Thesis submitted in fulfilment of the requirements for the degree Master of Technology Chemistry in the Faculty of (Science) Supervisor: Professor T.N. van der Walt Bellville campus Date submitted: October 2011 / It is important to quantify polymeric components in a coating because they greatly influence the performance of a coating. The difficulty associated with analysis of polymers by Fourier transform infrared (FTIR) analysis’s is that colinearities arise from similar or overlapping spectral features. A quantitative FTIR method with attenuated total reflectance coupled to multivariate/ chemometric analysis is presented. It allows for simultaneous quantification of 3 polymeric components; a rheology modifier, organic opacifier and styrene acrylic binder, with no prior extraction or separation from the paint. The factor based methods partial least squares (PLS) and principle component regression (PCR) permit colinearities by decomposing the spectral data into smaller matrices with principle scores and loading vectors. For model building spectral information from calibrators and validation samples at different analysis regions were incorporated. PCR and PLS were used to inspect the variation within the sample set. The PLS algorithms were found to predict the polymeric components the best. The concentrations of the polymeric components in a coating were predicted with the calibration model. Three PLS models each with different analysis regions yielded a coefficient of correlation R2 close to 1 for each of the components. The root mean square error of calibration (RMSEC) and root mean square error of prediction (RMSEP) was less than 5%. The best out-put was obtained where spectral features of water was included (Trial 3). The prediction residual values for the three models ranged from 2 to -2 and 10 to -10. The method allows paint samples to be analysed in pure form and opens many opportunities for other coating components to be analysed in the same way.

Les triterpènes sont un groupe important et structurellement diversifié de produits naturels issus du squalène. Les progestatifs et les glucocorticoïdes sont des triterpènes oxygénés ayant un squelette tétracyclique et reconnus pour leurs diverses propriétés thérapeutiques. Par ailleurs, l'érythrodiol et l'uvaol sont des triterpènes pentacycliques reconnus pour leurs effets bénéfiques dans l'alimentation humaine. Dans ce travail, l'interaction avec les membranes des vésicules lipidiques et la liaison à la sérum albumine humaine de ces molécules sont étudiées dans le but de mieux comprendre leurs propriétés pharmacologiques. Les vésicules lipidiques ont été caractérisées par DSC, spectroscopie Raman, FTIR et polarisation de fluorescence du DPH pour comprendre l'effet des molécules sélectionnées sur la fluidité membranaire.Également, nous avons étudié la liaison du cholestérol, d'une série de progestatifs et de glucocorticoïdes à l'albumine humaine par la spectroscopie de fluorescence.Les résultats ont démontré que les progestatifs, les glucocorticoïdes, l'érythrodiol et l'uvaol altèrent les propriétés physiques de la bicouche lipidique.Les progestatifs et les glucocorticoïdes démontrent un attachement modéré à l'albumine. Par ailleurs, la dydrogestérone présente la constante de liaison la plus importante. Enfin, notre étude a démontré que la constante de liaison du cholestérol à l'albumine est faible en comparaison avec les autres molécules étudiées. Notre étude a conduit à une connaissance approfondie des mécanismes moléculaires et des caractéristiques structurales impliqués dans l'interaction des triterpènes avec les protéines et les membranes synthétiques / The triterpenoids are a large and structurally diverse group of natural products derived from squalene. Progesterone derivatives and glucocorticoids are a group of oxygenated triterpenes having a tetracyclic skeleton and identified for their therapeutic properties. Whereas, erythrodiol and uvaol are pentacyclic triterpenes, known for their beneficial effects on human diet. In this thesis, we studied their interaction with the membranes of lipid vesicles and with human serum albumin to better understand their pharmacological properties. DSC, Raman spectroscopy, FTIR and fluorescence polarization of DPH were used to investigate the effect of triterpenes on the membrane fluidity. Besides, we used fluorescence spectroscopy to study the binding of cholesterol, a series of progesterone derivatives and another series of glucocorticoids to albumin. The results revealed that progesterone derivatives, glucocorticoids, erythrodiol and uvaol changed the physical properties of the bilayers. Progesterone derivatives and glucocorticoids have been proven to bind moderately to albumin. Dydrogesterone showed the highest binding constant. Finally, our study demonstrated that cholesterol exhibited a much weaker interaction with albumin compared to progesterone derivatives and glucocorticoids.Our work has led to a better understanding of triterpenes molecular mechanisms of their interaction with proteins and biological membranes and structural features controlling these interactions

