Indiana University-Purdue University Indianapolis (IUPUI) / In this thesis, we proposed and investigated the efficacy and safety of dental bleaching as a non-invasive aesthetic treatment option for stained arrested caries lesions (s-ACLs). Chapter 1 reports the suitability of this approach in extracted human teeth, as well as in a selected clinical case. Visual improvement in the color lighteness of the s-ACLs was observed and reported in photographs. In order to systematically study the impact of dental bleaching on the s-ACLs, we developed in vitro models simulating the development of metallic and non-metallic s-ACLs (Chapter 2). Human dental specimens were submitted to incipient caries-like lesion formation, followed by a 5-day cycling protocol based on remineralization and staining episodes. The created lesions were then bleached (simulating in-office/40% hydrogen peroxide). Color change was measured spectrophotometrically at baseline, after lesion creation, staining/remineralization cycling and bleaching; while mineral loss and lesion depth were quantified by transversal microradiography after staining/remineralization cycling. Metallic s-ACLs were darker, more remineralized and more difficult to bleach, compared to the non-metallic ones (p<0.05). In Chapter 3, we tested the efficacy and safety of different dental bleaching systems (simulating at-home/15% carbamide peroxide and in-office/40% hydrogen peroxide) using the in vitro models previously developed. Similar methods and outcomes were used, with the addition of demineralization after bleaching to simulate and test changes in caries susceptibility. At-home bleaching showed greater efficacy in color improvement compared to in-office (p<0.05), but also increased susceptibility to further demineralization (p<0.05), regardless of the type of stain. Overall, bleached non-metallic s-ACLs were more susceptible to demineralization compared to metallic ones (p<0.05). Within the limited laboratory testing conditions, we concluded that dental bleaching can improve the aesthetics of s-ACLs, with efficacy being dependent on the nature of the stain. At-home bleaching presented greater efficacy, but also raised some potential safety concerns, which should be further investigated in clinical conditions. In-office bleaching protocol showed to be an effective and safe procedure for the aesthetic treatment of stained arrested caries lesions.

The entire epidermis of patients with vitiligo exhibits accumulation of up to 10-3M concentrations of hydrogen peroxide (H₂O₂) (Schallreuter, Moore et al. 1999). Over the last decade our group and others have focused on the effect of H₂O₂-mediated oxidative stress on the function of many proteins and peptides due to oxidation of target amino acid residues in their structure including L-methionine, L-tryptophan, L-cysteine and seleno cysteine (Rokos, Beazley et al. 2002; Gillbro, Marles et al. 2004; Hasse, Kothari et al. 2005; Schallreuter, Chavan et al. 2005; Spencer, Chavan et al. 2005; Chavan, Gillbro et al. 2006; Elwary, Chavan et al. 2006; Gibbons, Wood et al. 2006; Schallreuter, Bahadoran et al. 2008; Shalbaf, Gibbons et al. 2008; Wood, Decker et al. 2009). Moreover, it was shown that patients with vitiligo possess up regulated wild type functioning p53 protein in their skin (Schallreuter, Behrens- Williams et al. 2003). The reason behind this up regulation has remained unclear (Schallreuter, Behrens-Williams et al. 2003). Therefore the aim of this thesis was to get a better understanding of these puzzling data. Along this project different techniques have been used including Western blot, dot blot, immuno precipitation, immuno fluorescence, EMSA and computer modelling. In this thesis we confirmed the previous result on up regulation of p53 in vitiligo and we showed that p90MDM2, the master regulator for p53 protein is not different in patients and healthy controls. Therefore we decided to test for expression of p76MDM2 which mediates the inhibition of p90MDM2-p53 binding. Our results show for the first time the presence and over expression of p76MDM2 protein in vitiligo compared to 3 healthy individuals. This result could provide an explanation, why up regulated p53 is not degraded in this disease. Since epidermal H₂O₂ accumulation has been extensively documented in vitiligo, we wanted to know whether other ROS could also contribute to the overall oxidative stress in this scenario. Therefore we turned our interest to nitric oxide (NO) and its possible effects on p53 protein. In order to elucidate this role in more detail, the expression levels of epidermal nitric oxide synthesase (iNOS) and the oxidation product of NO and O2 - i.e peroxynitrite (ONOO-) were investigated. Our data revealed over expression of iNOS and nitrated tyrosine residues, the foot print for ONOO-. Moreover, we show for the first time the presence of abundant nitration of p53 protein in vitiligo. In addition using purified p53 from E. coli strain (BL21/DE3) and mutant p53 protein from HT-29 cells (colon cancer cells), we show that nitration takes place in a dose and time