Kidney bean proteins and peptides can be developed to serve as an important ingredient for the formulation of high quality foods or therapeutic products that may positively impact on body function and human health. The main goal of this thesis was to determine the in vitro structural and functional characteristics of major proteins and enzymatic protein hydrolysate of red kidney bean (Phaseolus vulgaris). Selective aammonium sulfate precipitation of the kidney bean proteins yielded 88% globulin and 7% albumin.The globulin and albumin are glycoproteins that contained ~4% and 45% carbohydrate contents, respectively. Physicochemical and functional characteristics of the globulin fraction, such as, gelation concentration, foam stability, emulsion capacity, and emulsion stability were superior to those of albumin. Reducing SDS-PAGE revealed vicilin with molecular weight of ~45 kDa as the major globulin in kidney beans. Circular dichroism spectroscopy of the purified vicilin showed reductions in α-helix, and β-pleated sheet conformations upon addition of NaCl or changes in pH. Likewise, the tertiary structures as observed from the near-UV CD spectra were also changed by shifts in pH conditions and NaCl addition. Far UV-CD showed increased β-sheet content up till 60oC from room temperature, but a steady loss in the tertiary structure as temperature was further increased; however, β-sheet structure was still detectable at 80oC. Differential scanning calorimetry thermograms showed a prominent endothermic peak with denaturation temperature at around 90oC, attributed to thermal denaturation of vicilin. Alcalase hydrolysis of kidney bean globulin produced multifunctional peptides that showed potential antihypertensive properties because of the in vitro inhibition of activities of renin and angiotensin I converting enzyme as well as the antioxidant properties. The <1 and 5-10 kDa peptide fractions exhibited highest (p<0.05) renin inhibition and the ability to scavenge 2, 2-Diphenyl-1-picrylhydrazyl free radical, inhibit peroxidation of linoleic acid and reduce Fe3+ to Fe2+. Based on this study, incorporation of kidney bean globulin as an ingredient may be useful for the manufacture of high quality food products. Likewise, the kidney bean protein hydrolysates, especially the <1 kDa fraction represent a potential source of bioactive peptides for the formulation of functional foods and nutraceuticals.

Cheese is one of the most well known food products in the world dating back to the 8th century B.C. There are more than 2000 varieties of cheese that are manufactured all over the world. Feta cheese is a soft white cheese with a salty and slightly acidic taste, which has originated from Greece. Most of the feta cheese manufactured in Greece is consumed locally, the migration of greeks to other parts of the world led to a demand for feta cheese outside of Greece. The spreading of the popularity of feta cheese to other ethnic groups in different parts of the world resulted in the high demand for feta cheese worldwide. The modern and most efficient method of feta cheese production involves a membrane filtration method, known as ultrafiltration. The ultrafiltration process utilises pressure as a driving force to concentrate milk by removal of water and small dissolved molecules. Hollow fibre and spiral wound ultrafiltration membranes are the two types of membranes that are commonly used for cheese production. An extensive amount of research exists on the implementation of ultrafiltration to improve the efficiency of the cheese making process and the performance of the membranes. However, limited research has been conducted on the comparison of the hollow fibre and spiral wound membrane performance in the cheese making process. The objective of the research was to determine if the hollow fibre membranes used at Puhoi Valley Cheese can be replaced with spiral wound membranes without compromising the quality of cheese produced. In order to achieve the objective, feta cheese was produced using hollow fibre and spiral wound ultrafiltration pilot plants. The operating performances of the hollow fibre and spiral wound membrane units were compared. To ensure that the quality of cheese is maintained, the cheese manufactured on the pilot plant units was analysed in terms of composition, microbiology, texture and sensory properties. The cheese made using the hollow fibre membrane pilot plant was compared with the reference sample from Puhoi Valley Cheese as they use hollow fibre membranes to produce feta cheese. The cheese made from the spiral wound membrane unit was also compared to that made by the hollow fibre membrane pilot plant unit. The operating parameters such as the inlet and outlet pressure, pressure difference along the membrane, transmembrane pressure, flow rate, recycle rate (bleed off rate), temperature and the run time were recorded. The operating parameters of the hollow fibre and spiral wound runs were compared with the data from Puhoi Valley Cheese. The quality of cheese made on the hollow fibre and spiral wound pilot plant units were evaluated in terms of composition, texture, microbiology and sensory properties. The composition was defined by the fat, protein, total solids and salt contents. The fat content was determined by utilising the modified Schmid-Bondzynski-Ratzlaff method, protein by the Kjeldahl method, total solids by using the air drying oven and salt percentage by the volhard method. The texture of the cheese was determined by the fracturability and hardness from the compression curve generated using the single bite compression test. The microbiological testing was performed according to New Zealand testing methods for E.Coli, Staphylococcus aureus, coliforms and yeast and mould. The difference from the control method was utilised for sensory evaluation. The acid degree value method was used to determine the lipase activity in feta cheese. It was found from the composition, texture and sensory analysis that the cheese from the hollow fibre pilot plant was different from the cheese manufactured at Puhoi Valley Cheeses (PVC). The spiral wound cheeses were also found to be different to PVC cheese, however the spiral wound cheeses and the pilot plant hollow fibre cheese were the same. The differences between both the pilot plant cheeses and PVC cheese were in terms of the fat, salt, moisture contents and the lipase activity in the cheeses. The fat content in the hollow fibre and spiral wound pilot plant cheeses are lower in comparison to the PVC cheese. This difference in fat content is considered to be due to the difference in the fat to protein ratio of the milk concentrated on the pilot plant and the PVC ultrafiltration system. The lower fat content resulted in firmer cheese than PVC due to more cross linking between the protein strands in cheese. The salt content in the cheeses made using the hollow fibre and spiral wound pilot plants was lower than Puhoi Valley Cheese. This is considered to be due to the low ratio of brine volume to cheese volume used for salting the cheese. The salt content of brine decreases during brining; hence a low ratio of brine volume to cheese volume causes a significant decrease in brine concentration. The decrease in brine concentration decreases the salt intake of the cheese. As salt diffuses in the moisture diffuses out, lower salt content results in higher moisture content in the cheese. As mentioned, the moisture content of the hollow fibre pilot plant cheese was higher than the PVC cheese. The moisture content is inversely proportional to the total solids, hence higher moisture in pilot plant cheeses implies lower total solids than the PVC cheese. The lipase activity results showed that the hollow fibre and spiral wound pilot plant cheeses had higher lipase activity than the Puhoi valley cheese. The differences in lipase activity of the pilot plant cheeses and Puhoi Valley cheese were considered to be due to the incomplete inactivation of lipase present in milk during pasteurisation. The results from texture and sensory evaluation support the above mentioned differences. The microbiology results for all pilot plant cheeses were within the trigger limits set by Puhoi valley cheeses. The results from monitoring the operating parameters of both the pilot plant data show that the permeate flux decreases while the total solids in milk increase with time, which was also observed from the Puhoi Valley Cheese data. However, the rate of decrease of the permeate flux and the increase of the total solids in milk are dependent on the membrane area, feed volume, transmembrane pressure, pressure drop across the membrane and the flow characteristics. The rate of decrease in permeate flux and the rate of increase in the total solids of the hollow fibre runs and spiral wound runs are slightly different. The difference is due to the availability of larger membrane surface area and processing of larger feed volume of milk in the spiral wound runs. The transmembrane pressure and the pressure drop across the membrane were maintained as close as possible to Puhoi Valley Cheese. In conclusion, spiral wound membranes can be used to achieve the desired total solids concentration and successfully make the same feta cheese as the hollow fibre pilot plant. In order to make the same quality of feta cheese as Puhoi Valley Cheese using the spiral wound membrane pilot plant, the same composition of milk used for concentration at Puhoi Valley Cheese needs to be used on the spiral wound pilot plant unit. It is recommended that Puhoi Valley Cheeses should be replaced with spiral wound membranes if they are more economical in terms of cost than the hollow fibre membranes.

