O trabalho teve como objetivo avaliar a presença de fungos e aflatoxinas e ácido ciclopiazônico (ACP) em castanhas-do-brasil a campo e armazenadas, no solo e ar a fim de estabelecer vias de contaminação fúngica. A pesquisa de fungos foi feita pela técnica de semeadura em superfície em meio Ágar batata dextrose e Ágar Aspergillus flavus parasiticus e a determinação de micotoxinas, por cromatografia líquida de alta eficiência. As amostras a campo foram analisadas em: Dia 0; amostras na árvore, Dias 5, 10 e 15; em contato com solo 5, 10 e 15 dias, respectivamente. As armazenadas foram analisadas mensalmente por 11 meses. A campo, os fungos mais prevalentes foram: A. flavus em ouriços e amêndoas; Fusarium spp. em cascas. No solo foram isolados Penicillium spp. e Aspergillus flavus e no ar, Fusarium spp. e Penicillium spp. No armazenamento, em amêndoas, foi constatado A. flavus, Fusarium spp. e A. nomius; e em cascas, Fusarium spp. e A. flavus. Aflatoxinas e ACP não foram detectados. O aumento do tempo de contato das castanhas-do-brasil com o solo foi acompanhado de maior isolamento de A. flavus; sugerindo contaminação da castanha durante etapa a campo. A. nomius e A. parasiticus foram isolados apenas no armazenamento, indicando contaminação no estoque das amêndoas. / The objective of this present study was analyse, in the field and in the storage, mycobiota and the contamination by (aflatoxins and cyclopiazonic acid from Brazil nuts, soil and air samples to determine route of fungi contamination. The Brazil nuts mycobiota was determine by diluition plating method in Potate Dextrose Agar and Aspergillus flavus parasiticus agar and, micotoxins was done by high performance liquid chromatography. Field samples were collected in: day 0, samples still on the tree; days 5, 10 and 15, samples in contact with soil for 5, 10 and 15 days, respectively. The most prevalent fungi were Aspergillus flavus in fruit pods and nuts and Fusarium spp. in shells. Penicillium spp. and A. flavus were isolated from soil, and Fusarium spp. and Penicillium spp. from air. The storage samples were analysed montly during 11 months; and showed predominance A. flavus, Fusarium spp. e A. nomius in nuts, and Fusarium spp. and A. flavus in shells. Aflatoxins and cyclopiazonic acid were not detected. These findings indicate soil as the main source of fungal contamination of Brazil nuts. A. nomius e A. parasiticus were isolated only in storage, suggesting Brazil nuts contamination occurs in this period.

A própolis é uma substância resinosa coletada pelas abelhas de diversas partes das plantas, à qual têm sido atribuídas propriedades antimicrobiana, antioxidante, antinflamatória, antiviral, entre outras. Sua composição química depende de vários fatores, como a localização geográfica. Um novo tipo de própolis brasileira, denominada de própolis vermelha, por sua coloração intensa característica, foi identificada e coletada em região de mangue do Estado de Alagoas. O objetivo deste trabalho foi fracionar e isolar compostos com atividade antibacteriana dessa nova variedade de própolis. O extrato etanólico da própolis vermelha foi fracionado pela técnica de extração líquido-líquido, originando as frações hexânica (fr-Hex) e clorofórmica (fr-Clo). A fr-Clo apresentou alta atividade antibacteriana pelos testes de Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM), contra as bactérias Staphylococcus aureus, Streptococcus mutans e Actinomyces naeslundii. Dessa forma, a fr-Clo foi refracionada por coluna seca gerando sete sub-frações. Após a avaliação da atividade antibacteriana das sub-frações pelas técnicas de CIM e CBM, a sub-fração 3, a mais bioativa, foi purificada em coluna de Sephadex LH-20. Dessa purificação foram obtidas três sub-frações bioativas, as quais foram submetidas ao isolamento dos compostos pela técnica de Cromatografia Líquida de Alta Eficiência (CLAE) preparativa. Dois compostos foram isolados e denominados de composto 1 e composto 2. O composto 2 foi o mais potente para as atividades inibitória e bactericida e sua CIM para os três tipos de bactéria testados variou entre 15,6 e 31,2 µg/mL, enquanto para o composto 1, este parâmetro variou de 31,2 a 62,5 µg/mL. As CBM do composto 1 e do composto 2 variaram entre 125 e 250 µg/mL e entre 31,2 e 62,5 µg/mL, respectivamente. Por meio da técnica Ressonância Magnética Nuclear (RMN) foi possível identificar o composto 1 como pertencente à classe das isoflavanas e o composto 2 como uma chalcona (isoliquiritigenina). A forte atividade antibacteriana apresentada pelos compostos isolados da própolis vermelha torna este produto uma importante fonte de compostos antibacterianos naturais. / Propolis is a resinous material, collected by honeybees from several parts of plants, known for its antimicrobial, antioxidant, anti-inflammatory, antiviral properties, among others. Its composition varies according to several factors, such as the geographical location. A novel type of Brazilian propolis, named red propolis due to its intense characteristic color, was collected in a mangrove area in the State of Alagoas. This research aimed to fractionate and isolate the compounds in this new type of propolis that present antibacterial activity. The ethanolic extract of red propolis (EEP) was fractionated using the liquid-liquid extraction technique, yielding the hexanic (Hex-fr) and the chloroformic fractions (Chlo-fr). Chlo-fr showed high antibacterial activity determined by the minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests against Staphylococcus aureus Streptococcus mutans, and Actinomyces naeslundii. Thus, Chlo-fr was refractionated by Dry-Column Chromatography, yielding seven subfractions. After submitted to MIC and MBC techniques to assess their antibacterial activity, and subfraction 3, the most bioactive of them, was purified in Sephadex LH-20 column. After this purification, three bioactive subfractions were obtained, submitted to isolation of compounds using preparative High Performance Liquid Chromatography (HPLC). Two compounds were isolated and named compound 1 and compound 2. Compound 2 presented the highest inhibitory and bactericidal activities and its MIC for the three types of bacteria tested ranged from 15.6 to 31.2 µg/mL, whereas for compound 1, this parameter ranged from 31.2 to 62.5 µg/mL. MBC of compound 1 and compound 2 ranged from 125 to 250 µg/mL and from 31.2 to 62.5 µg/mL, respectively. Through Nuclear Magnetic Resonance tecnique (NMR), it was possible to identify the compound 1 as belonging to class of isoflavans and the compound 2 as a chalcone (isoliquiritigenin). Due to the strong antibacterial activity presented by the compounds isolated from red propolis, it can be concluded that this product is an important source of natural antibacterial compounds.

