Made available in DSpace on 2014-06-11T19:29:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-07-27Bitstream added on 2014-06-13T20:18:59Z : No. of bitstreams: 1 leandro_fz_me_araiq_prot.pdf: 1135475 bytes, checksum: f2d8e4f49663bf83eb7c7e55cb4d5a0c (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Atualmente há um grande interesse no estudo dos desreguladores endócrinos (substâncias estranhas aos organismos, que mimetizam ou bloqueiam a ação natural dos hormônios naturais), tanto em relação ao seu destino ambiental quanto à toxicologia dos mesmos. Neste trabalho, avaliou-se o Bisfenol A (BPA), um monômero do qual se produz o plástico policarbonato. Recentemente foi proposto que o BPA exibe atividade estrogênica em concentrações de 1 ng mL-1. Neste trabalho, desenvolveu-se um método para a determinação de BPA em águas naturais, fazendo-se uso de cartuchos de poliestireno-divinilbenzeno (PSDVB) para a extração em fase sólida (SPE) e cromatografia líquida de alta eficiência com um detector de fluorescência (HPLC/FLU) para a quantificação do composto. A recuperação do método foi avaliada utilizando-se amostras de água potável fortificada em 3 níveis de concentração: 0,06; 0,2 e 0,6 ng mL-1. A extração do analito (500 mL; n=4) resultou valores de recuperações entre 99 e 100% e CV entre 0,30 e 3,9%. Os limites de detecção e quantificação do método foram 0,02 e 0,06 ng mL-1, respectivamente. O mesmo estudo foi efetuado para água superficial sendo obtidos valores de recuperação entre 85,8 e 87% para níveis de fortificação similares aos utilizados para água potável, com CV entre 1,1 e 2,3%. O método validado foi aplicado com bom desempenho às amostras de água de entrada e saída da Estação de Tratamento de Água, onde constatou-se a presença de BPA entre 11,7 e 16,8 ng mL-1 na água bruta e 6,2 e 7,3 ng mL-1 na água potável. Desta forma, o monitoramento ambiental do BPA torna-se extremamente relevante e necessário. / Nowadays it has been increased the concern about endocrine disrupters (unknown substances for organisms that mimic or block the natural action of endogenous hormones), so much at relation of your environmental fate as toxicology about them. In this study, it had been evaluated Bisphenol A (BPA), a monomer, wich produces polycarbonate. Recently, it has been proposed that BPA exhibits estrogenic activity at 1 ng mL-1. At this work, it had been developed a method for determination of BPA at natural water with polistyrene-divinylbenzene (PSDVB) cartridge on phase solid extraction (SPE) and high performance chromatography liquid with fluorescence detection (HPLC/FLU) for the analyte's quantification. The method's recovery was evaluated with spiked drink water sample at three concentrations levels: 0,06; 0,2 and 0,6 ng mL-1. The analyte extraction (500 mL; n=4) gave recoveries between 99 and 100% with CV between 0,30 and 3,9%. The method detection and quantification limits were 0,02 and 0,06 ng mL-1, respectively. The same study was developed for superficial water with recoveries between 85,8% and 87% for similar spiked levels at the drink water with CV between 1,1 and 2,3%. The method developed was applied with good performance at water's samples of Water Treatment Plant, verifying the appearance of BPA between 11,7 and 16,8 ng mL-1 on sewage water and 6,2 until 7,3 ng mL-1 on drink water. Therefore the environmental monitoring of BPA is very important and indispensable.

