Orientadores: Maria Cecilia de Figueiredo Toledo / Tese (doutorado) Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-08T21:23:15Z (GMT). No. of bitstreams: 1 Arisseto_AdrianaPavesi_D.pdf: 1259436 bytes, checksum: 3d905ca51bf22b95fe5a0c5cb696018c (MD5) Previous issue date: 2007 / Resumo: Uma importante descoberta de pesquisadores suecos em abril de 2002 mostrou que acrilamida, uma substância provavelmente carcinogênica a seres humanos, pode ser formada em determinados alimentos que são submetidos a tratamento térmico em altas temperaturas. No presente estudo, 111 amostras de diferentes categorias de alimentos foram coletadas em supermercados, lojas de ¿fast-foods¿ e restaurantes da cidade de Campinas-SP, entre os meses de setembro de 2004 e abril de 2006, e analisadas para verificar a presença de acrilamida. As amostras foram selecionadas em função dos resultados divulgados por outros países e incluíram, além de produtos à base de batata, trigo e café, alimentos tipicamente brasileiros à base de mandioca e milho, processados em altas temperaturas. Os níveis de acrilamida foram determinados por cromatografia líquida de alta eficiência e espectrometria de massas em série (LC-MS/MS), conforme método previamente desenvolvido. Como a aplicação deste método em matrizes de cacau não apresentou resultados satisfatórios, o mesmo foi modificado através da inclusão de uma etapa de precipitação de proteínas e de alterações no procedimento de limpeza, o que resultou em melhor desempenho nesta matriz. As concentrações de acrilamida determinadas nas amostras analisadas confirmam que produtos à base de batata, tais como batatas fritas e batatas chips, biscoitos e café são os alimentos que apresentam os maiores níveis de acrilamida. Em etapa posterior, a ingestão potencial diária deste contaminante foi estimada combinando-se os dados analíticos de ocorrência de acrilamida obtidos no presente estudo com dados de consumo dos alimentos analisados, disponíveis para a população em geral e para uma população de adolescentes da cidade de Piracicaba-SP. Os valores médios de ingestão estimados para ambas as populações (0,14 e 0,12 µg/kg de peso corpóreo/dia, respectivamente) são inferiores aos valores relatados para populações de países da América do Norte e Europa (0,3 a 2 µg/kg de peso corpóreo/dia), o que pode ser parcialmente atribuído ao fato de que as estimativas de ingestão nestes países levaram em conta a contribuição de um maior número de alimentos. Dessa forma, é importante que mais amostras e grupos de alimentos sejam investigados para que, futuramente, possa ser calculada a contribuição da dieta total como fonte de acrilamida para a população brasileira e avaliados os possíveis riscos à saúde relacionados à exposição a este contaminante / Abstract: An important discovery of Swedish researchers in April 2002 showed that acrylamide, a probable carcinogen to humans, can be formed in certain foods which are submitted to thermal treatment at high temperatures. In the present study, 111 samples of different food categories were collected at supermarkets, fast-food restaurants and restaurants, in Campinas-SP, between September 2004 and April 2006, and analysed to verify the presence of acrylamide. The samples were selected according to results reported in other countries and included, beyond potato- and wheat-based products and coffee, typical Brazilian foods made from cassava and maize, and processed at high temperatures. The levels of acrylamide were determined by high performance liquid chromatography tandem mass spectrometry (LC-MS/MS), according to a method previously developed. As the application of this method in cocoa matrices did not presented satisfactory results, there was a need to modify it by the inclusion of a protein precipitation step and changes in the clean-up procedure, which improved its performance in this matrix. The concentrations of acrylamide determined in analysed samples confirm that potato-based products, such as French fries and potato chips, biscuits and coffee are the foods containing the highest levels of acrylamide. In a next step, the potential daily intake of this contaminant was estimated by combining analytical data on the occurrence of acrylamide obtained in the present study with data on food consumption for the analysed foods, available for the general population and for a population of adolescents from the city of Piracicaba-SP. The mean intakes estimated for both populations (0.14 e 0.12 µg/kg body weight/day, respectively) are below the values reported for populations from North American and European countries (0.3 to 2 µg/kg body weight/day), which may be partially attributed to the fact that intake estimates conducted in these countries took into account the contribution of a greater number of foods. In this way, it is important that more samples and food groups be investigated so that, in the future, it may be possible to calculate the contribution of the total diet as source of acrylamide for the Brazilian population, and to assess the possible risks to health with regard to the exposure to this contaminant / Doutorado / Doutor em Ciência de Alimentos

