PEREIRA, Ana Lucia Fernandes. Elaboração e secagem em spray dryer de bebida probiótica formulada a partir da fermentação do suco de caju. 2013. 114 f. : Tese (doutorado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Tecnologia de Alimentos, Fortaleza-CE, 2013 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-07T15:22:25Z No. of bitstreams: 1 2013_tese_alfpereira.pdf: 23321259 bytes, checksum: 8664c9c00f28dc1b79e6eb068f4d265d (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-07T15:22:45Z (GMT) No. of bitstreams: 1 2013_tese_alfpereira.pdf: 23321259 bytes, checksum: 8664c9c00f28dc1b79e6eb068f4d265d (MD5) / Made available in DSpace on 2016-07-07T15:22:45Z (GMT). No. of bitstreams: 1 2013_tese_alfpereira.pdf: 23321259 bytes, checksum: 8664c9c00f28dc1b79e6eb068f4d265d (MD5) Previous issue date: 2013 / The objective of this study was to develop a probiotic cashew apple juice ready to drink and in the dehydrated form through spray drying. The first stage of the study was the optimization of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, to optimize the proper inoculum amount and the fermentation time. The optimum conditions for probiotic cashew apple juice production were: initial pH 6.4, fermentation temperature of 30°C, inoculation level of 7.48 log CFU/mL (L. casei) and 16 h of fermentation process. Cashew apple juice showed to be as efficient as dairy products for L. casei growth. In a second stage, the stability of probiotic cashew apple juice stored for 42 days at 4°C was evaluated. Analyses were conducted in the non fermented cashew apple juice (control), and in the fermented juices with L. casei NRRL B-442, with 8% (w/v) of sucrose (sugar table), after fermentation, and without the addition of sugar. The viability of the probiotic bacteria, sugars and organic acids content, color, antioxidant and enzymatic activity, and sensory characteristics were evaluated during the storage. Viable cell counts increased in the probiotic cashew apple containing sucrose along the storage period. Moreover, the fermentation lead to the preservation of the ascorbic acid content, which had a less intense reduction in the fermented cashew apple juices compared to the non fermented sample. The antioxidant activity and total polyphenolic compounds of cashew apple juice had a similar trend. Browning reactions and nutritional breakdown caused by enzymes were minimized in the fermented samples during storage. In these samples, a higher reduction of the enzymatic activity of polyphenoloxidase and peroxidase activity was observed. During the storage, the increase in the chroma values indicated that yellowness was reinforced, being well accepted by consumers. The sensory attributes (aroma, flavor, acidity and color) of probiotic cashew apple juice were positively influenced by storage under refrigeration for 42 days. In the third stage of the research, the effects of dehydration by spray drying in cashew apple juice containing L. casei NRRL B-442 was assessed and the influence of storage temperature on the viability of L. casei NRRL B-442 and physical properties of the powder were evaluated during 35 days of storage. The drying agents used were: 20% (w/v) maltodextrin or 10% (w/v) maltodextrin + 10% (w/v) arabic gum. The powder of probiotic cashew apple juice showed satisfactory levels of L. casei survival, during drying. During storage, the addition of 10% (w/v) maltodextrin + 10% (w/v) arabic gum kept microbial viability within satisfactory levels when the powder was subjected to cooling at 4°C. However, greater differences in the reconstituted powder color and higher rehydration time were obtained in this condition. On the other hand, the addition