O ranelato de estrôncio (RS) diminui a reabsorção óssea e ao mesmo tempo age como um agente anabólico deste tecido. Este estudo foi realizado para avaliar os efeitos da ovariectomia e do tratamento com RS na reparação de defeitos ósseos em fêmures e nos componentes moleculares do osso em ratas. Vinte e sete ratas adultas foram submetidas a ovariectomia ou cirurgia Sham e, após trinta dias, defeitos ósseos em fêmures foram confeccionados e os animais divididos em três grupos: ovariectomizadas (OVZ), cirurgia Sham (SHAM) e ovariectomizadas + tratamento com 625 mg/kg/dia de RS (RS). A eutanásia foi realizada quatro semanas após a cirurgia do defeito ósseo. A reparação do defeito ósseo foi analisada por microtomografia computadorizada (?CT) e a composição química do tecido ósseo foi verificada por espectroscopia de infravermelho com transformada de Fourier (FTIR) e espectroscopia de energia dispersiva de raios-X (EDS). O grupo SHAM apresentou em média volume ósseo (BV) superior ao grupo OVZ (p=0,014) e o volume ósseo relativo (BV/TV) foi cerca de 8% superior ao grupo OVZ. A comparação entre SHAM e RS apresentou valores limítrofes para essas variáveis. A espessura trabecular (Tb.Th) no grupo RS foi significativamente maior que no grupo OVZ cerca de 1,8% (p = 0,049) e não diferiu em média do grupo SHAM. Não houve diferenças significativas nas relações que avaliam compostos inorgânicos sobre orgânicos nos grupos. A razão das áreas das bandas em 1057 e 1023 cm-1 para o cálculo do IC foi 1.024 no grupo SHAM, 1.015 no grupo RS e 1.108 no grupo OVZ. No estudo da banda de amida III foi observado nos animais ovariectomizados uma possível mudança na estrutura secundária das proteínas da matriz devido a uma diminuição das estruturas ?-hélice e random coil (RC) e aumento de ?-sheet. A maturidade do colágeno estimada pela relação de sub-bandas de amida I (1660/1690) foi menor no grupo RS. Neste grupo houve um aumento significativo de estrôncio (Sr) incorporado ao tecido ósseo e uma diminuição de cálcio (Ca) avaliados pela técnica do EDS. Os resultados demonstram que o tratamento sistêmico com RS promoveu a reparação óssea e melhorou a microarquitetura do osso neoformado no interior do defeito. No entanto, os resultados das análises com FTIR sugerem que o RS não reverteu as alterações na matriz orgânica ocasionadas pela ovariectomia e diminuiu as ligações cruzadas de colágeno maturas. / Strontium ranelate (SR) decreases bone resorption and at the same time acts as an anabolic agent in this tissue. This study was conducted to evaluate the effects of ovariectomy and treatment with SR on the repair of bone defects and molecular components of bones in femurs. Adult female rats (n=27) were subjected to ovariectomy or Sham surgery. Thirty days after surgery, a defect was made in the femur and the animals were divided into three groups: ovariectomy (OVZ), Sham surgery (SHAM) and ovariectomy plus treatment with SR, 625 mg / kg / day (SR). Euthanasia was performed four weeks after surgery to cause the bone defect. Repair in bone defect was assessed by computed microtomography (?CT) and the chemical composition of bone tissue was analyzed by Fourier transform infrared (FTIR) spectroscopy and energy dispersive X-ray spectroscopy (EDS). The SHAM group showed mean values for bone volume (BV) higher than those of the OVZ group (p = 0.014) and the relative bone volume (BV / TV) was about 8% higher than that of the OVZ group. Comparison between SHAM and SR showed borderline values for these variables. In the SR group, the average value for trabecular thickness (Tb.Th) was significantly higher (~1.8%) than that in the OVZ (p = 0.049) group and did not differ from that of the SHAM group. Significant difference was not observed in the relation between inorganic and organic compounds in the three groups. The ratio of the areas of the bands in 1057 and 1023 cm-1 for the calculation of IC was 1,024 in the SHAM group, 1,015 in the RS group and 1,108 in OVZ group. In the study of the amide III band, a change in the proportion of secondary structure of matrix proteins was observed in ovariectomized animals, possibly a decrease in of ?-helical and random coil (RC) and an increase in ?-sheet structures. The collagen cross-linking network maturity estimated by the relation between amide I subbands (1660/1690) showed the smaller number in the SR group. In this group, a significant increase in the amount of strontium (Sr) and a decrease in the amount of calcium (Ca) embedded to bone tissue, as assessed by the EDS technique, were also observed. The results demonstrate that systemic treatment with SR promoted bone repair and improved microarchitecture of the newly formed bone within the defect. However, results of analyses with FTIR suggest that the SR has not reversed the alterations caused by ovariectomy in the organic matrix and decreased cross-linking of collagen mature.