Although the presence of acrylamide in foods was detected a decade ago, public concern about this issue, and in general about food health, seems to have increased in recent years. Acrylamide is a toxic human carcinogen present mainly in foods from plant origin and subjected to transformation processes in which temperatures above 120 °C are reached, such as frying and baking. Foods that mostly contribute to the intake of acrylamide are: French fries and chips, coffee and coffee substitute, biscuits, bread, pastries, battered and breaded products, breakfast cereals ..., being children and adolescents the most exposed population. From a chemical point of view, acrylamide is formed mainly from the reaction, during thermal processing, between asparagine and reducing sugars, as an intermediate product of the Maillard reaction. In the last decade, both health authorities and the scientific community have made great efforts in scientifically establishing limits of toxicity as well as exploring strategies aimed at reducing acrylamide formation. The purpose of this doctoral thesis is framed in this last sense. On the one hand, our work has been focused on searching new strategies for acrylamide mitigation, in two different types of food: fried potatoes and battered products. The effect of an emerging frying technique, named hot air frying, as well as pre-frying treatments, were studied in fried potatoes. In the case of batters however, the strategy consisted in modifying their composition by adding a hydrocolloid, chitosan, after having tested its potential in model systems. Furthermore, the effect of chitosan addition on the physical properties of the batter formulations and on the quality of the final product was evaluated. Finally, acrylamide bioaccessibility of the food products with the highest acrylamide content was addressed. These food products were subjected to an in vitro gastrointestinal simulation in order to study how acrylamide content progressed throughout the digestion process. Results showed that air fried potatoes as well as chitosan addition to batter formulations reduced the formation of acrylamide by about 90 and 60 %, respectively. The modification of the characteristics of chitosan (molecular weight and deacetylation degree) conditioned its ability to reduce the formation of acrylamide. Furthermore, the incorporation of chitosan to batters reduced the absorption of fat during frying. Chitosan increased viscosity and consistency of batters but, the color and texture of the fried products were not significantly altered. The total replacement of wheat flour by rice flour allowed to obtain gluten-free batters with similar viscosity, when adding chitosan. This new formulation could be adjusted to gluten-free battered products. The simulation of in vitro digestion showed a significant increment of acrylamide after the gastric stage in a wide range of foods. However, acrylamide bioaccessibility (after the intestinal stage) was reduced until pre-digestion levels or even lower in chips and French fries. / Si bien es verdad que la presencia de acrilamida en alimentos se detectó hace más de una década, la preocupación por parte del consumidor, no sólo por este tema si en la alimentación saludable en general, parece haberse incrementado en los últimos años. La acrilamida es un tóxico posible carcinógeno humano presente principalmente en alimentos derivados de materias primas de origen vegetal y sometidos a procesos de transformación en los que se alcanzan temperaturas por encima de 120 ºC como la fritura y horneado. Los alimentos que contribuyen mayoritariamente a la ingesta de acrilamida son: patatas fritas (tipo French fries y chips), café y substituto de café, galletas, pan, bollería, rebozados y empanados, cereales de desayuno¿, siendo la exposición al tóxico superior por parte de población infantil y adolescente. Desde un punto de vista químico, la acrilamida se forma principalmente a partir de la reacción entre la asparagina y azúcares reductores durante el procesado térmico, como producto intermedio de las reacciones de Maillard. En la última década, tanto las autoridades sanitarias como la comunidad científica han puesto un gran empeño en