of 20% (w/v) maltodextrin provided better yield. In conclusion, cashew apple juice is a good substrate for the probiotic beverage production, and the condition of drying agents 10% maltodextrin + 10% arabic gum is adequate to maintain satisfactory levels of L. casei NRRL B-442 survival for 35 days, in the powder of probiotic cashew juice stored at 4°C. / O objetivo desta pesquisa foi elaborar um produto probiótico à base de suco de caju pronto para beber, como também, na forma desidratada obtida pela secagem por aspersão (spray drying). A primeira etapa da pesquisa consistiu em otimizar as condições de crescimento do Lactobacillus casei NRRL B-442 em suco de caju, a quantidade adequada de inóculo e o tempo de fermentação. As condições ótimas para produção do suco de caju probiótico foram: pH inicial de 6,4, temperatura de fermentação de 30°C, quantidade de inóculo de 7,48 log UFC/mL (L. casei) e 16 h de fermentação. O suco de caju mostrou ser tão eficiente quanto os produtos lácteos para o crescimento de L. casei. Em uma segunda etapa, foi avaliada a estabilidade da bebida probiótica de caju estocada por 42 dias a 4°C. Foram realizadas análises no suco de caju não fermentado (controle) e nos sucos fermentados com L. casei NRRL B-442, adicionado ou não de 8% (p/v) de sacarose depois da fermentação. Durante a estocagem, foram realizadas as determinações de viabilidade de L. casei NRRL B-442, conteúdo de açúcares e ácidos orgânicos, cor, atividade antioxidante e enzimática e aceitação sensorial. Foi observado que o número de células viáveis aumentou no suco de caju contendo sacarose ao longo da estocagem. Além disso, a fermentação proporcionou um efeito conservante no conteúdo de ácido ascórbico que teve uma redução menos intensa, com a estocagem, nos sucos fermentados, quando comparados com o controle. A atividade antioxidante e o conteúdo de polifenóis apresentaram similar tendência. Reações que reduzem o valor nutricional causadas por enzimas foram minimizadas nas amostras fermentadas durante a estocagem. Nessas amostras foi observada maior redução da atividade enzimática da polifenoloxidase e peroxidase. Durante a estocagem, o aumento do croma indicou que a cor amarela foi intensificada, sendo bem aceita pelos consumidores. Os atributos sensoriais (aroma, sabor, acidez e cor) do suco de caju probiótico foram positivamente influenciados pela estocagem sob refrigeração por 42 dias. Na terceira etapa da pesquisa, foi avaliado o efeito da desidratação por spray drying no suco de caju contendo L. casei NRRL B-442, além de avaliar a influência da temperatura de estocagem sobre a viabilidade de L. casei e nas propriedades físicas do pó, durante 35 dias de estocagem. Os agentes de secagem usados foram: 20% (p/v) de maltodextrina ou 10% (p/v) de maltodextrina + 10% (p/v) de goma arábica. O suco de caju probiótico desidratado por spray drying apresentou níveis satisfatórios de sobrevivência de L. casei NRRL B-442, durante a secagem. Durante a estocagem, a adição de 10% (p/v) de maltodextrina + 10% (p/v) de goma arábica manteve a viabilidade microbiana dentro de níveis satisfatórios quando o pó foi submetido à refrigeração a 4ºC. Entretanto, maiores diferenças na coloração do pó reconstituído e maior tempo de reidratação foram obtidos nesta condição. Já a adição de 20% (p/v) de maltodextrina proporcionou melhor rendimento. Em conclusão, o suco de caju pode ser utilizado como substrato para o desenvolvimento de bebida probiótica, e a condição dos agentes de secagem de 10% de maltodextrina + 10% de goma arábica mostra-se adequada para manter os níveis satisfatórios de L. casei NRRL B-442 por até 35 dias, no suco de caju probiótico desidratado estocado a 4°C.