dependent manner. On this basis we investigated the effect of both H₂O₂ and ONOO- on p53-DNA binding capacity employing EMSA, since this is the most acceptable technique to follow the binding between proteins and DNA. Our results revealed that ONOO- abrogated p53-DNA binding capacity at concentrations >300 μM, meanwhile oxidation of p53 protein with H₂O₂ at the same concentrations does not affect binding capacity. Importantly, a much higher p53- DNA binding capacity was observed after exposure to both ONOO- and H₂O₂. Taken together, p53 is regulated by both ROS (H₂O₂) and RNS (ONOO-). Next we identified the presence of phosphorylated and acetylated p53 in vitiligo. Phosphorylation of ser 9 and ser 15 residues of the protein are associated with over expressed ATM protein kinase, while acetylation of lys 373, 382 residues correlates with increased PCAF expression. We show that up regulated p53 is associated with over expressed p21 (cyclin dependent kinase inhibitor 1) and induced PCNA 4 expression. Hence, we can conclude that p53 in patients with vitiligo is up regulated, activated and functional. Finally we show up regulated BCL-2 supporting the long voiced absence of increased apoptosis in vitiligo. Given that patients with vitiligo have no increased risk for solar induced skin cancer and increased photo damage (Calanchini-Postizzi and Frenk 1987; Westerhof and Schallreuter 1997; Schallreuter, Tobin et al. 2002), despite the presence of increased DNA damage as evidenced by increased 8-oxoG levels in the skin and in the plasma, our findings suggest that both p53 and PCNA provide a powerful machinery to mediate DNA repair via hOgg1, APE1 and DNA polymerase ß (Shalbaf 2009). On this basis it is tempting to conclude that DNArepair is the overriding mechanism to combat oxidative stress in this disease.

Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) and superoxide (O2 • −) have been shown to serve as messengers in biological signal transduction, and many prokaryotic and eukaryotic proteins are now known to have their function controlled via ROS-mediated oxidation reactions occurring on critical cysteine residues. The tumour-suppressor protein p53 is involved in the regulation of a diverse range of cellular processes including apoptosis, differentiation, senescence, DNArepair, cell-cycle arrest, autophagy, glycolysis and oxidative stress. However, little is understood about the specific molecular mechanisms that allow p53 to discriminate between these various different functions. p53 is a multiple cysteine-containing protein and there is mounting evidence to suggest that redox-modification of p53 Cys residues participate in control of its biological activity. Furthermore, p53 activity has been linked to intracellular ROS levels. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) offers superior mass resolving power and mass measurement accuracy, which is beneficial for the study of intact proteins and the characterisation of their posttranslational modifications (PTMs). The primary goal of the work described in this thesis was to employ FT-ICR mass spectrometry to investigate the molecular details of p53 redox-regulation. The relative reactivity of each of the ten cysteine residues in the DNA-binding core domain of recombinant human p53 was characterised by treatment with the Cys-alkylating reagent N-ethylmaleimide (NEM) under various conditions. A combination of top-down and middle-down FT-ICR MS was used to unambiguously identify Cys182 and Cys277 as sites of preferential alkylation. These results were confirmed by site-directed mutagenesis. Interestingly, Cys182 and Cys277 have previously been implicated in p53 redox-regulation. Alkylation beyond these two residues was found to trigger rapid alkylation of the remaining Cys residues, presumably accompanied by protein unfolding. These observations have implications for the re-activation of mutant p53 with Cys-targeting compounds which result in the death of cancer-cells. Furthermore, the molecular interaction between p53 and the ROS hydrogen peroxide was investigated. p53 was found to form two disulfide bonds upon treatment with H2O2. An enrichment strategy was developed to purify oxidised p53 and top-down FT-ICR mass spectrometry revealed unambiguously that Cys176, 182, 238 and 242 were the oxidised residues. Interestingly, Cys176, 238 and 242 are Zn2+- binding residues suggesting that p53 contains a zinc-redox switch. The mechanism of H2O2 oxidation was investigated, and revealed that oxidation via an alternative pathway results in indiscriminate over-oxidation of p53. Moreover, Cys176, 238 or 242 was shown to act as a nucleophile, and the intracellular antioxidant glutathione (GSH) did not prevent oxidation of the Zn2+-binding Cys residues, providing further evidence for a role in p53 redox-regulation. This study has revealed hitherto unknown details regarding the chemistry of cysteine residues within the important tumour-suppressor protein p53. Furthermore, the analytical power of FT-ICR MS for the study of multiple Cys-containing proteins has been very clearly demonstrated.