O objetivo do presente trabalho foi avaliar diferentes tratamentos na remineralização do esmalte desmineralizado em situação de alto desafio cariogênico. Foram utilizados 55 fragmentos de esmalte bovino (4mmX4mmX2mm) planificados e polidos. Os espécimes foram cobertos com resina composta ficando exposta apenas a metade da superfície externa (2mmX4mm) e então, foram imersos em solução tampão de acetato (pH 4,7) por 43 horas para formar a lesão de desmineralização. Os espécimes foram divididos aleatoriamente em 5 grupos (n=11), de acordo com os tratamentos preventivos: 1- Controle (sem tratamento); 2- Verniz fluoretado a 5% Duraphat®; 3- Flúor-gel 1,23%; 4- MI PasteTM; e 5- MI Paste PlusTM. Os tratamentos foram realizados uma vez por semana durante 3 semanas. Nos intervalos das aplicações os espécimes foram submetidos a ciclos de pH (DES/RE) num total de 21 dias. Após a finalização da ciclagem os fragmentos foram seccionados longitudinalmente, sendo que em uma metade foi analisada a microdureza longitudinal, pelo cálculo da variação percentual da dureza e na outra metade foi avaliada a profundidade de desmineralização do esmalte através de microscopia óptica. Para a análise em FTIR, pó do esmalte foi obtido dos espécimes em três diferentes momentos: inicial sem desmineralização da superfície, após a desmineralização por 43 horas e após o tratamento. Para análise da microdureza longitudinal empregou-se a variação da dureza. Os dados foram analisados quanto a sua distribuição, porém como não foram normais realizou-se uma análise não paramétrica comparando grupo contra grupo pelo Mann Whitney test com &alpha;=5%. Para análise da profundidade de desmineralização, como os dados se apresentaram normais e homogêneos, realizou-se a análise de variância a 1 critério e teste Holm- Sidak para comparação das médias com &alpha;=5%. Para análise em FTIR fez-se apenas uma análise descritiva dos dados. Na análise dos dados pode-se observar que houve diferença estatística significante entre o grupo controle (G1) com o grupo flúor-gel (G3) (p<0,05), o qual apresentou menor perda de dureza e menor profundidade de desmineralização. Os grupos 2, 4 e 5 foram similares entre si e com o G1 para as duas propriedades analisadas. Na análise da absorbância nos diferentes comprimentos de onda pode-se observar um padrão similar em todas as amostras; a banda de fosfato inicial foi mais larga e posicionada em número de ondas menores. Após a desmineralização, esta banda ficou mais estreita e deslocada para número de ondas maiores. Esse comportamento se repetiu e foi similar para todos os grupo. Baseado nos resultados obtidos pode-se concluir que nenhum dos tratamentos preventivos realizados foi capaz de inibir o processo de desmineralização ou promover a remineralização em situações de alto risco a cárie, embora o flúor-gel tenha permitido uma evolução mais lenta da desmineralização / The aim of this study was to evaluate different treatments on remineralization of demineralized enamel in a high cariogenic challenge. 55 enamel fragments (4mmX4mmX2mm) flat and polished were used. The specimens were covered with composite exposed only half of the outer surface (2mmX4mm) and then were immersed in acetate buffer solution (pH 4.7) for 43 hours to form the demineralization lesion. The specimens were randomly divided into 5 groups (n = 11) according with the preventive treatments: 1 - Control (no treatment); 2 - 5% fluoride varnish Duraphat ®; 3 - Fluoride-gel 1.23%; 4 - MI PasteTM; and 5 - MI Paste PlusTM. The treatments were carried out once a week for 3 weeks. During intervals and among applications the specimens were subjected to pH cycles (DES / RE) for a total of 21 days. After finished of the cycling, the samples were longitudinally sectioned, and one half in the longitudinal microhardness was analyzed and the other half the depth of enamel demineralization using optical microscopy was evaluated. For data analyzed was calculate the percentage change in the hardness. For FTIR analysis, the enamel powder was obtained from specimens at three different times: without demineralization of the surface, after demineralization for 43 hours and after treatment. Data were analyzed for their distribution, but they were not normal there was a non-parametric analysis comparing group versus group by t test (Mann- Whitney test) with &alpha;= 5%. To analyze the depth of demineralization, as the data were normal and homogeneous, we performed analysis of variance criterion 1 and Holm-Sidak test for comparison of means with &alpha;= 5%. For FTIR was realized of descriptive analysis of the data. Data analyzed showed statistically significant difference between the control group (G1) with the fluoride gel group (G3) (p <0.05), which showed less loss of lower hardness and depth of demineralization. Groups 2, 4 and 5 were similar among them and with G1 for two properties analyzed. The analysis of the absorbance at different wavelengths can be observed a similar pattern in all samples; band initial phosphate was wider and placed on the number of smaller waves. After demineralization, this band became narrower and shifted to higher wave number. This behavior was similar for all groups and it is repeated for every group. Based on the results this study it can be concluded that none of the preventive treatment carried out was able to inhibit the demineralization or promote remineralization in high caries risk situations, although fluoride gel has allowed a slow evolution of demineralization