PEREIRA, Ana Lucia Fernandes. Elaboração e secagem em spray dryer de bebida probiótica formulada a partir da fermentação do suco de caju. 2013. 116 f. Tese (Doutorado em tecnologia de alimentos)- Universidade Federal do Ceará, Fortaleza-CE, 2013. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-07-07T17:55:40Z No. of bitstreams: 1 2013_tese_alfpereira.pdf: 24437687 bytes, checksum: 9e9963b2d25536d6bfb0cbe06acecac0 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-07-21T20:24:58Z (GMT) No. of bitstreams: 1 2013_tese_alfpereira.pdf: 24437687 bytes, checksum: 9e9963b2d25536d6bfb0cbe06acecac0 (MD5) / Made available in DSpace on 2016-07-21T20:24:58Z (GMT). No. of bitstreams: 1 2013_tese_alfpereira.pdf: 24437687 bytes, checksum: 9e9963b2d25536d6bfb0cbe06acecac0 (MD5) Previous issue date: 2013 / The objective of this study was to develop a probiotic cashew apple juice ready to drink and in the dehydrated form through spray drying. The first stage of the study was the optimization of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, to optimize the proper inoculum amount and the fermentation time. The optimum conditions for probiotic cashew apple juice production were: initial pH 6.4, fermentation temperature of 30°C, inoculation level of 7.48 log CFU/mL (L. casei) and 16 h of fermentation process. Cashew apple juice showed to be as efficient as dairy products for L. casei growth. In a second stage, the stability of probiotic cashew apple juice stored for 42 days at 4°C was evaluated. Analyses were conducted in the non fermented cashew apple juice (control), and in the fermented juices with L. casei NRRL B-442, with 8% (w/v) of sucrose (sugar table), after fermentation, and without the addition of sugar. The viability of the probiotic bacteria, sugars and organic acids content, color, antioxidant and enzymatic activity, and sensory characteristics were evaluated during the storage. Viable cell counts increased in the probiotic cashew apple containing sucrose along the storage period. Moreover, the fermentation lead to the preservation of the ascorbic acid content, which had a less intense reduction in the fermented cashew apple juices compared to the non fermented sample. The antioxidant activity and total polyphenolic compounds of cashew apple juice had a similar trend. Browning reactions and nutritional breakdown caused by enzymes were minimized in the fermented samples during storage. In these samples, a higher reduction of the enzymatic activity of polyphenoloxidase and peroxidase activity was observed. During the storage, the increase in the chroma values indicated that yellowness was reinforced, being well accepted by consumers. The sensory attributes (aroma, flavor, acidity and color) of probiotic cashew apple juice were positively influenced by storage under refrigeration for 42 days. In the third stage of the research, the effects of dehydration by spray drying in cashew apple juice containing L. casei NRRL B-442 was assessed and the influence of storage temperature on the viability of L. casei NRRL B-442 and physical properties of the powder were evaluated during 35 days of storage. The drying agents used were: 20% (w/v) maltodextrin or 10% (w/v) maltodextrin + 10% (w/v) arabic gum. The powder of probiotic cashew apple juice showed satisfactory levels of L. casei survival, during drying. During storage, the addition of 10% (w/v) maltodextrin + 10% (w/v) arabic gum kept microbial viability within satisfactory levels when the powder was subjected to cooling at 4°C. However, greater differences in the reconstituted powder color and higher rehydration time were obtained in this condition. On the other hand, the addition of 