Orientadores: Helena Teixeira Godoy, Elaine Conceição de Oliveira / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-23T09:32:28Z (GMT). No. of bitstreams: 1 Bochi_VivianCaetano_D.pdf: 14853461 bytes, checksum: 20196ff92ca0c3039ebb5aded05718ad (MD5) Previous issue date: 2013 / Resumo: A groselha do Ceilão (Dovyalis hebecarpa) é uma fruta exótica de coloração roxa quando madura. Não foram encontrados trabalhos que identifiquem os principais compostos fenólicos na espécie, ação antioxidante e efeitos biológicos. Sendo assim, além da otimização da extração, esse trabalho avaliou os principais compostos fenólicos na casca e polpa de D. hebecarpa por cromatografia líquida acoplada a espectrômetro de massas (CLAE-DAD-EM), o potencial antioxidante in vitro e o efeito sobre a resposta imunológica em camundongos. Com exceção da razão entre amostra e solvente, avaliada univariadamente, a otimização das variáveis de extração foi realizada utilizando planejamento experimental multivariado. O processo otimizado foi realizado em menor tempo (20 minutos), com menos solvente orgânico (20% de acetona) e com maior rendimento (aumento de 10% no teor total de compostos fenólicos e de 26% no teor total de antocianinas monoméricas) do que a metodologia inicial. A caracterização do perfil de antocianinas revelou que D. hebecarpa é fonte de compostos não acilados, sendo delfinidina-3-O-rutinosídeo e cianidina-3-O-rutinosídeo os majoritários. Casca e polpa possuem composição similar, no entanto, maiores concentrações são encontradas na parte externa do fruto. Amostras de dois anos consecutivos e em duas datas de amostragem foram recolhidas para avaliação de variações entre estações do ano. Considerando que a ação antioxidante se dá por diversos mecanismos, metodologias de FRAP, ABTS e ORAC foram empregadas. Os maiores resultados de atividade antioxidante foram obtidos com a metodologia de ORAC, indicando marcada atividade sequestradora de radicais peroxil dos compostos extraídos. De forma diferenciada para polpa e casca dos frutos, efeitos significativos sob os teores de antocianinas, de compostos fenólicos e capacidade antioxidante, foram observados devido à variação climática entre as datas. Adicionalmente foram avaliados parâmetros biométricos e composição nutricional. Alterações no peso e tamanho dos frutos (p<0,05), assim como, na composição nutricional (p<0,05) foram observadas. Esses resultados, possivelmente, devem-se as diferenças de disponibilidade de água e incidência solar entre as datas de amostragem. Os valores médios dos teores de compostos fenólicos e antocianinas encontradas são similares aos relatados para outras frutas vermelhas. Nos testes in vivo, camundongos C57Black6 foram divididos em 4 grupos que receberam por 5 dias uma dose diária de extrato bruto em alta, média e baixa dosagem (32µg, 16µg e 8µg equivalentes de cianidina-3-O-glucosídeo/animal, respectivamente). Para o grupo controle foi utilizada solução salina. Após 24 horas do fim do tratamento, os animais foram imunizados com OVA (ovalbumina), conforme metodologia padrão. No 14° dia, os animais foram sacrificados e os tecidos recolhidos para análise. Foram monitoradas a proliferação celular em baço e linfonodos, assim como, sua diferenciação em linfócitos T CD4+/CD8+ por citometria de fluxo. Os grupos tratados com CGCE apresentaram redução na proliferação celular em linfonodos, assim como na razão CD4+/CD8+. Sendo assim, os resultados indicam um possível efeito anti-inflamatório do extrato aquoso bruto de groselha do Ceilão, porém pesquisas adicionais de caracterização de citocinas e tratamentos prolongados são necessárias para confirmação dos resultados encontrados / Abstract: D. hebecarpa is a dark purple/red berry produced in Brazil as an exotic fruit with potential for large scale production and commercialization. Fruits are believed to contain high phenolic and anthocyanin concentration that provide color and defense to the plant. Moreover, these compounds have been extensively studied for their antioxidant activity and potential human health benefits. Thus, this work has optimized extraction prior to the skin and pulp anthocyanin profile characterization by HPLC-PDA-MS aiming to obtain the simplest and mildest conditions with maximum phenolic yield. Moreover, it was evaluated in vitro antioxidant capacity and the effect of water-based crude extract on immune system of mice. Solid-liquid ratio was determined in a linear experiment using ANOVA and Tukey (p<0.05). Acetone, ethanol and water were the extraction solvents evaluated by Simplex Lattice design. Time and acid concentration were evaluated using response surface methodology (RSM) to determine variables effect and their interactions. The optimized conditions achieved were: solid-liquid ratio of 1:120, acetone 20% with 0.35% formic acid, 20min, and no re-extraction. However, satisfactory results were obtained using just water as solvent. The optimized extraction used less organic solvent than the other conditions tested and showed higher yields than the initial ones (an increase of 10% and 26% of total phenolic compounds and total monomeric anthocyanins, respectively). The analytical HPLC chromatogram showed five major anthocyanins (Delphinidin-3-glucoside, Delphinidin-3-rutinoside, Cyanidin-3-glucoside, Cyanidin-3-rutinoside, and Petunidin-3-rutinoside) in addition to two minor pigments (Peonidin-3-rutinoside and Malvidin-3-rutinoside). Samples from two consecutive years were used for quantification purposes and antioxidant measurements. High concentration of phenolics and anthocyanins were detected in skin samples with the same anthocyanin profile of pulp part. FRAP, ABTS, and ORAC methodologies were used to evaluate antioxidant mechanisms. A strong peroxyl scavenger capacity was detected for D.hebecarpa samples by ORAC methodology which could indicate possibly health effects of fruit consumption. Significant variations