O ranelato de estrôncio (RS) diminui a reabsorção óssea e ao mesmo tempo age como um agente anabólico deste tecido. Este estudo foi realizado para avaliar os efeitos da ovariectomia e do tratamento com RS na reparação de defeitos ósseos em fêmures e nos componentes moleculares do osso em ratas. Vinte e sete ratas adultas foram submetidas a ovariectomia ou cirurgia Sham e, após trinta dias, defeitos ósseos em fêmures foram confeccionados e os animais divididos em três grupos: ovariectomizadas (OVZ), cirurgia Sham (SHAM) e ovariectomizadas + tratamento com 625 mg/kg/dia de RS (RS). A eutanásia foi realizada quatro semanas após a cirurgia do defeito ósseo. A reparação do defeito ósseo foi analisada por microtomografia computadorizada (?CT) e a composição química do tecido ósseo foi verificada por espectroscopia de infravermelho com transformada de Fourier (FTIR) e espectroscopia de energia dispersiva de raios-X (EDS). O grupo SHAM apresentou em média volume ósseo (BV) superior ao grupo OVZ (p=0,014) e o volume ósseo relativo (BV/TV) foi cerca de 8% superior ao grupo OVZ. A comparação entre SHAM e RS apresentou valores limítrofes para essas variáveis. A espessura trabecular (Tb.Th) no grupo RS foi significativamente maior que no grupo OVZ cerca de 1,8% (p = 0,049) e não diferiu em média do grupo SHAM. Não houve diferenças significativas nas relações que avaliam compostos inorgânicos sobre orgânicos nos grupos. A razão das áreas das bandas em 1057 e 1023 cm-1 para o cálculo do IC foi 1.024 no grupo SHAM, 1.015 no grupo RS e 1.108 no grupo OVZ. No estudo da banda de amida III foi observado nos animais ovariectomizados uma possível mudança na estrutura secundária das proteínas da matriz devido a uma diminuição das estruturas ?-hélice e random coil (RC) e aumento de ?-sheet. A maturidade do colágeno estimada pela relação de sub-bandas de amida I (1660/1690) foi menor no grupo RS. Neste grupo houve um aumento significativo de estrôncio (Sr) incorporado ao tecido ósseo e uma diminuição de cálcio (Ca) avaliados pela técnica do EDS. Os resultados demonstram que o tratamento sistêmico com RS promoveu a reparação óssea e melhorou a microarquitetura do osso neoformado no interior do defeito. No entanto, os resultados das análises com FTIR sugerem que o RS não reverteu as alterações na matriz orgânica ocasionadas pela ovariectomia e diminuiu as ligações cruzadas de colágeno maturas. / Strontium ranelate (SR) decreases bone resorption and at the same time acts as an anabolic agent in this tissue. This study was conducted to evaluate the effects of ovariectomy and treatment with SR on the repair of bone defects and molecular components of bones in femurs. Adult female rats (n=27) were subjected to ovariectomy or Sham surgery. Thirty days after surgery, a defect was made in the femur and the animals were divided into three groups: ovariectomy (OVZ), Sham surgery (SHAM) and ovariectomy plus treatment with SR, 625 mg / kg / day (SR). Euthanasia was performed four weeks after surgery to cause the bone defect. Repair in bone defect was assessed by computed microtomography (?CT) and the chemical composition of bone tissue was analyzed by Fourier transform infrared (FTIR) spectroscopy and energy dispersive X-ray spectroscopy (EDS). The SHAM group showed mean values for bone volume (BV) higher than those of the OVZ group (p = 0.014) and the relative bone volume (BV / TV) was about 8% higher than that of the OVZ group. Comparison between SHAM and SR showed borderline values for these variables. In the SR group, the average value for trabecular thickness (Tb.Th) was significantly higher (~1.8%) than that in the OVZ (p = 0.049) group and did not differ from that of the SHAM group. Significant difference was not observed in the relation between inorganic and organic compounds in the three groups. The ratio of the areas of the bands in 1057 and 1023 cm-1 for the calculation of IC was 1,024 in the SHAM group, 1,015 in the RS group and 1,108 in OVZ group. In the study of the amide III band, a change in the proportion of secondary structure of matrix proteins was observed in ovariectomized animals, possibly a decrease in of ?-helical and random coil (RC) and an increase in ?-sheet structures. The collagen cross-linking network maturity estimated by the relation between amide I subbands (1660/1690) showed the smaller number in the SR group. In this group, a significant increase in the amount of strontium (Sr) and a decrease in the amount of calcium (Ca) embedded to bone tissue, as assessed by the EDS technique, were also observed. The results demonstrate that systemic treatment with SR promoted bone repair and improved microarchitecture of the newly formed bone within the defect. However, results of analyses with FTIR suggest that the SR has not reversed the alterations caused by ovariectomy in the organic matrix and decreased cross-linking of collagen mature.