Orientador: Gilberto de Nucci / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-07T09:06:01Z (GMT). No. of bitstreams: 1 Sanfelice_AndreTassoDantas_M.pdf: 2681697 bytes, checksum: 0b507c57048f3b18ee152ffbbe878963 (MD5) Previous issue date: 2005 / Resumo: Objetivo: Avaliar a bioequivalência de uma formulação de ramipril 5 mg (formulação teste de ramipril dos laboratórios Biossintética Ltda. e a formulação de referência padrão Triatec@ da Aventis Pharma) em vinte e seis voluntários de ambos os sexos. Método: O estudo foi conduzido usando um estudo aberto, randomizado, cruzado de duas fases com um intervalo de washout de 2 semanas. As amostras de plasma foram obtidas sobre um período de 36 horas. As concentrações plasmáticas de ramipril e ramiprilato foram analisadas por cromatografia líquida acoplada à espectrometria de massa (LC-MS-MS), com modo de ionização electrospray positivo usando o monitoramento de múltiplas reações (MRM). Das curvas de concentração plasmática versus o tempo para ramipril e ramiprilato, os seguintes parâmetros farmacocinéticos foram obtidos: ASCo-36h ASCIast, ASC(}...infe Cmax. Resultados: O limite de quantificação foi 0.2 ng.mL-1 e 1.0 ng.mL-1 para ramipril e ramiprilato, respectivamente. A média geométrica e a respectivo intervalo de confiança de 90% para RamipriIJTriatec@ e Ramiprilatol Triatec@ foram respectivamente: 104.69".10 (90% IC= 93.210/0-117.59%) para Cmax" 102.49% (90% IC= 92.76%-113.24%) para ASClast" 102.97% (90% IC= 93.21%-113.74%) para ASCO-36h" 103.60% (90% IC= 93.56%-114.73%) para ASCinf;, 108.48% (90% IC= 98.86%-119.03%) para Cmax, 105.88% (90% IC= 101.55%-110.39%) para ASClast,105.81% (90% IC= 101.53%-110.27%) para ASCo.36h, 97.30% (90% IC= 90.17%-104.99%) para ASCinf. Conclusão: Desde que o IC de 90% para ASCtast, ASCO-36h, ASCO-inf e Cmax apresentou-se dentro do intervalo de 80-125% proposto pelo FDA (US), foi concluído que a formulação ramipril elaborada pelo laboratório Biosintética Ltda é bioequivalente para a formulação Triatec@ para taxa e velocidade de absorção / Abstract: Objective: To assess the bioequivalence of one ramipril 5 mg tablet formulation (ramipril test formulation from Laboratórios Biosintética Ltda and Triatec$ from Aventis Pharma standard reference formulation) in 26 healthy volunteers ofboth sexes. Methods: The study was conducted using an open, randomized, two-period crossover design with a 2-week washout interval. Plasma samples were obtained over a 36 h period. Plasma ramipril and ramiprilat concentrations were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using multiple reaction monitoring (MRM). From the ramipril and ramiprilat plasma concentration vs time curves, the following pharmacokinetic parameters were obtained: ASCO-36h ASClast, ASCo-infand Cmax. Results: The limit of quantification was 0.2 ng.mL-1 and 1.0 ng.mL-1 for ramipril and ramiprilat, respectively. The geometric mean and respective 90% confidence interval (CI) RamipriVTriatec@ and Ramiprilat/ Triatec@ percent ratios were respectively: 104.69% (90% CI= 93.21%-117.59%) for Crnax,1O2.49% (90% CI= 92.76%-113.24%) for ASCIast,1O2.97% (90% CI= 93.21%-113.74%) for ASCo-36h,103.60% (90% CI= 93.56%-114.73%) for ASCnf)08.48% (9001Ó CI= 98.86%-119.03%) for Crnax,1O5.88% (90% CI= 101.55%-110.39%) for ASClast,105.81% (90% CI= 101.53%-110.27%) for ASCO-36h, 97.30% (9001Ó CI= 90.17%-104.99%) for ASCinf. Conclusion: Since the 90% CI for ASClast, ASCO-36h, ASCO-inf and Cmax ratios were within the 80-125% interval proposed by the US FDA, it was concluded that ramipril formulation elaborated by Laboratórios Biosintética Ltda is bioequivalent to Triatec@ formulation for both the rate and the extent of absorption / Mestrado / Clinica Medica / Mestre em Clinica Medica