of anthocyanin, total phenolic content, and antioxidant capacity were observed in pulp and skim possibly as a result of weather conditions. Concomitantly, biometric parameters and nutritional composition seems to be affected as well (p<0.05). C57Black6 mice were used for in vivo evaluation of immuno-modulatory effect of water-based Ceylon gooseberry crude extract (CGCE). Animals were treated with high, medium, and low dosages of CGCE (32µg, 16µg, and 8µg cyanidin-3-O-glucose/animal/day, respectively) during 5 days. Saline solution was used as a control group. Animals were challenged after 24 hours from the last dose using a standardized immunization protocol with OVA and CFA. After euthanasia, blood, spleen, and lymph nodes were collected and analyzed. Cell proliferation using MTT and lymphocyte profile by flown citometry was determined in spleen and lymph nodes, as well as IgG levels in blood samples. Results indicate a possible anti-inflammatory effect. Since, there was an decrease in lymph node cell proliferation response as well as in CD4+/CD8+ ratio. In summary, D. hebecarpa is a rich source of anthocyanins with concentration levels as high as reported for other berry fruits. The strong antioxidant activity and some evidences of in vivo anti-inflammatory effects suggest this fruit as source for future scientific works aiming to evaluate possible health effects / Doutorado / Ciência de Alimentos / Doutora em Ciência de Alimentos

Orientador: Felix Guillermo Reyes Reyes / Dissertação (mestrado) - Universidade Estadual de Campiknas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-19T16:49:25Z (GMT). No. of bitstreams: 1 DelBianchi_Michelle_M.pdf: 2100554 bytes, checksum: 574b7310661133a3b473c024dee257a0 (MD5) Previous issue date: 2012 / Resumo: Edulcorantes sao compostos quimicos de origem sintetica ou natural que tem a propriedade de adocar alimentos, em substituicao total ou parcial a sacarose ou outros acucares e reduzem o teor calorico do produto resultante. A ingestao de adocantes artificiais no mundo tem crescido nos ultimos anos, com o objetivo principal de reduzir a ingestao calorica. O aumento global de doencas cronicas relacionadas com a ingestao de acucar, particularmente a obesidade e a diabetes, e tambem uma razao para o aumento do consumo de adocantes artificiais. O objetivo deste estudo foi avaliar a ingestao de adocantes artificiais atraves do consumo de adocantes de mesa liquidos e avaliar a possibilidade de consumo superior a sua ingestao diaria aceitavel (IDA) estabelecida por FAO/WHO comite de peritos em aditivos alimentares (JECFA). Inicialmente, uma pesquisa de campo foi realizada para avaliar qual marca de adocante de mesa liquido mais consumido no Brasil e as caracteristicas de uso do mesmo. Para tanto, um questionario foi enviado por correio eletronico, para individuos, com questoes relativas ao consumo de adocantes e, caso consumidos, aspectos de uso incluindo marca do adocante, ingestao e motivo de consumo. O questionario tambem continha perguntas sobre questoes socioeconomicas, demograficas e de saude. Em base a esta pesquisa, verificou-se quais sao as marcas de adocantes liquidos de mesa mais consumidas. Os edulcorantes presentes nesses adocantes mais consumidos sao: sacarina, ciclamato, aspartame e acessulfame-K. Foi realizada a quantificacao desses edulcorantes nos adocantes de maior consumo. Para a quantificacao de sacarina, aspartame e acessulfame-K foi utilizada a cromatografia liquida de alta eficiencia acoplada ao detector de arranjo de diodos (CLAE-DAD). A quantificacao de ciclamato foi realizada por espectrofotometria UV/Vis, em 314 nm, apos derivatizacao para N, Ndiclorociclohexilamina. Os metodos analiticos foram validados de acordo com os seguintes parametros de validacao: linearidade, faixa linear, especificidade, precisao (repetibilidade intra e inter dias), limite de deteccao (LOD) e limite de quantificacao (LOQ). Todos os metodos mostraram parametros de validacao de acordo com o guia para validacao de metodos analiticos estabelecidos pela Agencia Nacional de Vigilancia Sanitaria (ANVISA). A maioria das respostas ao questionario foram do genero feminio (70,2%) e provenientes da regiao Sudeste do Brasil (70,8%). Entre os entrevistados, 47% usavam adocantes liquidos de mesa. A maioria que relatou o uso de adocantes artificiais tinham entre 21 e 32 anos ou acima de 45 anos de idade. Destes, 83% afirmaram nao ter qualquer tipo de patologia e que a principal razao para o consumo de adocantes liquidos de mesa foi a preferencia em relacao ao acucar. Cerca da metade (54%) dos consumidores relataram sentir sabor desagradavel ou amargo quando consomem adocante e 32% nao sabiam qual era o edulcorante artificial contido no adocante. Os resultados da analise dos adocantes liquidos de mesa provenientes do mercado varejista indicou variacao significativa (p<0,05) na concentracao de edulcorante entre marcas e lotes da mesma marca. Com base nos resultados do questionario verificou-se qual e a marca mais consumida. Essa marca contem sacarina e ciclamato. Uma amostra dessa marca foi entregue a individuos adultos consumidores deste produto, para calcular o nivel de consumo dos edulcorantes artificiais. Foi verificado que a ingestao de sacarina e ciclamato atraves do consumo de adocantes liquidos de mesa representa, em media, 38,7% e 19,8% do valor de IDA do edulcorante, respectivamente. Considerando a variacao na concentracao de adocantes entre as marcas e lotes da mesma marca e que cada edulcorante artificial tem um valor de IDA estabelecido, recomenda-se que as concentracoes dos edulcorante em adocantes liquidos de mesa sejam apresentados na etiqueta da embalagem, para fornecer ao consumidor informacoes sobre seu consumo, e permitir controlar a sua exposicao total a estas substancias. Em geral, os resultados sugerem uma substituicao cada vez maior de acucar pelos adocantes artificiais, que indicam a