Embora a soja em grão seja pouco consumida pela população brasileira, seus derivados protéicos são utilizados como ingredientes em diversos alimentos e a tendência é um aumento significativo do seu uso com a aprovação pela FDA e pela ANVISA da alegação funcional referente ao consumo de proteína de soja. Em paralelo, um número crescente de pesquisas sobre as isoflavonas, fitoestrógenos presentes em quantidades significativas na soja, vem demonstrando diversos efeitos benéficos destes compostos, entre os quais a sua ação antioxidante, anticarcinogênica e hipocolesterolêmica. O objetivo foi estudar as interações entre isoflavonas e proteínas da soja, seu efeito na biodisponibilidade in vitro e in vivo e o status antioxidante das isoflavonas. Os resultados sugerem que a presença da proteína reduz a quantidade e leva a um retardo no tempo de absorção das isoflavonas em relação à administração na forma livre. O efeito sobre a capacidade antioxidante do plasma e sobre a atividade e expressão gênica das enzimas CAT, GPx e SOD divergiu para a suplementação de isoflavonas ou proteínas separadamente ou em associação. / Although soybeans are not commonly consumed by the Brazilian population, soybean products are used as ingredients by the food industry and there is an increased trend in their consumption due to the approval of FDA and ANVISA on the claim regarding the health benefits of soy proteins. Therefore, many studies involving isoflavones, the phytoestrogens found in significant quantities in soybeans, have demonstrated several health benefits of these compounds, such as antioxidant capacity, and anticarcinogenic and hypocholesterolemic effects. The objective of the present study was to evaluate the interaction between isoflavones and soy proteins as well as the effect of this interaction on the in vitro and in vivo bioavailability of isoflavones and on the antioxidant status. The results indicated that the association with protein decreased the rate and proportion of absorption of isoflavones when compared to the free form. The effect of isoflavones on the plasma antioxidant capacity and on the enzymatic activity and gene expression levels of CAT, GPx and SOD was different when isoflavones were combined with soy protein compared to isoflavones alone.

O mel é um alimento doce feito por abelhas a partir do néctar das flores. Estudos consideram o mel um adoçante natural, fonte de compostos antioxidantes, que quando incluídos na dieta tornam-se aliados contra danos oxidativos. No entanto, a composição do mel é muito variável, pois depende da fonte floral que o origina, das condições ambientais onde é produzido e do modo como é recolhido e processado. A região Sul distingue-se das demais regiões brasileiras em função do seu clima e vegetação. Além disso, o somatório dos principais estados da região Sul representa 49 % da produção de mel do país, destacando-se o estado do Rio Grande do Sul (RS) como o maior produtor. Uma recente pesquisa demonstrou que outro produto da colmeia, o pólen apícola, produzido na mesma região, apresentou maior capacidade antioxidante quando comparado ao pólen produzido em outras regiões do país, tornando oportuna a busca por méis com possíveis efeitos benéficos à saúde. Nesse contexto, o presente trabalho teve como objetivo caracterizar os méis de Apis mellifera, produzidos no estado do RS, quanto as propriedades físico-químicas e antioxidantes, e estabelecer correlações com a origem botânica. Para isso, foram obtidas 52 amostras, adquiridas na Casa do Mel, em Viamão, no RS, de diversas origens botânicas e diferentes locais do estado. As propriedades físico-químicas avaliadas foram: análise melissopalinológica, umidade, cinzas, condutividade elétrica, pH e acidez livre, cor, atividade diastásica, açúcares, hidroximetilfurfural (HMF) e reações qualitativas de Fiehe e Lund, de acordo com as análises preconizadas pelas normas brasileira e internacional. A avaliação das propriedades antioxidantes consistiu na identificação e quantificação dos compostos fenólicos por cromatografia líquida de alta eficiência; determinação da capacidade redutora do Folin-Ciocalteu; determinação de flavonoides totais; capacidade de absorção de radicais de oxigênio (ORAC); determinação do poder redutor do ferro (FRAP); e sequestro do radical estável DPPH. Os resultados para a análise físico-química caracterizaram os méis em: monoflorais (eucalipto, aroeira, quitoco, uva japão, flor do campo e laranjeira) e heterofloral; a umidade apresentou média de 18,3±0,7 % (máx. 20); o valor médio para cinzas foi de 0,3±0,2 % (máx. 0,6); a condutividade elétrica foi de 0,59±0,2 mS.cm-1 (máx. 0,8); o pH com média de 4,18±0,3 e a acidez livre com 32±9,8 mEq.Kg-1 (máx. 50); a cor variou do extra branco ao âmbar; a média da atividade diastásica foi de 16,4±11,0 °Gothe (mín. 8); as médias para frutose e glicose foram, respectivamente, 37,9±1,4 e 32±2,5 g.100-1, onde a razão frutose/glicose foi de 1,2±0,1; o teor médio de HMF foi de 5,6±5,8 mg.Kg-1 (máx. 60); e as reações de Fiehe e Lund apresentaram, respectivamente, resultados negativo e positivo para todas as amostras, indicando a pureza dos méis analisados. No que concerne à análise de antioxidantes, os ácidos gálico, cinâmico e o p-cumárico, e os flavonoides quercetina e miricetina, foram identificados. O teor de fenólicos e flavonoides totais foram de 61,3±18,3 mgEAG.Kg-1 e 0,7±0,7 mgQE.100g-1, respectivamente. A capacidade antioxidante pelo método ORAC foi de 7,8±4,3 mmolET.Kg-1; pelo ensaio FRAP foi de 1,2±0,6 &#181;molET.g-1; e pelo método DPPH (EC50) foi de 72,4±89,8 mg.ml-1. Os méis apresentaram-se de acordo com os parâmetros físico-químicos exigidos pela legislação brasileira e recomendações internacionais, para o controle de qualidade do mel. A capacidade antioxidante foi semelhante à encontrada na literatura científica. As espécies botânicas que apresentaram maior atividade antioxidante foram eucalipto e aroeira. O conteúdo de fenólicos totais não apresentou diferença estatística entre esses dois tipos de méis, no entanto o teor de flavonoides nos méis de aroeira foi maior (p<0,05) do que o referido nos méis de eucalipto. O ácido gálico foi o composto majoritário presente nos méis de eucalipto e aroeira, e o ácido