The self-assembling properties of a core-shell system are considered to be the most desirable characteristics that allow using this class of polymers in different applications. New hairy nanoparticles (HNPs) with hydrophobic polystyrene cores (PS Cores) and hydrophilic poly(2-hydroxyethylmethacrylate) (PHEMA) shells were synthesized by coupling polymerization methods. Living anionic polymerization in one-pot step was used to synthesize cross-linked polystyrene cores functionalized with hydroxyl groups and atom transfer radical polymerization (ATRP) was then carried out to prepare PHEMA hairs following the grafting form technique. The structural characterizations were carried out by FT-IR and NMR spectroscopy (1H NMR, 13C NMR, APT 13C NMR and 1H 13C HMQC). Dynamic light scattering measurements of obtained HNPs show small increase in the order of nanometers of their hydrodynamic radii after the grafting. Thermal properties were studied by TGA and DSC. The thermal stability of PS cores was affected by functionalization with the hydroxyl group. However, the stability of the PS core was not affected by grafting of PHEMA on their surfaces. DSC thermograms of the HNPs shows two distinct transition temperatures corresponding to glass transition temperatures (Tg) of a PS phase and of a PHEMA phase indicating the formation of a hydrophobic-hydrophilic phase separated system. SEM and AFM were utilized to study the morphologies and self-assembly of nanoparticles. The self-assembled HNPs morphologies were dependent on the solvents used. Complexes of the synthesized HNPs and R- or S-mandelic acid were prepared and characterized by circular dichroism (CD) and AFM. CD was used to study the induced chiral properties of the complexes. The CD spectra indicated the formation of enantiomeric chiral structures and the AFM images show toroidal self-assembled structures. Polymer blends of polystyrene functionalized with hydroxyl groups and PHEMA show different morphology and different thermal properties than the core-shell HNP system.

A formação de biofilme representa preocupação à indústria de alimentos pois é uma fonte crônica de contaminação. Encontrar estratégias eficientes para controlar o crescimento de microrganismos continua a ser um importante desafio. Uma delas é o uso dos ramnolipídeos (RLs), um biossurfatante produzido tipicamente por P. aeruginosa que apresenta potencial como agente antimicrobiano, anti-adesivo e dispersivo. Sua baixa toxicidade, biodegradabilidade, eficiência e especificidade em comparação aos surfatantes sintéticos podem torná-los promissores agentes de biocontrole. O presente estudo teve como objetivo estudar o potencial de uso de ramnolipídeos, em diferentes condições de concentração e temperatura, no controle e remoção de biofilmes de patógenos alimentares formados em meio de cultura e leite. Foram utilizadas Escherichia coli ATCC 43895, Listeria monocytogenes ATCC 19112, Staphylococcus aureus ATCC 8095, reconhecidos patógenos alimentares. Os biofilmes foram formados em placas de microtitulação de poliestireno nos meios de cultivo: caldo nutriente (CN), extrato de levedura com triptona de soja (TSYE) e matriz alimentar (leite) à 37 °C, por 24 h (E. coli) e 48 h (S. aureus e L. monocytogenes). Os biofilmes foram avaliados pela quantificação da biomassa, viabilidade celular, hidrofobicidade de superfície e análises qualitativa (microscopia eletrônica de varredura e de fluorescência) e quantitativa (caracterização da matriz polimérica). O ramnolipídeo foi submetido à análise físico-química de espalhamento dinâmico de luz (DLS), espalhamento de raios-X a baixo ângulo (SAXS). Os resultados obtidos para E. coli mostraram que a concentração de RL que mais removeu o biofilme foi 2 ‰, porém em temperaturas diferentes, para o CN à 25 °C e para o leite à 37 °C, com 33 ‰ e 80 ‰ de remoção, respectivamente. Para o biofilme de S. aureus em caldo nutriente os resultados mais eficientes foram à 25 °C, na concentração de 0,1 ‰ de RL e em leite 4 °C, na concentração de 0,05 ‰ de RL, com remoção de 35 ‰ e 89 ‰, respectivamente. O biofilme de L. monocytogenes em TSYE mostrou-se mais sensível à 37 °C, na concentração 0,5 ‰ de RL, o qual foi possível remover 35,3 ‰ da biomassa. Enquanto que em leite a 4 °C e 0,5 ‰ de RL, com remoção de 63,6 ‰ .Quanto à redução