Cheese is one of the most well known food products in the world dating back to the 8th century B.C. There are more than 2000 varieties of cheese that are manufactured all over the world. Feta cheese is a soft white cheese with a salty and slightly acidic taste, which has originated from Greece. Most of the feta cheese manufactured in Greece is consumed locally, the migration of greeks to other parts of the world led to a demand for feta cheese outside of Greece. The spreading of the popularity of feta cheese to other ethnic groups in different parts of the world resulted in the high demand for feta cheese worldwide. The modern and most efficient method of feta cheese production involves a membrane filtration method, known as ultrafiltration. The ultrafiltration process utilises pressure as a driving force to concentrate milk by removal of water and small dissolved molecules. Hollow fibre and spiral wound ultrafiltration membranes are the two types of membranes that are commonly used for cheese production. An extensive amount of research exists on the implementation of ultrafiltration to improve the efficiency of the cheese making process and the performance of the membranes. However, limited research has been conducted on the comparison of the hollow fibre and spiral wound membrane performance in the cheese making process. The objective of the research was to determine if the hollow fibre membranes used at Puhoi Valley Cheese can be replaced with spiral wound membranes without compromising the quality of cheese produced. In order to achieve the objective, feta cheese was produced using hollow fibre and spiral wound ultrafiltration pilot plants. The operating performances of the hollow fibre and spiral wound membrane units were compared. To ensure that the quality of cheese is maintained, the cheese manufactured on the pilot plant units was analysed in terms of composition, microbiology, texture and sensory properties. The cheese made using the hollow fibre membrane pilot plant was compared with the reference sample from Puhoi Valley Cheese as they use hollow fibre membranes to produce feta cheese. The cheese made from the spiral wound membrane unit was also compared to that made by the hollow fibre membrane pilot plant unit. The operating parameters such as the inlet and outlet pressure, pressure difference along the membrane, transmembrane pressure, flow rate, recycle rate (bleed off rate), temperature and the run time were recorded. The operating parameters of the hollow fibre and spiral wound runs were compared with the data from Puhoi Valley Cheese. The quality of cheese made on the hollow fibre and spiral wound pilot plant units were evaluated in terms of composition, texture, microbiology and sensory properties. The composition was defined by the fat, protein, total solids and salt contents. The fat content was determined by utilising the modified Schmid-Bondzynski-Ratzlaff method, protein by the Kjeldahl method, total solids by using the air drying oven and salt percentage by the volhard method. The texture of the cheese was determined by the fracturability and hardness from the compression curve generated using the single bite compression test. The microbiological testing was performed according to New Zealand testing methods for E.Coli, Staphylococcus aureus, coliforms and yeast and mould. The difference from the control method was utilised for sensory evaluation. The acid degree value method was used to determine the lipase activity in feta cheese. It was found from the composition, texture and sensory analysis that the cheese from the hollow fibre pilot plant was different from the cheese manufactured at Puhoi Valley Cheeses (PVC). The spiral wound cheeses were also found to be different to PVC cheese, however the spiral wound cheeses and the pilot plant hollow fibre cheese were the same. The differences between both the pilot plant cheeses and PVC cheese were in terms of the fat, salt, moisture contents and the lipase activity in the cheeses. The fat content in the hollow fibre and spiral wound pilot plant cheeses are lower in comparison to the PVC cheese. This difference in fat content is considered to be due to the difference in the fat to protein ratio of the milk concentrated on the pilot plant and the PVC ultrafiltration system. The lower fat content resulted in firmer cheese than PVC due to more cross linking between the protein strands in cheese. The salt content in the cheeses made using the hollow fibre and spiral wound pilot plants was lower than Puhoi Valley Cheese. This is considered to be due to the low ratio of brine volume to cheese volume used for salting the cheese. The salt content of brine decreases during brining; hence a low ratio of brine volume to cheese volume causes a significant decrease in brine concentration. The decrease in brine concentration decreases the salt intake of the cheese. As salt diffuses in the moisture diffuses out, lower salt content results in higher moisture content in the cheese. As mentioned, the moisture content of the hollow fibre pilot plant cheese was higher than the PVC cheese. The moisture content is inversely proportional to the total solids, hence higher moisture in pilot plant cheeses implies lower total solids than the PVC cheese. The lipase activity results showed that the hollow fibre and spiral wound pilot plant cheeses had higher lipase activity than the Puhoi valley cheese. The differences in lipase activity of the pilot plant cheeses and Puhoi Valley cheese were considered to be due to the incomplete inactivation of lipase present in milk during pasteurisation. The results from texture and sensory evaluation support the above mentioned differences. The microbiology results for all pilot plant cheeses were within the trigger limits set by Puhoi valley cheeses. The results from monitoring the operating parameters of both the pilot plant data show that the permeate flux decreases while the total solids in milk increase with time, which was also observed from the Puhoi Valley Cheese data. However, the rate of decrease of the permeate flux and the increase of the total solids in milk are dependent on the membrane area, feed volume, transmembrane pressure, pressure drop across the membrane and the flow characteristics. The rate of decrease in permeate flux and the rate of increase in the total solids of the hollow fibre runs and spiral wound runs are slightly different. The difference is due to the availability of larger membrane surface area and processing of larger feed volume of milk in the spiral wound runs. The transmembrane pressure and the pressure drop across the membrane were maintained as close as possible to Puhoi Valley Cheese. In conclusion, spiral wound membranes can be used to achieve the desired total solids concentration and successfully make the same feta cheese as the hollow fibre pilot plant. In order to make the same quality of feta cheese as Puhoi Valley Cheese using the spiral wound membrane pilot plant, the same composition of milk used for concentration at Puhoi Valley Cheese needs to be used on the spiral wound pilot plant unit. It is recommended that Puhoi Valley Cheeses should be replaced with spiral wound membranes if they are more economical in terms of cost than the hollow fibre membranes.

