.7 x 10(-8) M (n = 7).

A novel microbial biosensor system based on C. tropicalis yeast cells for selective determination of L-Ascorbic acid.

In this study, a novel microbial biosensor was developed for the selective determination of L-Ascorbic acid. In the construction of the microbial biosensor, lyophilized Candida tropicalis yeast cells were immobilized with o-aminophenol by forming a film layer on a platinum electrode surface using electropolymerization. L-Ascorbic acid was quantified on the basis of both amperometric and differential pulse voltammetry (DPV) methods using the biosensor. The measurements were made at +0.24 V (vs Ag/AgCl) for amperometric studies and between 0.0 V and +0.7 V for DPV studies based on the oxidation of L-Ascorbic acid to dehydro-L-Ascorbic acid by ascorbate oxidase which takes place within the catabolic metabolic pathway of C. tropicalis yeast cells. According to the results obtained from the two methods, the response of the biosensor depends linearly on L-Ascorbic acid concentration between 100 and 1500 µM. The detection limit was 62 µM and 59 µM for amperometric and DPV measurements, respectively. The response time of the microbial biosensor was 14 s and 5 s for DPV and amperometric measurements, respectively. In the optimization studies of the biosensor, some parameters such as the optimum amount of the microorganism, o-aminophenol concentration, pH and temperature were determined. For the characterization of the biosensor, reproducibility, storage stability and the effect of interferences were determined.

A novel biosensor based on activation effect of thiamine on the activity of pyruvate oxidase.

A biosensor based on pyruvate oxidase (POX) enzyme was developed for the investigation of the effect of thiamine (vitamin B(1)) molecule on the activity of the enzyme. The biosensor was prepared with a chemical covalent immobilization method on the dissolved oxygen (DO) probe by using gelatin and cross-linking agent, glutaraldehyde. POX catalyzes the degradation of pyruvate to acetylphosphate, CO(2) and H(2)O(2) in the presence of phosphate and oxygen. Thiamine is an activator for POX enzyme and determination method of the biosensor was based on this effect of thiamine on the activity of the enzyme. The biosensor responses showed increases in the presence of thiamine. Increases in the biosensor responses were related to thiamine concentration. Thiamine determination is based on the assay of the differences on the biosensor responses on the oxygenmeter in the absence and the presence of thiamine. The biosensor response depend linearly on thiamine concentration between 0.025 and 0.5 microM with 2 min response time. In the optimization studies of the biosensor the most suitable enzyme amount was found as 2.5 U cm(-2) and also phosphate buffer (pH 7.0; 50 mM) and 35 degrees C were obtained as the optimum working conditions. In the characterization studies of the biosensor some parameters such as activator and interference effects of some substances on the biosensor response and reproducibility were carried out.

Sensitive determination of L-lysine with a new amperometric microbial biosensor based on Saccharomyces cerevisiae yeast cells.

A new amperometric microbial biosensor based on Saccharomyces cerevisiae NRRL-12632 cells, which had been induced for lysine oxidase enzyme and immobilized in gelatin by a cross-linking agent was developed for the sensitive determination of L-lysine amino acid. To construct the microbial biosensor S. cerevisiae cells were activated and cultured in a suitable culture medium. By using gelatine (8.43 mg cm(-2)) and glutaraldehyde (0.25%), cells obtained in the logarithmic phase of the growth curve at the end of a 14 h period were immobilized and fixed on a pretreated oxygen sensitive Teflon membrane of a dissolved oxygen probe. The assay procedure of the microbial biosensor is based on the determination of the differences of the respiration activity of the cells on the oxygenmeter in the absence and the presence of L-lysine. According to the end point measurement technique used in the experiments it was determined that the microbial biosensor response depended linearly on L-lysine concentrations between 1.0 and 10.0 microM with a 1 min response time. In optimization studies of the microbial biosensor, the most suitable microorganism quantities were found to be 0.97x10(5)CFU cm(-2). In addition phosphate buffer (pH 7.5; 50 mM) and 30 degrees C were obtained as the optimum working conditions. In characterization studies of the microbial biosensor some parameters such as substrate specificity, interference effects of some substances on the microbial biosensor responses, reproducibility of the biosensor and operational and storage stability were investigated.

An amperometric microbial biosensor development based on Candida tropicalis yeast cells for sensitive determination of ethanol.