necessidade de se obter dados sobre a ingestao total de adocantes artificiais, a fim de avaliar o risco de que estas substancias apresentam para a saude dos consumidores / Abstract: Artificial sweeteners are chemical compounds of synthetic or natural origin that have sweetening properties, as full or partial replacement of sucrose or other sugars, and reduce the caloric content of the resulting product. The intake of artificial sweeteners in the world has grown in recent years, with the primary goal of reducing caloric intake. The overall increase of chronic diseases relating to sugar intake, particularly obesity and diabetes, is also a reason for the increased consumption of artificial sweeteners. The aim of this study was to evaluate the intake of artificial sweeteners through the consumption of liquid tabletop sweeteners and evaluate the possibility of consumption exceeding their Acceptable Daily Intake (ADI) established by the FAO/WHO Joint Expert Committee on Food Additives (JECFA). Initially, a field survey was undertaken to assess which liquid tabletop sweetener brand had the largest market share of the retail market in Brazil and the characteristics of usage. For this purpose, a questionnaire was submitted by electronic mail to individuals with questions relating to their consumption of artificial sweeteners, and if consumed, aspects of usage including brand, frequency and motivation of consumption. In addition, the questionnaire contained questions regarding socio-economic, demographic and health status. Based on the results of the survey the specific artificial sweeteners present in the most widely used liquid tabletop sweetener brands were identified and quantified. The artificial sweeteners present in the most widely consumed brands are: saccharin, cyclamate, aspartame and acesulfame-K. Quantification of saccharin, aspartame and acesulfame-K in the commercial products was determined by high performance liquid chromatography coupled to a diode array detector (HPLC-DAD). Quantification of cyclamate was carried out by spectrophotometry UV/Vis, at 314 nm, after derivatization to N, N diclorocyclohexilamine. The analytical methods employed were validated according to the following validation parameters: linearity, linear range, specificity, accuracy, precision (repeatability intra-and inter-day), limit of detection (LOD) and limit of quantification (LOQ). All the methods showed validation parameters in accordance to the Guide for the Validation of Analytical Methods established by the Brazilian Agency of Sanitary Surveillance (ANVISA). Most of the responses to the questionnaire came from females (70.2%), from the southeast region of Brazil (70.8%). Among the respondents, 47% used artificial tabletop sweeteners. The majority of the respondents who reported using artificial sweeteners were between 21 and 32 years old or above 45 years old. Among these 83% reported not having any type of pathology and that the primary reason for artificial sweetener consumption was the preference in relation to sugar. Approximately half (54%) of consumers reported an unpleasant or bitter aftertaste when consuming the sweetener and 32% did not know which artificial sweetener was in the sweetener they used. The results of the analysis of the liquid tabletop sweeteners samples from the retail market indicated there is a statistical variation (p<0.05) in the sweeteners concentration between brands and batches from the same brand. Based on the questionnaire results, it was verified the most consumed brand. The brand contains saccharin and cyclamate. A sample of this brand was provided to adult volunteers to estimate the level of consumption of those artificial sweeteners. It was found that the intake of saccharin and cyclamate through the consumption of the liquid tabletop sweetener represent, on average, 38.7% and 19.8% of their ADI values, respectively. Considering the variation found in the sweeteners concentration between brands and batches from the same brand and that each artificial sweetener has an ADI value established, it is recommended that the concentrations of the artificial sweeteners in the liquid tabletop sweeteners be shown on the packaging label, to allow the consumer to make informed decisions regarding their intake and to allow them to better control their total exposure to these substances. In general, the results suggest an increasing replacement of sugar by artificial sweeteners, which indicate the need to obtain up to date data on the total intake of artificial sweeteners in order to assess the risk that these substances present to the consumers health / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos

Orientador: Cesar Costapinto Santana / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-20T14:12:19Z (GMT). No. of bitstreams: 1 Nascimento_AnaCarolina_M.pdf: 2361694 bytes, checksum: bcd5c25d65fe1d7ac8a8666d6f13c023 (MD5) Previous issue date: 2012 / Resumo: O secnidazol corresponde à formulação 1-(hidroxipropil)-2-metil-5-nitroimidazol e possui espectro de atividade contra microorganismos anaeróbicos e eficácia no tratamento de amebíase, giardíase, tricomoníase e vaginose bacteriana. Ele é comercializado na forma racêmica, isto é, na proporção 1:1 dos seus enantiômeros R e S. Não é oficial em nenhuma farmacopeia. Estudos relatam que para alguns imidazóis o enantiômero R apresenta maior atividade biológica frente ao enantiômero S. Portanto a separação do secnidazol é importante para testes biológicos comparativos de efeitos colaterais. Inserindo-se neste contexto, foi desenvolvido este trabalho de pesquisa com o intuito de estudar a resolução enantiomérica do fármaco secnidazol pela técnica de cromatografia líquida de alta eficiência utilizando coluna recheada com fase estacionária tris(3,5-dimetilfenilcarbamato) de amilose. Experimentos de pulsos com soluções diluídas foram realizados variando a vazão de fase móvel de 1,0 a 