A vegetação nativa do sertão nordestino é rica em espécies forrageiras nos estratos herbáceo, arbustivo e arbóreo. Estudos têm revelado que acima de 70% das espécies botânicas da caatinga participam significativamente da composição da dieta dos ruminantes domésticos. Estrategicamente, as espécies lenhosas são fundamentais no contexto de produção e disponibilidade de forragem no semiárido Nordestino. Objetivou-se com este trabalho: (1) Quantificar a composição química bromatológica e os compostos fenólicos de leguminosas nativas; (2) Avaliar as características de degradação ruminal e o efeito biológico de plantas taniníferas baseado na técnica de produção de gás in vitro (bioensaio); (3) Estudar os efeitos de dietas constituídas de forrageiras nativas na síntese de nitrogênio microbiano in vitro com marcador 15N e nos parâmetros de fermentação pela técnica de produção gases. Os resultados quanto à composição química e quantificação dos compostos fenólicos das leguminosas taniníferas catingueira (Caesalpinea bracteosa), aroeira (Astronian urundeuva), leucena (Leucaena leucocephala), jureminha (Desmanthus virgatus) e do capim buffel (Cenchrus ciliaris L.) são apresentados. Estas plantas também foram avaliadas por incubação in vitro, juntamente com feno de alfafa (Medicago sativa) como controle, para avaliar o incremento de gás devido à adição de polietilenoglicol (PEG). As leguminosas apresentaram na produção de gases menor atividade biológica dos taninos com adição do PEG, sendo positivos os coeficientes de correlações entre o incremento na produção de gases na presença do PEG e os parâmetros de produção de metano, demonstrando o potencial destas plantas taniníferas para reduzir a produção de metano ruminal, com destaque para leucena e aroeira, com elevado teor proteic e menores teores de fibras. No capitulo 4 são apresentados os resultados do ensaio in vitro para avaliar os efeitos de dietas constituídas de forrageiras nativas da região de Caatinga do NE Brasileiro na síntese de nitrogênio microbiano, utilizando o marcador 15N; e os parâmetros de fermentação pela técnica in vitro de produção gases. As dietas foram constituídas com as espécies catingueira (Caesalpinea bracteosa), aroeira (Astronian urundeuva), leucena (Leucaena leucocephala), jureminha (Desmanthus virgatus) e capim buffel (Cenchrus ciliaris L.), oriundas de três coletas (Agosto de 2008, Março de 2009 e Agosto de 2009). Dois níveis (50% e 30%) de plantas foram usadas em cada dieta simulando sistema CBL (Caatinga, Capim buffel + Leucena). Em geral, as dietas experimentais apresentaram em sua composição química, elevado teor de proteína bruta baixa concentração de compostos fenólicos. Quando estas dietas foram avaliadas através da técnica de produção de gás in vitro, as dietas mostraram redução na emissão de metano, e os parâmetros fermentativos sugeriram que houve mudanças nas rotas de fermentação das dietas. O uso de leguminosas nativas da caatinga pode ser uma alternativa na região semi-árida, principalmente, oferecendo características favoráveis como: valor nutricional, potencial produtivo e rusticidade. No entanto, a viabilidade, no ponto de vista nutricional pode limitar o uso de algumas espécies, devido à alta concentração de compostos fenólicos, principalmente os taninos condensados. / The native vegetation of Northeastern region of Brazil is rich in forage species in the herbaceous, shrubby and arboreal strata. Studies have revealed that over 70% of botanical species from the caatinga participate significantly in the composition of the diet of domestic ruminants. Strategically, the woody species are fundamental in the context of production and availability of forage in such semi-arid area. The objectives of this work were: (1) to quantify the chemical composition and the phenolic compounds of native legumes form the semi-arid region of Pernambuco; (2) to assess the rumen degradation characteristics and biological effect of such tanninferous plants based on in vitro gas production technique (bioassay); (3) to study the effects of diets consist of the native forage on microbial nitrogen synthesis in vitro using 15N as marker and the parameters of fermentation by gas production technique. Chapter 4 presented the results of the in vitro test to assess the effects of diets consist of native forage of Caatinga region of Brazilian NE upon the synthesis of microbial nitrogen using the 15N as tracer, and fermentation parameters. Diets were formed by the species catingueira (Caesalpinea bracteosa), aroeira (Astronian urundeuva), leucena (Leucaena leucocephala), jureminha (Desmanthus virgatus) e capim buffel (Cenchrus ciliaris L.), from three collections (August 2008, March 2009- August 2009). Two levels (50 and 30%) of