A funcionalização da matriz celulósica é um ponto essencial para o aprimoramento dos dispositivos microfluídicos baseados em papel (µPADs). Ela permite minimizar o preparo de amostras e a interferência do usuário, principais fontes de erro no processo analítico. A oxidação da celulose durante uma hora com m-periodato de sódio e a imobilização química de enzimas a partir da formação de bases de Schiff (iminas), via a adição direta da enzima ao substrato oxidado sem a necessidade de outras etapas, é um processo rápido e de baixo custo, apresentando grande potencialidade de aplicação nos dispositivos microfluídicos em papel. A enzima glicose oxidase imobilizada na celulose, com a adição do estabilizante trealose, apresentou elevada atividade catalítica - de 31,9 ± 5,5 mmol L-1 para a enzima não imobilizada a 14,8 ± 2,0 mmol L-1 para a enzima imobilizada e com o estabilizante - além de apresentar maior homogeneidade de sinal, condições desejáveis em testes rápidos em papel. A confecção de dispositivos em papel via impressão em cera alia rapidez e baixo custo de produção, e o arranjo em camadas para originar dispositivos tridimensionais (3D) permite ampliar as funcionalidades dos dispositivos em duas dimensões, tal como o tratamento individualizado de camadas e o armazenamento de reagentes no próprio dispositivo. O método da adição de padrão para obtenção de curvas analíticas no próprio microchip em papel surge como alternativa às curvas analíticas externas, minimizando a manipulação e o preparo de amostras. O uso do ácido 2,2’-azino-bis (3-etilbenzotiazolina-6-sulfônico) - ABTS como indicador redox para as reações enzimáticas e o método de adição de padrão nos µPADs apresentou boa correlação com um modelo de crescimento e saturação de Michaelis-Menten (r2 = 0,8723) na faixa de 0 a 10 mmol L-1, e a utilização da faixa linear para quantificação de glicose (0 a 3 mmol L-1) apresentou grande correlação linear com a concentração estimada pelas curvas de adição de padrão (r2 = 0,959), demonstrando a potencialidade do método. A união da tecnologia desses dispositivos em papel com a de um software automatizado de reconhecimento de imagens (PAlizer) torna instantânea a obtenção de resultados, eliminando-se a necessidade de intervenção humana no processo, tornando os testes em papel mais robustos, reprodutíveis e rápidos. Com o contínuo aperfeiçoamento das funcionalidades e potencialidades dos dispositivos microfluídicos em papel espera-se que os testes diagnósticos de baixo custo atinjam àqueles que deles necessitam, contribuindo para a saúde da população. / Functionalization of a cellulosic matrix is essential for the success of the paper-based microfluidic analytical devices (µPADs). It allows minimization of sample preparation and user interference, both being major sources of errors in the analytical process. Cellulose oxidation with sodium m-periodate during one hour and the direct chemical immobilization of enzymes on it by Schiff-base (imines) formation, which is made by direct insertion of the enzyme on the oxidized substrate without subsequent steps, is a fast and low cost process of immobilization, presenting great potential of application in paper-based microfluidic analytical devices. The glucose oxidase enzyme immobilized on cellulose, with the addition of trehalose stabilizer presented enhanced catalytic activity - from 31.9 ± 5.5 mmol L-1 for the non-immobilized enzyme to 14.8 ± 2.0 mmol L-1 for the immobilized enzyme with the stabilizing agent - also presenting greater signal homogeneity, which are ideal characteristics in a paper-based rapid test. Wax printing is a simple, inexpensive and fast method by which micro-devices can be fabricated. Additionally, the stacking of layers originating tridimensional devices (3D) allow for the improvement of functionalities of 2-dimensional ones, such as individualized layer treatment and reagent storage at different layers in the same device. Standard addition to analytical curves in paper-based microchips is an alternative to external analytical curves, minimizing handling/sample preparation. The use of 2,2\'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid - ABTS redox indicator with the enzymatic reactions and the standard addition method in µPADs presented a good correlation in a growth and saturation Michaelis-Menten model (r2 = 0.8723), in the range of 0 to 10 mmol L-1, and the usage of the linear range to the glucose quantification (0 to 3 mmol L-1) presented a high linear correlation with the estimated concentration from the standard addition curves (r2 = 0.959), showing the potentiality of the method. The coupling of such paper-based devices to automated image analysis software, such as \'PAlizer\', turns the data acquisition process instantaneous, eliminating the need of human intervention during the process, making it more robust, reproducible and rapid. Expectations lie in improving the devices functions and potential so that these low-cost diagnostic devices can one day reach those who need them, contributing significantly to public health.