das células viáveis foi observado que para as bactérias Gram-positivas o tratamento mais efetivo foi à 4 °C com 0,05 ‰ de RL, nos meios CN e TSYEe 1 ‰ em leite. Para os biofilmes de E. coli a maior redução da viabilidade ocorreu em leite, após tratamento com RL 0,05 ‰ à 37 °C. As imagens de microscopia mostraram uma morfologia heterogênea na presença dos diferentes meios de cultivos, com destaque para os biofilmes de S. aureus (leite) e L. monocytogenes (TSYE), nos quais houve grande produção de matriz polimérica extracelular (MPE), e também apresentaram as maiores quantidades de carboidratos e proteínas. O tratamento com o ramnolipídeo reduziu a hidrofobicidade dos biofilmes. As análises de DLS e SAXS mostraram uma predominância em número de micelas com diâmetro entre 1-10 nm, independente das concentrações e temperaturas analisadas. De modo geral, a aplicação de ramnolipídeo promoveu remoção da biomassa celular como também redução de células viáveis presentes no biofilme. As evidências obtidas aqui, podem ser importantes subsídios para futuras investigações sobre as interações físico-químicas entre ramnolipídeos e a camada de biofilme visando aplicação como agentes sanitizantes em indústria de alimentos. / Biofilm formation is a concern to the food industry because it is a chronic source of contamination. Finding effective strategies to control the growth of microorganisms remains a major challenge. One strategy is the use of rhamnolipids (RLs), a biosurfactant typically produced by P. aeruginosa that has potential as antimicrobial, anti-adhesive and biofilm disrupting agent. RLs low toxicity, biodegradability, efficiency and specificity comparatively to synthetic surfactants, makes them promising biocontrol agents. This work aimed to study the potential use of rhamnolipid at different conditions of concentration and temperature, to control and removal of biofilms of food pathogens established in culture medium and milk. The bacterial strain utilized Escherichia coli ATCC 43895, Listeria monocytogenes ATCC 19112, Staphylococcus aureus ATCC 8095, are well-recognized food pathogens. The biofilms were formed in polystyrene microtiter plates in culture media: nutrient broth (NB), yeast extract and tryptone soya (TSYE) and in food matrix (milk) at 37 °C for 24 h (E. coli) and 48 h (S. aureus and L. monocytogenes). Biofilms were assessed by biomass quantification, cell viability, surface hydrophobicity, qualitative (scanning electron microscopy and fluorescence) and quantitative (characterization of polymer matrix) analysis. The rhamnolipid was subjected to physical and chemical analysis of dynamic light scattering (DLS) and X-ray small angle scattering (SAXS). E. coli biofilms were removed more efficiently using 2 ‰ RL, but at different temperatures for NB (25 °C) and milk (37 °C) showing 33 ‰ and 80 ‰ respectively. For the biofilm of S. aureus in NB the best results was obtained at 25 °C and 0.1 ‰ RL and in milk medium at 4 °C with 0.05 ‰ RL showing 35 ‰ and 89 ‰ of biofilm disruption, respectively. The biofilm of L. monocytogenes in TSYE was more sensitive to the treatment at 37 °C with 0.5 ‰ RL, removing 35.3 ‰ of the biofilm; while in milk at 4 °C and 0.5 ‰ RL, biofilm removal reached 63.6 ‰. Reduction on cell viability was more effective for Gram-positive bacteria at 4 °C with 0.05 ‰ RL, for NB and TSYE and at 1 ‰ in milk. For E. coli biofilms the largest reduction of viability occurred in milk after treatment with 0.05 ‰ RL at 37 °C. The microscopy images showed a heterogeneous morphology in the presence of different media, especially biofilms of S. aureus (milk) and L. monocytogenes (TSYE), in which there was a great production of extracellular polymeric matrix (EPM), and also the highest amounts of carbohydrates and protein. The treatment with RL reduced the hydrophobicity of biofilms. The DLS and SAXS analysis of RL showed a predominance of micelles with diameters between 1-10 nm, independent of the concentrations and temperatures utilized. In general, the application of rhamnolipid promoted a reduction in biofilm mass as well in cell viability. The evidences obtained can provide a basis for future research on the physical and chemical interactions between rhamnolipid and biofilm layer aiming their application as sanitizers in food industry.