Disease- and drug-related bone disorders are rapidly increasing in the population. The drugs which are used for the treatment of neurodegenerative diseases and metabolic derangements, may have negative or positive effects on bone tissues. In the first study, the possible side-effects of Carbamazepine and epileptic seizures on bone structure and composition were investigated by FTIR and synchrotron-FTIR microspectroscopy, AFM and micro- and nano-hardness analysis. The effects on the blood parameters, bone turnover and vitamin D metabolism were also investigated by ELISA and western blot analysis. The current study provides the first report on differentiation of the effects of both epileptic seizures and AED therapy on bones. Besides Carbamazepine treatment, seizures also caused a decrease in the strength of bone. The biochemical data showed that both the epileptic and drug-treated groups decreased vitamin D levels by increasing the vitamin D catabolism enzyme / 25-hydroxyvitamin D-24-hydroxylase. In the second study, the possible pleiotropic (positive) effects of cholesterol lowering drug / Simvastatin on bones were investigated by ATR-FTIR spectroscopy. The current study provides the first report on dose-dependent effects of simvastatin on protein structure and lipid conformation of bones. ATR-FTIR studies showed that although both high and low dose simvastatin strengthen bones, low dose simvastatin treatment is much more effective in increasing bone strength. Neural network analysis revealed an increased antiparallel and aggregated beta sheet and random coil in the protein secondary structure of high dose group implying a protein denaturation. Moreover, high dose may induce lipid peroxidation which limit the pleiotropic effects of high dose treatment on bones. This study clearly demonstrated that using low dose simvastatin is safer and more effective for bone health than high dose simvastatin treatment.

The two main sources of biomass for energy generation are purpose-grown energy crops and waste materials. Energy crops, such as Miscanthus and short rotation woody crops (coppice), are cultivated mainly for energy purposes and are associated with the food vs. fuels debate, which is concerned with whether land should be used for fuel rather than food production. The use of residues from agriculture, such as barley, canola, oat and wheat straw, for energy generation circumvents the food vs. fuel dilemma and adds value to existing crops. In fact, these residues represent an abundant, inexpensive and readily available source of renewable lignocellulosic biomass. In order to reduce industrys operational cost as well as to meet the requirement of raw material for biofuel production, biomass must be processed and handled in an efficient manner. Due to its high moisture content, irregular shape and size, and low bulk density, biomass is very difficult to handle, transport, store, and utilize in its original form. Densification of biomass into durable compacts is an effective solution to these problems and it can reduce material waste. Upon densification, many agricultural biomass materials, especially those from straw and stover, result in a poorly formed pellets or compacts that are more often dusty, difficult to handle and costly to manufacture. This is caused by lack of complete understanding on the natural binding characteristics of the components that make up biomass. An integrated approach to postharvest processing (chopping, grinding and steam explosion), and feasibility study on lab-scale and pilot scale densification of non-treated and steam exploded barley, canola, oat and wheat straw was successfully established to develop baseline data and correlations, that assisted in performing overall specific energy analysis. A new procedure was developed to rapidly characterize the lignocellulosic composition of agricultural biomass using the Fourier Transform Infrared (FTIR) spectroscopy. In addition, baseline knowledge was created to determine the physical and frictional properties of non-treated and steam exploded agricultural biomass grinds. Particle size reduction of agricultural biomass was performed to increase the total surface area, pore size of the material and the number of contact points for inter-particle bonding in the compaction process. Predictive regression equations having higher R2 values were developed that could be used by biorefineries to perform economic feasibility of establishing a processing plant. Specific energy required by a hammer mill to grind non-treated and steam exploded barley, canola, oat and wheat straw showed a negative power correlation with hammer mill screen sizes. Rapid and cost effective quantification of lignocellulosic components (cellulose, hemicelluloses and lignin) of agricultural biomass (barley, canola, oat and wheat) is essential to determine the effect of various pre-treatments (such as steam explosion) on biomass used as feedstock for the biofuel industry. A novel procedure to quantitatively predict lignocellulosic components of non-treated and steam exploded barley, canola, oat and wheat straw was developed using Fourier Transformed Infrared (FTIR) spectroscopy. Regression equations having R2 values of 0.89, 0.99 and 0.98 were developed to predict the cellulose, hemicelluloses and lignin compounds of biomass, respectively. The average absolute difference in predicted and measured cellulose, hemicellulose and lignin in agricultural biomass was 7.5%, 2.5%, and 3.8%, respectively. Application of steam explosion pre-treatment on agricultural straw significantly altered the physical and frictional properties, which has direct significance on designing new and modifying existing bins, hoppers and feeders for handling and storage of straw for biofuel industry. As a result, regression equations were developed to enhance process efficiency by eliminating the need for experimental procedure while designing and manufacturing of new handling equipment. Compaction of low bulk density agricultural biomass is a critical and desirable operation for sustainable and economic availability of feedstock for the biofuel industry. A comprehensive study of the compression characteristics (density of pellet and total specific energy required for compression) of ground non-treated and steam exploded barley, canola, oat and wheat straw obtained from three hammer mill screen sizes of 6.4, 3.2 and 1.6 mm at 10% moisture content (wb) was conducted. Four