20% (w/v) maltodextrin provided better yield. In conclusion, cashew apple juice is a good substrate for the probiotic beverage production, and the condition of drying agents 10% maltodextrin + 10% arabic gum is adequate to maintain satisfactory levels of L. casei NRRL B-442 survival for 35 days, in the powder of probiotic cashew juice stored at 4°C. / O objetivo desta pesquisa foi elaborar um produto probiótico à base de suco de caju pronto para beber, como também, na forma desidratada obtida pela secagem por aspersão (spray drying). A primeira etapa da pesquisa consistiu em otimizar as condições de crescimento do Lactobacillus casei NRRL B-442 em suco de caju, a quantidade adequada de inóculo e o tempo de fermentação. As condições ótimas para produção do suco de caju probiótico foram: pH inicial de 6,4, temperatura de fermentação de 30°C, quantidade de inóculo de 7,48 log UFC/mL (L. casei) e 16 h de fermentação. O suco de caju mostrou ser tão eficiente quanto os produtos lácteos para o crescimento de L. casei. Em uma segunda etapa, foi avaliada a estabilidade da bebida probiótica de caju estocada por 42 dias a 4°C. Foram realizadas análises no suco de caju não fermentado (controle) e nos sucos fermentados com L. casei NRRL B-442, adicionado ou não de 8% (p/v) de sacarose depois da fermentação. Durante a estocagem, foram realizadas as determinações de viabilidade de L. casei NRRL B-442, conteúdo de açúcares e ácidos orgânicos, cor, atividade antioxidante e enzimática e aceitação sensorial. Foi observado que o número de células viáveis aumentou no suco de caju contendo sacarose ao longo da estocagem. Além disso, a fermentação proporcionou um efeito conservante no conteúdo de ácido ascórbico que teve uma redução menos intensa, com a estocagem, nos sucos fermentados, quando comparados com o controle. A atividade antioxidante e o conteúdo de polifenóis apresentaram similar tendência. Reações que reduzem o valor nutricional causadas por enzimas foram minimizadas nas amostras fermentadas durante a estocagem. Nessas amostras foi observada maior redução da atividade enzimática da polifenoloxidase e peroxidase. Durante a estocagem, o aumento do croma indicou que a cor amarela foi intensificada, sendo bem aceita pelos consumidores. Os atributos sensoriais (aroma, sabor, acidez e cor) do suco de caju probiótico foram positivamente influenciados pela estocagem sob refrigeração por 42 dias. Na terceira etapa da pesquisa, foi avaliado o efeito da desidratação por spray drying no suco de caju contendo L. casei NRRL B-442, além de avaliar a influência da temperatura de estocagem sobre a viabilidade de L. casei e nas propriedades físicas do pó, durante 35 dias de estocagem. Os agentes de secagem usados foram: 20% (p/v) de maltodextrina ou 10% (p/v) de maltodextrina + 10% (p/v) de goma arábica. O suco de caju probiótico desidratado por spray drying apresentou níveis satisfatórios de sobrevivência de L. casei NRRL B-442, durante a secagem. Durante a estocagem, a adição de 10% (p/v) de maltodextrina + 10% (p/v) de goma arábica manteve a viabilidade microbiana dentro de níveis satisfatórios quando o pó foi submetido à refrigeração a 4ºC. Entretanto, maiores diferenças na coloração do pó reconstituído e maior tempo de reidratação foram obtidos nesta condição. Já a adição de 20% (p/v) de maltodextrina proporcionou melhor rendimento. Em conclusão, o suco de caju pode ser utilizado como substrato para o desenvolvimento de bebida probiótica, e a condição dos agentes de secagem de 10% de maltodextrina + 10% de goma arábica mostra-se adequada para manter os níveis satisfatórios de L. casei NRRL B-442 por até 35 dias, no suco de caju probiótico desidratado estocado a 4°C.

Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2014-09-05T15:25:03Z No. of bitstreams: 1 Dissertação LeandrodMF.pdf: 1496532 bytes, checksum: a3f1f8effe11a679e223ba5bdceeeb54 (MD5) / Approved for entry into archive by Fatima Cleômenis Botelho Maria (botelho@ufba.br) on 2014-09-05T15:56:02Z (GMT) No. of bitstreams: 1 Dissertação LeandrodMF.pdf: 1496532 bytes, checksum: a3f1f8effe11a679e223ba5bdceeeb54 (MD5) / Made available in DSpace on 2014-09-05T15:56:02Z (GMT). No. of bitstreams: 1 Dissertação LeandrodMF.pdf: 1496532 bytes, checksum: a3f1f8effe11a679e223ba5bdceeeb54 (MD5) / CNPq / Neste trabalho foi proposto um sistema de análises por injeção sequencial (SIA) para determinação espectrofotométrica da capacidade antioxidante em amostras de bebidas, a partir da geração “in situ” do radical derivado do reagente N-fenil-1,4-fenilenodiamino (NFFD•+). O radical NFFD•+ foi produzido em linha pela oxidação do NFFD empregando o Fe3+ como oxidante. Na otimização do sistema SIA foram avaliados os seguintes parâmetros: ordem de aspiração dos reagentes e amostra, estabilidade da solução de NFFD, pH da amostra, proporção NFFD/Fe3+, volumes das soluções dos reagentes e amostra e tempo de interrupção do fluxo. O ácido gálico foi empregado como padrão de calibração para os ensaios. Após as etapas de otimização univariada do método proposto obteve-se faixa linear de 10 a 40 mg L-1, com limites de detecção (3σ) de 1,0 mg L-1 e quantificação (10σ) de 3,3 mg L-1, respectivamente. Para avaliar o método proposto, foram realizados testes de reatividade do radical NFFD•+ frente a compostos fenólicos, tiólicos e de outra natureza. A redução do sinal analítico frente a inibição do radical derivado do NFFD foi mais pronunciada para ácido ascórbico, ácido tânico e cisteína, nesta ordem. A capacidade antioxidante com o radical derivado do NFFD foi avaliada em amostras de vinhos e chás, e os resultados foram concordantes com os métodos de referência (Folin-Denis, ABTS e DPPH), de acordo com o coeficiente de correlação, além do ensaio com radical DMPD•+, baseado no teste t. Assim a amina NFFD por ser utilizada para avaliação da atividade antioxidante em amostras de bebidas. Adicionalmente, as amostras de vinhos foram submetidas a testes de estabilidade temporal e avaliação do efeito do processo de ozonização em relação à variação da capacidade antioxidante / In this work was proposed a sequential injection system for spectrophotometric determination of antioxidant capacity in beverage samples with “in situ” radical generation from N-phenyl-1,4-phenylenediamine (NFFD•+). The radical NFFD•+ was produced by on line oxidation of NFFD employing Fe3+ as oxidant. The optimization of SIA system evaluated: aspiration order of reagents and sample, stability of the NFFD solution, sample pH, NFFD/Fe3+ proportion, reagents and sample volumes and stopped flow time. The gallic acid was used as standard of antioxidant compound for proposed sequential injection method. After chemical and physical univarieted optimization the linear range obtained in gallic acid concentration was from 10 to 40 mg L-1, with detection (3σ) and quantification (10σ) limits of 1.0 and 3.3 mg L-1, respectively. To evaluate selective of radical, studies of reactivity were performed with NFFD•+ and phenolic and thiol molecules and others compounds. The reducing capacity of NFFD•+ radical was observed preferentially for ascorbic acid, tannic acid and cysteine in this order. The antioxidant capacity with NFFD•+ radical was measured in samples of wines, infusions and teas. The results were agreement with the references methods (Folin-Denis, ABTS and DPPH), in accord with correlation coefficient, and additionally with the radical DMPD•+, based on t test. This way, the SIA spectrophotometric method exploring the radical NFFD•+ can be employed for the antioxidant capacity measured. Additionally, temporal stability and the ozonization effect in wine samples were evaluated using antioxidant capacity as parameter .