p-cumárico nos méis de uva japão. Por fim, as propriedades físico-químicas e biologicamente ativas dos méis foram influenciadas pela origem floral. / Honey is a sweet food made by bees from the nectar of flowers. Previous studies consider honey as a natural sweetener, source of antioxidant compounds that when included in the diet become allies against oxidative damage. However, honey composition is variable because it depends on the floral source, environmental conditions where it is produced and how it is collected and processed. The southern region is different from other Brazilian regions because of its climate and vegetation. In addition, the sum of the main states of the South is 49% of the country\'s honey production, and Rio Grande do Sul (RS) state is the biggest producer. A recent study showed that another product of the hive, bee pollen, derived from this region showed higher concentrations of antioxidant compounds when compared to other regions of the country, encouraging researches for honeys with possible beneficial health effects. In this sense, this study aims to characterize Apis mellifera honey produced in the state of RS, as the physicochemical and antioxidants properties, and establish correlations with the botanical origin. For this, 52 samples were obtained, collected from Honey House, in Viamão, RS, of various botanical origins and different parts of the state. The physicochemical properties evaluated were: melissopalinological analysis, moisture, ash, electrical conductivity, pH and free acidity, colour, diastase activity, sugars, hydroxymethylfurfural (HMF), and Fiehe and Lund reactions, according to the analytical standards established by the Brazilian and international recomendations. The evaluation of antioxidant properties consisted in the identification and quantification of the phenolic compounds by high-performance liquid chromatography; determining the reducing capacity of the Folin-Ciocalteu; determination of total flavonoid; ability to absorb oxygen radical (ORAC); determining the reducing power of iron (FRAP); and DPPH radical scavenging method. The results for the physicochemical analysis characterized the honeys in: monofloral (eucalyptus, mastic, quitoco, Japan grapes, field flower and orange) and heterofloral; the average to moisture was 18.3±0.7 % (max. 20%); the average of ashes was 0.3±0.2 % (max. 0.6%); the electrical conductivity was 0.59±0.2 mS.cm-1 (max. 0.8); the result to pH was 4.18±0.3 and free acidity was 32±9.8 mEq.Kg-1 (max. 50); the colour varied to extra white to amber; the average of diastase activity was 16.4±11.0 °Gothe (min. 8); the average of glucose and fructose determination were, respectively, 37.9±1.4 and 32±2.5 g.100-1; the ratio fructose/glucose was 1.2±0.1; the average content of HMF was 5.6±5.8 mg.Kg-1 (max. 60); and the Fiehe and Lund reactions presented, respectively, negative and positive results to all samples, indicating the purity of honeys analyzed. Regarding the analysis of antioxidants, gallic acid, cinnamic acid and p-coumaric acid, and flavonoids quercetin and myricetin, were identified. Flavonoid and phenolic total content were 61.3±18.3 mgEAG.Kg-1 and 0.7±0.7 mgQE.100g-1, respectively. ORAC antioxidant capacity was 7.8±4.3 mmolET.Kg-1; FRAP was 1.2±0.6 &#181;molET.g-1; and DPPH assay (EC50) was 72.4±89.8 mg.ml-1. Analyzed honeys presented all physicochemical parameters in accordance to honey quality control from Brazilian and international recommendations. The antioxidant capacity was similar to data from scientific literature. The botanical species with the highest antioxidant activity were eucalyptus and mastic. Total phenolic content showed no statistical difference between these two types of honeys, however flavonoid content of mastic honeys was higher (p<0.05) than eucalyptus honeys. Gallic acid was the major compound present in eucalyptus and mastic honeys, and p-coumaric acid in Japan grape honeys. Finally, physicochemical and biologically active properties honeys were influenced by honey floral source.

A incidência de doenças crônicas não transmissíveis como a diabetes tipo 2 e complicações cardiovasculares tem aumentado significativamente, e tem-se associado principalmente às mudanças nos hábitos alimentares tradicionais. O objetivo do presente estudo foi caracterizar diferentes cultivares de feijão, lupino e grãos da região dos Andes quanto a seus compostos fenólicos antioxidantes, capacidade antioxidante e potencial anti-hiperglicêmico e anti-hipertensivo in vitro. Dependendo do tipo de cultivar, o feijão é uma fonte promissora de taninos condensados, antocianinas, e flavonóis; enquanto que o lupino andino destacou-se pela presença de isoflavonas. Após o tratamento térmico, o feijão e lupino andino inibiram significativamente a enzima conversora da angiotensina I, relevante na prevenção da hipertensão, enquanto o milho roxo andino inibiu a &#945;-glicosidase, relevante na prevenção da hiperglicemia. Uma combinação apropriada de grãos tradicionais como parte da dieta poderia contribuir na modulação dos níveis de glicose e na prevenção das complicações relacionadas ao desequilíbrio óxido-redução. / Incidence of chronic diseases such as diabetes type 2 and related cardiovascular complications has increased significantly due mainly to current changes in traditional food dietary habits. The objective of this study was to characterize several bean and lupin cultivars along with grains from the Andean region in relation to their phenolic compounds, antioxidant capacity and anti-diabetes and anti-hypertension potential using in vitro assays. Depending on the cultivar, beans are interesting sources of condensed tannins, anthocyanins and flavonols, whereas major phenolic compounds in Andean lupins were isoflavones. Following thermal treatment, selected beans and Andean lupins inhibited significantly the hypertension relevant angiotensin I-converting enzyme and among Andean grains, the purple corn inhibited the hyperglycemia relevant &#945;-glucosidase. A good combination of traditional grains as a part of the overall diet can contribute to effective dietary strategies for managing Type 2 diabetes and associated complications linked to unbalanced cellular redox status.

