Este trabalho descreve a preparação, a caracterização e o uso de um biossensor de pasta de carbono modificado com a enzima acetilcolinesterase para a quantificação de carbamatos em alimentos, bem como o estudo das constantes de velocidade para a reação enzima/substrato (iodeto de acetiltiocolina) em solução. Inicialmente foram realizados testes de estabilidade, tanto para o substrato quanto para a enzima. Nestes testes, a absorção na região do UV-vis do substrato não apresentou diminuição significativa em 11 dias de análises, já a enzima apresentou uma grande perda de sua atividade com apenas três dias de preparo da solução. Na preparação do eletrodo de trabalho alguns parâmetros foram otimizados, tais como: quantidade de enzima e de ftalocianina de cobalto (CoPC) no eletrodo, bem como a porcentagem de glutaraldeído utilizada. A melhor resposta ocorreu para adição de 2,40x10-3g de enzima, 0,90x10-3g de CoPC (referentes à massa de 0,017g de pasta de carbono) e solução de glutaraldeido 1%. A seguir, realizou-se um experimento baseado na inibição da atividade da enzima, causada pela imersão do eletrodo na solução contendo o pesticida carbaril nas concentrações 5,00x10-5 e 1,00x10-4 mol L-1. Notou-se que, com o aumento da concentração do carbaril, houve aumento na inibição da atividade enzimática. Desta forma o eletrodo apresentou-se apto para determinação analítica de pesticidas. Estas medidas foram realizadas em meio de tampão fosfato 0,1 mol L-1, pH 7,4 e com tempo de incubação para o carbaril, metomil e aldicarbe foram de 8, 12 e 15 minutos, respectivamente. Os limites de detecção (LD) e quantificação (LQ) obtidos utilizando-se o biossensor amperométrico para o carbaril foram de 2,00x10-6 mol L-1 (0,40 mg L-1) e 6,70 x 10-6 mol L-1 (1,30 mg L-1), para o metomil de 1,88 x 10-7mol L-1 (30,45 micro g L-1) e 6,26 x 10-7 mol L-1 (0,10 mg / L-1) e para o aldicarbe de 1,10x10-6 mol L-1 (0,20 mg L-1) e 3,60x10-6 mol L-1 (0,70 mg L-1). Para a formulação comercial Lannate (metomil) os LD e LQ foram 2,13x10-7 mol L-1 (34,50 micro g L-1) e 7,09x10-7 mol L-1(0,12 mg L-1), respectivamente. As medidas de HPLC apresentaram LD e LQ de 1,58 x 10-8 mol L-1 (3,18 micro g L-1) e 5,27x10-8 mol L-1 (10,60 micro g L-1) para o carbaril e de 9,02 x 10-10 mol L-1 (0,15 micro g L-1) e 3,00 x 10-9 mol L-1 (48,60 micro g L-1) para o metomil. Testes de recuperação foram realizados usando ambas as técnicas para o carbaril e Lannate. As recuperações utilizando-se o biossensor mostraram-se eficientes, variando de 76,83 a 106,67% para o carbaril e de 78,00 a 96,50% para a Lannate, enquanto que nas medidas de HPLC, as recuperações foram de 78,00 a 108,33% para o carbaril e de 57,00 a 99,50% para o Lannate. A recuperação para o aldicarbe no tomate foi de 62,40 %. As análises da enzima em solução mostraram que a metodologia empregada neste estudo é adequada para a determinação das constantes de velocidade para a etapa lenta da reação AchE/AchI. Observou-se que os valores destas constantes são dependentes da concentração dos pesticidas fenitrothion (organofosforado) e carbaril (carbamato), em baixa concentração ambos apresentaram constantes de velocidade similares, mas com o aumento dessa concentração, o fenitrothion apresentou menor constante de velocidade em relação ao carbaril, sugerindo que este apresenta maior inibição da enzima e por conseqüência maior toxicidade no organismo. Esses resultados mostraram uma possível metodologia analítica para a quantificação destes pesticidas, obtendo-se os valores das constantes de velocidade enzimática e suas dependências com as concentrações dos pesticidas em solução. / This