the plants were used in each diet simulating CBL system (Caatinga + Buffel Grass + Leucena). In general, the experimental diets presented in their chemical composition high crude protein content and low concentration of phenolic compounds. When these diets were evaluated by the in vitro gas production technique, diets showed reduction of methane emission, and the fermentative parameters suggested that there have been changes of fermentation routes of diets. The use of legumes native to caatinga may be an alternative in the semi-arid region, primarily by offering favorable characteristics as: nutritional value, productive potential and homeliness. However, the viability, in nutritional point of view the use of some species can be limiting, because of high concentration of phenolic compounds in particular the condensed tannins.

As espécies nativas do semi-árido nordestino constituem uma importante fonte de proteína para o rebanho da região, especialmente durante a estação seca. Contudo, a composição química dessas plantas, especialmente teores de fibra, proteína bruta e compostos secundários, apresenta variações ao longo do ano, em função do ciclo fenológico. O conteúdo de taninos nas plantas pode variar de acordo com as condições climáticas e geográficas. Objetivou-se com o presente estudo: (i) avaliar a composição química de plantas de três Estados do nordeste brasileiro; (ii) verificar o efeito dos taninos na produção de gases; (iii) testar métodos de detanificação, para melhorar o valor nutritivo dessas plantas. O estudo foi dividido em três capítulos. No primeiro, foram avaliadas espécies nativas do semi-árido de Pernambuco, Bahia e Ceará por meio de análises químicas e técnica de produção de gases: angico - Anadenanthera macrocarpa, aroeira - Astronian urundeuva, catingueira - Caesalpinia bracteosa, jucazeiro - Caesalpinia ferre, jurema preta - Mimosa hostilis, leucena - Leucaena leucocephala, malva branca - Sida cordifolia, maniçoba - Manihot pseudoglaziovii, pau branco - Auxemma oncocalyx e sabiá - Mimosa caesalpinifolia. No segundo e terceiro capítulos, foram testados métodos de secagem e aplicação de soluções alcalinas (cinzas de madeira e hidróxido de cálcio) e de polietileno glicol para redução dos efeitos dos taninos. Os resultados obtidos pela avaliação da composição química nas três regiões demonstraram que as espécies estudadas representam uma importante reserva de forragem para os ruminantes e que o teor de compostos fenólicos deve ser monitorado, uma vez que algumas dessas espécies apresentam altos teores destes compostos. Os métodos de secagem testados não foram eficientes para minimizar a ação dos taninos e os tratamentos com soluções alcalinas e com PEG foram eficazes para reduzir a concentração dos taninos, quando medida pela análise química. O tratamento com PEG resultou uma resposta na diminuição da atividade dos taninos no bioensaio / The typical vegetation of the semi-arid region of northeast Brazil is considered an important source of protein for the animals, mainly during the dry season. However, the chemical composition of the plants, as fiber, protein and secondary compounds change during the year with the different stage of the plants. The tannin concentration varies according to climatic and geographic conditions. The aims of this study were: (i) to evaluate the chemical composition of plants of three regions in the northeast Brazil; (ii) to verify the effect of tannins on gas production; (iii) to use detanification methods to improve the nutritional value of those plants. The study contains three chapters. In the first chapter, the following species of plants from Pernambuco, Bahia and Ceara were evaluated by chemical analysis and gas production: angico - Anadenanthera macrocarpa, aroeira - Astronian urundeuva, catingueira - Caesalpinia bracteosa, jucazeiro - Caesalpinia ferre, jurema preta - Mimosa hostilis, leucena - Leucaena leucocephala, malva branca - Sida cordifolia, manicoba - Manihot pseudoglaziovii, pau branco - Auxemma oncocalyx and sabia - Mimosa caesalpinifolia. In the second and third chapters various detanification methods were tested, as drying processes and application of alkaline solutions (wood ash and calcium hydroxide) and polyethylene glycol application. The chemical analysis showed that all the plants studied could be used as animal feed, but the phenolics contents could be a detrimental factor and should be monitored. The drying methods were not efficient to minimize tannin effects as showed using the gas production. The treatment with wood ash, calcium hydroxide and polyethylene glycol resulted in a decrease of tannins concentration, but only polyethylene glycol treatment decreased tannins activity during the gas production.