establecer científicamente los límites de toxicidad y en la búsqueda de estrategias orientadas a reducir su formación. El cometido de esta tesis doctoral se enmarca en este último sentido. Por un lado, se ha trabajado en la búsqueda de nuevas estrategias para la reducción de acrilamida, en dos tipos de alimentos distintos: patatas fritas y masas de productos rebozados. En patatas fritas se estudió el efecto de una técnica emergente de fritura, concretamente la fritura por aire caliente, junto con tratamientos previos a la fritura. En masas de rebozados, en cambio, se optó por modificar la composición de estos, adicionando un hidrocoloide, el quitosano, después de haber ensayado su potencial en sistemas modelo. Además, se evaluó el efecto de la incorporación de quitosano sobre las propiedades físicas de la masa de rebozado y sobre la calidad del producto final. Por último y no por ello menos importante, se abordó la problemática de la acrilamida sobre la salud humana desde el punto de vista de su bioaccesibilidad. Para ello, se seleccionaron los productos con mayor contenido en acrilamida dietética y se sometieron a una simulación gastrointestinal in vitro con el fin de conocer los cambios experimentados por el tóxico durante este proceso y determinar su bioaccesibilidad. Los resultados mostraron que la fritura de patatas por aire caliente, así como la incorporación de quitosano a las masas de rebozado reducían la formación de acrilamida alrededor de un 90 y 60 %, respectivamente. La modificación de las características del quitosano (en cuanto a masa molecular y grado de desacetilación) condicionó su capacidad de reducir la formación de acrilamida. La incorporación de quitosano redujo la absorción de grasa durante la fritura de las masas. Si bien aumentó la viscosidad y consistencia de éstas, no se vio alterado significativamente el color y la textura del producto final frito. La substitución total de harina de trigo por harina de arroz permitió obtener masas de rebozado sin gluten con semejante viscosidad, al adicionar quitosano. Esta nueva formulación podría ser adaptada a productos rebozados sin gluten. La simulación de digestión in vitro mostró aumentos significativos de acrilamida tras la fase gástrica en una amplia gama de alimentos. Sin embargo, la bioaccesibildad de acrilamida (tras la fase intestinal) se redujo hasta niveles previos a la digestión o incluso menores, en patatas fritas tipo chips y tipo French fries. / Si bé és veritat que la presència d'acrilamida en aliments es va detectar fa més d'una dècada, la preocupació per part del consumidor, no només per aquest tema si no en l'alimentació saludable en general, sembla haver-se incrementat en els últims anys. L'acrilamida és un tòxic possible carcinogen per a l'èsser humà present principalment en aliments derivats de matèries primeres d'origen vegetal i sotmesos a processos de transformació en els quals s'assoleixen temperatures majors a 120 ºC, com el procés de fregida o al forn. Els aliments que contribueixen majoritàriament a la ingesta d'acrilamida són: creïlles fregides (tipus French fries i chips), cafè i substitut de cafè, galetes, pa, brioixeria, arrebossats i empanats, cereals d'esmorzar ..., sent l'exposició al tòxic superior per part de població infantil i adolescent. Des d'un punt de vista químic, l'acrilamida es forma principalment a partir de la reacció entre l'asparagina i sucres reductors durant el processat tèrmic, com producte intermedi de les reaccions de Maillard. En l'última dècada, tant les autoritats sanitàries com la comunitat científica han posat un gran esforç a establir científicament els límits de toxicitat i en la recerca d'estratègies orientades a reduir la seva formació. La comesa d'aquesta tesi doctoral s'emmarca en aquest últim sentit. D'una banda, s'ha treballat en la recerca de noves estratègies per a la reducció d'acrilamida, en dos tipus d'aliments diferents: creïlles fregides i masses de productes arrebossats. En creïlles fregides es va estudiar l'efecte d'una tècnica emergent de fregit, concretament el fregit per aire calent, juntament amb tractaments previs al fregit. En masses d'arrebossats, en canvi, es va optar per modificar la composició d'aquests, addicionant un hidrocol·loide, el quitosà, després d'haver assajat el seu potencial en sistemes model. A més, es va avaluar l'efecte de la incorporació de quitosà sobre les propietats físiques de la massa d'arrebossat i sobre la qualitat del producte final. Finalment i no per això menys important, es va abordar la problemàtica de l'acrilamida sobre la salut humana des del punt de vista de la seva bioaccesibilidad. Per a això, es van seleccionar els productes amb