Cheese is one of the most well known food products in the world dating back to the 8th century B.C. There are more than 2000 varieties of cheese that are manufactured all over the world. Feta cheese is a soft white cheese with a salty and slightly acidic taste, which has originated from Greece. Most of the feta cheese manufactured in Greece is consumed locally, the migration of greeks to other parts of the world led to a demand for feta cheese outside of Greece. The spreading of the popularity of feta cheese to other ethnic groups in different parts of the world resulted in the high demand for feta cheese worldwide. The modern and most efficient method of feta cheese production involves a membrane filtration method, known as ultrafiltration. The ultrafiltration process utilises pressure as a driving force to concentrate milk by removal of water and small dissolved molecules. Hollow fibre and spiral wound ultrafiltration membranes are the two types of membranes that are commonly used for cheese production. An extensive amount of research exists on the implementation of ultrafiltration to improve the efficiency of the cheese making process and the performance of the membranes. However, limited research has been conducted on the comparison of the hollow fibre and spiral wound membrane performance in the cheese making process. The objective of the research was to determine if the hollow fibre membranes used at Puhoi Valley Cheese can be replaced with spiral wound membranes without compromising the quality of cheese produced. In order to achieve the objective, feta cheese was produced using hollow fibre and spiral wound ultrafiltration pilot plants. The operating performances of the hollow fibre and spiral wound membrane units were compared. To ensure that the quality of cheese is maintained, the cheese manufactured on the pilot plant units was analysed in terms of composition, microbiology, texture and sensory properties. The cheese made using the hollow fibre membrane pilot plant was compared with the reference sample from Puhoi Valley Cheese as they use hollow fibre membranes to produce feta cheese. The cheese made from the spiral wound membrane unit was also compared to that made by the hollow fibre membrane pilot plant unit. The operating parameters such as the inlet and outlet pressure, pressure difference along the membrane, transmembrane pressure, flow rate, recycle rate (bleed off rate), temperature and the run time were recorded. The operating parameters of the hollow fibre and spiral wound runs were compared with the data from Puhoi Valley Cheese. The quality of cheese made on the hollow fibre and spiral wound pilot plant units were evaluated in terms of composition, texture, microbiology and sensory properties. The composition was defined by the fat, protein, total solids and salt contents. The fat content was determined by utilising the modified Schmid-Bondzynski-Ratzlaff method, protein by the Kjeldahl method, total solids by using the air drying oven and salt percentage by the volhard method. The texture of the cheese was determined by the fracturability and hardness from the compression curve generated using the single bite compression test. The microbiological testing was performed according to New Zealand testing methods for E.Coli, Staphylococcus aureus, coliforms and yeast and mould. The difference from the control method was utilised for sensory evaluation. The acid degree value method was used to determine the lipase activity in feta cheese. It was found from the composition, texture and sensory analysis that the cheese from the hollow fibre pilot plant was different from the cheese manufactured at Puhoi Valley Cheeses (PVC). The spiral wound cheeses were also found to be different to PVC cheese, however the spiral wound cheeses and the pilot plant hollow fibre cheese were the same. The differences between both the pilot plant cheeses and PVC cheese were in terms of the fat, salt, moisture contents and the lipase activity in the cheeses. The fat content in the hollow fibre and spiral wound pilot plant cheeses are lower in comparison to the PVC cheese. This difference in fat content is considered to be due to the difference in the fat to protein ratio of the milk concentrated on the pilot plant and the PVC ultrafiltration system. The lower fat content resulted in firmer cheese than PVC due to more cross linking between the protein strands in cheese. The salt content in the cheeses made using the hollow fibre and spiral wound pilot plants was lower than Puhoi Valley Cheese. This is considered to be due to the low ratio of brine volume to cheese volume used for salting the cheese. The salt content of brine decreases during brining; hence a low ratio of brine volume to cheese volume causes a significant decrease in brine concentration. The decrease in brine concentration decreases the salt intake of the cheese. As salt diffuses in the moisture diffuses out, lower salt content results in higher moisture content in the cheese. As mentioned, the moisture content of the hollow fibre pilot plant cheese was higher than the PVC cheese. The moisture content is inversely proportional to the total solids, hence higher moisture in pilot plant cheeses implies lower total solids than the PVC cheese. The lipase activity results showed that the hollow fibre and spiral wound pilot plant cheeses had higher lipase activity than the Puhoi valley cheese. The differences in lipase activity of the pilot plant cheeses and Puhoi Valley cheese were considered to be due to the incomplete inactivation of lipase present in milk during pasteurisation. The results from texture and sensory evaluation support the above mentioned differences. The microbiology results for all pilot plant cheeses were within the trigger limits set by Puhoi valley cheeses. The results from monitoring the operating parameters of both the pilot plant data show that the permeate flux decreases while the total solids in milk increase with time, which was also observed from the Puhoi Valley Cheese data. However, the rate of decrease of the permeate flux and the increase of the total solids in milk are dependent on the membrane area, feed volume, transmembrane pressure, pressure drop across the membrane and the flow characteristics. The rate of decrease in permeate flux and the rate of increase in the total solids of the hollow fibre runs and spiral wound runs are slightly different. The difference is due to the availability of larger membrane surface area and processing of larger feed volume of milk in the spiral wound runs. The transmembrane pressure and the pressure drop across the membrane were maintained as close as possible to Puhoi Valley Cheese. In conclusion, spiral wound membranes can be used to achieve the desired total solids concentration and successfully make the same feta cheese as the hollow fibre pilot plant. In order to make the same quality of feta cheese as Puhoi Valley Cheese using the spiral wound membrane pilot plant, the same composition of milk used for concentration at Puhoi Valley Cheese needs to be used on the spiral wound pilot plant unit. It is recommended that Puhoi Valley Cheeses should be replaced with spiral wound membranes if they are more economical in terms of cost than the hollow fibre membranes.