preset pressures of 31.6, 63.2, 94.7 and 138.9 MPa, were applied using an Instron testing machine to compress samples in a cylindrical die. It was determined that the applied pressure (60.4%) was the most significant factor affecting pellet density followed by the application of steam explosion pre-treatment (39.4%). Similarly, the type of biomass (47.1%) is the most significant factor affecting durability followed by the application of pre-treatment (38.2%) and grind size (14.6%). Also, the applied pressure (58.3%) was the most significant factor affecting specific energy required to manufacture pellets followed by the biomass (15.3%), pre-treatment (13.3%) and grind size (13.2%), which had lower but similar effect affect on specific energy. In addition, correlations for pellet density and specific energy with applied pressure and hammer mill screen sizes having highest R2 values were developed. Higher grind sizes and lower applied pressures resulted in higher relaxations (lower pellet densities) during storage of pellets. Three compression models, namely: Jones model, Cooper-Eaton model, and Kawakita-Ludde model were considered to determine the pressure-volume and pressure-density relationship of non-treated and steam exploded straws. Kawakita-Ludde model provided the best fit to the experimental data having R2 values of 0.99 for non-treated straw and 1.00 for steam exploded biomass samples. The steam exploded straw had higher porosity than non-treated straw. In addition, the steam exploded straw was easier to compress since it had lower yield strength or failure stress values compared to non-treated straw. Pilot scale pelleting experiments were performed on non-treated, steam exploded and customized (adding steam exploded straw grinds in increments of 25% to non-treated straw) barley, canola, oat and wheat straw grinds obtained from 6.4, 3.2, 1.6 and 0.8 mm hammer mill screen sizes at 10% moisture content (wb). The pilot scale pellet mill produced pellets from ground non-treated straw at hammer mill screen sizes of 0.8 and 1.6 mm and customized samples having 25% steam exploded straw at 0.8 mm. It was observed that the pellet bulk density and particle density are positively correlated. The density and durability of agricultural straw pellets significantly increased with a decrease in hammer mill screen size from 1.6 mm to 0.8 mm. Interestingly, customization of agricultural straw by adding 25% of steam exploded straw by weight resulted in higher durability (> 80%) pellets but did not improve durability compared to non-treated straw pellets. In addition, durability of pellets was negatively correlated to pellet mill throughput and was positively correlated to specific energy consumption. Total specific energy required to form pellets increased with a decrease in hammer mill screen size from 1.6 to 0.8 mm and also the total specific energy significantly increased with customization of straw at 0.8 mm screen size. It has been determined that the net specific energy available for production of biofuel is a significant portion of original agricultural biomass energy (89-94%) for all agricultural biomass.

The two main sources of biomass for energy generation are purpose-grown energy crops and waste materials. Energy crops, such as Miscanthus and short rotation woody crops (coppice), are cultivated mainly for energy purposes and are associated with the food vs. fuels debate, which is concerned with whether land should be used for fuel rather than food production. The use of residues from agriculture, such as barley, canola, oat and wheat straw, for energy generation circumvents the food vs. fuel dilemma and adds value to existing crops. In fact, these residues represent an abundant, inexpensive and readily available source of renewable lignocellulosic biomass. In order to reduce industrys operational cost as well as to meet the requirement of raw material for biofuel production, biomass must be processed and handled in an efficient manner. Due to its high moisture content, irregular shape and size, and low bulk density, biomass is very difficult to handle, transport, store, and utilize in its original form. Densification of biomass into durable compacts is an effective solution to these problems and it can reduce material waste. Upon densification, many agricultural biomass materials, especially those from straw and stover, result in a poorly formed pellets or compacts that are more often dusty, difficult to handle and costly to manufacture. This is caused by lack of complete understanding on the natural binding characteristics of the components that make up biomass. An integrated approach to postharvest processing (chopping, grinding and steam explosion), and feasibility study on lab-scale and pilot scale densification of non-treated and steam exploded barley, canola, oat and wheat straw was successfully established to develop baseline data and correlations, that assisted in performing overall specific energy analysis. A new procedure was developed to rapidly characterize the lignocellulosic composition of agricultural biomass using the Fourier Transform Infrared (FTIR) spectroscopy. In addition, baseline knowledge was created to determine the physical and frictional properties of non-treated and steam exploded agricultural biomass grinds. Particle size reduction of agricultural biomass was performed to increase the total surface area, pore size of the material and the number of contact points for inter-particle bonding in the compaction process. Predictive regression equations having higher R2 values were developed that could be used by biorefineries to perform economic feasibility of establishing a processing plant. Specific energy required by a hammer mill to grind non-treated and steam exploded barley, canola, oat and wheat straw showed a negative power correlation with hammer mill screen sizes. Rapid and cost effective quantification of lignocellulosic components (cellulose, hemicelluloses and lignin) of agricultural biomass (barley, canola, oat and wheat) is essential to determine the effect of various pre-treatments (such as steam explosion) on biomass used as feedstock for the biofuel industry. A novel procedure to quantitatively predict lignocellulosic components of non-treated and steam exploded barley, canola, oat and wheat straw was developed using Fourier Transformed Infrared (FTIR) spectroscopy. Regression equations having R2 values of 0.89, 0.99 and 0.98 