Estudos envolvendo o oxigênio molecular singlete (1O2) tem uma relevância biológica, uma vez que esta espécie, devido ao caráter eletrofílico, reage com moléculas ricas em elétrons como proteínas, lipídeos e DNA provocando danos que resultam em perdas de função e integridade celular. Em sistemas biológicos, a presença de outras espécies reativas de oxigênio e nitrogênio, dificultam a identificação de lesões específicas causadas por 1O2 .Neste contexto, este trabalho foi desenvolvido objetivando a síntese de endoperóxidos isotopicamente marcados com 17O para serem utilizados como fonte geradora limpa de 17[1O2] em estudos mecanísticos. A capacidade de geração de 17[1O2] pelo endoperóxido N,N\'-di(2,3-dihidroxipropil)-3,3\'-(1,4 naftilideno) dipropanamida 17O (DHPN17O2) foi confirmada utilizando o captador químico sulfato mono-{2-[10-(2-sulfoxi-etil)-antracen-9-il]-etil}éster de sódio e o nucleosídeo 2\'- desoxiguanosina. Os produtos isotopicamente marcados com 17O formados foram analisados por espectrometria de ressonância magnética nuclear e cromatografia líquida de alta eficiência acoplado ao espectrômetro de massa. Os lipídeos, em especial o colesterol ao reagir com o 1O2 geram hidroperóxidos de colesterol como produtos de oxidação primária e na presença de metais resulta em compostos de maior reatividade e toxicidade, como os radicais peroxila, que contribuem para a propagação da peroxidação lipídica. Neste trabalho, demonstramos que os hidroperóxidos de colesterol são capazes de gerar 1O2 na presença de metal através de medidas de luminescência, utilização de supressores e captador químico de 1O2. Os mecanismos de reação envolvidos foram estudados e determinados por espectrometria de massa acoplada a cromatografia líquida de alta eficiência. Por fim, a caracterização detalhada dos produtos formados por espectrometria de ressonância magnética nuclear e massa na reação do colesterol com 1O2 mostrou que além dos hidroperóxidos a reação também produz um aldeído, o 3&#946; -hidroxi-5&#946;-hidroxi-B-norcolestano-6&#946;-carboxialdeído. Até o momento, este composto havia sido identificado como um produto específico da ozonização do colesterol. Neste estudo, baseado nos estudos por reações de quimiluminescência, é proposto o mecanismo de formação deste aldeído em reações de oxidação de colesterol por 1O2 envolvendo intermediário dioxetano. / Studies involving singlet molecular oxygen (1O2) has biological relevance, once this species, due to its eletrophylic character, reacts with rich electron molecules such as proteins, lipids and DNA causing damages that result in loss of function and cellular integrity. In biological system, the presence of other reactive species of oxygen and nitrogen impair the identification of lesions caused by 1O2. In this context, this work was developed with the aim of synthesizing <SUP17O-labeleded endoperoxides to be used as a clean source of (1O2) in mechanistic studies. The ability of 17[1O2]generation by N,N\'-di(2,4-dihydroxypropyl)-1,4-naphthalene-dipropanamide labeled with 17O(DHNP17O2) was observed using the disodium salt of anthracene-9,10-diyldiethyl disulfate as a chemical trap and the nucleoside 2\'-deoxyguanosine. The products isotopically labeled with 17O were analyzed by nuclear magnetic resonance spectroscopy and high performance liquid chromatography coupled to a mass spectrometer. Lipds, in special the cholesterol, when reacting with singlet molecular oxygen generate cholesterol hydroperoxides as primary products and in the presence of metals result in compounds of higher reactivity and toxicity, such as peroxyl radicals which contribute to the propagation of lipid peroxidation. In this work, we demonstrated that cholesterol hydroperoxides are able to generate singlet molecular oxygen in the presence of metal by chemiluminescence measurements by testing the effect of singlet molecular oxygen quencher and by chemical trap. The involved reaction mechanisms were studied and determined by mass spectrometry coupled to the high performance liquid chromatography. Finally, we detailed characterization of the products formed in the reaction of cholesterol with 1O2 by nuclear magnetic resonance and mass spectroscopy showed that besides cholesterol hydroperoxides, the reaction also produces an aldehyde, 3&#946;hydroxy-5&#946;-hydroxy-B-norcholestan-6&#946;-carboxyaldehyde which had been identified as a specific product of cholesterol ozonization. In this study, based on the studies of chemiluminescence reactions, the mechanism of formation of this aldehyde in reaction of oxidation of cholesterol by 1O2 involving a dioxetane intermediate has been proposed.