Zugl.: Berlin, Techn. Univ., Diss., 2008

A deglutição é um processo coordenado e extremamente complexo, envolvendo contração e inibição de músculos localizados entre a boca e o estômago. Alterações neste sistema podem gerar disfagia, sinal comum de diversas doenças orgânicas, alterações neurológicas ou doenças neuromusculares, produzindo no paciente dificuldade na mastigação e deglutição de alimentos. Prover deglutição segura para indivíduos disfágicos é um desafio, contudo, esta pode ser facilitada se os alimentos tiverem a textura modificada e os líquidos forem espessados. Dessa maneira, este trabalho teve por objetivo desenvolver uma formulação de espessante alimentar, com valor nutricional agregado, para espessar diferentes líquidos, tornando-os com consistência adequada para pacientes disfásicos e avaliar sua ação em diferentes líquidos, testando a influência do tempo de espessamento e temperatura, sobre a estabilidade das viscosidades obtidas com as amostras. O ingrediente base usado para formulação e fonte de proteína foi o concentrado protéico de soro de leite (WPC), obtido experimentalmente, através da utilização das tecnologias de membranas, ultrafiltração (UF) e diafiltração (DF), através de três experimentos distintos. Inicialmente, 30 litros de soro em pó reconstituído, foram concentrados através da ultrafiltração, com redução do volume para cinco litros, a partir deste volume realizaram-se as diafiltrações. No primeiro experimento executaram-se quatro DF, duas de cinco litros e duas de 2,5 litros, obtendo-se WPC-1 com 56% de proteína. No segundo experimento também com quatro DF, executaram-se dois deles com 10 litros e dois com cinco litros, obtendo-se o WPC-2, com 71% de proteína. Para o terceiro experimento, os ciclos das diafiltrações foram aumentados para seis DF de cinco litros cada, obtendo-se o WPC-3, com 80% de proteína. Os concentrados obtidos foram liofilizados e caracterizados em relação a suas propriedades funcionais, sendo a solubilidade a mais importante por estar diretamente ligada à utilização em formulações alimentares de bebidas. Obteve-se solubilidade média de 70, 77 e 85% para WPC-1, 2 e 3 respectivamente. Pelas características obtidas de concentração de proteínas e percentual de solubilidade, o concentrado protéico obtido no terceiro experimento foi o selecionado para ser utilizado na formulação. Esta ficou constituída de 68% de concentrado protéico de soro de leite, 2% de mix de vitaminas e minerais e 30% do agente espessante (goma guar). Através de testes preliminares realizados com o agente espessante, determinou-se a porção do produto formulado necessária de ser adicionada aos líquidos para que os mesmos atingissem os níveis de consistência desejados, ou seja, 4,2 g para consistência de néctar (50 – 351 cP), 6,7 g para consistência de mel (351-1750 cP) e 9,2 g para consistência de pudim (> 1750 cP), tradicionalmente recomendadas para indivíduos disfásicos, segundo o National Dysphagia Diet Guidelines (NDD). Diferentes amostras (leite, sucos de abacaxi, de uva e de laranja) foram espessadas e realizadas medidas da viscosidade aparente, expressas em centipoise (cP), nos tempos pós-preparo: 10 minutos, 2 horas com taxa de cisalhamento (“shear rate”) de 50s-1 a 25ºC. As amostras foram armazenadas sob refrigeração e após 24 horas, novas medidas foram realizadas com taxa de cisalhamento de 50s-1 a 10ºC. Houve diferença estatística significativa entre as médias de viscosidade nos tempos de preparo de todos os níveis de consistência, demonstrando que o agente espessante utilizado continuou agindo, hidratando-se e aumentando a viscosidade com o passar do tempo. Também foi encontrada diferença significativa entre algumas amostras, com diferentes líquidos de diluição, quando comparadas entre si no mesmo nível de consistência. No entanto, as amostras apresentaram viscosidade dentro dos níveis sugeridos pela National Disfagic Diet, com exceção da consistência de pudim que, no tempo 10 minutos, permaneceu abaixo dos limites, adequando-se com o tempo, para o consumo de indivíduos disfágicos. / The swallowing is a coordinated and extremely complex process, involving contractions and inhibitions of muscles located between the mouth and the stomach. Alterations on this system can generate dysphagia, common sign of several organic diseases, neurological alterations or neuromuscular diseases, producing in the patient difficulty in the mastication and Swallowing of food .To provide safe swallowing for dysphagic individuals is a challenge, however, this can be facilitated, if the food has modified texture and if the liquids are thickened. In this way, the purpose of this work was to develop a formulation of food thickener, with aggregated nutritional value, to thicken different liquid foods, giving them an appropriate consistence for dysphagic individuals and to evaluate its action in different liquid foods, testing the influence of the time of thickening and temperature, over the stability of the viscosities obtained by the samples. As ingredient base for formulation and protein source was used whey protein concentrate (WPC), obtained experimentally, through the use of the technologies of membranes, the ultrafiltration (UF) and diafiltration (DF), through three different experiments. Initially, 30 liters of reconstituted powder serum, were concentrate through ultrafiltration, with reduction of the volume for cinco liters, starting from this volume the diafiltration took place. In the first experiment quatro DF were executed, two of cinco liters and two of 2.5 liters, obtained WPC-1 with 56% of protein. In the second experiment quatro DF were executed, two of them with 10 liters and two with cinco liters, obtaining WPC-2 with 71% of protein. For the third experiment, the cycles of the diafiltration were increased for 6 DF of 5 liters each, obtaining WPC-3 with 80% of protein.The obtained concentrates were liofilized and characterized in relation to its functional properties, being the solubility the most important for being directly linked to the use in alimentary formulations and drink. Average solubility of 70, 77 and 85% were obtained for WPC, 1, 2 and 3 respectively. Due to the obtained characteristics of protein concentrate and its solubility, the WPC obtained in the third experiment was selected for if used in the formulation. This was constituted of 68% of whey protein concentrate, 2% of mix of Vitamins and Minerals and 30% of the thickning agent (gum guar). Through preliminary tests accomplished with the thickening agent the amount of formulated product necessary to reach the desired consistence levels was determined, being 4.2 g for nectar consistence (50 – 351 cP), 6.7 g for honey consistence (351-1750 cP) and 9.2 g for pudding (> 1750 cP), traditionally recommended for dysphagic individuals according to National Dysphagia Diet Guidelines (NDD). Different samples (milk, pineapple juices, and grape and orange) were thickened and measurements of apparent viscosity were carried out, expressed in centipoise (cP), in the times after preparation: 10 minutes, 2 hour with shear rate of 50s-1 to 25±2ºC. The samples were stored under refrigeration and after 24 hours, new measurements were accomplished with shear rate of 50 s-1 to 10±2ºC. There were significant statistic difference among the average viscosity in the times of preparation of all the consistence levels, demonstrating that the thickening agent used continued acting, increasing the viscosity in the course of time. As well as significant differences among some samples when compared to each other in the same consistence level, caused by the different constituents of the drinks taken as sample. However, all samples presented viscosity inside the levels suggested by National Dysphagic Diet, except for the pudding consistence that, in the time of 10 minutes, was below these limits, fitting with time, being for this inside the levels suggested, appropriated for consumption by dysphagic individuals.