were developed to predict the cellulose, hemicelluloses and lignin compounds of biomass, respectively. The average absolute difference in predicted and measured cellulose, hemicellulose and lignin in agricultural biomass was 7.5%, 2.5%, and 3.8%, respectively. Application of steam explosion pre-treatment on agricultural straw significantly altered the physical and frictional properties, which has direct significance on designing new and modifying existing bins, hoppers and feeders for handling and storage of straw for biofuel industry. As a result, regression equations were developed to enhance process efficiency by eliminating the need for experimental procedure while designing and manufacturing of new handling equipment. Compaction of low bulk density agricultural biomass is a critical and desirable operation for sustainable and economic availability of feedstock for the biofuel industry. A comprehensive study of the compression characteristics (density of pellet and total specific energy required for compression) of ground non-treated and steam exploded barley, canola, oat and wheat straw obtained from three hammer mill screen sizes of 6.4, 3.2 and 1.6 mm at 10% moisture content (wb) was conducted. Four preset pressures of 31.6, 63.2, 94.7 and 138.9 MPa, were applied using an Instron testing machine to compress samples in a cylindrical die. It was determined that the applied pressure (60.4%) was the most significant factor affecting pellet density followed by the application of steam explosion pre-treatment (39.4%). Similarly, the type of biomass (47.1%) is the most significant factor affecting durability followed by the application of pre-treatment (38.2%) and grind size (14.6%). Also, the applied pressure (58.3%) was the most significant factor affecting specific energy required to manufacture pellets followed by the biomass (15.3%), pre-treatment (13.3%) and grind size (13.2%), which had lower but similar effect affect on specific energy. In addition, correlations for pellet density and specific energy with applied pressure and hammer mill screen sizes having highest R2 values were developed. Higher grind sizes and lower applied pressures resulted in higher relaxations (lower pellet densities) during storage of pellets. Three compression models, namely: Jones model, Cooper-Eaton model, and Kawakita-Ludde model were considered to determine the pressure-volume and pressure-density relationship of non-treated and steam exploded straws. Kawakita-Ludde model provided the best fit to the experimental data having R2 values of 0.99 for non-treated straw and 1.00 for steam exploded biomass samples. The steam exploded straw had higher porosity than non-treated straw. In addition, the steam exploded straw was easier to compress since it had lower yield strength or failure stress values compared to non-treated straw. Pilot scale pelleting experiments were performed on non-treated, steam exploded and customized (adding steam exploded straw grinds in increments of 25% to non-treated straw) barley, canola, oat and wheat straw grinds obtained from 6.4, 3.2, 1.6 and 0.8 mm hammer mill screen sizes at 10% moisture content (wb). The pilot scale pellet mill produced pellets from ground non-treated straw at hammer mill screen sizes of 0.8 and 1.6 mm and customized samples having 25% steam exploded straw at 0.8 mm. It was observed that the pellet bulk density and particle density are positively correlated. The density and durability of agricultural straw pellets significantly increased with a decrease in hammer mill screen size from 1.6 mm to 0.8 mm. Interestingly, customization of agricultural straw by adding 25% of steam exploded straw by weight resulted in higher durability (> 80%) pellets but did not improve durability compared to non-treated straw pellets. In addition, durability of pellets was negatively correlated to pellet mill throughput and was positively correlated to specific energy consumption. Total specific energy required to form pellets increased with a decrease in hammer mill screen size from 1.6 to 0.8 mm and also the total specific energy significantly increased with customization of straw at 0.8 mm screen size. It has been determined that the net specific energy available for production of biofuel is a significant portion of original agricultural biomass energy (89-94%) for all agricultural biomass.

This dissertation presents a study of the molecular mechanism underlying the highly efficient solar energy conversion processes that occur in the Photosystem I (PS I) reaction centers in plants and bacteria. The primary electron donor P700 is at the heart of solar energy conversion process in PS I and the aim is to obtain a better understanding of the electronic and structural organization of P700 in the ground and excited states. Static Fourier Transform Infra-Red (FTIR) difference spectroscopy (DS) in combination with site directed mutagenesis and Density Functional Theory (DFT) based vibrational frequency simulations were used to investigate how protein interactions such as histidine ligation and hydrogen bonding modulate this organization. (P700+-P700) FTIR DS at 77K were obtained from a series of mutants from the cyanobacterium Synechocystis sp. 6803 (S. 6803) where the amino acid residues near the C=O groups of the two chlorophylls of P700 where specifically changed. (P700+-P700) FTIR DS was also obtained for a set of mutants from C. reinhardtii where the axial ligand to A0-, the primary electron acceptor in PS I was modified. The FTIR DS obtained from these mutants provides information on the axial ligands, the hydrogen bonding status as well as the polarity of the environment of specific functional groups that are part of the chlorophyll molecules that constitute P700. Assignment of the FTIR bands to vibrational modes in specific types of environment is very difficult. In order to assist the assignment of the difference bands in experimental spectra DFT based vibrational mode frequency calculations were undertaken for Chl-a and Chl-a+ model molecular systems under different set of conditions; in the gas phase, in solvents using the Polarizable Continuum Model (PCM), in the presence of explicit solvent molecules using QM/MM methods, and in the presence of axial ligands and hydrogen bonds. DFT methods were also used to calculate the charge, spin and redox properties of Chl-a/Chl-a’ dimer models that are representative of P700, the primary electron donor in PS I.