Os flavonóides são de grande interesse na nutrição e medicina devido ao seu potencial antioxidante e possível efeito protetor à saúde, reduzindo doenças crônico-degenerativas tais como doenças cardiovasculares e câncer. Entretanto, para produzir o efeito biológico in vivo, é essencial que suficiente quantidade do composto alcance a célula alvo, ou seja, que apresente uma boa biodisponibilidade. Este trabalho teve como objetivo inicial realizar um screening de alimentos vegetais para avaliar seu potencial antioxidante na dieta alimentar e permitir a selecão de um alimento mais interessante para o estudo do mecanismo de absorção e metabolização de flavonóides, a fim de estabelecer sua biodisponibilidade. As frutas, hortaliças e polpas congeladas de frutas foram analisadas quanto à atividade antioxidante pelo método de co-oxidação do ácido linoléico/&#946;-caroteno e pela inibição da peroxidação do lipossomo catalisada pelo íon cobre. Dentre as amostras analisadas, aquelas contendo antocianinas em sua composição apresentaram os maiores valores de atividade antioxidante. As frações de flavonóides neutros e ácidos de repolho roxo, alface roxa, jambolão, amora silvestre, casca de maçã gala e polpa congelada de açaí, amora e acerola, a 10 e 50&#181;M equivalente em ácido gálico, apresentraram os maiores valores de atividade antioxidante em ambos os sistemas. Dentro desse grupo, os principais flavonóides de amora silvestre (Morus sp.) foram identificados por espectrometria UV/Vis e MS/MS como cianidina-3- glucosídeo (cyn-3-glu) e cianidina-3-glucoramnosídeo (cyn-3-glurha). As duas cianidinas glicosiladas apresentaram pico máximo de absorção em 15 min quando um extrato concentrado foi administrado a ratos Wistar por via oral. A cyn-3-glu e cyn-3-glurha foram encontradas inalteradas no sangue e em pequena quantidade na forma metabolizada. A área abaixo da curva de concentração vs tempo (AAC0-8h) foi de 2,76 ± 0,88 &#181;g.h/mL e 9,74 ± 0,75 &#181;g.h/g (b.u.) para plasma e rim, respectivamente. Assim, estimou-se que somente 0,11% da concentração inicial administrada de cianidina total foi absorvida em 8h. Apesar da baixa absorção, a elevação da concentração de antocianinas no sangue resultou em significativa elevação da atividade antioxidante do plasma. No trato gastrointestinal (estômago, intestino delgado e intestino grosso), cyn-3-glu e cyn-3-glurha foram encotradas na sua maioria nas formas glicosiladas, mas pequena quantidade da forma aglicona também foi encontrada no porção do intestino, provavelmente resultado de hidrólise pelas &#946;-glicosidases endógenas ou da microflora colônica. Ainda, estudos de fermentação in vitro mostraram que as cianidinas glicosiladas são completamente metabolizadas pela microflora colônica. Como demonstrado pelo sistema de intestino evertido, o transporte das antocianinas da amora silvestre (Morus sp.) parece envolver o transportador de glicose sódio dependente, através da interação dos glicosídeos com este. Esta conclusão é reforçada pelo efeito inibitório da absorção das duas cianidinas glicosiladas pela adição de D-glicose e floridzina e pela remoção do íon sódio do meio de incubação no segmento de intestino evertido. / Flavonoids are of great interest in nutrition and medicine because of their potent antioxidant activity and possible protective effects on human health, reducing the risk of chronic diseases such as cardiovascular diseases and cancer. To produce a biological effects in vivo, it is essential that sufficient quantities reach the target tissue. The initial objective of the present study was to make a screening of dietetic vegetable to evaluate their antioxidant potential in the diet and select one of the most promising to study the bioavailability of flavonoid. Fruits, vegetables and commercial frozen pulps (FP) consumed in the diet, were analyzed for antioxidant activities using two different methods, one that determines the inhibition of copper-induced peroxidation of liposome, and other based on the inhibition of the co-oxidation of linoleic acid and &#946;-carotene. The anthocyanin-rich samples showed the highest concentration dependent antioxidant activities in both systems. At both 10 and 50 &#181;M gallic acid equivalent addition level, the neutral and acidic flavonoids of red cabbage, red lettuce, black bean, mulberry, Gala apple peel, jambolao, acai frozen pulp (FP), blackberry FP and the acidic flavonoids of acerola FP showed the highest antioxidant activities in both systems. The major flavonoids from wild mulberry (Morus sp.) were identified by UV/Vis e MS/MS spectrometric methods as cyanidin-3-glucoside (cyn-3-glu) and cyanidin-3-glucorhamnoside (cyn-3-glurha). The two glycosilated forms showed maximum concentration at 15 min after oral administration for rats Wistar. Te cyn-3-glu and cyn-3-glurha were found unchanged in plasma but a little amount was found in the metabolized form. The area under the curve of concentration vs time (AAC0-8h) was 2.76 ± 0.88 &#181;g.h/mL and 9.74 ± 0.75 &#181;g.h/g (b.u.) for plasma and kidney, respectively. Then, the estimated uptake of total cyanidin was 0.11% of the initial concentration. In spite of the low absorption, the increase in its anthocyanin content in plasma resulted in significant increase in antioxidant activity (p<0.5). Cyn-3-glu e cyn-3-glurha were found unchanged at gastrointestinal tract (stomach, small and large intestine), but a low amount of the aglycone form was presented, probably due to the endogen or microbial &#946;-glycosidases hydrolyses. Moreover, in vitro fermentation showed that the two glycosylated cyanidins were totally metabolized by the rat colonic microflore. As showed by everted sacs system, anthocyanins from wild mulberry (Morus sp.) were transported across the enterocite by sodium dependent glucose transporter (SGLT1). This conclusion was evidenced because cyn-3-glu and cyn-3-glurha absorption were inhibited by the addition of D-glucose, phloridzin (a potent inhibitor of SGLT1) and remotion of sodium ion from the incubation medium.