Milhões de toneladas de resíduos são gerados pela agroindústria todos os anos, gerando problemas ambientais e econômicos; muitos destes resíduos, entretanto, são ricos em nutrientes e compostos funcionais, o que poderia tornálos subprodutos com valor agregado. A utilização de subprodutos possibilita enriquecimento dos alimentos processados, redução de descarte no meio ambiente e melhor aproveitamento dos recursos naturais. Os talos de beterraba são um exemplo de resíduo com potencial a ser explorado e, assim como os bulbos, são ricos em betalaína, composto responsável pela sua coloração vermelho-violeta. Esses pigmentos são aplicáveis na indústria alimentícia como corantes naturais, mas a sua estabilidade é um dos maiores desafios frente a sua utilização. Neste contexto, este trabalho tem como objetivo principal estudar a extração, a clarificação e a estabilização de betalaínas provenientes de talos de beterraba vermelha. Para tanto, foi inicialmente estudado a viabilidade da utilização dos talos de beterraba como fonte de betalaínas e, em sequência, a extração destes compostos via esmagamento. A utilização de diferentes processamentos com vistas à melhora da extração e o seu impacto na estabilidade das betalaínas presente no extrato foram investigados: o processamento por alta pressão (HPP do inglês High Pressure Processing) e o tratamento térmico com alta temperatura por curto tempo (HTST do inglês High Temperature Short Time). Por fim, foi realizado um estudo de clarificação do extrato por meio de processo de separação por membranas (PSM). A clarificação foi aplicada por microfiltração (MF) seguido por ultrafiltração (UF). Além disto, a MF também foi operada no modo diafiltração (DF). A redução da atividade da enzima peroxidase, as características físico-químicas, a cor e a taxa de degradação das betalaínas ao longo do armazenamento foram avaliados nos extratos antes e após a clarificação. Os resultados mostraram que os extratos obtidos via esmagamento possuíam em média 45% da quantidade de pigmentos presentes no extrato dos bulbos de beterraba. O processamento por HPP não foi considerado uma boa alternativa para melhorar a extração e também não contribuiu para o aumento na estabilidade dos pigmentos presentes nos extratos. O tratamento térmico HTST causou uma pequena degradação das betalaínas; porém, quando aplicado por 120 s a 85,7 °C nos extratos com alta concentração inicial, promoveu uma redução na taxa de degradação das betalaínas da amostra tratada de 1,2 para 0,4 mg de betanina/100 ml de extrato por dia após uma semana de armazenamento. No estudo de clarificação por PSM, os experimentos de MF e UF apresentaram reduções no fluxo do permeado ao longo do processo, além de elevada tendência ao fouling, em ambas as membranas. A aplicação da MF batelada seguida por UF ocasionou uma redução de 99,5% da atividade de peroxidase. Quando a MF foi operada em modo DF, a alimentação do processo foi modificada de duas bateladas de 2 L cada para uma batelada de 3 L, aproveitando melhor os pigmentos da alimentação. Além disto, a clarificação contribuiu para a estabilidade das betalaínas, uma vez que a taxa de degradação reduziu de 0,32 e 0,41 mg de betanina/100 ml de extrato por dia para 0,2 mg de betanina/100 ml de extrato por dia no 10° dia de armazenamento. Os processos de MF, em modo batelada e DF, seguidos por UF promoveram redução na turbidez e no teor de sólidos solúveis, além de extratos clarificados com cor mais intensa, luminosa e avermelhada. / Every year a huge amount of waste is generated by agricultural industry, becoming a serious environmental and economic problem; many of these wastes, however, contain nutrients and functional compounds, which could make them by-products with added value. The use of by-products allows the enrichment of processed foods and the reduction of discard in the environment, ensuring a better use of natural resources. The beet stalks are an example of waste with potential to be explored and, as the root, are rich in betalains, whose provide their red-violet color. These pigments are applicable in the food industry as natural dyes. The stability of these pigments, however, is one of the biggest challenges of using it. In this context, this work aims to study the extraction and clarification of betalains from red beet stalks. Initially it was studied the feasibility of using beet stalks like a betalain source and, after that, the extraction of these compounds through crushing was carried out. The utilization of different processing to improve the extraction and the impact on the betalain stability present in the extract were studied: the high pressure processing (HPP) and the thermal treatment high temperature short time (HTST). Lastly, a study of clarification of the extract using the membrane separation processes (PSM) was carried out. Clarification was first applied by microfiltration (MF) followed by ultrafiltration (UF). In addition, MF was also operated in diafiltration mode (DF). The peroxidase activity reduction, physico-chemical characteristics, color and degradation rate of betalains over the storage were evaluated in the extracts before and after the clarification. The results show that the stalks extract by crushing had about 45% of the pigments content in the extract from root of beet. HPP was not considered a suitable alternative to improve the extraction and did not contribute to the increase in the stability of the pigments present in the extracts. The thermal treatment HTST caused a small degradation of betalains. However, when it was applied for 120 s at 85.7 °C, in the extracts with high initial concentration, it promoted a reduction in the betalain degradation rate of the treated sample from 1.2 to 0.4 mg of betanin/100 ml of extract per day after one week of storage. In the PSM clarification study, the MF and UF experiments, showed permeated flux reduction, indicating high fouling tendency on both MF and UF membranes. The application of MF followed by UF caused a reduction of 99.5% in peroxidase activity. When MF was operated in DF mode, the process feed was modified from two batches of 2 L each to a 3 L batch, making better use of feed pigments. In addition, the clarification experiments contributed to the betalain stability, as the betalain degradation rate reduced from 0.32 and 41 mg of betanin/100 ml of extract per day to 0.2 mg of betanin/100 ml of extract per day on the 10th day of storage. The MF processes, batch and DF mode, followed by UF promoted a reduction in the turbidity and soluble solids content and the clarified extracts had color more intense, bright and reddish.