Functionalized nanoparticles provide a wide range of potential applications for Biological Mass Spectrometry (MS). Particularly, we have studied the effects of chromophore activity on the performance of gold nanoparticles (AuNPs) capped with substituted azo (-N=N-) dyes for analyte ion production in Laser Desorption Ionization Mass Spectrometry (LDI-MS) conditions. A series of aromatic thiol compounds were used as Self-Assembled Monolayers (SAM) to functionalize the surface of the AuNPs. Results indicate that AuNPs functionalized with molecules having an active azo chromophore provide enhanced analyte ion yields than the nanoparticles capped with the hydrazino analogs or simple substituted aromatic thiols. We have also conducted experiments using the azo SAM molecules on 2, 5, 20, 30, and 50 nm AuNPs exploring the changes of Relative Ion Yield (RIY) with increased AuNP diameters. Our results indicate that the role of the SAM to drive energy deposition decreases as the size of the AuNP increases. It was determined that 5 nm is the optimum size to exploit the benefits of the SAM on the ionization and selectivity of the AuNPs.

[1] Four specimens of zoned tourmaline from granitic pegmatites are characterised in detail, each having unusual compositional and/or morphologic features: (1) a crystal from Black Rapids Glacier, Alaska, showing a central pink zone of elbaite mantled by a thin rim of green liddicoatite; (2) a large (~25 cm) slab of Madagascar liddicoatite cut along (001) showing complex patterns of oscillatory zoning; and (3) a wheatsheaf and (4) a mushroom elbaite from Mogok, Myanmar, both showing extensive bifurcation of fibrous crystals originating from a central core crystal, and showing pronounced discontinuous colour zoning. Crystal chemistry and crystal structure of these samples are characterised by SREF, EMPA, and 11B and 27Al MAS NMR and Mössbauer spectroscopies. For each sample, compositional change, as a function of crystal growth, is characterised by EMPA traverses, and the total chemical variation is reduced to a series of linear substitution mechanisms. Of particular interest are substitutions accommodating the variation in [4]B: (1) TB + YAl ↔ TSi + Y(Fe, Mn)2+, where transition metals are present, and (2) TB2 + YAl ↔ TSi2 + YLi, where transition metals are absent. Integration of all data sets delineates constraints on melt evolution and crystal growth mechanisms. [2] Uncertainty has surrounded the occurrence of [4]Al and [4]B at the T-site in tourmaline, because B is difficult to quantify by EMPA and Al is typically assigned to the octahedral Y- and Z-sites. Although both [4]Al and [4]B have been shown to occur in natural tourmalines, it is not currently known how common these substituents are. Using 11B and 27Al MAS NMR spectroscopy, the presence of [4]B and [4]Al is determined in fifty inclusion-free tourmalines of low transition-metal content with compositions corresponding to five different species. Chemical shifts of [4]B and [3]B in 11B spectra, and [4]Al and [6]Al in 27Al spectra, are well-resolved, allowing detection of very small (< ~0.1 apfu) amounts of T-site constituents. Results show that contents of 0.0 < [4]B, [4]Al < 0.5 apfu are common in tourmalines containing low amounts of paramagnetic species, and that all combinations of Si, Al and B occur in natural tourmalines.