2,5 mL/min e as temperaturas de 20 a 35°C. Os resultados mostraram alta eficiência, com número de pratos superando 1000 e fatores de separação na ordem de 7,0. Os valores negativos de 'delta'H e 'delta'S* indicam que a adsorção dos enantiômeros da fase móvel na fase estacionária é entalpicamente favorável. Experimentos a altas concentrações (condições de sobrecarga) foram realizados com a finalidade de determinar as isotermas não-lineares pelo método da análise frontal e também os perfis de eluição sob estas condições. As isotermas de adsorção apresentaram comportamento não-linear e o modelo de Langmuir foi bem correlacionado aos dados experimentais de equilíbrio no intervalo de concentração analisado. A partir da metodologia shortcut foram obtidos os parâmetros operacionais da unidade leito móvel simulado. As purezas alcançadas para as correntes de extrato e refinado foram 85,50% e 72,50%, respectivamente / Abstract: The chemical formula of secnidazole is 1-(hydroxypropyl)-2-methyl-5-nitroimidazole. It acts activity against anaerobic microorganisms and is effective in the treatment of amebiasis, giardiasis, trichomoniasis, and bacterial vaginosis. It is marketed in the racemic form, that is, proportion 1:1 of their R and S-enantiomers. It's not officially recognized by any pharmacopoeia. Studies have reported that for some imidazoles the R-enantiomer has a higher biological activity than the S-enantiomer. For this reason the separation of secnidazole is important for comparative biological tests of side effects. It is in this context that this research was developed in order to study the enantiomeric resolution of drug secnidazole by the technique of high performance liquid chromatography using stationary phase column packed with amylose tris (3, 5- dimethylphenylcarbamate). Pulse experiments with dilute solutions were conducted by varying the mobile phase flow (1.0 to 2.5 mL/min) and temperature (20 to 35 °C). The results revealed high efficiency, with number of plates overcoming 1000 and selectivities in the order of 7.0. The negative values of 'delta'H and 'delta'S* indicates that the enantiomer adsorption from the mobile phase to stationary phase is enthalpically favorable. Experiments in overloaded conditions were realized to obtain the equilibrium adsorption isotherms by frontal analysis, as well overload elution profiles. The adsorption isotherms shown a nonlinear behavior and the Langmuir model was well correlated to equilibrium experimental data in the range of investigated concentration. From the shortcut method operating parameters were obtained to the simulated moving bed unit. The purities reached for the extract and raffinate lines were 85.50% and 72.50% , respectively / Mestrado / Desenvolvimento de Processos Biotecnologicos / Mestra em Engenharia Química

Orientador: Marcelo Alexandre Prado / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-21T00:42:45Z (GMT). No. of bitstreams: 1 Valerio_PedroPrates_M.pdf: 2326955 bytes, checksum: bec4ce7e7d6feaf8b1dae322ddf0d2c7 (MD5) Previous issue date: 2012 / Resumo: O crescimento do setor aquícola no Brasil se encontra em evidência. Em 2010 o volume produtivo foi maior que 479.390 toneladas, apresentando incrementos de 15,3%, em relação à produção de 2009, e 31,2%, em relação ao triênio 2008-2010. Neste mesmo ano de 2010, a aquicultura continental brasileira registrou volumes produtivos na ordem de 394.340 toneladas, das quais 63,4% foram representadas pela soma de tilápia e carpa. Na piscicultura nacional e mundial, principalmente em regiões tropicais, a tilápia representa uma das espécies mais indicadas para o cultivo intensivo. Dados da Balança Comercial de Pescados apontam que em 2005 o volume de exportação de tilápia brasileira já se aproximava de 314,8 toneladas. Em 2010 o montante de pescado brasileiro exportado se aproximou de US$ 263 milhões. Neste período os principais países importadores foram EUA, Europa e Ásia. A Comunidade Européia estabelece barreiras à exportação do pescado brasileiro, na busca pelo atendimento a exigências quanto à comprovação de teores de contaminantes, tais quais antibióticos, em produtos importados. Quanto ao cloranfenicol, a preocupação se relaciona à proibição de seu uso em animais destinados ao consumo humano, baseada em evidências toxicológicas como a ocorrência de anemia aplástica. Outro fator que contribui para a imposição de barreiras à exportação nacional é a falta de dados analíticos relacionados a pescados brasileiros. Para determinação analítica de Cloranfenicol, o método de cromatografia líquida com detecção por espectrometria de massas (CL-EM) é exigido em função de sua sensibilidade e seletividade. Métodos rápidos e eficientes se fazem necessários. Perante tais necessidades, este trabalho desenvolveu e validou uma metodologia analítica para determinação quantitativa de resíduos de cloranfenicol em pescado. O procedimento de extração apresentou inúmeras vantagens: é rápido, direto, apresenta baixo custo, e exclui etapas adicionais de limpeza (como extração em fase sólida). Ainda, utiliza pequenas quantidades de amostra e de solvente. Na etapa de separação e detecção foi empregada a técnica cromatografia líquida de alta eficiência, acoplada à espectrometria de massas em &quot;tandem¿ com ionização por &quot;electrospray¿ (CLAE-ESI/EM/EM). O método se adéqua à Comissão de Decisão 2002/657/EC quanto ao número de pontos de identificação requeridos pela para substâncias com tolerância zero. O tempo de corrida cromatográfica foi igual a 9 minutos. O método é linear na faixa entre 0,03µg/kg e 0,33µg/kg, além de preciso, e exato. Recuperações variaram entre 91,33 e 108,00%. A transição de quantificação 321.9>152.1 apresentou LQ igual a 0,03µg/kg, sendo o LMDR requerido pela Commission Decision 2003/181/EC = 0,30µg/kg / Abstract: The growth of the aquaculture industry in Brazil is in evidence. In 2010, domestic production was greater than 