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The present work reports the development of new methodologies to evaluate the in vitro and in vivo antioxidant capacity against peroxyl and hydroxyl radicals in five medicinal plant samples (Matricaria chamomilla L., Psidium guajava, Achyrocline satureoides, Baccharis genistelloides and Cymbopogon citratus) and seven phenolic compounds (rutin, quercetin, resveratrol, gallic acid, ferulic acid, caffeic acid and rosmarinic acid). The antioxidant capacity of the samples was analyzed by two independent methods. In the first one, a new approach was used to detect hydroxyl radicals indirectly using an electrochemical procedure, in which the radicals destroy a thiol self-assembled monolayer (SAM) on a gold electrode. This monolayer can block the electrochemical signal of a dissolved redox probe. When such an electrode with a SAM is exposed to free radicals, these radicals destroy the SAM and the electrochemical signal of a redox probe recovers to a degree proportional to the extent of dissolution of the SAM. In the second method, the evaluation of antioxidant capacity against peroxyl and hydroxyl radicals is based on the indirect detection of these reactive oxygen species (ROS) by fluorimetry (ex/em: 485/520 nm) employing 2 -7 -dichlorofluorescin diacetate (DCFH-DA) as a fluorescent probe. After the deacetilation reaction of DCFH-DA, it can be oxidized by ROS to the fluorescent compound DCF. The peroxyl radicals were produced at 37ºC by thermal decomposition of 2,2 -azobis (2 methylpropianamidine) dihydrochloride (ABAP), while hydroxyl radicals were generated by the Fenton reaction. In vivo assays were evaluated using zebrafish (Danio rerio) hepatocytes exposed to P. guajava and C. citratus extracts (better and worst in vitro results, respectively). The results obtained by the electrochemical and fluorimetric methods show that it is difficult to classify the tested plants according to their antioxidant capacity, since the results depended strongly on the extract concentration and composition as well as on the principle of the analytical method. However, there was a good correlation between the in vitro and in vivo assays for P. guajava and C. citratus extracts, since these results showed a reduction in the intracellular ROS concentration in the hepatocytes exposed to P. guajava extract. Furthermore, no antioxidant effect was observed in the assay with C. citrates extract, what is in agreement with the in vitro assays for this plant species. At last, the thesis intended to proof and to highlight the discrepant results obtained by independent methods for the same antioxidant species. Additionally, it aimed to compare the different methods according to the differences among the experimental procedures by using different ROS. Furthermore, the stability, reactivity and the half-life time of the free radicals are discussed. / O presente trabalho visa ao desenvolvimento de novas metodologias para a determinação da capacidade antioxidante in vitro e in vivo contra radicais peroxila e hidroxila em amostras de 5 plantas medicinais (Matricaria chamomilla L., Psidium guajava, Achyrocline satureoides, Baccharis genistelloides e Cymbopogon citratus) e sete compostos fenólicos (rutina, quercetina, resveratrol, ácido cafeico, ácido ferúlico, ácido gálico e ácido rosmarínico). A capacidade antioxidante das amostras foi determinada por dois métodos independentes. No primeiro, testou-se uma nova forma para a detecção de radicais hidroxila, usando-se um procedimento eletroquímico no qual os radicais destroem uma monocamada auto-organizada de hexanotiol sob um eletrodo de ouro. Esta monocamada consegue inibir o sinal eletroquímico de um par redox dissolvido, e, ao ser atacada por radicais livres, estes a destroem, e a recuperação do sinal eletroquímico do par redox se dá de forma proporcional à extensão de dissolução da monocamada. No segundo método, a determinação da capacidade antioxidante contra radicais peroxila e hidroxila ocorre através da detecção indireta destes radicais por fluorimetria (ex/em: 485/520 nm), empregando diacetato de 2 ,7 -diclorofluoresceína (DCFH-DA) como substrato. O DCFH-DA, após sofrer desacetilação, pode ser oxidado por espécies reativas de oxigênio (ERO), gerando fluorescência. Os radicais peroxila foram gerados através da termodegradação do reagente cloreto de 2,2 -azobis (2-metilamidinopropano) (ABAP) a 37ºC, enquanto os radicais hidroxila foram gerados pela reação de Fenton. Ensaios in vivo foram avaliados utilizando-se hepatócitos do peixe zebra (Danio rerio), expostos aos extratos das plantas P. guajava e C. citratus (melhor e pior desempenho in vitro, respectivamente). Os resultados obtidos pelos métodos eletroquímico e fluorimétrico demonstram que é extremamente difícil classificar as plantas de acordo com sua capacidade antioxidante, uma vez que o resultado obtido depende fortemente da concentração e da composição química dos extratos, além do princípio do método analítico empregado. Entretanto, obteve-se uma boa correlação entre os ensaios in vitro e in vivo feitos para os extratos das plantas P. guajava e C. citratus, já que os resultados obtidos mostraram uma redução da concentração de ERO intracelular em forma dose-dependete nos hepatócitos expostos a P. guajava, sem se observar efeito antioxidante no ensaio com C. citratus, a exemplo dos ensaios in vitro realizados. O presente trabalho pretende chamar a atenção para esses diferentes resultados obtidos, e compara os diferentes métodos levando em consideração as diferenças existentes entre as metodologias, assim como as diferenças existentes entre os radicais testados. Além disso, discute-se a estabilidade, a reatividade e tempo de meia-vida destas espécies.