Espumas à base de amido possuem limitações para utilização como embalagens na indústria alimentícia, principalmente, devido à alta hidrofilicidade e fragilidade. Para melhorar essas propriedades, torna-se necessário o uso de plastificantes e outros aditivos, como proteínas e fibras lignocelulósicas, ou ainda um aditivo fonte desses recursos, como a torta de gergelim obtida como resíduo do processamento do gergelim. Sendo assim, o objetivo deste trabalho foi desenvolver e caracterizar espumas à base de amido de mandioca adicionadas de 0 – 40 % (m/m) de resíduo de gergelim, produzidas pelo processo de expansão térmica, a fim de avaliar a influência do aditivo nas propriedades físicas, morfológicas e mecânicas dos materiais obtidos para utilização como embalagens para alimentos. A adição do resíduo resultou em espumas com menores espessuras e densidade aparente, bem como, maior capacidade de expansão dos materiais. Além disso, apresentaram melhores propriedades mecânicas e, em geral, menores capacidade de absorção de água e adsorção de água em baixas umidades relativas, em relação aos materiais compostos somente por amido. Embora as propriedades mecânicas das espumas sejam influenciadas pelas condições de umidade relativa de armazenamento, as espumas contendo 20 % de resíduo (20R) apresentaram uma combinação de propriedades no ensaio de tração (resistência à tração e módulo de elasticidade) que podem ser consideradas comparáveis às amostras de bandejas comercias de poliestireno expandido (EPS), nas três umidades relativas avaliadas (33, 53 e 90 %). Os resultados obtidos nos ensaios de flexão dessas espumas indicaram que as mesmas foram mais rígidas e menos flexíveis do que as amostras de EPS. A adição de 20 % de resíduo resultou em importantes melhorias nas propriedades físicas e mecânicas em comparação às espumas à base de amido de mandioca, além de apresentarem menos defeitos morfológicos do que os observados em espumas com maiores teores do resíduo (30 e 40 %). Foram realizados testes de aplicação das espumas 20R no acondicionamento de dois tipos de alimentos com diferentes teores de umidade: bolo (23 %) e brócolis (91 %), em comparação ao armazenamento em embalagens comerciais de EPS. A partir dos resultados obtidos para a variação de umidade, perda de massa e propriedades mecânicas na flexão das embalagens, foi observado que as espumas propostas não foram adequadas para o acondicionamento das amostras de brócolis, apresentando alta absorção de umidade e deformações após três dias de contato com o alimento. Por outro lado, as espumas 20R mostraram bom desempenho no acondicionamento das amostras de bolo, não apresentando mudanças significativas nas propriedades mecânicas no período de 3 a 9 dias de armazenamento. Dessa forma, o resíduo de gergelim em combinação com o amido de mandioca mostrou elevado potencial para o desenvolvimento de espumas que podem ser utilizadas como embalagens para alimentos com baixos teores de umidade, sendo necessários estudos mais detalhados, considerando principalmente aspectos de segurança para utilização de resíduos em produtos em contato direto com alimentos. / Starch based foams have limitations for use as food packaging, mainly due to high hydrophilicity and fragility. In order to improve these properties it is necessary the use of plasticizers and others additives, such as lignocellulosic fibers and proteins, or an additive source of these constituents like sesame cake, which it is obtained from sesame processing. In this way, the main objective of this work was to develop and characterize starch based foams added with 0 – 40 % (w/w) of sesame residue to evaluate the influence of this additive on physical, morphological and mechanical properties of materials for using as food packaging. The foams were produce by thermal expansion process. Residue addition resulted in foams with smaller thickness and density, as well as greater expansion capacity of the materials produced. In addition, foams incorporated with residue exhibited better mechanical properties and, in general, lower water capacity absorption and water adsorption at low relative humidities compared to foams made only with starch. Although mechanical properties of the foams are affected by storage humidity conditions, foams added with 20 % of residue (20R) showed