[1] Four specimens of zoned tourmaline from granitic pegmatites are characterised in detail, each having unusual compositional and/or morphologic features: (1) a crystal from Black Rapids Glacier, Alaska, showing a central pink zone of elbaite mantled by a thin rim of green liddicoatite; (2) a large (~25 cm) slab of Madagascar liddicoatite cut along (001) showing complex patterns of oscillatory zoning; and (3) a wheatsheaf and (4) a mushroom elbaite from Mogok, Myanmar, both showing extensive bifurcation of fibrous crystals originating from a central core crystal, and showing pronounced discontinuous colour zoning. Crystal chemistry and crystal structure of these samples are characterised by SREF, EMPA, and 11B and 27Al MAS NMR and Mössbauer spectroscopies. For each sample, compositional change, as a function of crystal growth, is characterised by EMPA traverses, and the total chemical variation is reduced to a series of linear substitution mechanisms. Of particular interest are substitutions accommodating the variation in [4]B: (1) TB + YAl ↔ TSi + Y(Fe, Mn)2+, where transition metals are present, and (2) TB2 + YAl ↔ TSi2 + YLi, where transition metals are absent. Integration of all data sets delineates constraints on melt evolution and crystal growth mechanisms. [2] Uncertainty has surrounded the occurrence of [4]Al and [4]B at the T-site in tourmaline, because B is difficult to quantify by EMPA and Al is typically assigned to the octahedral Y- and Z-sites. Although both [4]Al and [4]B have been shown to occur in natural tourmalines, it is not currently known how common these substituents are. Using 11B and 27Al MAS NMR spectroscopy, the presence of [4]B and [4]Al is determined in fifty inclusion-free tourmalines of low transition-metal content with compositions corresponding to five different species. Chemical shifts of [4]B and [3]B in 11B spectra, and [4]Al and [6]Al in 27Al spectra, are well-resolved, allowing detection of very small (< ~0.1 apfu) amounts of T-site constituents. Results show that contents of 0.0 < [4]B, [4]Al < 0.5 apfu are common in tourmalines containing low amounts of paramagnetic species, and that all combinations of Si, Al and B occur in natural tourmalines.

Alzheimer’s disease is an incurable neurodegenerative disorder linked to the amyloid β (Aβ) peptide, a 38-43 residue peptide. The detailed molecular disease mechanism(s) is (are) unknown, but oligomeric Aβ structures are proposed to be involved. In common for the papers in this thesis is interactions; interactions between Aβ(1-40) and selected molecules and metal ions. The purpose has been to find out more about the structural states that Aβ can adopt, in particular the β-sheet state, which probably is linked to the oligomeric structures. The methods used have been nuclear magnetic resonance (NMR), circular dichroism (CD) and fluorescence spectroscopy using Thioflavin T (ThT). Upon addition of SDS/LiDS detergent or Congo red (CR) to Aβ(1-40), the initial random coil/PII-helix state was transformed into β-sheet and, in the case of detergent, a final α-helical state. In contrast to SDS/LiDS and CR, the dimeric Affibody molecule locks monomeric Aβ(1-40) in a β-hairpin state. It was found that by truncating the flexible N-terminal end of the Affibody molecule its affinity to Aβ was improved. The aggregation of Aβ(1-40) was further studied in the presence of a β-cyclodextrin dimer by a kinetic assay using ThT. Although having a weak dissociation constant in the millimolar range, the β-cyclodextrin dimer modified the aggregation pathways of Aβ. Finally Aβ(1-40) was studied in presence of Cu2+ and Zn2+ at physiological and low pH. Cu2+ was observed to maintain its specific binding to Aβ when decreasing the pH to 5.5 while Zn2+ behaved differently. This could be of importance in the Alzheimer’s disease brain in which the environment can become acidic due to inflammation.        In conclusion the results show that Aβ(1-40) is very sensitive to its environment, responding by adopting different conformations and aggregating in aqueous solutions. The β-sheet state is induced by varying molecules with different properties, properties that govern the final Aβ state. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: Manuscript. Paper 5: Manuscript.