In the last decades, intensive research has been carried out in order to replace oil-based polymers with bio-based polymers due to growing environmental concerns. So far, most of the barrier materials used in food packaging are petroleum-based materials. The purpose of the barrier is to protect the packaged food from oxygen, water vapour, water and fat. The mechanical and barrier properties of coatings based on starch-plasticizer and starch-poly(vinyl alcohol) (PVOH)-plasticizer blends have been studied in the work described in this thesis. The plasticizers used were glycerol, polyethylene glycol and citric acid. In a second step, polyethylene coatings were extruded onto paperboard pre-coated with a starch-PVOH-plasticizer blend. The addition of PVOH to the starch increased the flexibility of the film. Curing of the film led to a decrease in flexibility and an increase in tensile strength. The flexibility of the starch-PVOH films was increased more when glycerol or polyethylene glycol was added than citric acid. The storage modulus of the starch-PVOH films containing citric acid increased substantially at high temperature. It was seen that the addition of polyethylene glycol or citric acid to the starch-PVOH blend resulted in an enrichment of PVOH at the surface of the films. Tensile tests on the films indicated that citric acid acted as a compatibilizer and increased the compatibility of the starch and PVOH in the blend. The addition of citric acid to the coating recipe substantially decreased the water vapour transmission rate through the starch-PVOH coated paperboard, which indicated that citric acid acts as a cross-linker for starch and/or PVOH. The starch-PVOH coatings containing citric acid showed oxygen-barrier properties similar to those of pure PVOH or of a starch-PVOH blend without plasticizer when four coating layers were applied on a paperboard. The oxygen-barrier properties of coatings based on a starch-PVOH blend containing citric acid indicated a cross-linking and increase in compatibility of the starch-PVOH blends. Polyethylene extrusion coating on a pre-coated paperboard resulted in a clear reduction in the oxygen transmission rate for all the pre-coating formulations containing plasticizers. The addition of a plasticizer to the pre-coating reduced the adhesion of polyethylene to pre-coated board. Polyethylene extrusion coating gave a board with a lower oxygen transmission rate when the paperboard was pre-coated with a polyethylene-glycol-containing formulation than with a citric-acid-containing formulation. The addition of polyethylene glycol to pre-coatings indicated an increase in wetting of the pre-coated paperboard by the polyethylene melt, and this may have sealed the small defects in the pre-coating leading to low oxygen transmission rate. The increase in brittleness of starch-PVOH films containing citric acid at a high temperature seemed to have a dominating effect on the barrier properties developed by the extrusion coating process. / Over the last few decades, industry and academia have made joint efforts to generate knowledge about renewable barrier materials in order to replace the oil-based barrier materials currently used in food packaging. This work has focused on the possibility of producing a material with high oxygen barrier properties including polyethylene as a moisture protection. The flexibility of starch films was increased by adding poly(vinyl alcohol) (PVOH) to the starch and the addition of a plasticizer to the starch-PVOH blend films further increase the flexibility of the films. The plasticizers used were glycerol, polyethylene glycol and citric acid. Curing of the films reduce their flexibility. The addition of citric acid to a starch-PVOH blend increased the compatibility of the starch-PVOH blend and affected the barrier properties of the coating layers containing citric acid. When a sufficient number of coating layers was applied, the starch-PVOH-citric-acid coatings showed oxygen-transmission-rate-values similar to those of the pure PVOH and of the starch-PVOH blend without plasticizers. Polyethylene extrusion coating on pre-coated paperboard resulted in a clear reduction in the oxygen transmission rate of all the pre-coating recipes based on starch-PVOH blends. The polyethylene extrusion coating showed a higher oxygen transmission rate for a board pre-coated with citric-acid-containing recipes than for a board pre-coated with polyethylene-glycol-containing recipes.