tensile properties (tensile strength and tensile elasticity modulus) that can be considered comparable to expanded polystyrene (EPS) commercial trays for different relative humidities evaluated (33, 53 and 90 %). Flexural properties indicated that starch foams produced was tougher and less flexible than EPS commercial trays. Addition of 20 % of sesame residue on cassava starch-based foams resulted in relevant improvements compared to foams without residue. Besides that, fewer morphological defects were noted, as observed in foams with higher residue content (30 and 40 %). Thus, 20R formulation was selected to carrying out applicability tests of starch-based foams in packaging of two types of food with different moisture content: cake (23 %) and broccoli (91 %) compared to EPS commercial packaging. From the results obtained for moisture variation, loss of weight and flexural properties of the foams, it was found that the material proposed was not applicable to package broccoli samples. Due to high water absorption by the foams alterations on the shape were noted in the materials after 3 days in contact with this food. Nonetheless, foams based on cassava starch and 20 % of sesame residue exhibited a good performance during storage time of cake samples (9 days). It was not observed any effect on flexural mechanical properties for 3 to 9 days of storage. Thereby, sesame cake added to cassava starch showed high potential to developing foams that could be used as packaging for low moisture food. Additionally, more detailed studies are necessary, especially considering safety aspects mainly for using agricultural residues directly in contact with food products.

Much of the aerosol research completed at the University of South Florida has revolved around evaluating industrial hygiene equipment and instrumentation in environmental chambers. Data collected during these studies has provided valuable baseline data on equipment and instrument performance under calm air conditions. A newly constructed wind tunnel now allows researchers to evaluate industrial hygiene instruments under moving air conditions. Because the wind tunnel is capable of producing wind velocities that a worker could encounter in the occupational setting, researchers may gain insight into instrument performance under simulated field conditions. Because aerosols can be introduced into the new wind tunnel testing section, researchers can also challenge industrial hygiene equipment and instrumentation with aerosols in sizes ranges that are of interest in public health. The purpose of this dissertation research was to develop a new wind tunnel to be used for aerosol research at the University of South Florida. Three specific aims had to be met for this study to be successful. They included: (1) designing a wind tunnel based on best practice information outlined in scientific literature, (2) constructing an operable wind tunnel to be used for aerosol research, and (3) characterizing wind tunnel performance by examining the wind tunnel velocity profile, turbulence intensity, and aerosol introduction/collection. The actual wind tunnel was constructed to a length of approximately 20 feet, a height of approximately 2 feet at its tallest point, and includes an entrance filter housing, a settling chamber, a contraction, a testing section, a diffuser, an exit filter housing, a fan, and exhaust duct. All components were designed and constructed using guidelines and best practices reported in the scientific literature. Velocity profile measurements were the first way that this wind tunnel was characterized. In order to successfully obtain measurements, the wind tunnel cross section was divided into 16 equal quadrants. Five measurements were taken for each quadrant at each wind velocity. Target wind velocities for this research were 0.5 m/s, 1.0 m/s, and 2.0 m/s. Actual average wind velocities of 0.48 m/s, 1.00 m/s, and 2.04 m/s. All were within established limits reported in the scientific literature. Turbulence intensity measurements were the second way that this wind tunnel was characterized. In order to successfully obtain measurements, the wind tunnel cross section was divided into 16 equal quadrants. Five measurements were taken for each quadrant at each wind velocity. Wind tunnels are typically designed to