The objectives of the research were to examine the effects of high dietary levels of fibrous feeds, and of supplementation with Roxazyme® G2 (RX), on the digestive metabolic and physiological responses of growing pigs fed maize-soybean diets. The nutrient and dietary fibre (DF) composition, the swelling and water-binding capacities of maize (MM), its hominy chop (HC) and cobs (MC), dehulled soybean (dSBM) and the hulls (SH), brewer’s grains (BG), lucerne hay (LH) and wheat bran (WB) were evaluated using standard procedures. Feed fibre fractions were isolated by simulating upper tract digestion in an Ankom® DaisyII Incubator, whereby each feed was digested in pepsin (porcine, 200 FIP-U/g, Merck No, 7190), followed by pancreatin (porcine, grade IV, Sigma No P-1750), with recovery of the fibrous residues. In a third step to complete the simulated pig gastro-intestinal digestion, the pepsin-pancreatin fibre extracts were digested by RX or Viscozyme L ® V2010 (VZ). Enzyme activity was measured as the coefficients of partial degradability (solubilisation) of the washed fibre extracts. The kinetics and products of fermentation of the DF were evaluated in an AnkomRF gas production system, using buffered faecal inoculum. Among the feed ingredients, dissimilar, fibre source-dependent activities between RX (0.02 to 0.12) and VZ (0.04-0.33) were observed. The lowest RX activities were observed on the maize and soybean derived fibres, with similarly low VZ activity on MC fibre. Variation in the activity of faecal microbial enzymes was similarly indicated by the variable production of fermentation gas (51.8-299.4 mL g-1 DM) and short chain fatty acids (SCFA) (2.3-6.0 mMol g-1 DM). Soy hull, dSBH, MM and HC fibres were highly fermentable, with low fermentability of BG, MC and WB fibres. The fibres differed in the composition of fermentation SCFA, whereby SH, LH and MC shifted fermentation to Ace, and BG, dSBM, WB, MM, HC favoured Pro, while MM and HC favoured But production. The same nutritional properties were similarly evaluated in complete diets which were formulated from the ingredients for growth, and metabolic trials. For the growth trial, a standard (STD) (control), 141 g total dietary fibre (TDF) kg-1 dry matter (DM) maize-soybean growing pig diet, and five iso-nutritive, 246 g TDF kg-1 DM nutritionally balanced diets were formulated. The high DF was achieved by partial replacement of the MM and dSBM in the STD diet with MC, SH, BG, LH or WB. The differences in RX and VZ activities and in the fermentation characteristics which were observed on the fibre extracts from the high fibre ingredients were reflected in the DF from the respective complete diets in which they were included. However, the fibre from the basal dietary ingredients reduced the absolute values and the variation in the activities of RX (0.03-0.06) and VZ (0.16-0.22), and similarly reduced the variation in gas (126.6-187.6 mL g-1 DM) and SCFA (4.1-5.4 mMol g-1 DM) production of the DF from the fibrous diets. Enzyme activities on the STD DF were low for RX (0.03) and high for VZ (0.25). The STD DF produced 205.3 mL gas g-1 DM, which was similar to SH DF, and higher than all the other diets. The STD DF produced 5.0-mMol SCFA g-1 DM, which was quantitatively, and not statistically higher than the other fibres. The composition of SCFA was similar across all diets, except for the high percent Ace, with low Pro by the SH DF. Compared to the STD, the high DF diets increased percent Ace, with reduced Pro and But. The STD, MC, SH, BG, LH and WB diets were each prepared in duplicate mixes, one of which was fortified with 200 mg RX kg-1 feed (as fed). Seventy-two intact Large White X Landrace, male, 32.0 ± 5.6 kg live weight (LW) pigs were allocated to the diets in two completely randomised weight blocks in a 2 (fibre source) X 2 (enzyme) factorial arrangement. The pigs were fed ad libitum for 10 weeks. Cumulative LW gain and feed intake were measured at different stages of growth, and at slaughter. Apparent total tract digestibility (ATTD) of nutrients was estimated at 65-70 kg LW, using 0.2% (as fed) chromium oxide as the indigestible marker. Ileal tissue was sampled 50 cm above the ileo-caecal valve, on which villi height and area, and crypt depth were evaluated by computerised image analysis. Blood was sampled at slaughter from the severed vena jugularis, 16 hours after feeding. Serum urea, creatinine, triglycerides, glucose, and total cholesterol were analysed chemically. The serum metabolome was further explored using Proton Nuclear Magnetic Resonance Spectroscopy (1H -NMRS). There was fibre X RX interaction for villi height, whereby the enzyme reduced the villi height in pigs on the SH, STD and WB diets, with an opposite effect on pigs on the MC, BG, LH diets. The soluble fibre content was negatively correlated with crypt depth. Chemical analysis did not detect differences in metabolite concentration between the STD and the high fibre diets. However, more serum cholesterol was observed in pigs fed the WB compared to the LH and MC diets. 