Made available in DSpace on 2014-06-11T19:35:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2003-08Bitstream added on 2014-06-13T21:07:15Z : No. of bitstreams: 1 romao_lpc_dr_araiq.pdf: 435919 bytes, checksum: 41d1a98abe8648f0ce971267eb617cda (MD5) / Um procedimento de ultrafiltração com fluxo tangencial e membrana polietersulfônica de porosidade 1 kDa foi utilizado na determinação da capacidade de complexação (CC) e constantes de estabilidade condicional (K) do íon cobre(II) para amostras de matéria orgânica natural, substâncias húmicas aquáticas e substâncias húmicas de solo de diferentes regiões do Brasil. As substâncias húmicas de solos apresentaram maiores valores de CC em relação à matéria orgânica natural e substâncias húmicas aquáticas. Os valores das capacidades de complexação das amostras de matéria orgânica aquática (1,25±0,07, 1,18±0,05 e 1,81±0,30) e das amostras de substâncias húmicas aquáticas (1,23±0,11, 1,12±0,05 e 1,44±0,04) foram similares, indicando que o procedimento de extração utilizando resina acrílica XAD-8 não modificou significativamente as características complexantes originais das amostras. Os valores das capacidades de complexação da matéria orgânica natural (MON) determinados por ultrafiltração para Cu(II) variaram de 0,17 a 1,99 mmol Cu(II)g-1 COT e os valores das constantes de estabilidade condicional variaram de 2,30 a 4,35 (log K1) e 1,36 a 2,60 (log K2) A validação da metodologia de ultrafiltração em fluxo tangencial foi feita comparando as médias das capacidades de complexação para cobre(II) determinadas por UF, e àquelas obtidas por eletrodo íon seletivo utilizando o teste de student t. Os resultados mostraram que não existem diferenças entre os valores das capacidades de complexação determinados por ambos os procedimentos. Resultados de experimentos de troca entre íons Cu(II) adicionados e espécies metálicas (Ni, Al e Fe) originalmente complexadas por substâncias húmicas, permitiram estabelecer a seguinte ordem decrescente de estabilidade relativa do complexo SH-metal: Fe<Al<<Ni. / An tangential-flow ultrafiltration (TF-UF) procedure a polyethersulfone membrane filter (cut-off 1 kDa) was used to determine the copper(II) complexation capacity (CC) and conditional stability constants (K) of the copper(II) ion from samples of aquatic organic matter, aquatic humic substances and soil humic substances (all from different regions of Brazil). The humic soil substances presented larger values of CC compared to the aquatic organic matter and aquatic humic substances. The complexation capacities from samples of aquatic organic matter (1,25±0,07, 1,18±0,05 and 1,81±0,30) and aquatic humic substances (1,23±0,11, 1,12±0,05 and 1,44±0,04) samples were similar, indicating that the extraction procedure using XAD-8 acrylic resin didn't, significantly, modify the original complexation characteristics of the samples. The values of complexation capacities of aquatic organic matter (AOM) by Cu(II) using ultrafiltration varied from 0,17 to 1,99 mmol Cu(II)g-1 TOC and the values of the conditional stability constants varied from 2,30 to 4,35 (log K1) and 1,36 to 2,60 (log K2). The validation of the tangential-flow ultrafiltration (TF-UF) methodology was done comparing the averages of complexation capacities determined by TF-UF and those obtained by selective ion electrode using the student test t. The results showed that there aren’t any differences between the values of complexation capacities for copper(II) determined by both the methods. Results of exchange experiments among Cu(II) ions added and metallic species (Ni, Al and Fe) originally complexed for humic substances allowed the establishment of the following decreasing order of relative stability of the compound HS-metal: Fe <Al << Ni.