<p>Sprovedena studija po prvi put izve&scaron;tava o prisustvu patulina, sekundarnog metabolita određenih vrsta plesni, u proizvodima od jabuka, kao i proceni rizika usled unosa patulina od strane odojčadi, dece, adolescenata i odrasle populacije u Republici Srbiji. Ukupno 356 uzoraka sokova i ka&scaron;ica za odojčad i malu decu (48 i 66, redom), sokova za decu (mala pakovanja sa cevčicom, 100) i sokova u porodičnom pakovanju (142), sakupljeno je sa trži&scaron;ta tokom tri godine i analizirano primenom tečne hromatografije sa ultraljubičastom detekcijom, metodom koja je predhodno validirana. Prisustvo patulina je detektovano u 44% i 17% uzoraka sokova i ka&scaron;ica za odojčad i malu decu, redom, u količinama nižim od zakonskog ograničenja od 10 &mu;g/kg (maksimalno 8,3 i 7,7 &mu;g/kg, redom). Udeo kontaminiranih uzoraka među sokovima za decu iznosio je 43%, sa najvi&scaron;om koncentracijom patulina od 30,2 &mu;g/kg ispod maksimalno dozvoljenog nivoa od 50 &mu;g/kg. Patulin je detektovan u 51% sokova u porodičnom pakovanju, sa 0,7% uzoraka iznad zakonske granice od 50 &mu;g/kg (prosečna koncentracija 4,3 &mu;g/kg). Sokovi od jabuka su pokazali značajno vi&scaron;i udeo kontaminiranih uzoraka (74% u odnosu na 28%), kao i vi&scaron;i prosečni sadržaj patulina (6,4 u odnosu na 2,1 &mu;g/kg) u poređenju sa sokovima od me&scaron;anog voća. Procena rizika usled unosa patulina od strane odojčadi, dece, adolescenata i odrasle populacije u Republici Srbiji, sprovedena primenom determinističkog i probabilističkog pristupa, uključujući rezultate biodostupnosti patulina, pokazala je da je odnos između procenjenog dnevnog unosa i toksikolo&scaron;ke referentne doze za patulin od 0,4 &mu;g/kg telesne mase, koji se označava kao &ldquo;hazard quotient&rdquo;, znatno ispod 1, &scaron;to ukazuje na tolerantan nivo izloženosti i nepostojanje razloga za zabrinutost za zdravlje populacije. Dodatno, analiza ostataka pesticida i toksičnih metala potvrdila je bezbednost proizvoda od jabuka na trži&scaron;tu. Međutim, identifikacija vi&scaron;estrukih ostataka pesticida je razlog za aktivan pristup i pažljivo planiranje i sprovođenje monitoringa bezbednosti hrane, posebno hrane za odojčad i malu decu, kao najosetljivije populacione grupe.</p> / <p>This study reports for the first time the occurrence of patulin, a secondary metabolite of certain fungi, in apple-based food, as well as risk assessment related to patulin intake by infants, children, adolescents and adults in Serbia. In total, 356 samples of infant fruit juices (48), infant pur&eacute;e (66), juices for children (small package with straw, 100), and juices in family package (142) were collected from the market over three years (2013&ndash;15) and analysed using validated method based on liquid chromatography with ultraviolet detection. Patulin was found in 44% of infant juices and 17% of infant pur&eacute;e, with all values below the legal limit of 10 &mu;g/kg (maximum 8.3 and 7.7 &mu;g/kg, respectively). The proportion of contaminated samples among fruit juices for children was 43%, with the highest patulin concentration at 30.2 &mu;g/kg, not exceeding the maximum allowed level of 50 &mu;g/kg. Patulin was found in 51% of juices in family package, with 0.7% of the samples in excess of the legal limit of 50 &mu;g/kg (mean 4.3 &mu;g/kg). Apple juices showed significantly higher percentage of contaminated samples (74% versus 28%), as well as higher mean patulin content (6.4 versus 2.1 &mu;g/kg) when compared with the multifruit ones. Risk assessment of patulin intake by Serbian infants, children, adolescents and adults, conducted by deterministic and probabilistic approaches and including the bioaccessibility results, revealed a ratio between exposure and toxicological reference dose for patulin of 0,4 &mu;g/kg body weigth, called hazard quotients, well below 1, indicating a tolerable exposure level and no health concern. Furthermore, analysis of pesticide residues and toxic metals confirmed safety of apple products on the market. However, identification of multiple pesticide residues is a reason for an active attitude and carefully planned and conducted monitoring of food safety, expecialy in the case of food for infants and young children, as they are the most susceptible population group.</p>