1H-NMRS indicated that feeding pigs the WB diet increased serum Cys and His, while supplementation of RX increased serum formate, glucose, and urea. There was diet X enzyme interaction for fructose, glucose, Arg, Cys, Ser, and Trp, whereby RX increased the levels in pigs on MC and WB, with an opposite effect in pigs on the other diets. There was large DF source-dependent variation among diets in ATTD of DM (0.80-0.85), organic matter (OM) (0.81-0.87), gross energy (GE) (079-0.85) and CP (0.81-0.85), whereby, relative to the STD diet, high DF reduced the ATTD of DM (all diets except SH), organic matter (OM) and energy and CP (all diets except the MC). Positive correlation was observed between fermentability and the ATTD digestibility of DM, OM, energy, ADF, NDF, and fat. Negative correlation was observed between the swelling capacity and the ATTD of DM, OM, energy and protein, between DF solubility and DM, OM, protein, ADF and NDF, and between water binding capacity and ATTD of DM and OM, energy and NDF. At slaughter, there was similarly large, and DF source-dependent variation among the high fibre diets in feed intake (2.31-2.71 kg as fed day-1), live weight gain (0.75-0.86 kg day-1), and feed: gain ratio (2.73-3.00). Corresponding values for the STD diet were 2.44 kg day-1, 0.83 kg day-1and 2.86 kg day-1, respectively. Relative to the STD, LH reduced feed intake and live weight gain, and MC increased the feed: gain ratio. Predictions based on the in vitro fermentability of DF and feed intake suggested that due to poor fermentability, and or restriction of feed intake, relative to a standard fibre diet, high dietary levels of MC, WB and BG may reduce fermentation in the lower gut, while similar dietary levels of SH and LH may result in substantial increases in fermentation. At 50 kg LW, the fermentability of DF was positively correlated with feed intake and with weight gain, while water binding capacity and solubility of DF were negatively correlated with feed intake. At slaughter, the solubility of DF was negatively correlated with feed intake and feed: gain ratio. Large variation among the high fibre diets was also observed in the slaughter weight (89.2-96.8 kg), dressing % (68.6-76.4), meat colour (80.4-82.3), lean % (69.5-71.2), and fat % (10.1-12.6). In comparison, pigs on the STD diet scored 94.7 kg slaughter weight, 75.1% dressing, 81.6 cm carcass length, 82.5 meat colour, 68.4% lean, and 15.0% fat. Relative to the STD, LH reduced dressing and fat %. Lucerne hay and WB increased the lean%. For the metabolic trial, two iso-nutritive, mixed high fibre (319 g TDF kg-1 DM), nutritionally balanced diets were formulated to contain DF of high (HF) versus low (LF) fermentability. The diets had similar content of soluble DF and similar swelling and water binding capacities. Viscozyme was more active than RX on both the HF (0.20 versus 0.04) and the LF (0.17 versus 0.07) DF. The combination of RX and VZ statistically increased the enzyme activity on the HF (0.25) and quantitatively increased enzyme activity on the LF (0.18) DF, suggesting additive or synergistic effects. More gas was produced by the HF (159.5 mL g-1 DM) compared to the LF DF (96.6 mL g-1 DM). More SCFA were produced by HF (5.0 mMol g-1 DM), compared to the LF DF (3.6 mMol g-1 DM). Compared to the STD, HF DF increased percent Ace, with reduced Pro and But. The LF DF increased percent Ace, with quantitative, and not statistical reduction of Pro and But. In a metabolic trial, the HF and LF diets, and their duplicates containing 0.270 g RX kg-1 DM of feed (as fed) were fed ad libitum to eight ileum T-cannulised, intact Large White X Landrace male pigs weighing 65.0 ± 5.1 kg. The diets were allocated to the pigs in a duplicate 4 x 4 Latin Square design, in a 2 (enzyme) x 2 (fermentability) factorial arrangement. Each period consisted of two weeks of adaptation followed by five days of sampling. The ileal digesta was collected in each period and was similarly subjected to the fermentation test. Apparent ileal digestibility (AID) and ATTD were determined using 0.2% (as fed) chromium oxide as the indigestible marker. N excretion in faeces and urine were measured, and N retention was calculated. Blood was sampled by vena jugularis puncture on the last day of each period. Two blood samples were collected, the first 15 hours after removal from feed (15-hour serum), and the second 3 hours after re-introduction to feed (3-hour serum). Serum metabolites were evaluated by both chemical analyses and by 