The use of two online sensors, the S::CAN and the FLD, was used to evaluate the treatment of synthetic wastewater by UV/H₂O₂ advanced oxidation. The UV dose and intensity for the flow through reactor was also determined using H₂O₂ actinometry. The total power of the lamps in the reactor was determined to be 139 W which, with the absorbance of the water, produced a UV dose of 39.4 mJ/cm². It was determined that the S::CAN was unable to differentiate between treatment of a particular water, however it could differentiate between different test waters. The FLD was very sensitive and was able to detect minute variations in treatment of the test water.

Orientador: Alessandra Nara de Souza Rastelli / Resumo: O objetivo deste estudo in vitro foi avaliar a eficácia de diferentes técnicas de clareamento dental, através das análises de dureza e contéudo mineral do esmalte dental clareado. Diferentes protocolos foram testados após manchamento dos espécimes com (café), como segue: G1: Whiteness HP Maxx (FGM); G2: Whiteness HP Maxx (FGM) + LED Azul - Twin Flex Evolution (MMOptics); G3: Whiteness HP Maxx (FGM) + LED Violeta - Bright Max Whitening (MMOptics); G4: Peróxido de Hidrogênio a 10% (FGM); G5: Peróxido de Hidrogênio a 10% (FGM) + Led Violeta - Bright Max Whitening (MMOptics); G6: Peróxido de Carbamida a 22% (FGM); G7: Peróxido de Carbamida a 22% (FGM) + LED Violeta - Bright Max Whitening (MMOptics) e G8: LED Violeta - Bright Max Whitening (MMOptics). Foram obtidos oitenta (n=80) espécimes de (4x4x2mm) da face vestibular de incisivos bovinos, após foram seccionados ao meio em máquina de corte, separando cada espécime em duas metades, uma metade foi utilizada como espécime sem procedimento clareador (controle) e a outra metade como espécime com procedimento clareador (tratado). Foi realizada análise em profundidade no esmalte, afim de verificar as possíveis alterações na superficíe (S) e subsuperficie (P), assim, cento e sessenta (n=160) espécimes foram avaliados quanto microdureza Knoop (n =10) e espectroscopia Raman (n = 5). Os diferentes protocolos para clareamento dental foram aplicados de acordo com os fabricantes dos produtos e dos equipamentos. Os espécimes permaneceram em s... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this in vitro study was to evaluate the effectiveness of different dental bleaching techniques through microhardness analysis and mineral content of bleached dental enamel. Different protocols were tested after spotting the specimens with (coffee), as follows: G1: Whiteness HP Maxx (FGM); G2: Whiteness HP Maxx (FGM) + Blue LED - Twin Flex Evolution (MMOptics); G3: Whiteness HP Maxx (FGM) + Violet Led - Bright Max Whitening (MMOptics); G4: 10% Hydrogen Peroxide (FGM); G5: 10% Hydrogen Peroxide (FGM) + Violet Led - Bright Whitening (MMOptics); G6: 22% carbamide peroxide (FGM); G7: 22% Carbamide Peroxide (FGM) + LED Violet - Bright Max Whitening (MMOptics) and G8: Violet LED - Bright Max Whitening (MMOptics). Eighty (4x4x2mm) specimens were collected from the bovine incisal vestibular surface, after being sectioned in the cutting machine, separating each specimen into two halves, one half was used as a specimen without bleaching procedure (control) and the other half as a specimen with bleaching procedure (treated). To verify the possible changes in surface (S) and subsurface (P), one hundred and sixty (n = 160) specimens were evaluated for Knoop microhardness (n = 10) and Raman spectroscopy (n = 5). The different dental bleaching protocols were applied according to the manufacturers of the products and equipment. The specimens remain in artificial saliva at 37 ° C throughout the experiment period and are replaced weekly. Changes in bovine dental enamel were eva... (Complete abstract click electronic access below) / Mestre