Orientador: Mary Rosa Rodrigues de Marchi / Banca: Manoel Lima de Menezes / Banca: Eny Maria Vieira / Resumo: Atualmente há um grande interesse no estudo dos desreguladores endócrinos (substâncias estranhas aos organismos, que mimetizam ou bloqueiam a ação natural dos hormônios naturais), tanto em relação ao seu destino ambiental quanto à toxicologia dos mesmos. Neste trabalho, avaliou-se o Bisfenol A (BPA), um monômero do qual se produz o plástico policarbonato. Recentemente foi proposto que o BPA exibe atividade estrogênica em concentrações de 1 ng mL-1. Neste trabalho, desenvolveu-se um método para a determinação de BPA em águas naturais, fazendo-se uso de cartuchos de poliestireno-divinilbenzeno (PSDVB) para a extração em fase sólida (SPE) e cromatografia líquida de alta eficiência com um detector de fluorescência (HPLC/FLU) para a quantificação do composto. A recuperação do método foi avaliada utilizando-se amostras de água potável fortificada em 3 níveis de concentração: 0,06; 0,2 e 0,6 ng mL-1. A extração do analito (500 mL; n=4) resultou valores de recuperações entre 99 e 100% e CV entre 0,30 e 3,9%. Os limites de detecção e quantificação do método foram 0,02 e 0,06 ng mL-1, respectivamente. O mesmo estudo foi efetuado para água superficial sendo obtidos valores de recuperação entre 85,8 e 87% para níveis de fortificação similares aos utilizados para água potável, com CV entre 1,1 e 2,3%. O método validado foi aplicado com bom desempenho às amostras de água de entrada e saída da Estação de Tratamento de Água, onde constatou-se a presença de BPA entre 11,7 e 16,8 ng mL-1 na água bruta e 6,2 e 7,3 ng mL-1 na água potável. Desta forma, o monitoramento ambiental do BPA torna-se extremamente relevante e necessário. / Abstract: Nowadays it has been increased the concern about endocrine disrupters (unknown substances for organisms that mimic or block the natural action of endogenous hormones), so much at relation of your environmental fate as toxicology about them. In this study, it had been evaluated Bisphenol A (BPA), a monomer, wich produces polycarbonate. Recently, it has been proposed that BPA exhibits estrogenic activity at 1 ng mL-1. At this work, it had been developed a method for determination of BPA at natural water with polistyrene-divinylbenzene (PSDVB) cartridge on phase solid extraction (SPE) and high performance chromatography liquid with fluorescence detection (HPLC/FLU) for the analyte's quantification. The method's recovery was evaluated with spiked drink water sample at three concentrations levels: 0,06; 0,2 and 0,6 ng mL-1. The analyte extraction (500 mL; n=4) gave recoveries between 99 and 100% with CV between 0,30 and 3,9%. The method detection and quantification limits were 0,02 and 0,06 ng mL-1, respectively. The same study was developed for superficial water with recoveries between 85,8% and 87% for similar spiked levels at the drink water with CV between 1,1 and 2,3%. The method developed was applied with good performance at water's samples of Water Treatment Plant, verifying the appearance of BPA between 11,7 and 16,8 ng mL-1 on sewage water and 6,2 until 7,3 ng mL-1 on drink water. Therefore the environmental monitoring of BPA is very important and indispensable. / Mestre

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-09-05T19:49:54Z No. of bitstreams: 1 michelecamposalves.pdf: 4465106 bytes, checksum: 619948227c4d2d419bdbeb66a9cad785 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-09-06T11:36:14Z (GMT) No. of bitstreams: 1 michelecamposalves.pdf: 4465106 bytes, checksum: 619948227c4d2d419bdbeb66a9cad785 (MD5) / Made available in DSpace on 2017-09-06T11:36:14Z (GMT). No. of bitstreams: 1 michelecamposalves.pdf: 4465106 bytes, checksum: 619948227c4d2d419bdbeb66a9cad785 (MD5) Previous issue date: 2013-08-12 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Atualmente, existe um interesse crescente