A funcionalização da matriz celulósica é um ponto essencial para o aprimoramento dos dispositivos microfluídicos baseados em papel (µPADs). Ela permite minimizar o preparo de amostras e a interferência do usuário, principais fontes de erro no processo analítico. A oxidação da celulose durante uma hora com m-periodato de sódio e a imobilização química de enzimas a partir da formação de bases de Schiff (iminas), via a adição direta da enzima ao substrato oxidado sem a necessidade de outras etapas, é um processo rápido e de baixo custo, apresentando grande potencialidade de aplicação nos dispositivos microfluídicos em papel. A enzima glicose oxidase imobilizada na celulose, com a adição do estabilizante trealose, apresentou elevada atividade catalítica - de 31,9 ± 5,5 mmol L-1 para a enzima não imobilizada a 14,8 ± 2,0 mmol L-1 para a enzima imobilizada e com o estabilizante - além de apresentar maior homogeneidade de sinal, condições desejáveis em testes rápidos em papel. A confecção de dispositivos em papel via impressão em cera alia rapidez e baixo custo de produção, e o arranjo em camadas para originar dispositivos tridimensionais (3D) permite ampliar as funcionalidades dos dispositivos em duas dimensões, tal como o tratamento individualizado de camadas e o armazenamento de reagentes no próprio dispositivo. O método da adição de padrão para obtenção de curvas analíticas no próprio microchip em papel surge como alternativa às curvas analíticas externas, minimizando a manipulação e o preparo de amostras. O uso do ácido 2,2’-azino-bis (3-etilbenzotiazolina-6-sulfônico) - ABTS como indicador redox para as reações enzimáticas e o método de adição de padrão nos µPADs apresentou boa correlação com um modelo de crescimento e saturação de Michaelis-Menten (r2 = 0,8723) na faixa de 0 a 10 mmol L-1, e a utilização da faixa linear para quantificação de glicose (0 a 3 mmol L-1) apresentou grande correlação linear com a concentração estimada pelas curvas de adição de padrão (r2 = 0,959), demonstrando a potencialidade do método. A união da tecnologia desses dispositivos em papel com a de um software automatizado de reconhecimento de imagens (PAlizer) torna instantânea a obtenção de resultados, eliminando-se a necessidade de intervenção humana no processo, tornando os testes em papel mais robustos, reprodutíveis e rápidos. Com o contínuo aperfeiçoamento das funcionalidades e potencialidades dos dispositivos microfluídicos em papel espera-se que os testes diagnósticos de baixo custo atinjam àqueles que deles necessitam, contribuindo para a saúde da população. / Functionalization of a cellulosic matrix is essential for the success of the paper-based microfluidic analytical devices (µPADs). It allows minimization of sample preparation and user interference, both being major sources of errors in the analytical process. Cellulose oxidation with sodium m-periodate during one hour and the direct chemical immobilization of enzymes on it by Schiff-base (imines) formation, which is made by direct insertion of the enzyme on the oxidized substrate without subsequent steps, is a fast and low cost process of immobilization, presenting great potential of application in paper-based microfluidic analytical devices. The glucose oxidase enzyme immobilized on cellulose, with the addition of trehalose stabilizer presented enhanced catalytic activity - from 31.9 ± 5.5 mmol L-1 for the non-immobilized enzyme to 14.8 ± 2.0 mmol L-1 for the immobilized enzyme with the stabilizing agent - also presenting greater signal homogeneity, which are ideal characteristics in a paper-based rapid test. Wax printing is a simple, inexpensive and fast method by which micro-devices can be fabricated. Additionally, the stacking of layers originating tridimensional devices (3D) allow for the improvement of functionalities of 2-dimensional ones, such as individualized layer treatment and reagent storage at different layers in the same device. Standard addition to analytical curves in paper-based microchips is an alternative to external analytical curves, minimizing handling/sample preparation. The use of 2,2\'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid - ABTS redox indicator with the enzymatic reactions and the standard addition method in µPADs presented a good correlation in a growth and saturation Michaelis-Menten model (r2 = 0.8723), in the range of 0 to 10 mmol L-1, and the usage of the linear range to the glucose quantification (0 to 3 mmol L-1) presented a high linear correlation with the estimated concentration from the standard addition curves (r2 = 0.959), showing the potentiality of the method. The coupling of such paper-based devices to automated image analysis software, such as \'PAlizer\', turns the data acquisition process instantaneous, eliminating the need of human intervention during the process, making it more robust, reproducible and rapid. Expectations lie in improving the devices functions and potential so that these low-cost diagnostic devices can one day reach those who need them, contributing significantly to public health.