O objetivo deste estudo clínico randomizado duplo cego boca-dividida foi avaliar, in vivo, a efetividade do condicionamento ácido prévio do esmalte e de géis clareadores a base de peróxido de hidrogênio (PH) de baixa concentração, associados a fontes de luz híbrida (LH), tendo como variáveis de reposta: sensibilidade dental, fluxo sanguíneo pulpar, grau de mudança e estabilidade de cor ao longo de 6 meses. Trinta e seis voluntários que se encaixaram nos critérios de inclusão e exclusão foram selecionados e randomizados em 3 grupos (n=12), de acordo com o protocolo clareador a ser utilizado. Dentro de cada grupo, um dos hemiarcos foi aleatoriamente selecionado para a realização do condicionamento ácido do esmalte, previamente a aplicação do gel clareador. Os géis que compunham os três protocolos clareadores foram: PH a 10% (experimental) associado a LH a base de LED violeta + laser de diodo (experimental); PH a 15% associado a LH a base de LED azul + laser de diodo; e PH a 35% sem ativação com fonte de luz. A sensibilidade foi avaliada com a escala VAS; a fluxometria pulpar, com o sistema Laser Doppler; e a cor pelo cálculo do ?E via espectrofotômetro. Os dados foram tratados pela análise de variância de medidas repetidas (ANOVA), Tukey e teste de Friedman (p<0,05). Com relação à sensibilidade, foram encontradas diferenças significativas na interação tempo e grupo, mas não com relação ao ácido. Quanto à vascularização pulpar, o efeito de grupo não foi rejeitado, sendo o grupo de PH a 35% o que apresentou as menores medianas para as variações percentuais, porém os efeitos de ácido e tempo não foram significativos. Quanto à cor, diferenças significantes foram encontradas entre os grupos, mas não com relação ao uso do ácido, sendo o PH a 35% o que promoveu maior aumento do ?E. Conclui-se assim que o condicionamento ácido não foi capaz de favorecer a efetividade do clareamento em função do grau de mudança e estabilidade de cor, bem como não aumentou a sensibilidade e o fluxo sanguíneo da polpa dental. A concentração dos agentes clareadores foi determinante para o grau de mudança de cor e demonstrou influenciar a sensibilidade e o fluxo sanguíneo pulpar. / The aim of this double blind split-mouth randomized clinical study was to evaluate, in vivo, the effectiveness of prior acid etching on enamel and low concentration bleaching gels based on hydrogen peroxide (HP) associated with hybrid light (HL) sources, having as response variables: dental sensitivity, dental pulp vascularization and degree of color change and stability over 6 months. Thirty-six volunteers in agreement with the inclusion and exclusion criteria were selected and randomly divided into 3 groups (n = 12) according to the bleaching protocol to be used. Within each group, one of the sides was randomly selected for the execution of acid etching prior to the gel application. The three bleaching gels were: 10% HP (experimental) associated with HL-based violet LED/laser (experimental); 15% HP associated with HL-based blue LED/laser; and 35% HP without light activation. Sensitivity was evaluated with the VAS scale; pulp flowmetry by the use of the Laser Doppler system; and efficacy was evaluated by calculating the ?E via a spectrophotometer. Data were analyzed by analysis of variance for repeated measures (ANOVA), Tukeys and Friedman tests (p<0.05). Regarding sensitivity, significant differences were found in the interaction between the application time and group but not regarding etching. For dental pulp flowmetry, the group effect was not rejected, where the 35% HP showed the lowest medians for the percentile variations, but the effects of etching and time were not significant. For color, significant differences were found between the groups, but not between the sides with and without acid etching, where the 35% HP gel promoted the highest increase in ?E. It is therefore concluded that the etching was not able to favor the effectiveness of whitening due to color variation and stability, as well as not increasing sensitivity or pulp blood flow. The concentration of bleaching agents determined the degree of color change and influenced the sensitivity and blood flow of the dental pulp.