nas potenciais atividades de proteção à saúde do chá verde (Camellia sinensis), que é caracterizado pela presença de grandes quantidades de polifenois, sendo a maioria deles representados por catequinas. Galato de epigalocatequina (EGCG) é o componente mais abundante e ativo, sendo geralmente utilizado como biomarcador junto com a cafeína (CAF) e o ácido gálico (AG). EGCG é pouco absorvido no tratogastrointestinal e está sujeito ao metabolismo de primeira passagem pelo fígado quando administrado por via oral, forma mais comumente utilizada para o chá verde. Um sistema transdérmico poderia então ser um mecanismo de liberação alternativo. Dessa forma, o presente trabalho teve como objetivo: desenvolver e validar métodos por cromatografia em fase líquida de alta eficiência (CLAE) para a quantificação simultânea de EGCG, CAF e AG em cápsulas e emulsões contendo extrato seco de chá verde e determinar na emulsão a liberação de EGCG in vitro e sua permeação cutânea em pele humana, utilizando modelo ex vivo. Os métodos utilizaram CLAE de fase inversa e detecção em arranjo de fotodiodo. A separação foi atingida utilizando as seguintes condições: coluna octadecilsilano; fase móvel composta por água, etanol, acetato de etila e ácido acético (84:12:3:1, v/v/v/v); temperatura do compartimento para a coluna de 35 °C; e um sistema de fluxo por gradiente, alternando o fluxo entre 0.7 e 1.4 mL.min-1. Os compostos foram separados em 35 min. Os métodos foram simples, seletivos, precisos, exatos e rápidos; e mostraram a confiabilidade necessária para que fossem utilizados no controle de qualidade das formulações testadas. O método para a emulsão provou ser adequado para os ensaios de liberação in vitro e de permeação ex vivo. A taxa de liberação in vitro de EGCG foi de 8896.01 mg.cm-2, seguindo o modelo de pseudo-primeira ordem, também conhecido como modelo de Higuchi. O teste de permeação ex vivo mostrou que EGCG não é capaz de exercer suas atividades biológicas sistemicamente quando utilizado a partir da emulsão testada, permanecendo apenas no estrato córneo. / Currently, there is an increased interest in potential health-protective activities of green tea (Camellia sinensis), which is characterized by the presence of polyphenols huge amounts, being the majority of them catechins. Epigallocatechin 3- gallate (EGCG) is the most abundant and active component, been generally used as biomarker together with caffeine (CAF) and gallic acid (GA). EGCG is poorly absorbed in the gastrointestinal tract and is subject to first-pass metabolism by the liver when administered orally, the most widely used forms of green tea. A transdermal system could then be an alternative delivery mechanism. In this light, the present work aimed: to develop and validate high performance liquid chromatography (HPLC) methods for simultaneous quantification of EGCG, CAF and GA in emulsions and capsules containing dry extract green tea and to determine in emulsion the in vitro EGCG release and its human skin cutaneous permeation using ex vivo model. The methods used reversed phase HPLC and photodiode array detection. Separation was achieved using the follow conditions: octadecylsilyl column; mobile phase composed by water, ethanol, ethyl acetate and acetic acid (84:12:3:1, v/v/v/v); column temperature of 35 °C; and flow gradient syst em, alternating flow between 0.7 and 1.4 mL.min-1. The compounds were separated within 35 min. The methods were simple, selective, precise, accurate and fast; and provide the reliability required for them to be used for quality control of tested formulations. The emulsion method proved to be suitable both for in vitro release and ex vivo permeation. EGCG in vitro release rate was found to be 8896.01 μg.cm–2, following pseudo-first-order model, also known as Higuchi’s model. Ex vivo permeation testing showed that EGCG is not able to exert its biological activities systemically when used from tested emulsion, remaining only in the stratum corneum.