major contingut en acrilamida dietètica i es van sotmetre a una simulació gastrointestinal in vitro per tal de conèixer els canvis experimentats pel tòxic durant aquest procés i determinar la seva bioaccesibilidad. Els resultats van mostrar que el fregit de creïlles per aire calent, així com la incorporació de quitosà a les masses d'arrebossat reduïen la formació d'acrilamida al voltant d'un 90 i 60%, respectivament. La modificació de les característiques del quitosà (quant a massa molecular i grau de desacetilación) va condicionar la seva capacitat de reduir la formació d'acrilamida. La incorporació de quitosà va reduir l'absorció de greix durant el fregit de les masses. Si bé va augmentar la viscositat i consistència d'aquestes, no es va veure alterat significativament el color i la textura del producte final fregit. La substitució total de farina de blat per farina d'arròs va permetre obtenir masses d'arrebossat sense gluten amb semblant viscositat, en addicionar quitosà. Aquesta nova formulació podria ser adaptada a productes arrebossats sense gluten. La simulació de digestió in vitro va mostrar augments significatius d'acrilamida després de la fase gàstrica en una àmplia gamma d'aliments. No obstant això, la bioaccesibildad d'acrilamida (després de la fase intestinal) es va reduir fins a nivells previs a la digestió o fins i tot menors, en creïlles fregides tipus chips i tipus French fries. / Sansano Tomás, M. (2017). Evaluation of acrylamide in foods and development of some strategies for its reduction [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/86160 / TESIS

The diploma thesis deals with the Influence of the Enzyme L-asparagine and the Inorganic salts (NaCl, CaCl2, NaHCO3 and NH4HCO3) on the elimination of the acrylamide in food-stuffs and a simulated model cereal matrix. The acrylamide belongs to the probable carcinogenic compounds which is incipient in the course of thermal processing of food, which are rich in the reducing sugars and amino acids as L-asparagine. Because of L-asparagine is the natural component of cereals and simultaneously is dominant antecedent incipient acrylamide, the way of the elimination by enzyme L-asparaginase (or the combination of L-asparaginase and salt) leads to the reduced level of acrylamide in a final product. The L- asparagine and salts were used on food-stuffs and a simulated model cereal matrix. It was found that individual used substances (except for NH4HCO3) cause the reduction of acrylamide production even about 90 % without change in the sensory properties of final product.

Targeted covalent inhibitors (TCIs) are irreversible enzyme inhibitors that are designed to first bind to a targeted enzyme’s active site reversibly using non-covalent interactions between the molecular scaffold of the inhibitor and the surrounding amino acid residues of the enzyme’s binding site. They then form a covalent bond between the inhibitor’s electrophilic warhead and a nucleophilic amino acid residue located inside of the binding pocket. Cysteine (Cys), a redox-sensitive thiol, is found in many enzyme active sites and is used as the target for many current TCIs in clinical application. Electrophilic warheads such as acrylamides and chloroacetamides are known to readily undergo thiol-addition, and although they are commonly used in the development of enzyme inhibitors, few previous studies have explored the mechanism of thiol-addition and the intrinsic reactivities of these moieties. In this work, a robust kinetic assay was developed to perform mechanistic studies of thiol-addition to the electrophilic warhead derivatives N-phenylacrylamide (NPA), N-acryloylpiperidine (AcrPip), and N-phenylchloroacetamide (NPC). By reacting these warhead derivatives with thiol nucleophiles having various pKa values, we were able to construct Brønsted-type plots, resulting in shallow positive βNucRS- values for NPA, AcrPip and NPC (βNucRS- = 0.07 ± 0.04, 0.11 ± 0.03, and 0.21 ± 0.07, respectively), meaning that these electrophiles are relatively insensitive to thiolate nucleophilicity. However, while the trend in their reactivity across thiolate nucleophilicity is similar, their intrinsic reactivity was found to be vastly different. In conjunction with the Brønsted-type plot, temperature, ionic strength, and kinetic isotope effects were studied to afford information about the rate-limiting transition state and elucidate the mechanism of thiol-addition. NPA and AcrPip were found to undergo very similar thiol-additions, consistent with the microscopic reverse of the E1cbrev elimination, whereas NPC follows an SN2 type addition, consistent with the intuitive mechanism of addition to a haloacetamide.