Different branchs of industry need to use ethanol in their production and some progress and not only the industry also to determine ethanol sensitively, accurately, fast and economical is very important. For the sensitive determination of ethanol a new amperometric biosensor based on Candida tropicalis cells, which contains alcohol oxidase enzyme, immobilized in gelatin by using glutaraldehyde was developed. In the study, before the microbial biosensor construction C. tropicalis cells were activated and cultured in a culture medium. By using gelatine and glutaraldehyde (0.1%) C. tropicalis cells obtained in logarithmic phase were immobilized and fixed on a pretreated teflon membrane of a dissolved oxygen probe. Ethanol determination is based on the assay of the differences on the respiration activity of the cells on the oxygenmeter in the absence and the presence of ethanol. The microbial biosensor response was depend linearly on ethanol concentration between 0.5 and 7.5 mM with 2 min response time. In the optimization studies of the microbial biosensor the most suitable microorganism amount were found as 4.42 mg cm(-2) and also phosphate buffer (pH:7.5; 50 mM) and 30 degrees C were obtained as the optimum working conditions. In the characterization studies of the microbial biosensor some parameters such as substrate specificity, interference effects of some substances on the biosensor response, operational and storage stability were carried out.

MWCNT-cysteamine-Nafion modified gold electrode based on myoglobin for determination of hydrogen peroxide and nitrite.

In this work, a novel amperometric biosensor of hydrogen peroxide (H2O2) was developed based on the immobilization of myoglobin (Mb) on the surface of the multi-walled carbon nanotube (MWCNT) -Nafion-cysteamine (CA) modified gold electrode (Au) and its electrocatalytic activity was used for the determination of nitrite (NO2(-)). In the optimization studies, the best MWCNT and myoglobin amount were investigated. It was discovered at the experiments for the optimization of the working conditions that the buffer at this study as 50.0mM, pH7.0 phosphate buffer (PBS) and working temperature as 30°C for the H2O2 biosensor. It was determined at the characterization studies on the biosensor that linear results are obtained between the ranges of 0.1µM to 70.0µM for H2O2 concentration and 1-250µM for NO2(-). The reproducibility of the biosensor was determined both H2O2 and nitrite. From the experiments, average value, standard deviation (SD) and coefficients of variation (CV%) were calculated to be 10.02±0.43µM, and 4.29% for 10.0µM H2O2 (n=6) and 52.0±2.1µM, and 3.89% for 50.0µM nitrite (n=8), respectively. At the same time the sample was analyzed for NO2(-) in drinking and mineral waters.

A microbial biosensor based on Lactobacillus delbruecki sp. bacterial cells for simultaneous determination of lactic and pyruvic acid.

The aim of this study was mainly to develop a microbial biosensor for the simultaneous determination of lactic acid and pyruvic acid. In developing biosensor, lyophilised Lactobacillus delbruecki sp. bacterial cells were immobilised with polypyrrole on a platin electrode surface using electropolymerization method. Lactate concentration was determined based on the differences in amperometric responses at cathodic peak (+0.2V) of potassium ferricyanide, whereas pyruvate concentration was determined using the differences at anodic peak (+0.1V). The response of biosensor showed linearity between 0.1 and 1.0mM for both of two substrates. Optimisation studies were carried out for amount of microorganism, pyrrole concentration, pH and temperature. In the characterisation studies, substrate specificity, interference effect of some substances on the biosensor response, and storage stability were established.

A new pyruvate oxidase biosensor based on 3-mercaptopropionic acid/6-aminocaproic acid modified gold electrode.

In the biosensor construction, 3-mercaptopropionic acid (3-MPA) and 6-aminocaproic acid (6-ACA) were used for forming self-assembled monolayer (SAM) on a gold disc electrode and pyruvate oxidase was immobilized on the modified electrode surface by using glutaraldehyde. Biosensor response is linearly related to pyruvate concentration at 2.5-50 µM, detection limit is 1.87 µM and response time of the biosensor is 6 s for differential pulse voltammograms. From the repeatability studies (n = 6) for 30.0 µM pyruvate revealed that the average value ([Formula: see text]), standard deviation (S.D) and coefficient of variation (CV %) were calculated to be 31.02 µM, ± 0.1914 µM and 0.62%, respectively.

Cysteamine-palladium complex ([Pd(µ-OAc)(ppy)]2, ppy:2-phenylpyridine, PhMe)-modified peroxidase biosensor immobilized on a gold electrode.