Os alimentos fornecem não somente nutrientes essenciais necessários para a vida, mas também compostos bioativos com propriedades biológicas ditas promotoras da saúde, tais como atividades antioxidante, antiinflamatória e anticarcinogênica de nutrientes como as vitaminas C, A e E, e de compostos fenólicos como os flavonóides. De maneira geral, a capacidade antioxidante de frutos e hortaliças está relacionada aos teores destes compostos, que impedem a ação dos radicais livres e são o foco de interesse deste trabalho. Desta forma, o objetivo deste trabalho foi identificar fontes potenciais de compostos bioativos através da determinação dos teores de flavonóides, ácido elágico, vitamina C, capacidade antioxidante e inibição de &#945;-glicosidase e &#945;-amilase de frutas e polpas de frutas nativas. Foram analisadas 16 frutas e 6 polpas congeladas de frutas nativas, e destas, camu-camu, cupuaçu, coquinho azedo e cagaita mostraram-se excelentes fontes de vitamina C. Camu-camu, cambuci, uxi e tucumã e as polpas de cambuci, cagaita, coquinho azedo e araçá foram as amostras que apresentaram melhor capacidade antioxidante. Os flavonóides de maior incidência entre as amostras foram quercetina e caempferol. O ácido elágico foi detectado apenas em umbu, camu-camu, cagaita, araçá e cambuci e o teor variou de 218 a 512 mg/100 g de amostra seca. Cambuci e cagaita foram as amostras mais eficientes em inibir &#945;-glicosidase e &#945;-amilase. Em suma, este trabalho mostrou que as frutas nativas brasileiras também podem ser consideradas boas fontes de compostos bioativos. / Foods provide not only essential nutrients for life, but also bioactive compounds with health-promoting properties such as antioxidant, anti-inflammatory and hypocholesterolemic activities, which have been related with vitamins A, C and E and phenolics compounds such as flavonoids. In general, antioxidant capacity of fruits and vegetables is linked to the amount of these compounds, which are effective against free radicals. Therefore, the aim of this work was to identify potential sources of bioactive compounds through determination of flavonoids, ellagic acid and vitamin C contents, antioxidant capacity and &#945;-lucosidase and &#945;-amylase inhibition by native fruits and commercial frozen pulps. In this study, 16 native fruits and 6 commercial frozen pulps were analyzed, and camu-camu, cupuaçu, coquinho azedo and cagaita showed themselves as excellent vitamin C sources. Camu-camu, cambuci, uxi and tucumã and commercial frozen pulps of cambuci, cagaita, coquinho azedo and araçá were the samples that presented the highest antioxidant capacity. Quercetin and kaempferol derivatives were the main flavonoids present in most of the samples. Ellagic acid was detected only in umbu, camu-camu, cagaita, araçá and cambuci and the content ranged from 218 to 512 mg/100 g (DW). Cambuci and cagaita showed the highest &#945;-glucosidase and &#945;-amylase inhibitory activity. According to our results, native Brazilian fruits can also be considered as good sources of bioactive compounds.