have the lowest turbulence intensity possible, generally below 10%. The overall average turbulence intensities for this wind tunnel at wind velocities of 0.5 m/s, 1.0 m/s, and 2.0 m/s were 9%, 10%, and 8% respectively. Overall turbulence intensity measurements were at or below 10%. Isokinetic sampling was the final method used to characterize this wind tunnel by collecting and detecting aerosols traveling through the wind tunnel testing section. The wind tunnel was operated at wind velocities of 0.5 m/s, 1.0 m/s, and 2.0 m/s with isokinetic sampling flow rates of 15.4 L/min, 30.9 L/min, and 61.7 L/min respectively. Monodisperse fluorescent polystyrene latex spheres were used as the test aerosol because they are uniform in size and shape and can be detected by fluorometry. The Blaustein Multi-Jet Atomizer (BLAM) was used to generate monodisperse fluorescent polystyrene latex aerosol 0.5 µm and 2.0 µm particles from liquid suspensions. The Vilnius Aerosol Generator (VAG) was used to generate monodisperse fluorescent polystyrene latex aerosol of 6.0 µm and 12.0 µm particles from dry powders. Nitrogen gas was used for delivering test aerosols into the wind tunnel. Five experimental runs were completed for each particle size and wind velocity for a total of 60 experimental runs. Fluorescence was detected in all 60 samples with average mass concentrations ranging from 0.000050 ng/ml to 0.002703 ng/ml. Based on velocity profile measurements, turbulence intensity measurements, and isokinetic sampling, the performance of University of South Florida wind tunnel was found to be excellent, indicating that it was designed and constructed appropriately. The wind tunnel can now successfully be used by researchers interested in evaluating industrial hygiene sampling equipment with aerosols ranging from 0.5 µm to 12.0 µm in moving air with velocities ranging from 0.5 m/s to 2.0 m/s.

This work investigates the fabrication, characterization and testing of Opuntia ficus-indica mucilage nanofibers to be utilized in water filtration systems. These mucilage nanofibers are formed using different polymers through a process called electrospinning. The polymers used to promote the formation of nanofibers are poly vinyl alcohol (PVA) and polystyrene (PS). The mucilage is a jelly like substance extracted from the pads of the cactus plant. It is a mixture of proteins, complex polysaccharides and monosaccharaides. It is an inexpensive, non-toxic, biodegradable and biocompatible material which is present in abundance. The mucilage extracted from the pads is mixed with acetic acid to form the mucilage solution. The mucilage solution is then mixed by volume with co-spinning polymers, PVA and PS. PVA is a synthetic polymer that is water-soluble, and this work considers two types of PVA differentiated based upon molecular weight, such as low molecular weight PVA and high molecular weight PVA. Polystyrene is a synthetic polymer extracted from a monomer styrene, and it is inexpensive, biodegradable, and abundant. The polystyrene, in its solid form, is further decomposed using a solvent called D-Limonene. D-Limonene is a biodegradable, non-toxic solvent formed from the citrus extract of orange peelings. The PVA and PS solutions are mixed in several different volume ratios with the mucilage solutions. These solutions were electrospun and consistent nanofibers were obtained using the low molecular weight PVA solutions and the polystyrene solutions. The fibers and polymeric solutions were characterized by scanning electron microscopy (SEM), contact angle measurements, viscosity, and FTIR. Resulting mucilage nanofiber membranes were characterized by atomic fluorescence spectrometry (AFS) filtration testing. In addition, a life cycle analysis using the SimaPro software was performed to understand the environmental impact of solutions used to fabricate the mucilage nanofiber membranes. Characterization results confirm the formation of PVA:mucilage and PS:mucilage nanofibers. Filtration testing of the nanofiber membranes indicates better performance with membranes formed by PS: mucilage solutions as compared to PVA: Mucilage solutions. Overall, this work has shown that natural materials, such as cactus mucilage, can be synthesized with polymeric solutions to form environmentally friendly water filters.