A new peroxidase biosensor was developed using cysteamine-palladium complex-modified gold electrode. The principle of the measurements is based on monitoring increase in the oxidation potential of palladium complex (at + 0.47 V vs Ag/AgCl) using amperometric detection. In the optimization studies of the biosensor, effects of enzyme amount, palladium complex amount, and duration of SAM formation on biosensor responses were investigated to optimize the bioactive layer. The biosensor has a fast response time of less than 10 s to hydrogen peroxide (H2O2), with a linear range of 5.0 × 10(- 6) to 150 × 10(- 6) M and a detection limit of 3.38 × 10(- 6) M.

A novel non-competitive amperometric immunosensor by using thiourea-glutaraldehyde-modified gold electrode for immunoglobulin M detection.

A novel non-competitive amperometric immunosensor based on a self-assembled monolayer (SAM) of thiourea modified by a polymeric Schiff's base of glutaraldehyde on gold electrode has been developed for determination of IgM. Alkaline phosphatase (ALP)-conjugated monoclonal anti-mouse immunoglobulin M (IgM) antibody was selectively bound to IgM molecules onto the surface of the electrode. Electrochemical response arising from the catalytic reaction of alkaline phosphatase enzyme. Its reaction with various phosphates such as p-aminophenyl phosphate and p-nitrophenyl phosphatase (p-NPP) generates the electrochemically active products p-aminophenol (p-AP) and p-nitrophenol (p-NP), respectively.

Voltammetric determination of epinephrine by White rot fungi (Phanerochaete chrysosporium ME446) cells based microbial biosensor.

The lyophilized biomass of White rot fungi (Phanerochaete chrysosporium ME446) was immobilized in gelatine using glutaraldehyde crosslinking agent on a Pt working electrode. The fungal cells retained their laccase activity under entrapped state. The immobilized cells were used as a source of laccase to develop amperometric epinephrine biosensor. The catalytic action of the laccase in the biosensor released an epinephrinequinone as a result of redox activity, thereby causing an increase in the current. The optimal working conditions of the biosensor were carried out at pH 4.5 (50 mM acetate buffer containing 100 mM K(3)Fe(CN)(6)), and 20°C. The sensor response was linear over a range of 5-100 µM epinephrine. The detection limit of the biosensor was found to be 1.04 µM. In the optimization and characterization studies of the microbial biosensor some parameters such as effect of fungi and gelatine amount, percentage of glutaraldehyde on the biosensor response and substrate specificity were carried out. In the application studies of the biosensor, sensitive determination of epinephrine in pharmaceutical ampules was investigated.

Preparation and optimization of a bienzymic biosensor based on self-assembled monolayer modified gold electrode for alcohol and glucose detection.

The aim of this project was to develop a bienzymic biosensor, which was based on co-immobilization of alcohol oxidase and glucose oxidase on the same electrode by formation of self-assembled monolayer (SAM) for selective determination of ethanol and glucose. In the biosensor construction the enzymes and the mediator, tetrathiafulvalene (TTF), were immobilized with cross-linking agents glutaraldehyde and cysteamine by forming a self-assembled monolayer (SAM) on a gold disc electrode. Amounts of ethanol and glucose were amperometrically detected by monitoring current values at reduction potential of TTF(+), 0.1V. Decreases in biosensor responses were linearly related to glucose concentrations between 0.1 and 1.0 mM and ethanol concentrations between 1.0 and 10 mM. Limits of detection of the biosensor for ethanol and glucose were calculated to be 0.75 and 0.03 mM, respectively. In the optimization studies of the biosensor some parameters such as optimum pH, optimum temperature, enzyme amount, effect of TTF concentration and duration of SAM formation were investigated.

Do copper ions activate tyrosinase enzyme? A biosensor model for the solution.

Some metal ions play a cofactor role for the activity of tyrosinase enzyme and one of them is copper ion. In this study an amperometric biosensor was developed in order to investigate the effect of the copper ions on the activity of tyrosinase enzyme. In the construction of the biosensor tyrosinase enzyme was immobilized on a Clark-type dissolved oxygen probe which was covered with a oxygen sensitive teflon membrane, by using a chemical covalent immobilization method based on gelatine and bifunctional reagent, glutaraldehyde. The principle of the measurement was based on the determination of the differentiation of dissolved oxygen level in the enzymatic reaction catalyzed by tyrosinase in the absence and the presence of copper ions. Differences between the dissolved oxygen concentrations were related to copper ion concentration which was added in to the reaction medium. The biosensor response depends linearly on copper ion concentration between 2.5-20.0microM with a response time 1min. The detection limit of the biosensor is 0.95microM. In the optimization studies of the biosensor, the most suitable amounts of tyrosinase, gelatin and glutaraldehyde ratio were determined to be 69.0U/cm(-2), 4.21mg/cm(-2), and 2.5%, respectively. In the optimization studies of the biosensor, phosphate buffer (pH 7.0 ,50mM) and 30 degrees C were detected to be working conditions. For the characterization of the biosensor some parameters such as reproducibility, thermal and pH stability were carried out.