1H-NMRS, as described for the growth trial. Roxazyme did not affect the fermentation characteristics of the ileal digesta. In similar proportion to the fermentability of the PP digesta, the HF ileal digesta was more fermentable (65.4 mL gas g-1 DM and 6.1 mMol SCFA g-1 DM) than the LF ileal digesta (46.7 mL gas g-1 DM and 4.4 mMol SCFA g-1 DM SCFA). Prediction based on the in vitro fermentability of DF and feed intake suggested the HF diet could support one half times more fermentation in the lower gut compared to the LF diet. The HF diet had higher AID of DM (62.5 vs. 58.6), OM (65.6 vs. 62.1), energy (64.4 vs. 61.0), fat (85.8 vs. 81.7) and ash (41.8 vs. 32.7). The AID of HO-Pro, Met and Val were higher for the LF diet. There was diet X enzyme interaction on the AID of Met, whereby the RX reduced the AID of met in the LF diet, and not that of the HF diet. The ATTD was higher for the HF diet for DM (74.2 vs. 68.4), NDF (64.7 vs. 57.4), and ADF (35.1 vs. 21.0). There was positive correlation between the fermentability of DF and the AID DM, OM, ash, ash, fat and energy. The solubility of DF was negatively correlated with the AID of DM, OM, ash, fat, ADF and energy, and with the ATTD of DM, OM, ash, fat, energy, NDF, and ADF. Negative correlation was also observed between the swelling capacity of DF and the AID of protein, Trp and Lys. The solubility of DF was positively correlated with Ser, Ala, Val, Iso-Leu and His. There was diet X enzyme interaction for urea in the 15- hour serum, whereby RX tended to reduce the urea in the LF diet, while it increased that of the HF diet. Fermentability negatively correlated with urea in the 15- hour serum, and positively correlated with serum glucose in the 3-hour serum. In the 3-hour sample, 1H-NMRS indicated higher fucose, Pro and cholesterol in the LF diet. 1H-NMRS also indicated fermentability x RX interaction for Ser, Tyr, Lys, creatine, and possibly, glucose or fructose, glycerol or Gly and His or Arg, whereby RX increased the levels in the LF diets, with opposite effect in the HF diet. In conclusion, enzyme activities and fermentability were higly variable among different DF sources, and the effects were evident in the fibrous complete diets. The results of the in vitro studies supported the application of the methods to formulate fermentable insoluble fibre-rich, maize-soybean-mixed co-product diets. Correlation analyses suggested that DF fermentability, and solubility, swelling and water binding capacities explained significant proportions of the variances of the metabolic and physiological responses of the pigs to different feeds. Predictions based on the in vitro fermentability of DF and feed intake suggested that a strategy whereby pig diets are enriched in DF after the feedstuffs are screened on DF fermentability could substantially increase fermentation in the lower gut. Overall, the results suggested that productivity can be maintained in growing pigs fed diets containing up to twice the standard levels of DF, provided producers target co-product feeds that contain highly fermentable DF. The use of RX to improve nutrient digestion and to stimulate gut fermentation was not justified. / Environmental Sciences / Ph.D. (Environmental Sciences)

The HIV-1 pandemic affects millions of people worldwide with approximately 70% of those affected residing in sub-Saharan Africa (SSA) relying on traditional medicines for treatment. The key aim of the study was to isolate and characterise an active phyto-pharmaceutical ingredient (API) from L. trigonus for use as a vaginal microbicide. The aerial parts of L. trigonus were oven-dried at 80°C, ground and then extracted with boiling water for 30 minutes. Aqueous extracts were screened using an HIV-1 neutralization assay in TZM bl cells. Chromatographic and spectroscopic techniques were used to purify, isolate and identify the API. The API (BP36-117-26464C) was identified as a polymeric macromolecule with IC50 = 0.04 μg/ml against HIV-1 HXB 2 subtype B. This activity is comparable to the ARV drug, enfuvirtide (IC50 = 0.02 μg/ml). The API consists of galacturonic acid polymer and a mixture of seven compounds. Its mode of action may involve inhibiting virus attachment. The activity of this precipitate (BP36-117-26464C) tested against HIV-1 subtype C pseudovirions and shown to compare favorably with that of enfuvirtide (T20). The water-soluble nature of this API and its mode of action identified it as a potential microbicide. In the current form, the precipitate (API) would be difficult to develop as an oral treatment for HIV, as high-molecular weight agents often have poor bioavailability following oral administration. However, large molecules with potent anti-HIV activity are ideal for topical use and potent development as a microbicide. / Life and Consumer Sciences / M.Sc (Life Sciences)