work describes the development, characterization and utilization of a carbon paste biosensor based in the acetylcholinesterase enzyme for carbamates determinations in foodstuff, as well as the study of rate constants for enzyme/substrate reaction in solution. Stability tests were initially performed for both the substrate and the enzyme. In these tests, the signal for UV-vis adsorption for the substrate shows no inhibition during 11 days while for the enzyme it has been demonstrated that a considerable loss of activity occurs after three days from the solution preparation. In the electrode preparation, some experimental parameters were optimized, such as the amount of enzyme and the content of cobalt ftalocyanine (CoPC) in the electrode, as well as the employed percentage of glutaraldehide. The highest analytical signals were obtained for the addition of 2.40x10-3 g enzyme, 0.90x10-3 g CoPC (related to the massa of 0,017g of carbon paste) and a 1% glutaraldehide solution. The next step was to carry out an experiment based in the inhibition of enzyme activity by the pesticide. For this, the biosensor was immersed in 5.00x10-5 e 1.00x10-4 mol L-1 carbaryl solutions. It was observed that, by increasing the carbaryl concentration, the electrochemical signal of the sensor was inhibited proportionally. This was indicative that the sensor was adequate to be used in carbaryl monitoring and analytical determinations. The analytical determinations of carbamate pesticides were performed in 0.1 mol L-1 phosphate buffer, pH 7,4, with incubation time of 8, 12 and 15 minutes for carbaryl, metomil and aldicarb, respectively. The detection (LD) and quantification (LQ) limits obtained with the biosensor were 2.00x10-6 mol L-1 (0.40 mg L-1) and 6.70 x 10-6 mol L-1 (1.30 mg L-1) for carbaryl, 1.88x10-7mol L-1 (30.45 micro g L-1) and 6.26x10-7 mol L-1 (0.10 mg / L-1) for metomil and 1.10x10-6 mol L-1 (0.20 mg L-1) and 3.60x10-6 mol L-1 (0.70 mg L-1) for aldicarb. For the commercial formulation of metomil, Lannate, LD and LQ obtained were 2.13x10-7 mol L-1 (34.50 microg L-1) and 7.09x10-7 mol L-1(0.12 mg L-1), respectively. The HPLC measurements showed LD and LQ of 1.58x10-8 mol L-1 (3.18micro g L-1) and 5.27x10-8 mol L-1 (10.60 micro g L-1) for carbaryl and 9.02x10-10 mol L-1 (0.15 micro g L-1) and 3.00x10-9 mol L-1 (48.60 micro g L-1) for metomil. Recovering tests were also done with both analytical techniques for carbaryl and Lannate. The obtained recoveries using the biosensor were in the range of 76.83 to 106.67% for carbaryl and 78.00 to 96.50% for Lannate, while using the HPLC, the recoverings were 78.00 a 108.33% for carbaryl and 57.00 to 99.50% for Lannate. The recovering of aldicarb in tomatoes, with HPLC, were 62.40 %. The study of the enzymatic reaction in solution showed that the employed methodology allows to obtain the rate constant values for the rate determining step of the AchE/AchI reaction. It was observed that these rate constant values were strongly dependent in the pesticide concentrations for fenitrothion (organofosforous) and carbaryl (carbamate). At low concentration levels of the pesticide in the electrolyte, all the rate constants showed similar values but, when the pesticide concentration was raised, fenitrothion was found to exert a more powerful inhibition action for the enzyme activity than carbaryl, thus suggesting its higher toxic character. These results showed the development of a possible analytical methodology for quantification of these pesticides, by calculating the rate constant value and its dependence to the pesticide concentration in solution.