Este estudo in vitro avaliou a variação do pH de géis clareadores à base de peróxido de hidrogênio de diferentes concentrações e seu efeito sobre o esmalte bovino submetido à escovação simulada. Fragmentos de esmalte de dentes bovinos com 1,5cm x 0,5cm x 0,4cm, receberam o tratamento clareador e a escovação em uma metade, ficando a outra como controle; foram divididos aleatoriamente em nove grupos (n=10): G1 = controle; G2 = Whiteness HP 35% - 3 sessões (3x15); G3 = Whiteness HP 35% - 3 sessões (1x45); G4 = Lase Peroxide 35% - 1 sessão (4x730) + ativação com luz híbrida LED/Laser; G5 = Lase Peroxide 25% - 1 sessão (4x730) + ativação com luz híbrida LED/Laser; G6 = Lase Peroxide 15% - 1 sessão (4x730) + ativação com luz híbrida LED/Laser; G7 = Whitegold Office 35% - 3 sessões (1x45); G8 = Whiteness HP Blue Calcium 35% - 3 sessões (1x40); G9 = Whiteness HP Blue Calcium 20% - 3 sessões (1x50). A rugosidade aritmética (Ra) inicial, após clareamento e após escovação foi determinada pela média (µm) de 3 leituras com o rugosímetro Hommel Tester T 1000. Os valores de pH foram determinados pelo peagômetro digital Sentron Model 1001 nos tempos inicial e final independente do número de aplicações ou de sessões. Os espécimes foram armazenados em saliva artificial por 7 dias, submetidos à 100.000 ciclos de escovação simulada. Após 24 horas o desgaste superficial foi determinado (µm) novamente com o rugosímetro Hommel Tester T1000. Os dados da análise de pH foram submetidos a ANOVA um critério e teste de Tukey (p<0,05) com diferenças estatísticas. O teste de Kruskal Wallis e Dunn (p<0,05) aplicado sobre a variação da rugosidade e desgaste apontou diferenças estatísticas entre os grupos. Os géis clareadores apresentaram uma tendência de diminuição dos valores de pH do tempo inicial para o tempo final. Os géis clareadores apresentaram um tendência de diminuição dos valores de pH do tempo inicial para o tempo final, assim como os géis com valores de pH mais próximos dos valores ácidos tendem a levar a superfície do esmalte à uma maior susceptibilidade ao aumento da rugosidade e desgaste superficial, quando submetido à escovação simulada. / This in vitro study evaluated the variation of the pH of bleaching gels, roughness and the wear on bovine enamel, after various in office bleaching protocols and brushing. Ninety fragments of enamel, measuring 15mm x 5mm, were randomly divided into nine groups (n=10) according to bleaching procedure: G1: control; G2: 35% hydrogen peroxide (HP) (Whiteness HP, FGM) 3x15, 3 sessions with one week between each; G3: 35% HP (Whiteness HP, FGM) 1x45, in 3 sessions; G4: 35% HP (Lase Peroxide, DMC) + hybrid light (HL) (LED/Diode laser, Whitening Lase II, DMC Equipments), 4x730 (6 of HL activation), in 1 session; G5: 25% HP (Lase Peroxide II, DMC) + HL, 4x730 (6 of HL activation), in 1 session; G6: 15% HP (Lase Peroxide Light, DMC) 4x730 (6 of HL activation), in 1 session; G7: 35% HP (Whitegold Office, Dentsply) 3x15, in 3 sessions; G8: 35% HP (Whiteness HP Blue Calcium, FGM) 1x40, in 3 sessions; G9: 20% HP (Whiteness HP Blue Calcium, FGM) 1x50, in 3 sessions. The pH values were determined using a pH meter (Sentron Model 1001, Sentron) during the initial and final gel application times. The surface roughness (Ra) was evaluated with a rugosimeter (Hommel Tester T 1000) before and after bleaching, and after brushing (100,000 strokes). The rugosimeter was used to evaluate wear surface after 24 hours brushing. The data were submitted to one-way ANOVA and Tukey tests for pH values, Kruskall Wallis and Dunns test in relation to wear and surface roughness (p<0.05). It could be concluded that the pH values tended to decrease from the initial to final bleaching. After tooth brushing, bleaching procedures with highly acidic products provided a significant increase in enamel wear and surface roughness.