A deglutição é um processo coordenado e extremamente complexo, envolvendo contração e inibição de músculos localizados entre a boca e o estômago. Alterações neste sistema podem gerar disfagia, sinal comum de diversas doenças orgânicas, alterações neurológicas ou doenças neuromusculares, produzindo no paciente dificuldade na mastigação e deglutição de alimentos. Prover deglutição segura para indivíduos disfágicos é um desafio, contudo, esta pode ser facilitada se os alimentos tiverem a textura modificada e os líquidos forem espessados. Dessa maneira, este trabalho teve por objetivo desenvolver uma formulação de espessante alimentar, com valor nutricional agregado, para espessar diferentes líquidos, tornando-os com consistência adequada para pacientes disfásicos e avaliar sua ação em diferentes líquidos, testando a influência do tempo de espessamento e temperatura, sobre a estabilidade das viscosidades obtidas com as amostras. O ingrediente base usado para formulação e fonte de proteína foi o concentrado protéico de soro de leite (WPC), obtido experimentalmente, através da utilização das tecnologias de membranas, ultrafiltração (UF) e diafiltração (DF), através de três experimentos distintos. Inicialmente, 30 litros de soro em pó reconstituído, foram concentrados através da ultrafiltração, com redução do volume para cinco litros, a partir deste volume realizaram-se as diafiltrações. No primeiro experimento executaram-se quatro DF, duas de cinco litros e duas de 2,5 litros, obtendo-se WPC-1 com 56% de proteína. No segundo experimento também com quatro DF, executaram-se dois deles com 10 litros e dois com cinco litros, obtendo-se o WPC-2, com 71% de proteína. Para o terceiro experimento, os ciclos das diafiltrações foram aumentados para seis DF de cinco litros cada, obtendo-se o WPC-3, com 80% de proteína. Os concentrados obtidos foram liofilizados e caracterizados em relação a suas propriedades funcionais, sendo a solubilidade a mais importante por estar diretamente ligada à utilização em formulações alimentares de bebidas. Obteve-se solubilidade média de 70, 77 e 85% para WPC-1, 2 e 3 respectivamente. Pelas características obtidas de concentração de proteínas e percentual de solubilidade, o concentrado protéico obtido no terceiro experimento foi o selecionado para ser utilizado na formulação. Esta ficou constituída de 68% de concentrado protéico de soro de leite, 2% de mix de vitaminas e minerais e 30% do agente espessante (goma guar). Através de testes preliminares realizados com o agente espessante, determinou-se a porção do produto formulado necessária de ser adicionada aos líquidos para que os mesmos atingissem os níveis de consistência desejados, ou seja, 4,2 g para consistência de néctar (50 – 351 cP), 6,7 g para consistência de mel (351-1750 cP) e 9,2 g para consistência de pudim (> 1750 cP), tradicionalmente recomendadas para indivíduos disfásicos, segundo o National Dysphagia Diet Guidelines (NDD). Diferentes amostras (leite, sucos de abacaxi, de uva e de laranja) foram espessadas e realizadas medidas da viscosidade aparente, expressas em centipoise (cP), nos tempos pós-preparo: 10 minutos, 2 horas com taxa de cisalhamento (“shear rate”) de 50s-1 a 25ºC. As amostras foram armazenadas sob refrigeração e após 24 horas, novas medidas foram realizadas com taxa de cisalhamento de 50s-1 a 10ºC. Houve diferença estatística significativa entre as médias de viscosidade nos tempos de preparo de todos os níveis de consistência, demonstrando que o agente espessante utilizado continuou agindo, hidratando-se e aumentando a viscosidade com o passar do tempo. Também foi encontrada diferença significativa entre algumas amostras, com diferentes líquidos de diluição, quando comparadas entre si no mesmo nível de consistência. No entanto, as amostras apresentaram viscosidade dentro dos níveis sugeridos pela National Disfagic Diet, com exceção da consistência de pudim que, no tempo 10 minutos, permaneceu abaixo dos limites, adequando-se com o tempo, para o consumo de indivíduos disfágicos. / The swallowing is a coordinated and extremely complex process, involving contractions and inhibitions of muscles located between the mouth and the stomach. Alterations on this system can generate dysphagia, common sign of several organic diseases, neurological alterations or neuromuscular diseases, producing in the patient difficulty in the mastication and Swallowing of food .To provide safe swallowing for dysphagic individuals is a challenge, however, this can be facilitated, if the food has modified texture and if the liquids are thickened. In this way, the purpose of this work was to develop a formulation of food thickener, with aggregated nutritional value, to thicken different liquid foods, giving them an appropriate consistence for dysphagic individuals and to evaluate its action in different liquid foods, testing the influence of the time of thickening and temperature, over the stability of the viscosities obtained by the samples. As ingredient base for formulation and protein source was used whey protein concentrate (WPC), obtained experimentally, through the use of the technologies of membranes, the ultrafiltration (UF) and diafiltration (DF), through three different experiments. Initially, 30 liters of reconstituted powder serum, were concentrate through ultrafiltration, with reduction of the volume for cinco liters, starting from this volume the diafiltration took place. In the first experiment quatro DF were executed, two of cinco liters and two of 2.5 liters, obtained WPC-1 with 56% of protein. In the second experiment quatro DF were executed, two of them with 10 liters and two with cinco liters, obtaining WPC-2 with 71% of protein. For the third experiment, the cycles of the diafiltration were increased for 6 DF of 5 liters each, obtaining WPC-3 with 80% of protein.The obtained concentrates were liofilized and characterized in relation to its functional properties, being the solubility the most important for being directly linked to the use in alimentary formulations and drink. Average solubility of 70, 77 and 85% were obtained for WPC, 1, 2 and 3 respectively. Due to the obtained characteristics of protein concentrate and its solubility, the WPC obtained in the third experiment was selected for if used in the formulation. This was constituted of 68% of whey protein concentrate, 2% of mix of Vitamins and Minerals and 30% of the thickning agent (gum guar). Through preliminary tests accomplished with the thickening agent the amount of formulated product necessary to reach the desired consistence levels was determined, being 4.2 g for nectar consistence (50 – 351 cP), 6.7 g for honey consistence (351-1750 cP) and 9.2 g for pudding (> 1750 cP), traditionally recommended for dysphagic individuals according to National Dysphagia Diet Guidelines (NDD). Different samples (milk, pineapple juices, and grape and orange) were thickened and measurements of apparent viscosity were carried out, expressed in centipoise (cP), in the times after preparation: 10 minutes, 2 hour with shear rate of 50s-1 to 25±2ºC. The samples were stored under refrigeration and after 24 hours, new measurements were accomplished with shear rate of 50 s-1 to 10±2ºC. There were significant statistic difference among the average viscosity in the times of preparation of all the consistence levels, demonstrating that the thickening agent used continued acting, increasing the viscosity in the course of time. As well as significant differences among some samples when compared to each other in the same consistence level, caused by the different constituents of the drinks taken as sample. However, all samples presented viscosity inside the levels suggested by National Dysphagic Diet, except for the pudding consistence that, in the time of 10 minutes, was below these limits, fitting with time, being for this inside the levels suggested, appropriated for consumption by dysphagic individuals.