479,390 tons, showing an increase of 15.3% compared to 2009 production, and of 31.2% compared to the period 2008-2010. In the same year, the Brazilian continental aquaculture recorded production volumes of around 394,340 tonnes, of which 63.4% were represented by the sum of tilapia and carp. Tilapia is one of the most suitable species for intensive fish farming. In 2005 the export volume of Brazilian Tilapia had already approached 314.8 tons. . In 2010 Brazil exported U.S. $ 263 million in fish. The main importers were U.S.A., Europe and Asia. The European Community establishes barriers to Brazilian exports of fish. There is a need of meeting requirements of laboratory tests in order to show levels of antibiotics in certain products. The concern in relation to chloramphenicol relates to the prohibition of its use in animals intended for human consumption, due to toxicological factors such aplastic anemia. Another factor that contributes to the imposition of trade barriers is the lack of available data concerning its presence in Brazilian fishes. For the determination of chloramphenicol in fishes, the method LC-MS (liquid chromatography with detection by mass spectrometry) is required due to its sensitivity and selectivity. Methods for the analysis of chloramphenicol should be related to speed and efficiency. Given these needs, this study developed and validated an analytical method for quantitative determination of chloramphenicol residues in fish. The extraction procedure has shown several advantages: is fast, direct, is inexpensive and eliminate additional cleanup steps (i.e. solid phase extraction). What is more, requires low amounts of sample and of solvents. For the separation and detection steps, HPLCESI/ MS/MS has been employed. The method is adequate for the number of identification points required by the Commission Decision 2002/657/EC for substances with zero tolerance,. In chromatography, the run time was equal to 9 minutes. The method is linear in the range between 0.03µg/kg and 0.33µg/kg. It is also precise, and accurate. Recoveries ranged between 91.33 and 108.00%. The quantification transition 321.9>152.1 showed a LOD and LOQ of 0.03µg/kg. The MLPR required by Commission Decision 2003/181/EC is of 0.30mg / kg / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos

Orientador: Isabel Cristina Sales Fontes Jardim / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-04T03:19:20Z (GMT). No. of bitstreams: 1 Faria_LeonardoJardimdaSilva_M.pdf: 547743 bytes, checksum: c807d29f792ecf6c57e17ca59816fdfe (MD5) Previous issue date: 2004 / Mestrado / Quimica Analitica / Mestre em Química

>Magister Scientiae - MSc / In South Africa, it is estimated that approximately 70% of the population frequently make use of traditional medicinal plants for their health care needs. The use of Lessertia frutescens by the various cultural groups in South Africa dates back to the earlier civilizations and continues to be used today to treat a multitude of ailments. To get the best results from a medicinal plant, one would need to ensure that the crude material is of good quality through interventions like being properly grown, well dried and correctly processed. This would add a measure of quality assurance, which will contribute towards the safety and efficacy aspect of herbal medicine. The aim of this study was to investigate what impact a particular season of harvest and the time in storage would have on the flavonoid and triterpenoid marker levels of Lessertia frutescens. To achieve this, the following was investigated: (1) storage variation of Lessertia frutescens leaves by comparing the results obtained from the High Performance Liquid Chromatography (HPLC) analysis of the flavonoids and triterpenoids, (2) seasonal variation of Lessertia frutescens leaves by comparing the results obtained from the HPLC analysis of the flavonoids and triterpenoids, (3) leaf and stem variation of Lessertia frutescens by comparing the results obtained from HPLC analysis of the flavonoids and triterpenoids. The hypotheses were: (1) the stored sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of the freshly prepared sample, (2) the sample that was harvested during the summer season would indicate higher levels of the biomarkers of flavonoids and triterpenoids than the other three seasons, (3) the leaf sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of the stem sample. An Agilent 1200 series HPLC was used for the determination of the flavonoids sutherlandin A and sutherlandin D as well as the triterpenoids sutherlandioside B and sutherlandioside D. Results show that for both sutherlandin A (summer: 3.67 ± 2.88 mg/ml; storage: 4.07 ± 2.88 mg/ml) and D (summer: 4.10 ± 1.06 mg/ml; storage: 4.25 ± 1.06 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the storage samples. For both sutherlandioside B (summer: 3.01 ± 0.39 mg/ml; storage: 2.82 ± 0.39 mg/ml) and D (summer: 5.82 ± 0.42 mg/ml; storage: 4.66 ± 0.42 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the fresh summer samples. For the seasonal comparison, results show that for sutherlandin A (summer: 3.67 ± 12.49 mg/ml; autumn: 4.75 ± 12.49 mg/ml; winter: 4.23 ± 12.49 mg/ml; spring: 6.56 ± 12.49 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the spring sample. For sutherlandin D (summer: 4.10 ± 10.32 mg/ml; autumn: 6.37 ± 10.32 mg/ml; winter: 5.25 ± 10.32 mg/ml; spring; 6.08 ± 10.32 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the autumn sample. For both sutherlandioside B (summer: 3.01 ± 7.19 mg/ml; autumn: 2.15 ± 7.19 mg/ml; winter: 2.89 ± 7.19 mg/ml; spring: 1.47 ± 7.19 mg/ml) and D (summer: 5.82 ± 14.48 mg/ml; autumn: 3.33 ± 14.48 mg/ml; winter: 4.23 ± 14.48 mg/ml; spring: 2.50 ± 14.48 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the autumn sample. For the summer leaf/stem comparison, results show that for sutherlandin A (leaf: 3.67 ± 8.18 mg/ml; stem: 4.67 ± 8.18 