Este trabalho descreve a preparação, a caracterização e o uso de um biossensor de pasta de carbono modificado com a enzima acetilcolinesterase para a quantificação de carbamatos em alimentos, bem como o estudo das constantes de velocidade para a reação enzima/substrato (iodeto de acetiltiocolina) em solução. Inicialmente foram realizados testes de estabilidade, tanto para o substrato quanto para a enzima. Nestes testes, a absorção na região do UV-vis do substrato não apresentou diminuição significativa em 11 dias de análises, já a enzima apresentou uma grande perda de sua atividade com apenas três dias de preparo da solução. Na preparação do eletrodo de trabalho alguns parâmetros foram otimizados, tais como: quantidade de enzima e de ftalocianina de cobalto (CoPC) no eletrodo, bem como a porcentagem de glutaraldeído utilizada. A melhor resposta ocorreu para adição de 2,40x10-3g de enzima, 0,90x10-3g de CoPC (referentes à massa de 0,017g de pasta de carbono) e solução de glutaraldeido 1%. A seguir, realizou-se um experimento baseado na inibição da atividade da enzima, causada pela imersão do eletrodo na solução contendo o pesticida carbaril nas concentrações 5,00x10-5 e 1,00x10-4 mol L-1. Notou-se que, com o aumento da concentração do carbaril, houve aumento na inibição da atividade enzimática. Desta forma o eletrodo apresentou-se apto para determinação analítica de pesticidas. Estas medidas foram realizadas em meio de tampão fosfato 0,1 mol L-1, pH 7,4 e com tempo de incubação para o carbaril, metomil e aldicarbe foram de 8, 12 e 15 minutos, respectivamente. Os limites de detecção (LD) e quantificação (LQ) obtidos utilizando-se o biossensor amperométrico para o carbaril foram de 2,00x10-6 mol L-1 (0,40 mg L-1) e 6,70 x 10-6 mol L-1 (1,30 mg L-1), para o metomil de 1,88 x 10-7mol L-1 (30,45 micro g L-1) e 6,26 x 10-7 mol L-1 (0,10 mg / L-1) e para o aldicarbe de 1,10x10-6 mol L-1 (0,20 mg L-1) e 3,60x10-6 mol L-1 (0,70 mg L-1). Para a formulação comercial Lannate (metomil) os LD e LQ foram 2,13x10-7 mol L-1 (34,50 micro g L-1) e 7,09x10-7 mol L-1(0,12 mg L-1), respectivamente. As medidas de HPLC apresentaram LD e LQ de 1,58 x 10-8 mol L-1 (3,18 micro g L-1) e 5,27x10-8 mol L-1 (10,60 micro g L-1) para o carbaril e de 9,02 x 10-10 mol L-1 (0,15 micro g L-1) e 3,00 x 10-9 mol L-1 (48,60 micro g L-1) para o metomil. Testes de recuperação foram realizados usando ambas as técnicas para o carbaril e Lannate. As recuperações utilizando-se o biossensor mostraram-se eficientes, variando de 76,83 a 106,67% para o carbaril e de 78,00 a 96,50% para a Lannate, enquanto que nas medidas de HPLC, as recuperações foram de 78,00 a 108,33% para o carbaril e de 57,00 a 99,50% para o Lannate. A recuperação para o aldicarbe no tomate foi de 62,40 %. As análises da enzima em solução mostraram que a metodologia empregada neste estudo é adequada para a determinação das constantes de velocidade para a etapa lenta da reação AchE/AchI. Observou-se que os valores destas constantes são dependentes da concentração dos pesticidas fenitrothion (organofosforado) e carbaril (carbamato), em baixa concentração ambos apresentaram constantes de velocidade similares, mas com o aumento dessa concentração, o fenitrothion apresentou menor constante de velocidade em relação ao carbaril, sugerindo que este apresenta maior inibição da enzima e por conseqüência maior toxicidade no organismo. Esses resultados mostraram uma possível metodologia analítica para a quantificação destes pesticidas, obtendo-se os valores das constantes de velocidade enzimática e suas dependências com as concentrações dos pesticidas em solução. / This work describes the development, characterization and utilization of a carbon paste biosensor based in the acetylcholinesterase enzyme for carbamates determinations in foodstuff, as well as the study of rate constants for enzyme/substrate reaction in solution. Stability tests were initially performed for both the substrate and the enzyme. In these tests, the signal for UV-vis adsorption for the substrate shows no inhibition during 11 days while for the enzyme it has been demonstrated that a considerable loss of activity occurs after three days from the solution preparation. In the electrode preparation, some experimental parameters were