In dieser Arbeit wird die Entwicklung und Charakterisierung neuer „smarter“ Redoxhydrogele mit drei verschiedenen funktionellen Eigenschaften und deren erfolgreicher Einsatz zur elektrochemischen Kontaktierung von Oxidoreduktasen beschrieben. Diese neuen Redoxpolymere 1. tragen kovalent integrierte Redoxzentren umgeben von einer hydrophilen Polymermatrix, 2. reaktive Kopplungsgruppen für den Aufbau selbstassemblierter Polymerschichten auf Elektrodenoberflächen und 3. lassen sich in ihrer Redoxaktivität durch Verwendung „intelligenter“ Polymere über externe Stimuli kontrollieren. Die Redoxhydrogele wurden nach dem Vorbild eines Baukastensystems in einfachen Ein-Stufen-Synthesen synthetisiert. Dazu wurden verschiedene Redoxzentren (Ferrocen, 1,10-Phenanthrolin-5,6-dion und 4-Carboxy-2,5,7-Trinitro-9-fluorenon), reaktive Kopplungsgruppen (Epoxy-, Amino-, Thiol- oder Disulfidfunktionen) und Polymermatrices (Poly-(N-Isopropylacrylamid) (PNIPAM) und Poly(ethylenglykolmethacrylat) (PEGMA)) in unterschiedlichen Zusammensetzungen miteinander copolymerisiert. Die Polymere wurden in Form von dünnen Polymerfilmen über die wiederholenden Funktionalitäten auf Elektrodenoberflächen aufgebracht und physiko- und elektrochemisch charakterisiert. Durch die erstmals gezeigte, derartige Ankopplung der Polymere, entstehen dreidimensionale, hydrophile selbstassemblierte Polymerschichten. Die Elektronentransferwege sind kurz und der Elektronentransfer effizient. Diese Polymer-modifizierten Elektroden wurden für die Kontaktierung von zwei exemplarisch ausgewählten Oxidoreduktasen eingesetzt, die Nicotinsäureamid-adenin-dinucleotid-abhängige Glucosedehydrogenase (NAD-GDH), welche ein freibewegliches Coenzym und die Pyrrolochinolinchinon-abhängige Glucosedehydrogenase (PQQ-GDH), welche ein prosthetisches Coenzym verwenden. Die Redoxaktivitäten des PNIPAMFoxy- und PEGMA-Fc-Polymers ließen sich durch externe Stimuli in Form von Temperatur und Calciumkonzentrationen kontrollieren. Ein Modell für die Komplexierung der Calciumionen durch die PEG-Seitenketten unter Ausbildung Kronenether-ähnlicher Strukturen und der daraus resultierenden Steigerung des Elektronentransfers wurde gezeigt. / This work describes the development and characterization of new, smart redox polymeres with three functionalities and their use in electrochemical wiring of oxidoreductases. These polymers 1. bear redox-active sites surrounded by hydrophilic polymeric matrix 2. surface-reactive groups to create self-assembled monolayers on electrodes 3. ionic-tunable redox activities by using stimuli-responsive polymers The syntheses of the redoxpolymers were resolved in simple one-step approaches using a building block system. Different mediators (ferrocene, 1,10-phenanthroline-5,6-dione and 4-carboxy-2,5,7-trinitro-9-fluorenone), reactive anchoring groups (groups epoxide, amine, thiol and disulfide) and polymer matrices (poly-(N-isopropylacrylamide) (PNIPAM), poly(ethylene