The study of quadrupolar nuclei using NMR spectroscopy in the solid state significantly increased in popularity from the end of the 20th century, with the introduction of specific methods to acquire spectra free from the effects of the quadrupolar interaction, that results in broadened lineshapes that cannot be completely removed by spinning the sample at the magic angle (MAS), unlike most of the other interactions present in the solid state. The first technique which allows, without any specific hardware, the removal of this broadening has been the Multiple-Quantum MQMAS experiment. The method quickly gained a popularity within the NMR community, with numerous successful applications published. However, the multiple-quantum filtration step in this experiment relies on severely limits sensitivity, restricting application to the most sensitive nuclei. Extending the applicability of MQMAS to less receptive nuclei requires the use of signal improvement techniques. There are multiple examples of such approaches in the literature, but most of these require additional optimisation that may be time-consuming, or simply impossible, on less receptive nuclei. This work introduces a novel signal improvement technique for MQMAS, called FAM-N. Its optimisation is solely based on density matrix simulations using SIMPSON, implying no additional experimental optimisation is required, while improving the signal in MQMAS spectra by equivalent or higher amounts than other common methods. In order to prove the applicability of this method on virtually any system, FAM-N has been investigated by simulation, and tested experimentally using a number of model samples, as well as samples known to be challenging to study by NMR. This work also explores other aspects of NMR spectroscopy on quadrupolar nuclei. Adiabatic inversion of the satellite populations can be performed to improve the central transition signal in static or MAS spectra. A range of methods has been tested and compared, with particular attention given to hyperbolic secant-shaped pulses, for which its performance have been described. Finally, cross-polarisation from a spin I = 1/2 nucleus to a quadrupolar nucleus has been investigated. After reviewing the theory for the static case, simulations have been performed under MAS in order to identify the conditions for efficient magnetisation transfer, with applications in spectral editing or for the combination with MQMAS.

This thesis describes applications of advanced multinuclear solid-state nuclear magnetic resonance (NMR) experiments to the characterisation of industrially-relevant catalyst materials. Experiments on γ-Al₂O₃ introduce the use of solid-state NMR spectroscopy for the investigation of disordered solids. The existence of Al(V) sites on the surface of this material is demonstrated, showing that removal of adsorbed H₂O may facilitate a rearrangement effect in γ-Al₂O₃ that promotes the formation of these Al environments. A range of aluminium oxide-based supported metal catalysts has been investigated. Studies of these systems by ¹H and ²⁷Al solid-state NMR spectroscopy indicate that a metal-support interaction (MSI) exists between surface cobalt oxide crystallites and the γ-Al₂O₃ support, and is strongest for materials containing small, well dispersed Co oxide crystallites. It is shown that the hygroscopic nature of γ-Al₂O₃ allows the extent of the MSI to be visualised by ¹H MAS NMR, by observing the extent of the proton-metal oxide interaction resulting from the presence of adventitious adsorbed H₂O. The surface/bulk chemistry of Co spinel aluminate materials is also investigated. ¹H, ²⁹Si, ²⁷Al and ¹⁷O solid-state NMR techniques are used to gain insight into the structural nature of silicated alumina catalysts. The combination of isotopic enrichment and dynamic nuclear polarisation (DNP) surface-enhanced NMR spectroscopy can provide a definitive and fully quantitative description of the surface structure of Si-γ-Al₂O₃ (1.5 wt% Si), and the role of adventitious surface water is highlighted. Analysis of silicated aluminas prepared by “sequential grafting” and “single shot” approaches shows that silica growth on γ-Al₂O₃ follows two distinct morphologies. ¹⁷O gas exchange enrichment is also shown to be successful in facilitating ¹⁷O solid-state NMR studies of these materials. It is demonstrated that double (²⁹Si and ¹⁷O) enrichment of Si-γ-Al₂O₃ (1.5 wt% Si) can facilitate access to ²⁹Si-¹⁷O 2D correlation experiments, even at low silica loading. An exploratory investigation of Ti-alumina model catalysts has also been carried out using ¹H, ²⁷Al and ¹⁷O solid-state NMR spectroscopy. These studies indicate that Ti-γ-Al₂O₃ and Ti-Al M50 may be structurally distinct materials.