A novel cyclopentadienylruthenium(II) thiolato Schiff base, [Ru(SC6H4NC(H)C6H4OCH2CH2SMe)(&eta / 5-C2H5]2 was synthesized and deposited as a selfassembled monolayer (SAM) on a gold electrode. Effective electronic communication between the Ru(II) centers and the gold electrode was established by electrostatically cycling the Shiff base-doped gold electrode in 0.1 M NaOH from -200 mV to +600 mV. The SAMmodified gold electrode (Au/SAM) exhibited quasi-reversible electrochemistry. The integrity of this electro-catalytic SAM, with respect to its ability to block and electro-catalyze certain Faradaic processes, was interrogated using Cyclic and Osteryoung Square Wave voltammetric experiments. The formal potential, E0', varied with pH to give a slope of about - 34 mV pH-1. The surface concentration, &Gamma / , of the ruthenium redox centers was found to be 1.591 x 10-11 mol cm-2. By electrostatically doping the Au/SAM/Horseradish peroxidase at an applied potential of +700 mV vs Ag/AgCl, a biosensor was produced for the amperometric analysis of hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide. The electrocatalytic-type biosensors displayed typical Michaelis-Menten kinetics with their limits of detection of 6.45 &mu / M, 6.92 &mu / M and 7.01 &mu / M for hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide respectively.

A novel cyclopentadienylruthenium(II) thiolato Schiff base, [Ru(SC6H4NC(H)C6H4OCH2CH2SMe)(&eta / 5-C2H5]2 was synthesized and deposited as a selfassembled monolayer (SAM) on a gold electrode. Effective electronic communication between the Ru(II) centers and the gold electrode was established by electrostatically cycling the Shiff base-doped gold electrode in 0.1 M NaOH from -200 mV to +600 mV. The SAMmodified gold electrode (Au/SAM) exhibited quasi-reversible electrochemistry. The integrity of this electro-catalytic SAM, with respect to its ability to block and electro-catalyze certain Faradaic processes, was interrogated using Cyclic and Osteryoung Square Wave voltammetric experiments. The formal potential, E0', varied with pH to give a slope of about - 34 mV pH-1. The surface concentration, &Gamma / , of the ruthenium redox centers was found to be 1.591 x 10-11 mol cm-2. By electrostatically doping the Au/SAM/Horseradish peroxidase at an applied potential of +700 mV vs Ag/AgCl, a biosensor was produced for the amperometric analysis of hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide. The electrocatalytic-type biosensors displayed typical Michaelis-Menten kinetics with their limits of detection of 6.45 &mu / M, 6.92 &mu / M and 7.01 &mu / M for hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide respectively.

A novel cyclopentadienylruthenium(II) thiolato Schiff base,[Ru(SC6H4NC(H)C6H4OCH2CH2SMe)(η5-C2H5]2 was synthesized and deposited as a selfassembled monolayer (SAM) on a gold electrode. Effective electronic communication between the Ru(II) centers and the gold electrode was established by electrostatically cycling the Shiff base-doped gold electrode in 0.1 M NaOH from -200 mV to +600 mV. The SAMmodified gold electrode (Au/SAM) exhibited quasi-reversible electrochemistry. The integrity of this electro-catalytic SAM, with respect to its ability to block and electro-catalyze certain Faradaic processes, was interrogated using Cyclic and Osteryoung Square Wave voltammetric experiments. The formal potential, E0', varied with pH to give a slope of about - 34 mV pH-1. The surface concentration, Γ, of the ruthenium redox centers was found to be 1.591 x 10-11 mol cm-2. By electrostatically doping the Au/SAM/Horseradish peroxidase at an applied potential of +700 mV vs Ag/AgCl, a biosensor was produced for the amperometric analysis of hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide. The electrocatalytic-type biosensors displayed typical Michaelis-Menten kinetics with their limits of detection of 6.45 μM, 6.92 μM and 7.01 μM for hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide respectively / Magister Scientiae - MSc

Magister Scientiae - MSc / A novel cyclopentadienylruthenium(II) thiolato Schiff base, [Ru(SC6H4NC(H)C6H4OCH2CH2SMe)(η5-C2H5]2 was synthesized and deposited as a selfassembled monolayer (SAM) on a gold electrode. Effective electronic communication between the Ru(II) centers and the gold electrode was established by electrostatically cycling the Shiff base-doped gold electrode in 0.1 M NaOH from -200 mV to +600 mV. The SAMmodified gold electrode (Au/SAM) exhibited quasi-reversible electrochemistry. The integrity of this electro-catalytic SAM, with respect to its ability to block and electro-catalyze certain Faradaic processes, was interrogated using Cyclic and Osteryoung Square Wave voltammetric experiments. The formal potential, E0', varied with pH to give a slope of about - 34 mV pH-1. The surface concentration, Γ, of the ruthenium redox centers was found to be 1.591 x 10-11 mol cm-2. By electrostatically doping the Au/SAM/Horseradish peroxidase at an applied potential of +700 mV vs Ag/AgCl, a biosensor was produced for the amperometric analysis of hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide. The electrocatalytic-type biosensors displayed typical Michaelis-Menten kinetics with their limits of detection of 6.45 M, 6.92 M and 7.01 M for hydrogen peroxide, cumene hydroperoxide and tert-butylhydroperoxide respectively. / South Africa