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the stem sample. For the sutherlandin D (leaf: 4.10 ± 4.81 mg/ml; stem: 3.31 ± 4.81 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the summer leaf sample. For both the sutherlandioside B (leaf: 3.01 ± 4.24 mg/ml; stem: 3.62 ± 4.24 mg/ml) and D (leaf: 5.82 ± 0.42 mg/ml; stem: 5.80 ± 0.42 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the stem samples.Results demonstrate that the production of secondary metabolites are influenced by environmental factors like seasonal harvesting, as indicated by the variation in the chemical constituent composition of Lessertia frutescens depending on the season collected in. Moreover, the storage of Lessertia frutescens for a period of one year resulted in an increase of two of the four constituents being monitored. There was slight variations in the chemical constituents, depending on whether the leaf or stem material of Lessertia frutescens was being used. Finally, the type of chemical constituent being monitored was also important in the consideration of this study. Therefore, this study can be seen as a starting point to further investigations of these aspects, which are of clinical, pharmacological and economic importance.

Natural products are complex mixtures of compounds with therapeutic effects which are often reported to be due to the synergistic action of multiple and sometimes unknown components. Consequently, standardization of these products is complex and a lack of effective quality control (QC) criteria in most countries has led to marketing of commercial products with questionable quality, safety and efficacy (QSE). The aim of this study was therefore to develop qualitative and quantitative analytical methods for use in the QC of Ginkgo biloba solid oral dosage forms. Initially, a micellar electrokinetic chromatography (MEKC) method was developed for the identification of the flavonol glycosides, rutin and quercitrin as well as 3 flavonol aglycones, quercetin, kaempferol and isorhamnetin in crude extracts of 4 Ginkgo biloba solid oral dosage forms using ultraviolet (UV) detection. A reversed-flow cyclodextrin-modified MEKC method was subsequently developed for the simultaneous determination of the aforementioned flavonols as well as ginkgolide A, B, C, J and bilobalide (all positive markers) in Ginkgo commercial products. A non-aqueous capillary electrophoresis (CE) method was also developed for fingerprinting the presence of ginkgolic acids (negative markers) in Ginkgo biloba leaf extracts, which are purported to be associated with toxic properties. This method was also applied to 2 Ginkgo biloba commercial products. Since the flavonols have strong UV absorbing chromophores, a reversed phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated using photo-diode-array (PDA) detection which was then successfully applied to fingerprint commercially available Ginkgo biloba solid oral dosage forms as well as quantify the relevant flavonol markers present in these extracts. Sample preparation was simple, rapid and cost efficient with minimal clean-up and the employment of a minibore column which requires low mobile phase flow rates contributed to the economy of the method. Unlike the conventional QC approach, samples were not hydrolyzed and direct determination of 2 intact flavonol glycosides, together with the usual aglycone markers was facilitated which provided maximal content information for fingerprint comparisons. On the other hand, terpene trilactones possess poor chromophores and an alternative detection method to UV was required in order to obtain suitable sensitivity. RP-HPLC with evaporative light scattering detection (ELSD) was selected for quantification of these non-volatile constituents in Ginkgo dosage forms and this method was deemed suitable for the routine QC analysis of these positive markers in commercial products. Since approximately 33 flavonoids have been identified in Ginkgo biloba leaf extracts, baseline separation using UV/PDA detection normally requires complex gradient programs and long analysis times. In addition, unequivocal identification of the flavonoids with similar UV spectra and elution times cannot be guaranteed. A liquid chromatographic tandem mass spectrometric (LC-MS-MS) method was therefore developed and validated in order to ensure accurate quantification of the selected flavonol marker compounds in Ginkgo commercial products. LC-MS-MS analysis of Ginkgo extracts revealed, in addition to rutin, the possible presence of other quercetin analogues, quercetin-3-Orhamnoside-7-O-glucoside or quercetin-3-O-glucoside-7-O-rhamnoside, previously unreported in Ginkgo biloba leaf extracts or dosage forms. In terms of evaluating the most suitable analytical method for QC, CE shows exceptional potential in the future analysis of Ginkgo biloba dosage forms while HPLC-PDA and HPLC-ELSD are currently the most affordable and practical instruments for the routine analysis of the flavonols and terpenoids, respectively. LC-MS-MS proved to be pivotal for the accurate identification and quantification of the flavonols due to interference by other flavonoid compounds with similar retention times and UV spectra to the peaks of interest. All quantitative and qualitative results revealed large discrepancies in the marker content between the products regardless of which batch was analysed and product labels disclosed little relevant information. Although currently not required by most regulatory agencies, some of the usual quality criteria applied to orthodox medicines was evaluated. In particular, dissolution analysis, disintegration, tablet hardness and weight uniformity were assessed and revealed similar inconsistencies. This thesis emphasises that implementation of effective QC criteria is long overdue and is essential to ensure consistent product QSE of commercially available Ginkgo biloba solid oral dosage forms.