Made available in DSpace on 2016-12-08T15:50:04Z (GMT). No. of bitstreams: 1 PGMS14DA022.pdf: 1661890 bytes, checksum: 3147ca0fadcd842b04a10dab73754a64 (MD5) Previous issue date: 2014-05-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The total content of phosphorus (P) is distributed in the soil forms with different degrees of lability, being the distribution of the forms affected by type soils and soil manegement. The work consisted of three studies, being that the first study quantified the stock of the forms of inorganic and organic P in the surface horizon and in 42 profiles soil of six orders in the Santa Catarina State. After collection, the samples were subjected to chemical fractionation of P Hedley. The soils most weathered had higher stock of P total, while the soils less weathered showed higher levels of labile organic P. In the second study was conducted fractionation analysis of P and P nuclear magnetic resonance (NMR) in soil agroecosystems with high organic matter content. The areas were evaluated in apple orchard, native grass and pine plantation. The results of chemical fractionation and NMR of P, showed that the conversion of natural ecosystems to areas cultivated altered the distribution of P forms in soil, where the fractions organic were reduced in culture without the addition of phosphate fertilizer systems. The third study involved the assessment of forms of P in soils with different clay contents that subjected the applications of liquid swine manure. Soil samples were collected in three regions of Santa Catarina State and subjected to chemical fractionation of P. The results showed that the addition of liquid swine manure for long term increased the content of all forms of P evaluated by fractionation, mainly in inorganic forms, with more accumulated in the topsoil. In soils with sandy texture there accumulation of labile P, and in soils clay there accumulation the of forms lower lability / O conteúdo total de fósforo (P) do solo é distribuido em formas com diferentes graus de labilidade. A distribuição das formas de P podem ser afetadas pelo tipo de solo e o manejo adotado durante seu uso. O trabalho foi composto de três estudos. O primeiro estudo quantificou o estoque das formas de P inorgânicas e orgânicas no horizonte superficial e nos perfis de 42 solos de seis ordens no Estado de Santa Catarina (SC). Após a coleta, as amostras foram submetidas ao fracionamento químico de P de Hedley. No segundo estudo foi realizado o fracionamento químico de P e análise de ressonância magnética nuclear de P (RMN) em amostras de solo de áreas de pomar de maçieira, campo nativo e florestamento de pinus com alto teor de matéria orgânica. O terceiro estudo consistiu na avaliação das formas de P em solos com diferentes teores de argila que foram submetidos a aplicações de dejetos líquidos de suínos. As amostras de solo foram coletadas em três regiões do Estado de SC e submetidas ao fracionamento químico de P. No estudo 1 os solos mais intemperizados apresentaram maiores estoque de P total enquanto os solos menos intemperizados apresentaram maiores teores de P lábil e P orgânico. No segundo estudo os resultados do fracionamento químico e RMN de P mostraram que a conversão de ecossistemas naturais em áreas cultivadas alterou a distribuição das formas de P no solo, onde as frações orgânicas foram reduzidas em sistemas de cultivo sem a adição de fertilizantes fosfatados. Os resultados do terceiro estudo mostraram que a adição de dejetos líquidos de suínos por longo prazo aumentou o teor em todas as formas de P avaliadas pelo fracionamento, principalmente em formas inorgânicas, sendo mais acumulado na camada superficial do solo. Nos solos com textura arenosa houve acúmulo de P principalmente nas frações lábeis e em solos de textura argilosa houve acúmulo de P principalmente em formas de menor labilidade