Yes / A low concentration poly(acrylamide) sensor has been developed which uses the segmental mobility of another polymer probe with a covalently attached fluorescent marker. Interpolymer complexation with poly(acrylic acid) leads to reduced segmental mobility which can be used to determine the concentration of polymer in solution. This technique could be useful in detecting the runoff of polymer dispersants and flocculants in fresh water supplies following water purification processes. / Funding for the research was kindly provided by the Engineering and Physical Sciences Research Council (EPSRC).

<p> In part I of this dissertation, several cationic polyacrylamides were tested under different conditions for their ability to improve the retention of fines in papermaking. A dynamic drainage jar was used to simulate the turbulence encountered in the papermaking process. Several factors, including temperature, the amount and intensity of turbulence, the additive concentration and the presence of impurities were found to affect fines retention with polymers present. A polymer made by Nalco Chemicals proved to be superior to a commonly used polymer, Percol 292 for a standard fine paper pulp. It was thought that further retention improvements might be possible by tailoring the charge density and molecular weight of polyacylamide retention aides for the specific papermaking system they are intended for. Novel approaches to retention such as those employing combinations of an anionic polymer, a cationic polymer and zirconium oxychloride were thought to show promise as well.</p> <p> In part II of this dissertation several broad polyacrylamide molecular weight standards were prepared by inverse suspension and solution processes on pilot plant equipment at the McMaster Institute for Polymer Production Technology. They were characterized by laser light scattering and viscometry at McMaster, and externally by other methods. Although the polyacrylamides prepared compare favourably to currently available commercial standards when both are analysed by SEC, further analysis must be done to be certain of the molecular weight averages.</p> <p> A relationship is presented to provide for simpler and more accurate light scattering analysis in the future. This relationship relating Mw to the second Virial coefficient may be used to eliminate some uncertainty in the often scattered plots encountered when calculating molecular weights for polyacrylamides analysed by light scattering.</p> / Thesis / Master of Engineering (MEngr)

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No description available.

The neuronal cytoskeleton is vital for normal growth and differentiation of the nervous system, as well as for maintenance of the normal intracellular environment. Without it, major functional deficits occur due to interference with cellular transport of membrane components, proteins and neurotransmitter substances and as a result, inadequate maintenance of the distal axon occurs. Through the study of both nervous tissue and primary neuronal culture, specific cytoskeletal markers have been found to predominate in axonal or dendritic processes, as well as in different stages of neuronal development. In vitro study of neuroblastoma cell lines has also been utilized to develop hypotheses of neuronal development. These hypotheses attempt to explain the appearance of certain cytoskeletal elements, such as phosphorylated neurofilament proteins, in relationship to functional maturity of the neuron. We used SY5Y human neuroblastoma cells as an in vitro model of neurotoxicity to investigate cytoskeletal changes that may occur from the exposure of the nervous system to a known neurotoxicant. Cells were differentiated with either retinoic acid (RA) or dibutyryl cyclic adenosine monophosphate (dbcAMP) and 3-isobutyl-1- methyl-xanthine (IBMX). Differentiation was based morphologically on the appearance of neuritic processes in a majority (>50%) of the cells. Using the peroxidase-antiperoxidase technique, cells were labeled with monoclonal antibodies to cytoskeletal proteins (phosphorylated neurofilament, microtubule associated protein 2, vimentin and low molecular weight neurofilament protein) either specific for axonal markers or linked to stages in neuronal development. Staining patterns were compared to undifferentiated cells using the same protocol. There were no differences in staining patterns found between methods of differentiation or between differentiated cells and undifferentiated controls. Axonal markers of differentiation, defined as phosphorylated neurofilament immunopositivity, were only detected in cells exposed to retinoic acid for 9 days. Once these studies were completed, both differentiated and undifferentiated cultures were exposed to acrylamide as an example of a neurotoxicant with known cytoskeletal effects. Cells were fixed and stained after the observation of cellular swelling 24 hours post acrylamide treatment. In spite of obvious alterations in morphology in unstained cells in culture, the cytoskeletal staining pattern was unchanged after acrylamide treatment. We conclude that there is no difference in the cytoskeletal immunoreactivity of SY5Y neuroblastoma cells when differentiated with RA or dbcAMP/IBMX after three days. Retinoic acid differentiated cells, however, do develop immunoreactivity to axonal markers of differentiation after nine days of treatment. We also conclude that acrylamide does not affect the cytoskeletal structure of SY5Y neuroblastoma cells in undifferentiated or in RA or dbc AMP differentiated cells at the time and concentration tested. / Master of Science