In biological systems, several metabolic and environmental factors are responsible for the production of reactive oxygen (ROS) and nitrogen (RNS) species. The exacerbated production of these reactive species or the significant decrease in the effectiveness of the defenses against them causes the redox imbalance, with consequent damage to biological macromolecules, which is associated to the emergence and progression of several diseases. Among the exogenous antioxidants, phenolic compounds of plants have been highlighted by their ability to scavenge various reactive species. Among the most studied plants, Curcuma longa has many beneficial properties for health, which are mainly associated with the phenolic compounds present in the rhizome, known as curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin), but the benefits are more particularly attributed to curcumin. Thus, the objective of this study was to investigate the antioxidant capacity of C. longa extracts and to determine their contents in curcuminoids, investigating the antioxidant action of the extract and of each isolated curcuminoid against ROS and RNS. The identification and quantification of curcuminoids, the total phenols content analysis and the antioxidant capacity in terms of scavenging of the 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH•) and the FRAP method (ferric reducing antioxidant power) were carried out for methanol (CM), defatted methanol (CHM), ethanol (EC) and hexane (CH) extracts, besides commercial extract (ExtFarC) of C. longa. The elimination capacity of ROS and RNS was performed for CE and isolated curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin). The CM, CHM and CE had shown the three curcuminoids in their compositions, while, for the ExtFarC, curcumin was the predominant compound. The CE and ExtFarC had shown the higher total phenols content and antioxidant activity by the FRAP method, besides the lower IC50 values for to the DPPH• radical. The use of high performance liquid chromatography (HPLC) with spectrophotometric detection (DPPH•-HPLC-UV) indicated that curcumin and demethoxycurcumin had the greater potential for capture of DPPH•, as observed by the reduction of their peaks in HPLC, at equivalent contact time. In the experiments related to hypochlorous acid (HOCl), nitric oxide (•NO) and peroxynitrite (ONOO‾), the curcuminoids had shown a direct efficiency toward their elimination, similarly to the positive control (quercetin). For CE, despite showing higher IC50 values against these species, in comparison to the isolated compounds, it presented lower IC50 values, when compared to other plants’ extracts, studied by the same methods. Although curcumin is the target of many therapeutic studies for a number of diseases, the present data show that the three curcuminoids can play essential roles against ROS and RNS, and thus, may be considered promising in the prevention and treatment of diseases related to oxidative stress. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Em sistemas biológicos, diversos fatores metabólicos e ambientais são responsáveis pela produção de espécies reativas de oxigênio (EROs) e de nitrogênio (ERNs). Quando a produção é exacerbada ou quando há uma diminuição significativa na eficácia das defesas contra essas espécies, ocorre o desequilíbrio redox, causando danos a macromoléculas biológicas, o que está associado ao surgimento e progressão de várias doenças. Dentre os antioxidantes exógenos, os compostos fenólicos de plantas têm se destacado pela capacidade de sequestrar diversas espécies reativas. Das plantas mais estudadas, a Curcuma longa apresenta inúmeras propriedades benéficas para a saúde, que são principalmente associadas aos compostos fenólicos presentes no