Orientador: Andréa Abi Rached Dantas / Resumo: Após o clareamento dental, radicais livres permanecem na estrutura dental interferindo negativamente na adesão, além disso também geram redução da resistência dental à fratura. Sugere-se que substâncias antioxidantes podem ser capazes de reverter tais efeitos. Assim, esse estudo buscou avaliar os efeitos da solução de alfatocoferol em diferentes concentrações sobre a resistência à fratura, formação de camada híbrida e resistência de união adesiva de dentes tratados endodonticamente e clareados. Foram realizados 3 experimentos. Experimento 1: 60 incisivos bovinos receberam tratamento endodôntico e foram distribuídos em 6 grupos (n=10): G1 - coroas apenas restauradas; G2 - coroas clareadas e restauradas; G3 - coroas clareadas e aplicação de solução de ascorbato de sódio 10%; G4 ao G6: coroas clareadas e aplicação das soluções de alfatocoferol nas concentrações de 15%, 20% e 25%, respectivamente. Os espécimes foram submetidos ao teste de resistência à fratura em máquina de ensaios mecânicos (EMIC). Experimento 2: 60 incisivos bovinos foram tratados endodonticamente, distribuídos e tratados, de acordo com o grupo, de forma idêntica ao experimento 1. Os espécimes foram seccionados na junção cemento-esmalte e no sentido vestíbulo-lingual, lixados e polidos, passaram por processo de desproteinização com ácido clorídrico 18% e hipoclorito de sódio 5%. Em seguida, foram moldados com silicona de adição e vasados com resina epóxi. Os modelos obtidos foram analisados em microscópio eletr... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: After dental bleaching, free radicals remain at the dentin, which causes reduces at adhesive and fracture resistance. To minimize these undesirable effects, the use of antioxidants to neutralize free radicals before the final restoration is recommended. This study evaluated the effects of alphatocopherol solution at different concentrations on the fracture resistance, hybrid layer and shear bond strength of endodontically treated teeth subjected to tooth bleaching. Three experiments were done. Experiment 1: 60 bovine incisors were endodontically treated and then divided into 6 groups (n=10): G1 - the teeth immediately restored with composite resin; G2 - the teeth were bleached and immediately restored; G3 - the teeth received treatment similar to that used for G2, but with 10% sodium ascorbate gel applied after the bleaching protocol; G4 to G6: the teeth received treatment similar to that used for G3, rather than sodium ascorbate, it was applied alphatocopherol at concentrations of 15%, 20% and 25%, respectively. Teeth were subjected to a fracture resistance test in an electromechanical testing machine (EMIC). Experiment 2: 60 bovine incisors were endodontically treated, distributed and treated identically to experiment 1. The specimens were sectioned at the cementum-enamel junction and in the vestibular-lingual direction, sanded and polished, and then it was made the deproteinization with 18% hydrochloric acid and 5% sodium hypochlorite. Then, they were molded with addition / Mestre

Atualmente, o setor sucroenergético está se tornando cada vez mais competitivo, o que de fato é incentivado, principalmente, pela representativa demanda e exigências do mercado externo. Ao se analisar a produção do açúcar refinado, podem-se encontrar barreiras para a sua ampla aceitação nacional e internacional devido ao método de clarificação empregado, uma vez que este é realizado através da queima de enxofre elementar em câmaras de combustão. O uso do sulfito acima dos limites permitidos pode ocasionar potenciais problemas de saúde pública. Neste sentido, o Brasil tem realizado estudos sobre a adoção de novas tecnologias para a substituição da sulfitação, sem alteração da qualidade do açúcar refinado a ser produzido. Partindo deste pressuposto, fez-se a avaliação do uso do peróxido de hidrogênio (H2O2) em solução comercial (35% v/v) como reagente alternativo ao dióxido de enxofre para obtenção de açúcar refinado com menor cor ICUMSA. Para isso, na calda de açúcar do tipo VHP, foram testadas doses distintas de peróxido de hidrogênio, aliadas a diferentes pH e temperaturas frente a um Delineamento composto central composto central, permitindo a otimização e o acompanhamento das transformações decorrentes do processo. Com isso, obteve-se como melhores condições