En la presente Tesis Doctoral se han usado diferentes estrategias para incorporar compuestos activos no volátiles y volátiles en películas biodegradables a base de almidón (S) procedente de diferentes fuentes, quitosano (CH) y mezclas CH:S. Se evaluaron las propiedades funcionales de las películas y las cinéticas de liberación del compuesto activo debido a la incorporación de los polifenoles del tomillo (TE) y del ácido tánico (TA). Además, las propiedades funcionales y cinéticas de liberación de películas de almidón obtenidos por casting o por termoconformado también fueron evaluados en función de la adición de eugenol (E) en forma libre o previamente encapsulado con diferentes materiales (proteína de suero de leche (WP) o lecitinas (LE) junto con maltodextrinas como agente coadyuvante). Los productos microencapsulados en polvo fueron obtenidos por atomización. El efecto de la incorporación de ácido oleico (OA) en los encapsulados, como soporte para el E, y CH, como estabilizador potencial de las capsulas también fue estudiado. Las diferentes formulaciones fueron caracterizadas antes y después del secado en términos de eficiencia de encapsulación (EE), estabilidad térmica, cinéticas de liberación y capacidad antioxidante y antimicrobiana. Los resultados mostraron que TE confirió a las películas de S y CH actividad antioxidante. Los polifenoles (TE y TA) interaccionaron con las cadenas de CH y actuaron como agentes de entrecruzamiento, mejorando el comportamiento mecánico de las películas y reduciendo la velocidad de liberación y la cantidad de polifenol liberado de las películas en agua y en disoluciones acuosas de etanol. El efecto opuesto fue observado cuando el TE fue incorporado en la matriz de S pura. Todas las películas con TE fueron más oscuras, más rojizas y menos transparentes, especialmente las matrices de S:TE, lo que sugiere que los compuestos de TE fueron escasamente encapsulados. Éstas películas mostraron la liberación más rápida y la mayor proporción de TE liberado. Se observó una buena EE en los polvos sin CH (87-98%). Sin embargo, el empleo de CH provocó una disminución de la EE en los polvos formulados con WP y con LE (22% y 46%, respectivamente). Las formulaciones mostraron un comportamiento similar en cuanto a la liberación de E en simulantes alimentarios de diferente polaridad, donde prácticamente el contenido de E fue liberado a una velocidad similar. El efecto antibacteriano de polvos sin CH contra E. coli fue coherente con su contenido de eugenol. Un efecto adicional positivo de OA se detectó en la actividad antilisteria de este polvo. El proceso de moldeo por termocompresión de las películas provocó importantes pérdidas de E (80-65%), minimizadas cuando se utilizó OA en las microcápsulas. En las películas con E no encapsulado, se promovió la retención de E debido a la formación de complejos S-E. La presencia de E microencapsulado aumentó las discontinuidades en la matriz polimérica, produciendo películas menos resistentes y elásticas. Las propiedades barrera se mejoraron al agregar microencapsulados a las películas, a excepción de las microcápsulas con OA. La liberación de E de las películas se vio afectada por la microcápsula, su composición y el simulante alimentario. En la película de S obtenida por casting, la adición de polvo E microencapsulado en las dispersiones formadoras de película modificó la microestructura de las películas, que fueron menos resistentes y menos elásticas con un contenido reducido de humedad, y una menor transparencia y permeabilidad al oxígeno en comparación con las películas formuladas con E no encapsulado. La adición de E microencapsulado con OA promovió la actividad antioxidante de las películas, especialmente en los simulantes alimenticios menos polares. Las películas S-EOA-LE fueron efectivas para prevenir la oxidación del aceite de girasol durante 53 días de almacenamiento a 30°C. / In the present Doctoral Thesis, different strategies have been used to incorporate non-volatile (polyphenols from thyme extract) and a volatile active compound (eugenol) into biodegradable films prepared with starch (S) of different sources or chitosan (CH) or with mixtures of CH:S. The functional properties and release kinetics of casted films prepared with pea starch or CH or CH:S blends were evaluated as affected by the incorporation of polyphenols from an aqueous thyme extract (TE) and tannic acid (TA), a polyphenol which was used as a cross-linking agent. Moreover, the functional properties and release kinetics of corn starch films, obtained by casting or by thermo-compression moulding, were evaluated as a function of the addition of eugenol (E) in free form or pre-encapsulated with different wall materials. In order to encapsulate E, whey protein isolate (WP) or soy lecithin (LE), as wall material, and maltodextrin (MD), as drying coadjuvant, were selected. Microencapsulate powders were obtained by spray-drying. The effect of the addition of oleic acid (OA), as eugenol carrier, and CH, as a potential capsule stabilizer, was also analysed. The different formulations were characterized before and after drying, in terms of encapsulation efficiency, thermal stability, release kinetics and antioxidant capacity and antimicrobial activity. Results showed that TE provided pea starch and chitosan films with antioxidant activity. Polyphenols (TE and TA) interacted with CH chains and acted as cross-linkers, thus improving the tensile behaviour of films and reducing the release rate and the amount of polyphenol released from the films in water and ethanol aqueous solutions. The opposite effect was observed when TE was incorporated into the pure S matrix. All the films became darker, more reddish and less transparent when TE was incorporated, and this effect was more marked in pure S matrices, which suggests that the TE compounds were poorly encapsulated. Thus, S:TE films showed the fastest delivery rate and the highest delivery ratio of TE. A good encapsulation efficiency (EE) of E was observed in the CH-free powders (87-98%). However, the use of CH provoked a marked EE decrease in both WP and LE powders (22% and 46%, respectively). The formulations exhibited similar E release behaviour in food simulants of different polarity, where practically the total E content was delivered at a similar rate. The antibacterial effect of CH-free powders against E. coli was also coherent with their eugenol content. An additional positive effect of OA was detected in terms of the antilisterial action of this powder. The process of thermo-compression moulding of the films led