A mediated polyphenol oxidase biosensor immobilized by electropolymerization of 1,2-diamino benzene.

A biosensor based on a partially purified polyphenol oxidase (PPO) enzyme was developed by using electropolymerization of [(2,2'-bipyridine)(chloro)(p-cymene)rutenium(II)]chloride] mediator complex and 1,2-diamino benzene (DAB) on a screen printing Pt electrode (1mm diameter). The electropolymerization was carried out at +0.7V for 45min in phosphate buffer (50mM, pH 7.0) which contained 14.0U/10mL polyphenole oxidase, 200mM DAB and 2.5mM Ru-mediator complex solutions. Measurement is based on the detection of the oxidation current of the Ru-mediator complex that related to the enzymatic reaction catalyzed by PPO at +0.65V. The phosphate buffer (50mM, pH 7.0 containing 0.1M KCl) and 30 degrees C were established as being the optimum working conditions. Under the optimum experimental conditions a linear calibration curve was obtained between 5 and 100microM catechol concentration. The detection limit of the biosensor is 2.385microM. In the characterization studies of the biosensor some parameters such as effect of Ru-mediator types on the biosensor response, substrate specificity, reproducibility and storage stability were studied. From the experiments, the average value (x), standard deviation (SD) and coefficient of variation (CV%) were found to be 48.75microM,+/-1.56microM, and 3.2% respectively for 50microM catechol standard.

An inhibition type amperometric biosensor based on tyrosinase enzyme for fluoride determination.

In this study, a new biosensor based on the inhibition of tyrosinase for the determination of fluoride is described. To construct the biosensor tyrosinase was immobilized by using gelatine and cross-linking agent glutaraldehyde on a Clark type dissolved oxygen (DO) probe covered with a teflon membrane which is sensitive for oxygen. The phosphate buffer (50mM, pH 7.0) at 30 degrees C were established as providing the optimum working conditions. The method is based on the measurement of the decreasing of dissolved oxygen level of the interval surface that related to fluoride concentration added into reaction medium in the presence of catechol. Inhibitor effect of fluoride results in decrease in dissolved oxygen concentration. The biosensor response depends linearly on fluoride concentration between 1.0 and 20 microM with a response time of 3 min. In the characterization studies of the biosensor some parameters such as reproducibility, substrate specificity and storage stability were carried out. From the experiments, the average value (x), Standard deviation (S.D) and coefficient of variation (C.V %) were found as 10.5 microM, +/-0.57 microM, 5.43%, respectively for 10 microM fluoride standard.

Whole cell immobilized amperometric biosensor based on Saccharomyces cerevisiae for selective determination of vitamin B1 (thiamine).

A new amperometric whole cell biosensor based on Saccharomyces cerevisiae immobilized in gelatin was developed for selective determination of vitamin B1 (thiamine). The biosensor was constructed by using gelatin and crosslinking agent glutaraldehyde to immobilize S. cerevisiae cells on the Teflon membrane of dissolved oxygen (DO) probe used as the basic electrode system combined with a digital oxygen meter. The cells were induced by vitamin B1 in the culture medium, and the cells used it as a carbon source in the absence of glucose. So, when the vitamin B1 solution is injected into the whole cell biosensor system, an increase in respiration activity of the cells results from the metabolic activity and causes a decrease in the DO concentration of interval surface of DO probe related to vitamin B1 concentration. The response time of the biosensor is 3 min, and the optimal working conditions of the biosensor were carried out as pH 7.0, 50mM Tris-HCl, and 30 degrees C. A linear relationship was obtained between the DO concentration decrease and vitamin B1 concentration between 5.0 x 10(-3) and 10(-1) microM. In the application studies of the biosensor, sensitive determination of vitamin B1 in the vitamin tablets was investigated.