Linfomas são cânceres que se iniciam a partir da transformação maligna de um linfócito no sistema linfático. Os linfomas são divididos em duas categorias principais: os linfomas de Hodgkin e todos os outros linfomas, denominados linfomas não-Hodgkin (LNH). Os pacientes com LNH são comumente tratados com radioterapia apenas ou combinada com quimioterapia utilizando-se de anticorpo monoclonal anti-CD20, principalmente o rituximab (MabThera&reg). O uso de anticorpos monoclonais (Acm) conjugados à quelantes bifuncionais radiomarcados com radionuclídeos metálicos ou lantanídeos é uma realidade de tratamento para portadores de LNH pelo princípio de radioimunoterapia (RIT). Este estudo concentrou-se nas condições de conjugação do anticorpo monoclonal rituximab (MabThera&reg) com grupamentos quelantes bifuncionais DOTA e DTPA. Na marcação dos Acm conjugados com lutécio-177, foram estudadas as condições de pré-purificação do Acm, condições de conjugação, determinação de número de quelantes acoplados à molécula do anticorpo, purificação do anticorpo conjugado, radiomarcação do anticorpo conjugado, com lutécio-177, purificação do anticorpo marcado, a ligação específica in vitro dos compostos marcados às células Raji, e distribuição biológica em camundongos BALB/c sadios. As três metodologias empregadas na pré-purificação do anticorpo (diálise, cromatografia de exclusão molecular com coluna Sephadex G-50 e ultrafiltração) demonstram-se eficientes e proporcionaram recuperação da amostra superior a 90%. A metodologia de ultrafiltração foi considerada a mais simples e prática, podendo ser aplicada a procedimentos rotineiros de produção de radiofármacos. Além disso, proporcionou a recuperação final de amostra de 97% em microlitros. Nas conjugações do anticorpo com os quelantes DOTA e DTPA em razões molares diferentes do Acm:quelante, observou-se número de grupamentos quelantes acoplados à molécula do Acm proporcional à razão molar estudada. Quando foi avaliada a influência de condições diferentes de conjugação no número de quelantes acoplados à molécula do Acm, não foram observadas diferenças significativas, com resultados de pureza radioquímica (PR) inferior a 80% em todas as condições estudadas. Na comparação de métodos de purificação do Acm conjugado, a abordagem inédita apresentada neste estudo, na qual a cromatografia de exclusão molecular foi combinada com a ultrafiltração resultou em maior eficiência na purificação e preservação da estrutura do anticorpo. Nos estudos de radiomarcação do anticorpo conjugado com DOTA e DTPA, os imunoconjugados de DTPA apresentaram, de forma geral, maior eficiência de marcação com resultados reprodutíveis quando comparados com os imunoconjugados de DOTA, considerando-se as diferentes razões molares utilizadas. As metodologias cromatográficas empregadas no controle de pureza radioquímica do composto radiomarcado proporcionaram a discriminação das diferentes espécies radioquímicas no meio de marcação. A metodologia de purificação do composto conjugado e radiomarcado utilizada proporcionou a obtenção de compostos com alta pureza radioquímica, 97,4&plusmn;1,3% (DOTA 1:50) e 98,7&plusmn;0,2% (DTPA 1:50). Nos estudos de ligação específica às células tumorais Raji, o anticorpo conjugado com quelante DTPA nas razões molares de 1:50 e 1:20 apresentaram perfil semelhante de ligação, com aumento da porcentagem de ligação específica proporcional à concentração celular, enquanto que o imunoconjugado na razão molar de 1:10 apresentou alta porcentagem de ligação não específica. Os resultados obtidos nos estudos de biodistribuição in vivo do anticorpo conjugado e radiomarcado nem sempre se mostraram compatíveis com a biodistribuição de anticorpos radiomarcados íntegros. No caso do quelante DOTA, o imunoconjugado obtido a partir da razão molar 1:20, apresentou melhores características de biodistribuição. No caso do quelante DTPA, a razão molar utilizada pareceu refletir diretamente no clareamento sanguíneo do anticorpo e todas as razões molares utilizadas apresentaram instabilidade in vivo. / Lymphomas are malignancies or cancers that start from the malign transformation of a lymphocyte in the lymphatic system. Lymphomas are divided in two major categories: Hodgkin lymphoma and non-Hodgkin lymphoma (NHL). Patient with NHL are generally treated with radiotherapy alone or combined with immunotherapy using monoclonal antibody rituximab (MabThera&reg). Currently, monoclonal antibodies (Mab) conjugated with bifunctional chelate agents and radiolabeled with metallic or lanthanides radionuclides are a treatment reality for patients with NHL by the principle of radioimmunotherapy (RIT). This study focused on the conditions of conjugation of Acm rituximab (MabThera&reg) with bifunctional chelating agents DOTA and DTPA and labeling with 177-luthetium. Various parameters were studied: method of Acm purification, conditions of Acm conjugation and the determination of the number of chelate coupled to the Acm, the purification of the conjugated Mab, labeling conditions with lutetium-177, purification of the radiolabeled immunoconjugate, radiochemical purity (RP), in vitro specific binding determination to Raji cells (Human Burkitt) and biological distribution performed in normal BALB/c mouse. The three methodologies employed in pre-purification of Acm (dialysis, size exclusion chromatograph and ultrafiltration) demonstrated to be efficient; they provided sample recovery exceeding 90%. However, the methodology of ultrafiltration resulted in greater sample recovery and in microliters. The number of chelate attached to the Mab molecule was proportional to the molar ratio studied. When the influence of different conditions of conjugation in the number of chelate bounded to the Mab was studied, no notable differences were observed. The RP < 80% was observed in all the methods applied. Purification of the conjugated antibody by different methods showed that the innovative combination of Sephadex and ultrafiltration methods resulted in higher efficiency of purification. The optimized conditions for purification of the conjugated antibody preserved the protein integrity. Radiolabelling studies of DOTA and DTPA immunoconjugated showed that DTPA derivatives presented, in general, radiochemical yield superior than DOTA conjugated Mab, considering the different molar ratios studied. The chromatographic methods employed in the RP determination were efficient to separate the different radiochemical species presented in the reaction medium. The methodology used in the purification of the labeled Mab resulted in labeled compounds with high radiochemical purity, 97.4&plusmn;1.3% (DOTA 1:50) and 98.7&plusmn;0.2% (DTPA 1:50). Considering specific cell binding assays (Raji cells), the Mab conjugated to DTPA at 1:50 and 1:20 molar ratios presented similar results, and the percent of cell binding were proportional to the cell concentration, whereas the cell binding for 1:10 molar ratio showed high percent of nonspecific cell binding. The results of in vivo biodistribution studies of labeled Mab not always were compatible with the biodistribution of intact radiolabelled antibody. The DOTA immunoconjugated produced at 1:20 molar ratio, showed better performance in biodistribution studies. In the case of DTPA immunoconjugated, the blood clearance seems to be influenced by the molar ratio applied and the immunoconjugated produced with DTPA chelate at different molar ratio resulted in high in vivo instability compounds.