to important E losses (80-65%), which were minimised when using OA in the microcapsules (EOA-WP or EOA-LE). In the films containing non-encapsulated E, the retention of E was promoted due to the formation of E-starch complexes. The presence of microencapsulated eugenol increased the presence of discontinuities in the polymeric matrix, thus yielding films that were mechanically less resistant and stretchable, except for E-LE-S films, which became more resistant to break. The barrier properties were improved when adding microencapsulates to the films, except for OA-based microcapsules. Eugenol release from the films was affected by microcapsule, their composition and the food simulant. In starch film obtained by casting, the addition of microencapsulated eugenol powder in starch film-forming dispersions modified film microstructure, yielding less resistant and less elastic films with reduced moisture content, transparency and oxygen permeability as compared to casted films formulated with non-encapsulated eugenol. The addition of eugenol microencapsulated with OA promoted the preservation of the antioxidant activity of the films, especially in less polar food simulants. S-EOA-LE films were effective in preventing sunflower oil oxidation during 53 days of storage at 30¿C. / En la present Tesi Doctoral s'han usat diferents estratègies per a incorporar compostos actius no volàtils i volàtils en pel·lícules biodegradables a base de midó (S) procedent de diferents fonts, quitosano (CH) i mescles CH:S. Es van avaluar les propietats funcionals de les pel·lícules i les cinètiques d'alliberament del compost actiu a causa de la incorporació dels polifenoles del timó (TE) i de l'àcid tànnic (TA). A més, les propietats funcionals i cinètiques d'alliberament de pel·lícules de midó obtinguts per càsting o per termoconformat també van ser avaluats en funció de l'addició d'eugenol (E) en forma lliure o prèviament encapsulat amb diversos materials (proteïna de sèrum de llet (WP) o lecitines (LE) junt amb maltodextrines com a agent coadjuvant). Els productes microencapsulats en pols van ser obtinguts per atomització. L'efecte de la incorporació d'àcid oleic (OA) en els encapsulats, com a suport per al E, i CH, com estabilitzador potencial de les càpsules també va ser estudiat. Les diferents formulacions van ser caracteritzades abans i després de l'assecat en termes d'eficiència d'encapsulació (EE), estabilitat tèrmica, cinètiques d'alliberament i capacitat antioxidant i antimicrobiana. Els resultats van mostrar que es va conferir a les pel·lícules de S i CH activitat antioxidant. Els polifenols (TE i TA) van interaccionar amb les cadenes de CH i van actuar com a agents d'entrecreuament, millorant el comportament mecànic de les pel·lícules i reduint la velocitat d'alliberament i la quantitat de polifenol alliberat de les pel·lícules en aigua i en dissolucions aquoses d'etanol. L'efecte oposat va ser observat quan el TE va ser incorporat en la matriu de S pura. Totes les pel·lícules amb TE van ser més fosques, més rogenques i menys transparents, especialment les matrius de S:TE, la qual cosa suggerix que els compostos de TE van ser escassament encapsulats. Estes pel·lícules van mostrar l'alliberament més ràpid i la major proporció de TE alliberat. Es va observar una bona EE en les pols sense CH (87-98%). No obstant, la incorporació de CH va provocar una disminució de l'EE en les pols formulats amb WP i amb LE (22% i 46%, respectivament). Les formulacions van mostrar un comportament semblant quant a l'alliberament de E en simulants alimentaris de diferent polaritat, on pràcticament el contingut de E va ser alliberat a una velocitat semblant. L'efecte antibacterià de pols sense CH contra E. coli va ser coherent amb el seu contingut d'eugenol. Un efecte addicional positiu d'OA es va detectar en l'activitat antilisteria d'esta pols. El procés de modelat per termocompresió de les pel·lícules va provocar importants pèrdues de E (80-65%) , minimitzades quan es va utilitzar OA en les microcàpsules. En les pel·lícules amb E no encapsulat, es va promoure la retenció de E a causa de la formació de complexos S-E. La presència de E microencapsulat va augmentar les discontinuïtats en la matriu polimérica, produint pel·lícules menys resistents i elàstiques. Les propietats barrera es van millorar a l'agregar microencapsulats a les pel·lícules, a excepció de les microcàpsules amb OA. L'alliberament de E de les pel·lícules es va veure afectada per la microcàpsula, la seua composició i el simulant alimentari. En la pel·lícula de S obtinguda per càsting, l'addició de pols E microencapsulat en les dispersions formadores de pel·lícula va modificar la microestructura de les pel·lícules, que van ser menys resistents i menys elàstiques amb un contingut reduït d'humitat, i una menor transparència i permeabilitat a l'oxigen en comparació amb les pel·lícules formulades amb E no encapsulat. L'addició de E microencapsulat amb OA va promoure l'activitat antioxidant de les pel·lícules, especialment en els simulants alimentaris menys polars. Les pel·lícules S-EOA-LE van ser efectives per a previndre l'oxidació de l'oli de gira-sol durant 53 dies d'em / Talón Argente, E. (2018). INCORPORATION OF ACTIVE COMPONENTS IN BIOPOLYMER-BASED FILMS FOR FOOD USE [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/101283 / TESIS

Diploma thesis deals with currant (Ribes L.) and substances which currants contain especially anthocyanin pigments, vitamin C and antioxidants. In theoretical part is described basic characteristic about plant, including varieties of currants. The following chapters include a list of active substances which the fruit contains and also the list of some possibilities of processing and utilization of currant especially in the food industry. Big attention is given to anthocyanins, their characteristics, utilization in the food industry, factors that affect their stability and especially methods of determination. In conclusion, the theoretical part describes importance and methods of determination of vitamin C and antioxidant capacity. Selected nutritional parameters were determined in fifteen known and in three unknown varieties of currants. Anthocyanins were measured spectrophotometrically and determined by HPLC in color samples. By pH differential method was evaluated a content of monomeric and polymeric pigments. By HPLC method were identified individual anthocyanin pigments contained in fruits. Also, content of ascorbic acid was determined by HPLC in all varieties. Antioxidant capacity of white currants measured by DPPH was compared.