optimized, such as the amount of enzyme and the content of cobalt ftalocyanine (CoPC) in the electrode, as well as the employed percentage of glutaraldehide. The highest analytical signals were obtained for the addition of 2.40x10-3 g enzyme, 0.90x10-3 g CoPC (related to the massa of 0,017g of carbon paste) and a 1% glutaraldehide solution. The next step was to carry out an experiment based in the inhibition of enzyme activity by the pesticide. For this, the biosensor was immersed in 5.00x10-5 e 1.00x10-4 mol L-1 carbaryl solutions. It was observed that, by increasing the carbaryl concentration, the electrochemical signal of the sensor was inhibited proportionally. This was indicative that the sensor was adequate to be used in carbaryl monitoring and analytical determinations. The analytical determinations of carbamate pesticides were performed in 0.1 mol L-1 phosphate buffer, pH 7,4, with incubation time of 8, 12 and 15 minutes for carbaryl, metomil and aldicarb, respectively. The detection (LD) and quantification (LQ) limits obtained with the biosensor were 2.00x10-6 mol L-1 (0.40 mg L-1) and 6.70 x 10-6 mol L-1 (1.30 mg L-1) for carbaryl, 1.88x10-7mol L-1 (30.45 micro g L-1) and 6.26x10-7 mol L-1 (0.10 mg / L-1) for metomil and 1.10x10-6 mol L-1 (0.20 mg L-1) and 3.60x10-6 mol L-1 (0.70 mg L-1) for aldicarb. For the commercial formulation of metomil, Lannate, LD and LQ obtained were 2.13x10-7 mol L-1 (34.50 microg L-1) and 7.09x10-7 mol L-1(0.12 mg L-1), respectively. The HPLC measurements showed LD and LQ of 1.58x10-8 mol L-1 (3.18micro g L-1) and 5.27x10-8 mol L-1 (10.60 micro g L-1) for carbaryl and 9.02x10-10 mol L-1 (0.15 micro g L-1) and 3.00x10-9 mol L-1 (48.60 micro g L-1) for metomil. Recovering tests were also done with both analytical techniques for carbaryl and Lannate. The obtained recoveries using the biosensor were in the range of 76.83 to 106.67% for carbaryl and 78.00 to 96.50% for Lannate, while using the HPLC, the recoverings were 78.00 a 108.33% for carbaryl and 57.00 to 99.50% for Lannate. The recovering of aldicarb in tomatoes, with HPLC, were 62.40 %. The study of the enzymatic reaction in solution showed that the employed methodology allows to obtain the rate constant values for the rate determining step of the AchE/AchI reaction. It was observed that these rate constant values were strongly dependent in the pesticide concentrations for fenitrothion (organofosforous) and carbaryl (carbamate). At low concentration levels of the pesticide in the electrolyte, all the rate constants showed similar values but, when the pesticide concentration was raised, fenitrothion was found to exert a more powerful inhibition action for the enzyme activity than carbaryl, thus suggesting its higher toxic character. These results showed the development of a possible analytical methodology for quantification of these pesticides, by calculating the rate constant value and its dependence to the pesticide concentration in solution.

Darbo tikslas buvo ištirti nuo pirolo chinolinchinono (PQQ) priklausomų oksidoreduktazių veikimą homogeninėje ir heterogeninėje aplinkose ir jų pagrindu sukurti naujas bioelektrokatalizines sistemas. Tuo tikslu buvo kurtos impedimetrinės ir bioamperometrinės sistemos su nuo PQQ priklausomomis alkoholio (tADH ir mADH), gliukozės (GDH) ir fruktozės (FDH)dehidrogenazėmis naujų elektrodinių medžiagų pagrindu, parenkant optimalų fermento imobilizavimo būdą ir elektronų pernašos (EP) mediatorius. Fermentai ir sistemos tirtos klasikinės elektrochemijos, impedanso spektroskopijos, spektrofotometrijos, atominės jėgos mikroskopijos metodais. Nustatytos tADH ir mADH pKa reikšmės ir EP keliai, pagrindinės bioelektrocheminių sistemų charakteristikos, įvertinta heterogeninės aplinkos įtaka fermentų specifiškumui. Sukurtas imobilizavimo būdas leidžiantis išlaikyti GDH aktyvumą iki 9-ių mėnesių. Pirmą kartą parodyta, jog fermentai gali tiesiogiai re-oksiduotis ant poli - N - (N, N‘- dietil ditiokarbamoil etil amidoetil) – anilino ir grafito produktų. Nustatyta, jog 2-(3-nitro(fenil)amino)- cikloheksa-2,5-dien-1,4-dionas iki 10 kartų pagreitina FDH katalizuojamą fruktozės oksidaciją. Pirmą kartą parodyta, kad bioamperometrinės sistemos su FDH gali oksiduoti D(-)tagatozę. Tyrimų rezultatai pritaikyti