glycol methacrylate) (PEGMA)) were copolymerized in different compositions. The polymers were anchored to electrode surfaces via the repetitive functionalities and physico- and electrochemical characterized. This kind of anchoring of the redoxpolymers was shown for the first time and three-dimensional hydropilic self-assembled polymer monolayers are created. The electron transfer pathways are short and the electron tranfer efficient. The polymer-modified electrodes were applied for wiring two oxidoreductases, nicotinamide-adenine-dinucleotide-dependent glucose dehydrogenase (NAD-GDH) with a diffusing coenzyme and the pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GDH) with a prosthetic coenzyme. The redox activities of PNIPAMFoxy and PEGMA-Fc-SS are tuneable with external stimuli like temperature and calcium concentrations. A model for the complexation of calcium by PEG side chains and the explanation of the resulting effects was shown.

Surfactants are molecules that contain a water-soluble and afat-soluble part. They have important functions in productssuch as detergents, cosmetics, pharmaceuticals and foods aswell as in many industrial processes. Surfactants are used onvery large scale, which makes it important to decrease theirimpact on the environment. This can be done by starting withnatural materials, by improving the synthetic methods and byreducing the use of limited resources such as energy andorganic solvents. This thesis focuses on lipase-catalyzed synthesis ofsurfactants based on natural products. It also includesfunctional studies of the produced surfactants; as antioxidantsin oils, or as surfactants to solubilize pharmaceuticals. Unsaturated fatty acid esters of ascorbic acid weresynthesized with catalysis by Candida antarctica lipase B int-amyl alcohol and in ionic liquids. High yields ofascorbyl oleate were obtained in an ionic liquid that wasdesigned to improve the solubility of the fatty acid, when thereaction was performed under vacuum. Ascorbyl oleate wasamorphous and was a better antioxidant than ascorbyl palmitatein rapeseed oil. Polyethylene glycol (PEG) stearate, PEG 12-hydroxystearateand a series of PEG 12-acyloxy-stearates were synthesized in avacuum-driven, solvent-free system usingC. antarcticalipase B as catalyst. Critical micelleconcentration and solubilization capacity were determined forthe PEG 12-acyloxy-stearates. Their effects on living cellswere evaluated in studies of hemolysis and transepithelialelectrical resistance. Several PEG1500 12-acyloxy-stearateswere excellent solubilizers for pharmaceutical use and hadnegligible negative effects on living cells even at highconcentrations. Enzymatic and chemo-enzymatic methods offer uniquepossibilities to synthesize surfactants of high purity. Pureand well-defined surfactants enable new applications and areimportant for the understanding of surfactantstructure-function relationships.