L'acrylamide, connu comme étant une molécule cancérigène, apparait lors de traitement thermique à haute température (>120°C) d'aliments riches en asparagine et en sucre. La friture profonde génère ce composé à des teneurs proche de 0,44 ppm pour les chips de plantain. Or les aliments à base de plantain sont largement consommés dans les pays du sud. Ces travaux de recherche ont pour objectif de mieux comprendre l'accumulation de l'acrylamide lors de l'élaboration de produits frits à base de plantain. Pour y parvenir la teneur en précurseurs (asparagine) a d'abord été obtenue dans différentes variétés de bananes, mais aussi durant le mûrissage d'une variété de plantain d'exportation (Harton). Ces résultats ont permis d'identifier l'asparagine comme étant limitant pour la formation d'acrylamide dans le plantain. Ensuite, une étude cinétique du couplage entre la température et l'activité en eau a été entreprise en système fermé. Le domaine d'étude consistait en 4 températures de traitement (140, 160, 180 et 200°C) et 3 activités en eau (0,43 ; 0,9 ; 0,97). La cinétique de formation et d'élimination de l'acrylamide a été traduite par la somme de 2 cinétiques d'ordre 1. Les paramètres cinétiques ont été identifiés et montrent que la température est le paramètre qui influence le plus la cinétique de l'acrylamide. Le modèle cinétique a dans une autre étape été couplé à un modèle traduisant les transferts d'énergie et de vapeur durant la friture. Le modèle a bien simulé les teneurs en eau et en acrylamide de deux produits frits à base de plantain « tajadas » (produit épais) et « tostones » (produit mince). Les coulages entre transfert et réaction sont discutés en vue d'imaginer des stratégies de réduction de l'acrylamide. Il a été démontré que la formation de l'acrylamide se fait presque exclusivement dans la zone hygroscopique périphérique (croûte). De ce fait, la conduite de la friture en températures variables est recommandée et permet de réduire d'au moins 50% la teneur en acrylamide. La sélection variétale et l'état de maturité du plantain sont les moyens en amont les plus souples pour contrôler la teneur en acrylamide dans le produit final. Des prétraitements par immersion se sont avérés efficaces induisant une réduction significative de l'acrylamide. En revanche, la formulation de molécules antioxydantes ne semble pas avoir d'effet réducteur. / Acrylamide, a molecule known as a carcinogen, appears during heat treatment of asparagine and sugars rich foods at high temperature (> 120 ° C). Deep frying produces this compound at levels close to 0.44 ppm for plantain chips, these foods being widely consumed in developing countries. The aim of the present work is to better understand the accumulation of acrylamide during the process of plantain-based products. In this context, the level of precursors (asparagine) was obtained firstly in different varieties of bananas and also during ripening of an export plantain variety (Harton). These results allowed identifying asparagine as a limiting factor of acrylamide formation in plantain. Afterwards, a kinetic study of the coupling between temperature and water activity was undertaken in a closed system. The field of study consisted of four temperatures of treatment (140, 160, 180 and 200 ° C) and three water activities (0.43, 0.9, 0.97). The kin etics of formation and elimination of acrylamide was translated by the sum of 2 order 1 kinetics. The kinetic parameters were identified and showed that temperature is the parameter that influences the most acrylamide kinetic. The kinetic model, in a further step, was coupled to a model describing energy and steam transfers during frying. The model simulated well water and acrylamide content in two plantain-based fried products: "Tajada" (produced thick) and "tostones" (thin product). The coupling between transfers and reaction were discussed in order to identify strategies to reduce acrylamide. It was demonstrated that acrylamide formation occurs almost exclusively in the hygroscopic and peripheral area (crust). Thus, conducting frying process at varying temperatures is recommended and can reduce by at least 50% the acrylamide content. The varietal selection and the ripeness degree of plantain are more flexible ways to control upstream the levels of acrylamide in the final product. Immersion pretreatments were also effective to reduce significantly acrylamide. However, the formulation of antioxidant molecules did not appear to have reduction effect.