kuriant alkoholių, angliavandenių ir anglies monoksido stebėjimo sistemas. / The aim of this work was to study the action of pyrroloquinoline quinone (PQQ) dependent oxidoreductases in homogeneous and heterogeneous ambiences and to create new bioelectrocatalytic systems based on these enzymes. Bioelectrochemical systems with PQQ dependent alcohol (sADH and mADH), glucose (GDH) and fructose (FDH) dehydrogenases were constructed by using new electrode materials, enzyme immobilization techniques and electron transfer (ET) mediators. Enzymes and systems were studied by different electrochemical methods and atomic force microscopy. pKa values and ET pathways in bioelectrochemical systems were determined for sADH and mADH. The main characteristics of systems and influence of heterogeneous ambience to the specificity of the enzymes were determined. The GDH immobilization method, which ensures enzyme activity up to 9 months, was created. The direct ET from reduced enzymes active sites to poly(N-(N’,N’-diethyldithiocarbamoylethylamidoethyl)aniline) and graphite oxidation products was revealed for the first time. It was observed that 2-(3-nitro(phenyl)amino)- ciclohexa-2,5-dien-1,4-dione is the most effective mediator for FDH. The ability of bioamperometric systems with FDH to oxidize D(-)tagatose was determined for the first time. It was shown, that bioamperometric systems based on PQQ dependent enzymes can be applied for detection of alcohols, carbohydrates and carbon monoxide.

Darbo tikslas buvo ištirti nuo pirolo chinolinchinono (PQQ) priklausomų oksidoreduktazių veikimą homogeninėje ir heterogeninėje aplinkose ir jų pagrindu sukurti naujas bioelektrokatalizines sistemas. Tuo tikslu buvo kurtos impedimetrinės ir bioamperometrinės sistemos su nuo PQQ priklausomomis alkoholio (tADH ir mADH), gliukozės (GDH) ir fruktozės (FDH)dehidrogenazėmis naujų elektrodinių medžiagų pagrindu, parenkant optimalų fermento imobilizavimo būdą ir elektronų pernašos (EP) mediatorius. Fermentai ir sistemos tirtos klasikinės elektrochemijos, impedanso spektroskopijos, spektrofotometrijos, atominės jėgos mikroskopijos metodais. Nustatytos tADH ir mADH pKa reikšmės ir EP keliai, pagrindinės bioelektrocheminių sistemų charakteristikos, įvertinta heterogeninės aplinkos įtaka fermentų specifiškumui. Sukurtas imobilizavimo būdas leidžiantis išlaikyti GDH aktyvumą iki 9-ių mėnesių. Pirmą kartą parodyta, jog fermentai gali tiesiogiai re-oksiduotis ant poli - N - (N, N‘- dietil ditiokarbamoil etil amidoetil) – anilino ir grafito produktų. Nustatyta, jog 2-(3-nitro(fenil)amino)- cikloheksa-2,5-dien-1,4-dionas iki 10 kartų pagreitina FDH katalizuojamą fruktozės oksidaciją. Pirmą kartą parodyta, kad bioamperometrinės sistemos su FDH gali oksiduoti D(-)tagatozę. Tyrimų rezultatai pritaikyti kuriant alkoholių, angliavandenių ir anglies monoksido stebėjimo sistemas. / The aim of this work was to study the action of pyrroloquinoline quinone (PQQ) dependent oxidoreductases in homogeneous and heterogeneous ambiences and to create new bioelectrocatalytic systems based on these enzymes. Bioelectrochemical systems with PQQ dependent alcohol (sADH and mADH), glucose (GDH) and fructose (FDH) dehydrogenases were constructed by using new electrode materials, enzyme immobilization techniques and electron transfer (ET) mediators. Enzymes and systems were studied by different electrochemical methods and atomic force microscopy. pKa values and ET pathways in bioelectrochemical systems were determined for sADH and mADH. The main characteristics of systems and influence of heterogeneous ambience to the specificity of the enzymes were determined. The GDH immobilization method, which ensures enzyme activity up to 9 months, was created. The direct ET from reduced enzymes active sites to poly(N-(N’,N’-diethyldithiocarbamoylethylamidoethyl)aniline) and graphite oxidation products was revealed for the first time. It was observed that 2-(3-nitro(phenyl)amino)- ciclohexa-2,5-dien-1,4-dione is the most effective mediator for FDH. The ability of bioamperometric systems with FDH to oxidize D(-)tagatose was determined for the first time. It was shown, that bioamperometric systems based on PQQ dependent enzymes can be applied for detection of alcohols, carbohydrates and carbon monoxide.