Целью выпускной квалификационной работы является разработка потенциометрического подхода для определения антиоксидантной емкости с использованием комплексов железа с 2,2’-бипиридином и его производными. В работе были проведены исследования электрохимического поведения комплексов железа с 2,2’-бипиридином и его производными методом циклической вольтамперометрии. Проведен расчет условных констант устойчивости выбранных комплексных соединений. Была исследована антиоксидантная емкость ряда фенольных антиоксидантов, обладающих различной растворимостью, новым разработанным потенциометрическим подходом с использованием в качестве модели окислителя бипиридиновых комплексов. Проведены исследования на многокомпонентных объектах – экстрактов травянистых растений. Модифицированным спектрофотометрическим методом было доказано стехиометрическое протекание реакций комплексов хлорида железа (III) c 2,2’-бипиридином и его синтетическими производными с модельными антиоксидантами. Были выявлены перспективные комплексные соединения с производными бипиридина для дальнейшего исследования антиоксидантных свойств потенциометрическим способом. / The aim of the final qualifying work is to develop a potentiometric approach for determining the antioxidant capacity using iron complexes with 2,2’-bipyridine and its derivatives. The electrochemical behavior of iron complexes with 2,2’-bipyridine and its derivatives was studied by cyclic voltammetry. The conditional stability constants of the selected complex compounds are calculated. The antioxidant capacity of a number of phenolic antioxidants with different solubility was investigated by a newly developed potentiometric approach using bipyridine complexes as an oxidizer model. Studies have been carried out on multicomponent objects – extracts of herbaceous plants. The modified spectrophotometric method proved the stoichiometric course of reactions of iron (III) chloride complexes with 2,2’-bipyridine and its synthetic derivatives with model antioxidants. Promising complex compounds with bipyridine derivatives have been identified for further investigation of antioxidant properties by a potentiometric method.

Motivation In the black population of South Africa, cardiovascular disease (CVD) is rapidly increasing due to urbanisation. Stroke is usually accompanied by a prothrombotic haemostatic profile. Changing lifestyle factors that accompany the urbanisation process could have a negative impact on the haemostatic profile of black South Africans. Elevated levels of pro-coagulant factors, von Willebrand factor (vWF), fibrinogen and fibrin D-dimer have been reported in the black population, which could increase the black population’s susceptibility to CVD. However, low levels of plasminogen activator inhibitor-1 (PAI-1) previously reported in the black population could contribute towards a pro-fibrinolytic state, which may counteract the hypercoagulant state. This may have a beneficial effect on the haemostatic profile of the black population. More investigation into the haemostatic profile of black South Africans is therefore needed to determine if an altered haemostatic profile exists in this group, and if so, to what extent these alterations may relate to cardiovascular dysfunction. This study included markers of both the coagulation (vWF, fibrinogen, fibrin D-dimer) and fibrinolytic (PAI-1, fibrin D-dimer and fibrinolytic potential) systems in an attempt to investigate the haemostatic profile of the black population of South Africa, and for comparison purposes that of the white population as well. The relationship of these markers’ with selected markers of cardiovascular function was also examined to determine if they could possibly contribute to an increase in cardiovascular risk, especially in the black population. Aims The aims of this study were to first compare coagulation and fibrinolysis markers in the black and white populations of South Africa. Furthermore, to determine if associations exist between the selected components of the haemostatic system and markers of cardiovascular function, especially in the black population of South Africa, who tends to be at a higher cardiovascular risk due to altered metabolic and haemostatic profiles. Methodology The Sympathetic activity and Ambulatory Blood Pressure in Africans (SABPA) study was a prospective cohort study that consisted of 409 participants at baseline (2008-2009) that were equally distributed according to both ethnicity (200 black; 209 white) and gender (black, 101 men, 99 women; white, 101 men, 108 women). At follow-up (2011/2012) the cohort totalled 359 participants (170 black, 88 men and 82 women; 189 white, 93 men and 96 women). Data from baseline measurements were used for the first two manuscripts (chapters 2 and 3), while followup data was used for the third manuscript (chapter 4). vWF, fibrinogen, PAI-1, fibrin D-dimer, CLT, serum peroxides, glutathione, glutathione peroxidase and reductase activity were determined, and ambulatory blood pressure and the retinal vessel calibres were measured. The groups were stratified by ethnicity as specified by statistical interaction terms. T-tests and chi-square tests were used to compare means and proportions, respectively. Pearson and partial regression analyses were used to determine correlations between the components of the haemostatic system and cardiovascular function markers. This was followed by multiple linear regression analyses to investigate whether independent associations exist between the variables in both ethnic groups. P-values ≤0.050 were deemed significant. Results and conclusion of each manuscript The first manuscript (chapter 2) compares the haemostatic profiles of the black and white population to determine whether ambulatory blood pressure is related to components of the haemostatic system. The black participants displayed a prothrombotic profile with significantly higher vWF, fibrinogen, PAI-1, fibrin D-dimer and a longer CLT than their white counterparts. Furthermore, partial and multiple linear regression analyses showed a positive association of systolic and diastolic blood pressure with fibrin D-dimer in the black population, while a negative association existed between ambulatory blood pressure and CLT in the white population. These associations suggest that fibrin D-dimer may contribute, at least in part, to the high prevalence of hypertension in the black population. The second manuscript (chapter 3) determined associations between markers of the haemostatic and oxidant-antioxidant systems in the black and white populations. In addition to the prothrombotic profile that exists in the black population, this group also had significantly higher serum peroxides (oxidative stress) and lower glutathione peroxidase activity (antioxidant) levels. Multiple linear regression analyses indicated positive associations between fibrinogen and serum peroxides in both populations. In the white population, an additional positive association was found between serum peroxide and CLT. In the black population, vWF and CLT were negatively associated with GPx activity. The results suggest that there are ethnic-specific relationships between the haemostatic and oxidant-antioxidant systems. The third manuscript (chapter 4) investigated the relationships between the retinal vessel calibres and components of the haemostatic system in the black and white population. The investigation focussed specifically on arteriolar diameters in the lower median, since a narrow arteriolar diameter is known to be associated with elevated blood pressure. In both ethnic groups, a narrower arteriolar calibre was accompanied by narrower venular calibres. Independent positive associations were found between the central retinal vein equivalent (CRVE) and fibrinogen in the black population, as well as vWF and CLT in the white population. In addition, independent negative associations were found between the central retinal artery equivalent and CLT in the black population and with vWF in the white population. The results suggest that haemostatic alterations are linked to early vascular changes that may differ between ethnicities. General conclusion Ethnic-specific relationships between the components of the haemostatic system and measures of cardiovascular function are evident. The prothrombotic profile that is observed in the black population, together with the adverse associations of the haemostatic components with blood pressure, a compromised oxidant-antioxidant profile, and retinal vessel calibres may contribute, at least in part, to the high cardiovascular and cerebrovascular risk evident in this population group. / PhD (Physiology), North-West University, Potchefstroom Campus, 2015