o pH 7,5 e 10, temperaturas entre 50 e 70 °C e dosagens entre 500 e 797,6 ppm de H2O2. A análise a partir da cinética química da peroxidação da calda permitiu observar maiores reduções da cor ICUMSA em geral nos tempos de 50 a 75 minutos e nos 30 primeiros minutos para os pontos extremos de pH, temperatura e dose de H2O2. A degradação de sacarose não foi um fator expressivo quanto ao tempo, pois a mesma na maioria dos casos foi degradada nos 5 primeiros minutos e após esse tempo se mantinha sem muitas alterações, sendo mais vulnerável as condições de pH 3,32 e 11,68, altas temperaturas (83,6 °C) e máxima dosagem H2O2 (1000 ppm) aplicados. Além de favorecer a redução da cor ICUMSA da calda, o H2O2 também se mostrou como um bom agente antimicrobiano, principalmente quando associado às altas temperaturas. Agindo com maior intensidade na diminuição da carga bactériana do que na diminuição da carga fungica. A rede neural artificial (RNA) mostrou um bom ajuste e indicou a variável °Brix (teor de sólidos solúveis) como a que apresentou maior influência na redução da cor ICUMSA e a variável tempo a que menor influenciou na redução de cor. / Nowadays, the sugar-energy industry is becoming increasingly competitive, which indeed is encouraged mainly by representative demand and requirements of foreign markets. When analyzing the production of refined sugar can be found barriers to their widespread international acceptance due to the clarification method employed, since this is performed by burning elemental sulfur in combustion boilers. The use of sulfite above the permitted limits can result in potential public health problems. In this regard, Brazil has conducted studies on the adoption of new technologies to replace the sulfite, without changing the quality of the produced refined sugar. On that basis, it was done the evaluation of the use of hydrogen peroxide (H2O2) in commercial solution (35% v/v) as an alternative reagent to sulfur dioxide to obtain refined sugar with less ICUMSA color. For this the sugar liquor VHP different doses of hydrogen peroxide were tested, together with different pH and temperatures outside a central composite design, enabling optimization and monitoring of changes resulting from the process. Thus there was obtained as the best pH conditions 7.5 to 10, temperatures between 50 and 70 °C and dosages between 500 and 797.6 ppm H2O2. The analysis from the chemical kinetics of the peroxidation of the liquor has observed greater reductions in ICUMSA color in general in the time of 50 minutes to 75 minutes and in the first 25 minutes to the extremes of pH, temperature and H2O2 dose. As for sucrose degradation was not a significant factor, since it in most cases was first degraded within 5 minutes and after this time remained without many changes, being more vulnerable conditions (3.32 and 11.68), high temperatures (83.6 °C) and H2O2 maximum dosage (1000 ppm) applied. In addition to further reduce color ICUMSA H2O2 Liquor also showed such a good antimicrobial agent, particularly when combined with high temperatures. Acting with more intensity in the decrease of bacteria than the reduction of fungi. The artificial neural network (ANN) showed good fit and indicated the variable ° Brix (soluble solids) as the one with the greatest influence in reducing the ICUMSA color and variable time that less influenced the color reduction.

Triacetone triperoxide has been synthesised for a range of experimental studies towards the reactivity, stability and gas phase metal complexation of the explosive. Reaction with transition metals was performed and a series of molten TATP experiments conducted at 120 °C. The effect of a molten state on the degradation, conformation and reactivity of TATP has been studied with the formation of diacetone diperoxide, acetone and acetic acid degradation products observed. The complexation of TATP with a range of metals has been studied by electrospray ionisation-mass spectrometry, yielding [M+Cat]+ ions for all of the alkali metals. The first observations of a [TATP+Ag]+ adduct have been made. The formation of [2TATP+Li+LiX]+ (X = Br, Cl) and [2TATP+Ag]+ sandwich complexes was also observed. Collision cross sections (CCS) for the lithium-containing complexes of TATP were measured by travelling wave ion mobility-mass spectrometry, comparing well to computationally determined structures.