Orientador: Helena Teixeira Godoy / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-03T14:50:05Z (GMT). No. of bitstreams: 1 Prado_MarceloAlexandre_D.pdf: 10679979 bytes, checksum: 271e49a76278f93376a9b985c59b1111 (MD5) Previous issue date: 2003 / Resumo: O uso de corantes artificiais pelas indústrias de alimentos em todo o mundo é bastante difundido, isto porque os corantes permitem suplementar ou repor a coloração perdida durante o processamento e ou estocagem, e assim garantir a aceitabilidade do produto frente ao consumidor, sendo utilizados ainda como um importante instrumento para garantir a uniformidade dos produtos em linhas de produção de larga escala. Do ponto de vista da saúde pública, existem diferentes opiniões quanto à inocuidade dos diversos corantes artificiais utilizados em alimentos. Muitos estudos mostram que esses aditivos podem causar uma série de males à saúde da população quando consumidos de forma incorreta, seja por abusos da indústria ou exagero no consumo. O fato é que, técnicas analíticas para a determinação desses corantes devem ser desenvolvidas, e principalmente validadas, para garantir a segurança alimentar dos produtos que ingerimos. No presente trabalho foram desenvolvidos e validados dois métodos para a determinação de corantes artificiais em bebidas alcoólicas, utilizando duas diferentes técnicas, a cromatografia a líquido de alta eficiência (CLAE) e a eletroforese capilar (EC). Os métodos foram desenvolvidos para a separação simultânea dos onze corantes artificiais permitidos para uso em alimentos no Brasil. No método por CLAE, para a separação dos corantes, utilizou-se coluna de fase reversa e eluição por gradiente, com fase móvel composta por água/metanol. Na EC a separação ocorreu utilizando um capilar de sílica, com 73 cm de comprimento efetivo, preenchido com uma solução tampão composta por fosfato (10mmol/L) e dodecil sulfato de sódio (l0mmol/L), a pH 11, com aplicação de voltagem de 25 kV. Nos dois métodos, a detecção dos corantes foi feita na região do visível e a quantificação através de curvas de calibração externa. Os limites de detecção obtidos ficaram na faixa de 0,1 a 0,4 µ tg/mL e 0,4 a 2,5 µ tg/mL, enquanto os limites de quantificação foram de 0,2 a 1,3 µ tg/mL e 1,3 a 7,1 µ tg/mL para a CLAE e EC, respectivamente. As taxas de recuperação, em dois níveis de concentração, para todos os corantes foram de 95,2 a 103,2% para a CLAE, e de 92,6 a 104,0% para a EC. Os valores de repetibilidade calculado para padrões e amostras demonstraram a boa precisão para os dois métodos desenvolvidos. As metodologias propostas e validadas foram aplicadas em 45 amostras de bebidas alcoólicas de diferentes fabricantes brasileiros, sendo: 6 aguardentes aromatizadas, 9 coolers, 7 aperitivos, 3 coquetéis, 8 licores e 12 vinhos tinto. Não houve diferença significativa entre os dados obtidos pelos dois métodos. Em todas as amostras analisadas, os teores de corantes artificiais encontrados estavam em conformidade com a legislação brasileira / Abstract: The use of synthetic dyes for the food industries in the whole world is sufficiently spread out, it is because the colors allow to supplemental or to replace the lost coloration during the processing and or storage, and thus to guarantee the acceptability of the product front to the consumer, being used still as an important instrument to guarantee the uniformity of the products in the production. Of the point of view of the public health, there are different opinions about the safety of the different synthetic dyes used in foods. Much of the studies show that these additives can be dangerous for the health of the population when consumed inadequately, either for abuses of the industry or exaggerate in the consumption. The fact is that, analytical methods for the determination of these colors must be developed, and mainly validated, to guarantee the alimentary security of the products that we ingest. In the present work they had been developed and validated two methods for synthetic dyes determination in alcoholic beverages, using two different techniques, the high performance liquid chromatography (HPLC) and capillary eletrophoresis (CE). The methods had been developed for the simultaneous separation of the eleven synthetic dyes allowed for use in foods in Brazil. In the method for HPLC, for the separation of the synthetic dyes, a reverse phase column was used with gradient elution system composed by water/methanol. In the EC method the separation occurred using a silica capillary with 73 cm of effective length, filled with buffer phosphate solution (l0mmol/L) with sulpfate dodecyl sodium (SDS) (10mmol/L), at pH 11, with application 25kV of voltage. The detection and quantification were done made in same manner for the two methods, using absorption in the visible region and external standardization, respectively. The detection limits were 0.1 to 0.4 µ g/rnL and 0.4 to 2.5 µ g/rnL, while quantification limits were 0.2 to 1.3 µ g/rnL and 1.3 to 7.1 µ g/rnL for HPLC and CE, respectively. Recovery percentage at two levels of concentration for all the synthetic dyes were of the order of 95.2 to 103.2% for HPLC, and of92.6 to 104.0% for CE. The values of repeatability calculated for standards and samples demonstrated the precision of the two methods. The proposed and validated methods were used to analyses 45 alcoholic beverage samples of different Brazilian manufacturers, being: 6 perfumed spirits, 9 coolers, 7 bitters, 3 cocktails, 8 liquors and 12 red wines. The data obtained were the two methods did not present significant difference. It was observed that the limits permitted by Brazilian Legislation for the use of these synthetic dyes were respected in the analyzed samples / Doutorado / Mestre em Ciência de Alimentos