Motivation In the black population of South Africa, cardiovascular disease (CVD) is rapidly increasing due to urbanisation. Stroke is usually accompanied by a prothrombotic haemostatic profile. Changing lifestyle factors that accompany the urbanisation process could have a negative impact on the haemostatic profile of black South Africans. Elevated levels of pro-coagulant factors, von Willebrand factor (vWF), fibrinogen and fibrin D-dimer have been reported in the black population, which could increase the black population’s susceptibility to CVD. However, low levels of plasminogen activator inhibitor-1 (PAI-1) previously reported in the black population could contribute towards a pro-fibrinolytic state, which may counteract the hypercoagulant state. This may have a beneficial effect on the haemostatic profile of the black population. More investigation into the haemostatic profile of black South Africans is therefore needed to determine if an altered haemostatic profile exists in this group, and if so, to what extent these alterations may relate to cardiovascular dysfunction. This study included markers of both the coagulation (vWF, fibrinogen, fibrin D-dimer) and fibrinolytic (PAI-1, fibrin D-dimer and fibrinolytic potential) systems in an attempt to investigate the haemostatic profile of the black population of South Africa, and for comparison purposes that of the white population as well. The relationship of these markers’ with selected markers of cardiovascular function was also examined to determine if they could possibly contribute to an increase in cardiovascular risk, especially in the black population. Aims The aims of this study were to first compare coagulation and fibrinolysis markers in the black and white populations of South Africa. Furthermore, to determine if associations exist between the selected components of the haemostatic system and markers of cardiovascular function, especially in the black population of South Africa, who tends to be at a higher cardiovascular risk due to altered metabolic and haemostatic profiles. Methodology The Sympathetic activity and Ambulatory Blood Pressure in Africans (SABPA) study was a prospective cohort study that consisted of 409 participants at baseline (2008-2009) that were equally distributed according to both ethnicity (200 black; 209 white) and gender (black, 101 men, 99 women; white, 101 men, 108 women). At follow-up (2011/2012) the cohort totalled 359 participants (170 black, 88 men and 82 women; 189 white, 93 men and 96 women). Data from baseline measurements were used for the first two manuscripts (chapters 2 and 3), while followup data was used for the third manuscript (chapter 4). vWF, fibrinogen, PAI-1, fibrin D-dimer, CLT, serum peroxides, glutathione, glutathione peroxidase and reductase activity were determined, and ambulatory blood pressure and the retinal vessel calibres were measured. The groups were stratified by ethnicity as specified by statistical interaction terms. T-tests and chi-square tests were used to compare means and proportions, respectively. Pearson and partial regression analyses were used to determine correlations between the components of the haemostatic system and cardiovascular function markers. This was followed by multiple linear regression analyses to investigate whether independent associations exist between the variables in both ethnic groups. P-values ≤0.050 were deemed significant. Results and conclusion of each manuscript The first manuscript (chapter 2) compares the haemostatic profiles of the black and white population to determine whether ambulatory blood pressure is related to components of the haemostatic system. The black participants displayed a prothrombotic profile with significantly higher vWF, fibrinogen, PAI-1, fibrin D-dimer and a longer CLT than their white counterparts. Furthermore, partial and multiple linear regression analyses showed a positive association of systolic and diastolic blood pressure with fibrin D-dimer in the black population, while a negative association existed between ambulatory blood pressure and CLT in the white population. These associations suggest that fibrin D-dimer may contribute, at least in part, to the high prevalence of hypertension in the black population. The second manuscript (chapter 3) determined associations between markers of the haemostatic and oxidant-antioxidant systems in the black and white populations. In addition to the prothrombotic profile that exists in the black population, this group also had significantly higher serum peroxides (oxidative stress) and lower glutathione peroxidase activity (antioxidant) levels. Multiple linear regression analyses indicated positive associations between fibrinogen and serum peroxides in both populations. In the white population, an additional positive association was found between serum peroxide and CLT. In the black population, vWF and CLT were negatively associated with GPx activity. The results suggest that there are ethnic-specific relationships between the haemostatic and oxidant-antioxidant systems. The third manuscript (chapter 4) investigated the relationships between the retinal vessel calibres and components of the haemostatic system in the black and white population. The investigation focussed specifically on arteriolar diameters in the lower median, since a narrow arteriolar diameter is known to be associated with elevated blood pressure. In both ethnic groups, a narrower arteriolar calibre was accompanied by narrower venular calibres. Independent positive associations were found between the central retinal vein equivalent (CRVE) and fibrinogen in the black population, as well as vWF and CLT in the white population. In addition, independent negative associations were found between the central retinal artery equivalent and CLT in the black population and with vWF in the white population. The results suggest that haemostatic alterations are linked to early vascular changes that may differ between ethnicities. General conclusion Ethnic-specific relationships between the components of the haemostatic system and measures of cardiovascular function are evident. The prothrombotic profile that is observed in the black population, together with the adverse associations of the haemostatic components with blood pressure, a compromised oxidant-antioxidant profile, and retinal vessel calibres may contribute, at least in part, to the high cardiovascular and cerebrovascular risk evident in this population group. / PhD (Physiology), North-West University, Potchefstroom Campus, 2015