Stimuli responsive coatings offer a versatile method by which to manipulate interfacial interactions of proteins in a desired way. However, there exists little guidance as to how the structure of a responsive polymer coating influences adsorption of proteins. In this dissertation, the adsorption behavior of immuglobulin G (IgG) on poly (N-isopropylacryamide) (PNIPAAm) hydrogel coatings was investigated as a function of film thickness. PNIPAAm exhibits a hydrophilic to hydrophobic transition above a critical temperature of ~32°C in aqueous solutions. In this research, through the use of quartz crystal microbalance with dissipation (QCM-D) it was observed that the adsorption was thickness dependent and became non-reversible as the temperature was decreased. Interestingly, QCM-D results also suggested a similar amount of protein adsorption on both hydrated and dehydrated PNIPAAm surfaces. A rigid film analysis using Sauerbrey equation revealed a multi-layer formation on the collapsed PNIPAAm coatings. Although it is allegedly reported that PNIPAAm favors adsorption above the critical temperature due to hydrophobic interactions, there have been several studies that reported adsorption of proteins below the critical temperature. To better understand the QCM-D results, hydrodynamic shear force assays in a spinning disk configuration were performed in order to quickly measure and quantify adhesion of polystyrene (PS) probe spheres (10μm) to the PNIPAAm coatings in both the solvated (hydrophilic) and collapsed (hydrophobic) state. The influence of polymer coating thickness, polymer chain cross-link density, microsphere concentration and adsorption time on the adhesion characteristics of the coatings was investigated in relation with volume phase transition of the polymer coatings. A series of experiments on quantification of the temperature dependent adhesion of proteins adsorbed on surface attached PNIPAAm coatings of thicknesses was performed as the surface chemistry was switched from hydrophilic to hydrophobic. First, adhesion of polystyrene (PS) microspheres on PNIPAAm coatings was quantified in order to have a guideline for temperature dependent adhesion performance of these coatings. PS particles were subjected to a range of detachment shear stresses through hydrodynamic flow in a spinning disk configuration. These experiments provide an indirect method to determine the force of adhesion since it is proportional to the hydrodynamic force. Model protein, IgG, was then linked to PS microspheres and the mean adhesion strength of the IgG coated PS microspheres were determined through the detachment shear stresses. The influence of PS deposition time, PS bead concentration, PNIPAAm coating thickness and PNIPAAm cross-link density on the adhesion strength were addressed. The results indicated that in the collapsed state, the adhesion of bare hydrophobic PS microspheres depends strongly on coating thickness. For hydrophilic charged PS microspheres the adhesion was always higher on the hydrated PNIPAAm surfaces and appeared not to be strongly affected by the increase in PNIPAAm coating thickness. The adhesion of IgG was higher on the collapsed PNIPAAm surfaces and the adhesion trend did not significantly change as the PNIPAAm film thickness was increased. For PNIPAAm coatings with the cross-link density reduced by factor of 10, the adhesion was again higher on the collapsed PNIPAAm surface and scaled linearly with thickness. Moreover, the influence of thickness became prominent at the higher thickness values (165 nm-185 nm). In addition, the adhesion of carboxylated microspheres on PNIPAAm did not reach equilibrium and increased linearly with microsphere deposition time. A study on the sensing characteristics of PNIPAAm coatings in response to heavy metal ions was also conducted in this dissertation. The temperature-dependent swelling behavior of poly(N-isopropylacrylamide) and tripeptide Gly-Gly-His/poly(NIPAAm) conjugate hydrogel coatings were investigated using a quartz crystal microbalance with dissipation (QCM-D) while in contact with NaCl, ZnCl2, NiCl2, and CuCl2 solutions. To fabricate the tripeptide conjugated gels, precursor gels of poly(NIPAAm-co-3-aminopropylmethacrylamide[3.5 mole%]) were synthesized via free radical polymerization. The metal binding tripeptide, Gly-Gly-His, was subsequently synthesized in the gel via a Merrifield solid phase peptide synthesis (SPPS) technique, in which the amino group of the copolymer gel provided a functional site to support peptide synthesis. It was found that the logarithm of the transition temperature of the tripeptide Gly-Gly-His/poly(NIPAAm) conjugate hydrogel was proportional to the ionic strength, showing two distinct regions at low and high ionic strengths for the divalent ions. In the low ionic strength regime, the salting out constants were 0.08 M-1, 0.07 M-1, and 0.06 M-1 for Cu2+, Ni2+, and Zn2+, respectively, which follows the known trend for binding of the ions to Gly-Gly-His. In the high ionic strength region, when the metal-ion binding sites in the tripeptide conjugate hydrogel were saturated, the salting out constants were similar to the salting out constants associated with pure poly(NIPAAm).