Submitted by Maria Medeiros (maria.dilva1@ufcg.edu.br) on 2018-07-09T12:54:25Z No. of bitstreams: 1 THAÍS MARIA ALVES MARINHO - DISSERTAÇÃO (PPGCEMat) 2016.pdf: 4526314 bytes, checksum: 93577e66a4279c33e84cb662d3ce4735 (MD5) / Made available in DSpace on 2018-07-09T12:54:25Z (GMT). No. of bitstreams: 1 THAÍS MARIA ALVES MARINHO - DISSERTAÇÃO (PPGCEMat) 2016.pdf: 4526314 bytes, checksum: 93577e66a4279c33e84cb662d3ce4735 (MD5) Previous issue date: 2016-03-08 / Capes / Com o ascendente desenvolvimento de novas tecnologias, a procura por novos sistemas de sensoriamento vem aumentando, pois há uma necessidade de obter uma melhor precisão nos resultados de análises biológicas, com baixo custo e em tempo real, diante disso, os biossensores são dispositivos capazes de proporcionar esse tipo de resultado. Doenças ligadas a insuficiência renal necessitam desse tipo de precisão e rapidez, que é caracterizada pela concentração de ureia no sangue ou na urina, eliminado assim uma série de procedimentos laboratoriais. Logo, para montagem de um biossensor de ureia é preciso um biocomponente seletivo e específico, que possa identificar a ureia sem nenhuma interferência, como é o caso da enzima. A enzima capaz de catalisar a hidrolise de ureia é a uréase, formando como produto da reação o gás carbônico e a amônia. Para ser aplicado em um biossensor, a enzima precisa ser imobilizada, pois dessa forma confere uma maior estabilidade operacional e um maior armazenamento da mesma. Essa imobilização pode ser realizada por meio de polímeros. A quitosana é um polímero natural que vêm ganhando destaque para esta aplicação, diante de suas propriedades como baixo custo, abundância na natureza, fácil processamento, capacidade de formação membrana, entre outras. Diante do exposto, o objetivo desse trabalho é imobilizar a enzima urease utilizando como matriz a quitosana e quitosana/glutaraldeído, avaliando diferentes condições de preparação das membranas. As membranas foram depositadas sobre um transdutor que utilizou como suporte fitas de aço inox, eletrodepositadas com antimônio. Logo após, foram realizadas caracterizações por microscopia óptica (MO) e microscopia eletrônica de varredura (MEV), para avaliar as superfícies das membranas. Também, foram realizados testes de bioresposta dos eletrodos, para avaliar a efetividade da enzima imobilizada, através das respostas de sensibilidade, seletividade, estabilidade, tempo de resposta, faixa de linearidade, especificidade e repetibilidade dos biossensores. E foi observado que, a enzima imobilizada com quitosana e com adição do reticulante glutaraldeído, proporcionou melhores respostas, com mais estabilidade operacional e maior armazenamento da enzima, melhorando assim o tempo de vida da mesma. / With the upward development of new technologies, demand for new sensing systems is increasing because there is a need for better accuracy in the results of biological analyzes, with low cost and in real time, before that, the biosensors are devices capable of provide that kind of result. Diseases related to renal insufficiency require such precision and rapidity, which is characterized by the concentration of urea in blood or urine, thus eliminating a series of laboratory procedures. Therefore, for assembling a urea biosensor is needed a selective and specific bio-component that can identify the urea without any interference, as in the case of the enzyme. The enzyme capable of catalyzing the hydrolysis of urea is urease, forming a reaction product of carbon dioxide and ammonia. To be used in a biosensor, the enzyme must be immobilized, as this way confer greater operational stability and a longer storage thereof. This immobilization can be performed by means of polymers. Chitosan is a natural polymer that become increasingly more important for this application, on its properties such as low cost, abundance in nature, easy processing capacity of membrane formation, among others. Given the above, the objective of this work is to immobilize the enzyme urease as a matrix using chitosan and chitosan / glutaraldehyde, evaluating different conditions of preparation of membranes. Membranes were deposited on a transducer used as a support of stainless steel strips, electrodeposited with antimony. Soon after, characterizations were performed by optical microscopy (OM) and scanning electron microscopy (SEM) to evaluate the surfaces of the membranes. Also, the bioresponse electrode tests were performed to evaluate the effectiveness of the immobilized enzyme, through sensitivity responses, selectivity, stability, response time, linearity range, specificity and reproducibility of biosensors. It was observed that the immobilized enzyme with chitosan and crosslinking addition of glutaraldehyde gave the best responses with more operational stability and increased storage of the enzyme, thereby improving the lifetime thereof.