<p>Surfactants are molecules that contain a water-soluble and afat-soluble part. They have important functions in productssuch as detergents, cosmetics, pharmaceuticals and foods aswell as in many industrial processes. Surfactants are used onvery large scale, which makes it important to decrease theirimpact on the environment. This can be done by starting withnatural materials, by improving the synthetic methods and byreducing the use of limited resources such as energy andorganic solvents.</p><p>This thesis focuses on lipase-catalyzed synthesis ofsurfactants based on natural products. It also includesfunctional studies of the produced surfactants; as antioxidantsin oils, or as surfactants to solubilize pharmaceuticals.</p><p>Unsaturated fatty acid esters of ascorbic acid weresynthesized with catalysis by Candida antarctica lipase B in<i>t</i>-amyl alcohol and in ionic liquids. High yields ofascorbyl oleate were obtained in an ionic liquid that wasdesigned to improve the solubility of the fatty acid, when thereaction was performed under vacuum. Ascorbyl oleate wasamorphous and was a better antioxidant than ascorbyl palmitatein rapeseed oil.</p><p>Polyethylene glycol (PEG) stearate, PEG 12-hydroxystearateand a series of PEG 12-acyloxy-stearates were synthesized in avacuum-driven, solvent-free system using<i>C. antarctica</i>lipase B as catalyst. Critical micelleconcentration and solubilization capacity were determined forthe PEG 12-acyloxy-stearates. Their effects on living cellswere evaluated in studies of hemolysis and transepithelialelectrical resistance. Several PEG1500 12-acyloxy-stearateswere excellent solubilizers for pharmaceutical use and hadnegligible negative effects on living cells even at highconcentrations.</p><p>Enzymatic and chemo-enzymatic methods offer uniquepossibilities to synthesize surfactants of high purity. Pureand well-defined surfactants enable new applications and areimportant for the understanding of surfactantstructure-function relationships.</p>

The research presented in this dissertation describes the creation and characterization of a novel antibody-vesicle conjugate modified with polyethylene glycol (PEG) that possesses enhanced binding to and uptake by inflammation-activated endothelial cells with improved storage stability and longer shelf-life and potential reduction in immunogenic potential compared to previous designs. Targeted drug delivery provides an effective means of delivering therapeutic concentrations of a drug to the site or organ of action. The drug is delivered using a niosome, a vesicle with an aqueous core and a bilayer membrane composed of non-ionic surfactants and cholesterol. Antibodies that recognize specific cell antigens are attached to the niosome to complete the targeting molecule, an immuno-niosome (IN). When functionalized PEG, a water soluble, biologically inert polymer, is attached to proteins, it can protect the protein, increasing its half-life in vivo. The immuno-niosome synthesis process is modified to include PEG incorporation into the niosome membrane, a process known as PEGylation. Since the vasculature connects the entire body, immuno-niosomes targeted to endothelial cells were used. When endothelial cells are activated during disease, stress and injury, certain receptors are expressed and upregulated. One such receptor, CD44, is upregulated in response to vascular inflammation associated with atherosclerosis. The research hypothesis is that the addition of polyethylene glycol to the drug delivery vesicle (immuno-niosome) using cyanuric chloride linking chemistry will improve colloidal stability and binding performance of the PEGylated immuno-niosomes to endothelial cell surface receptors expressed during an inflammatory response. The research presented in this dissertation provides the following evidence to support this hypothesis: Construction and characterization of a modified drug delivery vesicle using non-ionic surfactants conjugated with PEG and functionalized with antibodies against endothelial cell surface receptors (PEGylated immuno-niosomes) improves the colloidal stability over previously designed vesicles. Binding of PEGylated CD44-IN to activated endothelial cells is improved over previously designed vesicles. Binding of PEGylated CD44-IN to activated endothelial cells under physiological conditions (flow) is demonstrated. Uptake of PEGylated immuno-niosomes by activated or injured endothelial cells is demonstrated using confocal microscopy.