Polymeric systems near their glass transition are known to exhibit heterogeneous dynamics that evolve both over space and time, yet many of the underlying principles of these dynamics are still poorly understood. In this thesis, experimental single molecule studies aimed at uncovering the dynamics of polystyrene near its glass transition temperature are described. In a first approach, the influence of temperature on the timescales associated with dynamic heterogeneity – also referred to as exchange times – are identified by following the dynamics of a fluorescent perylene diimide probe embedded in a high-molecular weight polystyrene host. No clear influence on the lifetime of dynamics is found in the temperature regime Tg to Tg + 10 K. In a second study, heterogeneous dynamics are investigated in the context of molecular weight and fragility. In a similar experimental approach to that of the first study, two fluorescent dyes are utilized to report on the rotational dynamics of low- to high-molecular weight polystyrene hosts. In accordance with previous reports, the stretching exponent, β, is found to be correlated with the system’s molecular weight, even on a single molecule level. However, no clear correlation with the system’s exchange time was found. In a final study, several single molecule approaches aimed at uncovering the dynamics in confined polystyrene films are described. As no evidence for previously-described mobile surface molecules has been found, this final chapter is meant to provide a basis for future single molecule studies in confined systems.

Coupled two-dimensional HPLC systems were developed for the separation of complex sample matrices. Low molecular weight polystyrene oligomers were used as model compounds for the development of these systems since the sample dimensionality of oligostyrenes could be defined and classified according to two sample characteristics. That is, oligostyrenes could be classed as a two-dimensional sample, where one sample dimension is the variation in the number of monomers that make up oligomeric units determining the weight of the polymer, with a second sample dimension being the variation in stereoisomerism of each oligomer. During this study it was found that by combining two separation dimensions consisting of C18 and carbon clad zirconia (CCZ) phases separation of oligostyrenes according to molecular weight and stereochemistry was possible in coupled two-dimensional systems. The C18 phase separated the sample according to molecular weight, while the CCZ phase separated the sample according to the stereochemistry. The stereoisomer separations of the oligostyrenes reported on the CCZ surface were far superior to those previously reported in the literature. The efficiency of the separation process in a coupled two-dimensional system was then evaluated by studying the variation in band variance of a test probe. The results of the study indicated that the performance of a two-dimensional separation process was highly dependent upon the mobile phase compatibility, even when solvents were completely miscible and of similar polarities. Extracts from 17 species of Australian native plants were examined for xanthine oxidase inhibitory activity, the enzyme responsible for the formation of the disease gout. Chromatographic separation was conducted on plant extracts found to possess significant inhibitory activity against xanthine oxidase, with an extract from the species Clerodendrum floribundum R. Br. found to possess the greatest activity of the species examined. A two-dimensional separation of a crude extract from Clerodendrum floribundum R. Br. was conducted using one of the developed HPLC systems, to illustrate the use of such a system for the separation of a non-model complex sample mixture / Doctor of Philosophy (PhD)

[Truncated abstract] Chapter 1 provides an introduction to the 'sarcophagine' class of ligands and the field of metallopolymers. The synthesis, stereochemistry, physical properties and functionalisation of 'sarcophagines' and their metal complexes are discussed. A brief overview of the burgeoning field of metallopolymers is given with specific mention of the synthetic routes to pendant metallopolymers, and how these could be employed to prepared cage amine containing metallopolymers. Chapter 2 deals with the synthesis, characterisation and reactivity of cinnamylamino and styrylamido derivatives of the cage amines [Co((NH2)2sar)]3+, [Co((CH3)(NH2)sar)]3+ and [Cu((NH2)2sar)]2+. The cinnamylamino derivatives were prepared using reductive alkylation of the aforementioned amines with cinnamaldehyde. Procedures