High performance multifunctional epoxy resin systems are becoming increasingly important as matrix materials for the advanced composites used in aerospace, electronics, automotive and other industries. In a composite based on epoxy resin systems, a three-dimensional network of the matrix is formed around the reinforcing fibre as a result of the chemical reaction between the resin and the curing agent. This chemical process, known as curing, is an important event to he considered in the production of composite components made up of these resin systems. Two process parameters namely viscosity and chemical conversion are of paramount significance in the production of composite materials Curing studies of the resin systems based on these two parameters, would therefore assume great importance in deciding the performance reliability of the end product. The objectives of the present investigation are 1. to study the cure kinetics of three thermoset resin systems, viz., i) epoxy novolac (EPIT)/ diamino diphenyl methane{DDM), ii) trigylcidyl para- ammo phenol (TGPAP)/toluene diamine (TDA) and iii) tetraglycidyl diamino diphenyl methane (TGDDM)/pyridine diamine(PDA) using the cure kinetic models based on chemical conversion (α), Theological conversion (β) and viscosity. 2.to develop a correlation between a and viscosity (η) and modify an existing autocatalytic model based on α, to the viscosity domain and 3.to investigate the cure behaviour of these systems in terms of the TTT cure diagram and its associated models. EPN/DDM, TGPAP/PDA and TGDDM/PDA resin systems were chosen for the studies to represent a range of functionalities, The cure was monitored using differential scanning calorimetry (DSC), fourier transform infrared (FTIR) and dynamic mechanical analysis (DMA) techniques by following the changes in enthalpy, functional groups and rheology, respectively. The kinetic parameters namely, order of reaction and activation energy were estimated from dynamic DSC data using the methods of Freeman-Carroll and Ellerstein using nth rate expression. Barton, Kissinger and Osawa methods were employed to find out the activation energy from the peak/equal conversion at different heating rates. Isothermal DSC data were also analyzed using nth order model and it was observed that the data could be fitted satisfactorily only for higher temperatures The results obtained from the analysis of both dynamic and isothermal DSC data using nth order model clearly indicate that this model is inadequate for describing the cure behavior. The isothermal DSC data was analyzed by the autocatalytic models of Hone and Kamal Good correlation was observed with Hum and Kamal models up to 60-70%, 25% and 45% conversions for EPN/DDM, TGPAP/TDA and TCDDM/PDA systems respectively. However, the parameters m and n in Kamal model were found to be temperature dependent for EPN/DDM and TCPAP/TDA systems. The limited applicability of the autocatalytic models IK attributed to the counter-effect offered by the intra-molecular bonding taking place. The primary amine and epoxy groups conversions obtained from FTIR were analyzed using autocatalytic model and the kinetic parameters were calculated. The reactivity ratio of the primary amine and the secondary amine with epoxy was found to be dependent on temperature in agreement with the recent findings reported m the literature. The existing models that relate the cure kinetics and the rheological changes, are dual Arrhenius nth order model and autocatalytic model The nth order kinetic model was used to evaluate the kinetic parameters using the viscosity data at different cure temperatures under isothermal conditions As the storage modulus, G' is proportional to the chemical cross links and becomes significant only after the g<4 point, it was used to follow the changes in conversion known as rheoconversion after the gel point The rheoconversion was found by normalizing the G' data with G1^, the storage modulus of the fully cured resin It was used to study the cuie kinetics using an autocatalytic model The kinetic parameters such as rate constant, acceptation and retardation parameters were evaluated and that temperature dependence was established. While the existing models relate viscosity and conversion only up to gel point the new proposed model, termed VISCON model takes into account the changes up to vitrification. The relation so developed is used to modify the autocatalytic cure model based on chemical conversion. The parameters appearing in this model were evaluated using Levenberg-Marquardt error minimization algorithm. The kinetic parameters obtained are comparable with the values estimated using the DSC data. All the models cited above represent the microkinetic aspects. The models based on the information of TTT cure diagrams, however, represent the macrokinetic aspects of the cure, as they are based on the cure stages such as gelation and vitrification TTT diagram relates the cure characteristics like cure temperature, cure time, Ta and, indirectly, chemical conversion Hence the ultimate properties of the composite could he predicted and established with the help of the models based on TTT cure diagrams The changes in the storage modulus, G1 and loss modulus, G", were followed to identify the gel and vitrification points of the resin systems at different cure temperatures Gel point and vitrification point were used to generate gelation and vitrification hues in the construction of TTT cure diagrams for EPN/DDM, TGPAP/TDA and TGDDM/PDA resin systems Theoretical TTT diagrams were generated and IBO-T, contours were established using the TTT diagram-based models The cure schedule for the resin systems investigated could be determined from the TTT diagram and the respective rheological data.