Water is becoming increasingly scarce as the demand for fresh water continues to rise. One potential new water resource is purified produced water. Produced water is generated during oil and gas production, and it is often contaminated with emulsified oil, high levels of salt, and particulate matter. Produced water purification using polymer membranes has been investigated, but its implementation is limited by membrane fouling. This study focused on the preparation and application of poly(ethylene glycol) (PEG) hydrogels as fouling-resistant coatings for commercial reverse osmosis (RO) membranes. To prepare fouling-resistant coatings for RO membranes, three series of copolymer hydrogel networks were synthesized using poly(ethylene glycol) diacrylate (PEGDA) as the crosslinker and acrylic acid (AA), 2-hydroxyethyl acrylate (HEA), or poly(ethylene glycol) acrylate (PEGA) as comonomers, and their transport properties were evaluated. The hydrogels have high water uptake and high water permeability, and crosslink density strongly influences water uptake and water permeability. For example, a 100 mol% PEGDA hydrogel contained 61% water by volume, but 80PEGA, which has essentially the same chemical composition but lower crosslink density, contained 72% water by volume. Hydrogel water permeability ranged from 10 to 26 (L [mu]m)/(m² hr bar) and correlates well with water uptake; high water uptake often leads to high water permeability. Additionally, the copolymers have hydrophilic surfaces with a low affinity for oil, based on contact angle measurements using n-decane in water. Commercial RO membranes (AG RO membrane from GE Water and Process Technologies) were coated with PEG hydrogels, and the desalination and fouling resistance properties of the coated membranes were tested. The water flux of coated membranes and a series-resistance model were used to estimate coating thickness; the coatings were approximately 2 [mu]m thick. NaCl rejection for both uncoated and coated membranes was 99.0% or greater. As determined by zeta potential measurements, both uncoated and coated RO membranes are negatively-charged, but coated membranes are less negatively-charged than uncoated RO membranes. Model oil/water emulsions, prepared with either a cationic or an anionic surfactant, were used to probe membrane fouling. In the absence of oil, surfactant charge, and therefore, electrostatic interactions play a significant role in membrane fouling. In the presence of DTAB, a cationic surfactant, the AG RO membrane water flux immediately dropped to 30% of its initial value, but in the presence of SDS, an anionic surfactant, its water flux gradually decreased to 74% of its initial value after 24 hours. However, in both cases, coated membranes exhibited less flux decline than uncoated membranes. Coated membranes also experienced little fouling in the presence of an n-decane/DTAB emulsion. After 24 hours, the water flux of a PEGDA-coated AG RO membrane was 73% of its initial value, while the water flux of an AG RO membrane fell to 26% of its initial value. Conversely, both coated and uncoated membranes fouled significantly in the presence of an n-decane/SDS emulsion, indicating that oil fouling is controlled both by electrostatic and hydrophobic interactions. Overall, this work provides answers to some of the fundamental questions posed regarding the viability of using modified membranes for produced water treatment. / text

The development of a chip-based sensor array composed of individually addressable polystyrene-polyethylene glycol and agarose microspheres has been demonstrated. The microspheres are selectively arranged in micromachined cavities localized on silicon wafers. These cavities are created with an anisotropic etch and serve as miniaturized reaction vessels and analysis chambers. The cavities possess pyramidal pit shapes with trans-wafer openings that allow for both fluid flow through the microreactors/analysis chambers as well optical access to the chemically sensitive microspheres. Identification and quantification of analytes occurs via colorimetric and fluorescence changes to receptor and indicator molecules that are covalently attached to termination sites on the polymeric microspheres. Spectral data is extracted from the array efficiently using a charge-coupled device (CCD) allowing for the near-real-time digital analysis of complex fluids. The power and utility of this new microbead array detection methodology is demonstrated here for the analysis of complex fluids containing a variety of important classes of analytes including acids, bases, metal cations, sugars and antibody reagents. The application of artificial neural network analyses to the microbead array is demonstrated in the context of pH measurements. To assess the utility of the analysis and gain an understanding of the molecular level design of the sensor, parameters such as the choice of the indicator dyes, array size, data pre-processing techniques, as well as different network types and architectures were evaluated. Additionally, the development of miniaturized chromatographic systems localized within individual polymer microspheres and their incorporation into an array is reported. The integrated chromatographic and detection concept is based on the creation of distinct functional layers within the microspheres. Such beads have been incorporated into the array platform and used for speciation and concentration determination of aqueous metal cation solutions. / text

No description available.