were developed to isolate the complexes without causing unwanted additions to the double bond. The cinnamylamino derivatives displayed unexpected reactivity towards a range of reducing agents, resulting in unexpected reduction of the double bond and cleavage of the cinnamyl group, but ultimately in the preparation of phenylpropylamino derivatives of [Co((NH2)2sar)]3+ and [Co((CH3)(NH2)sar)]3+. Attempts to rationalise the reactivity of the double bond have been explored based upon the physical properties and reactivity of the double bond. The styrylamido derivatives were prepared by treatment of the cage amines with 4-vinylbenzoyl chloride, and the complexes isolated in a similar manner to those of the cinnamylamino derivatives to ensure the amide linkage remained intact. Most of the complexes have been structurally characterised. ... Both the 2-thienyl and 3-thienyl derivatives of [Co((NH2)2sar)]3+ and [Co((CH3)(NH2)sar)]3+ have been prepared using reductive alkylation with the respective carboxaldehydes of thiophene. One of the optically pure isomers has been prepared. The complexes have been fully characterised including structural characterisation. Polymerisation of the thiophene-cage amine complexes was investigated under a range of chemical and electrochemical conditions, though polymerisation was never observed. Cleavage of the thienyl groups was observed when ceric ammonium nitrate in nitric acid was used as the oxidant. The attachment of oligothiophenes and mixed pyrrole-thiophene oligomers to cage amines were investigated using reductive alkylation and various pyrrole ring-forming reactions about the apical amino groups, though none of the desired complexes were isolated, reasons for the lack of reactivity were discussed. An efficient synthesis of N-(4-benzoic acid)- 2,5-di(2-thienyl)pyrrole was developed and was shown to the electropolymerisable, albeit the polymer films were non-conducting. Attempts to couple N-(4-benzoic acid)- 2,5-di(2-thienyl)pyrrole with a cage amine via its acid chloride were complicated by decomposition reactions, the nature of one of these products is discussed. Chapter 5 presents investigations into the preparation of simple complexes containing multiple cage amines using alkylation and acylation procedures with aromatic substrates. The complexes were found to exhibit some interesting electrochemical and chemical properties, demonstrating that even simple multiple cage amine species can display complicated and interesting behaviour.

Hybrid macrocycles, which chimerically integrate multiple chemical compositions and architectures, provide an effective way to impart new properties to polymers that are not found in their linear or homocyclic analogues. This dissertation addresses the incorporation of hydrophilic blocks into hydrophobic polymer, as either a poly(dimethyl siloxane)-block-poly(oxyethylene) (PDMS-POE) tadpole with a hydrophobic head and a hydrophilic tail or as a diblock poly(styrene)-block-diethylene glycol (PS-DEG) hydrophobic-hydrophilic macrocycle. The supramolecular association properties of both kinds of cycles were studied: the PDMS-POE tadpoles in forming micelles, and the PS-DEG macrocycles in threading with linear polymer to form polyrotaxanes. For the PDMS-POE macrocycle, linear alpha,omega-dihydroxy PDMS was cyclized under dilute conditions with dichloromethylhydrosilane as a linking group to produce hydrosilane-functionalized cyclic PDMS. This was joined to alpha-methoxy,omega-allyl POE via a free radical hydrosilylation reaction to produce the hybrid tadpole macrocycle, which was analyzed by GPC, DSC, and 1H, 13C, and 29Si NMR spectroscopy. Supramolecular aggregation consisting of the formation of micelles under both polar and nonpolar conditions was studied by surface tensiometry and quasielastic light scattering. For the PS-DEG macrocycle, linear alpha,omega-dihydroxy PS was prepared by ATRP polymerization of styrene, followed by reaction with KOH to give hydroxyl endgroups. The linear PS was then cyclized under dilute conditions with diethylene glycol ditosylate, and the product was analyzed by GPC, MALDI-TOF MS, DSC, and 1H, 13C and DOSY NMR spectroscopy. The macrocycle was then statistically threaded with linear PS to give the supramolecular structure poly(styrene)-rotaxa-cyclo[poly(styrene)-block-diethylene glycol]. Characterization was performed with DOSY NMR to verify that the product was threaded, and 1H NMR was collected to determine that the product was 13% macrocycle by weight. DSC showed only one Tg, indicating that the linear and cyclic species were present in the same phase.