No description available.

<p>This study firstly reports the development, characterisation, and application of thick-film acetylcholinesterase (AChE) biosensors based on a gold electrode modified with a mercaptobenzothiazole (MBT) self-assembled monolayer and either poly(omethoxyaniline) (POMA) or poly(2,5-dimethoxyaniline) (PDMA) in the presence of polystyrene(4-sulphonic acid) (PSSA). The Au/MBT/POMA-PSSA/AChE and Au/MBT/PDMA-PSSA/AChE biosensors were then applied to successfully detect standard organophosphorous and carbamate pesticides in a 0.1 M phosphate buffer, 0.1 M KCl (pH 7.2) solution. Secondly, it reports the construction of the Au/MBT/PANI/AChE/PVAc thick-film biosensor for the determination of certain organophosphate and carbamate pesticide solutions in selected aqueous organic solvent solutions.</p>

<p>This study firstly reports the development, characterisation, and application of thick-film acetylcholinesterase (AChE) biosensors based on a gold electrode modified with a mercaptobenzothiazole (MBT) self-assembled monolayer and either poly(omethoxyaniline) (POMA) or poly(2,5-dimethoxyaniline) (PDMA) in the presence of polystyrene(4-sulphonic acid) (PSSA). The Au/MBT/POMA-PSSA/AChE and Au/MBT/PDMA-PSSA/AChE biosensors were then applied to successfully detect standard organophosphorous and carbamate pesticides in a 0.1 M phosphate buffer, 0.1 M KCl (pH 7.2) solution. Secondly, it reports the construction of the Au/MBT/PANI/AChE/PVAc thick-film biosensor for the determination of certain organophosphate and carbamate pesticide solutions in selected aqueous organic solvent solutions.</p>

Magister Scientiae - MSc / The main hypothesis in this study is the development of a nanocomposite mediated amperometric biosensor for detection of hydrogen peroxide. The aim is to combine the electrochemical properties of both polyaniline and ferrocenium hexafluorophosphate into highly conductive nano composites capable of exhibiting electrochemistry in non acidic media; shuttling electrons between HRP and GCE for biosensor applications. / South Africa

Philosophiae Doctor - PhD / This study firstly reports the development, characterisation, and application of thick-film acetylcholinesterase (AChE) biosensors based on a gold electrode modified with a mercaptobenzothiazole (MBT) self-assembled monolayer and either poly(omethoxyaniline) (POMA) or poly(2,5-dimethoxyaniline) (PDMA) in the presence of polystyrene(4-sulphonic acid) (PSSA). The Au/MBT/POMA-PSSA/AChE and Au/MBT/PDMA-PSSA/AChE biosensors were then applied to successfully detect standard organophosphorous and carbamate pesticides in a 0.1 M phosphate buffer, 0.1 M KCl (pH 7.2) solution. Secondly, it reports the construction of the Au/MBT/PANI/AChE/PVAc thick-film biosensor for the determination of certain organophosphate and carbamate pesticide solutions in selected aqueous organic solvent solutions. / South Africa