Here we report the identification and approximate quantification of cytochrome P450 (CYP) proteins in human liver microsomes as determined by nano-LC-MS/MS with application of the exponentially modified protein abundance index (emPAI) algorithm during database searching. Protocols based on 1D-gel protein separation and 2D-LC peptide separation gave comparable results. In total, 18 CYP isoforms were unambiguously identified based on unique peptide matches. Further, we have determined the absolute quantity of two CYP enzymes (2E1 and 1A2) in human liver microsomes using stable-isotope dilution mass spectrometry, where microsomal proteins were separated by 1D-gel electrophoresis, digested with trypsin in the presence of either a CYP2E1- or 1A2-specific stable-isotope labeled tryptic peptide and analyzed by LC-MS/MS. Using multiple reaction monitoring (MRM) for the isotope-labeled tryptic peptides and their natural unlabeled analogues quantification could be performed over the range of 0.1-1.5 pmol on column. Liver microsomes from four individuals were analyzed for CYP2E1 giving values of 88-200 pmol/mg microsomal protein. The CYP1A2 content of microsomes from a further three individuals ranged from 165 to 263 pmol/mg microsomal protein. Although, in this proof-of-concept study for CYP quantification, the two CYP isoforms were quantified from different samples, there are no practical reasons to prevent multiplexing the method to allow the quantification of multiple CYP isoforms in a single sample.

BACKGROUND: Acrylamide is a common dietary exposure that crosses the human placenta. It is classified as a probable human carcinogen, and developmental toxicity has been observed in rodents. OBJECTIVES: We examined the associations between prenatal exposure to acrylamide and birth outcomes in a prospective European mother-child study. METHODS: Hemoglobin (Hb) adducts of acrylamide and its metabolite glycidamide were measured in cord blood (reflecting cumulated exposure in the last months of pregnancy) from 1,101 singleton pregnant women recruited in Denmark, England, Greece, Norway, and Spain during 2006-2010. Maternal diet was estimated through food-frequency questionnaires. RESULTS: Both acrylamide and glycidamide Hb adducts were associated with a statistically significant reduction in birth weight and head circumference. The estimated difference in birth weight for infants in the highest versus lowest quartile of acrylamide Hb adduct levels after adjusting for gestational age and country was -132 g (95% CI: -207, -56); the corresponding difference for head circumference was -0.33 cm (95% CI: -0.61, -0.06). Findings were similar in infants of nonsmokers, were consistent across countries, and remained after adjustment for factors associated with reduced birth weight. Maternal consumption of foods rich in acrylamide, such as fried potatoes, was associated with cord blood acrylamide adduct levels and with reduced birth weight. CONCLUSIONS: Dietary exposure to acrylamide was associated with reduced birth weight and head circumference. Consumption of specific foods during pregnancy was associated with higher acrylamide exposure in utero. If confirmed, these findings suggest that dietary intake of acrylamide should be reduced among pregnant women.