rizoma, conhecidos como curcuminoides (curcumina, desmetoxicurcumina e bisdesmetoxicurcumina), porém os benefícios são mais particularmente atribuídos à curcumina. Dessa forma, o objetivo do estudo foi avaliar a capacidade antioxidante de extratos de C. longa e determinar seus curcuminoides constituintes, investigando a ação antioxidante do extrato e de curcuminoides isolados contra as EROs e ERNs. A identificação e quantificação de curcuminoides, o conteúdo total de fenóis e a capacidade antioxidante, em termos de sequestro do radical 2,2-difenil-1-picrilhidrazila (DPPH•) e pelo método de FRAP (do inglês, ferric reducing antioxidant power), foram realizados para os extratos metanólico (CM), metanólico desengordurado (CHM), etanólico (CE), hexânico (CH) e para o extrato comercial (ExtFarC) de C. longa. A capacidade de eliminação das EROs e ERNs foi realizada para o CE e para os curcuminoides (curcumina, desmetoxicurcumina e bisdesmetoxicurcumina). Os extratos CM, CHM e CE apresentaram, em suas composições, os três curcuminoides, enquanto o ExtFarC apresentou a curcumina como componente majoritário. O extrato etanólico (CE) e o ExtFarC apresentaram maior conteúdo total de fenóis e atividade antioxidante pelo método de FRAP e menores valores de IC50 frente ao radical DPPH•. Cromatografia líquida de alta eficiência (CLAE), com detecção espectrofotométrica (DPPH•-CLAE-UV) indicou que a curcumina e a desmetoxicurcumina apresentaram maior potencial de captura de DPPH•, observado pela redução de seus picos em CLAE, em tempos de contato equivalentes. Nos experimentos de capacidade de eliminação das EROs e ERNs, os curcuminoides apresentaram atividade semelhante ao controle positivo (quercetina) frente ao ácido hipocloroso (HOCl), óxido nítrico (•NO) e peroxinitrito (ONOO‾), mostrando-se eficientes de forma direta contra essas espécies. O extrato CE, apesar de exibir maiores valores de IC50 para essas espécies, quando comparado aos compostos isolados, apresentou valores de IC50 inferiores, em comparação com extratos de outras plantas estudados pelos mesmos métodos. Apesar de a curcumina ser o alvo de estudos com finalidade terapêutica para inúmeras doenças, os dados evidenciam que os três curcuminoides têm papel potencial contra as EROs e ERNs, desta forma são promissores na prevenção e tratamento de doenças relacionadas ao estresse oxidativo.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-03-24T19:51:42Z No. of bitstreams: 2 Dissertação - Márcio Fernando Cardoso Zago - 2014.pdf: 2344826 bytes, checksum: 785eb9d33a3c85c094d7e3cfc111730d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-03-24T19:54:15Z (GMT) No. of bitstreams: 2 Dissertação - Márcio Fernando Cardoso Zago - 2014.pdf: 2344826 bytes, checksum: 785eb9d33a3c85c094d7e3cfc111730d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-03-24T19:54:16Z (GMT). No. of bitstreams: 2 Dissertação - Márcio Fernando Cardoso Zago - 2014.pdf: 2344826 bytes, checksum: 785eb9d33a3c85c094d7e3cfc111730d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-05-29 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The jaboticaba (Myrciaria cauliflora Berg) is a much appreciated fruit and nutritional importance mainly found in south-central Brazil. Its peel is a by-product discarded in Brazilian agribusiness. The concern about the environmental impacts and the high rate of losses and waste leads to the need to establish means for recovery of such waste. This study aimed to produce flour with peel Jaboticaba, develop biscuit cookie type - enriched with Jaboticaba skin flour instead of wheat flour and oatmeal, analyze technological aspects as much flour as biscuits made from mathematical models and analyze the chemical and functional aspects as much flour as the elaborate cookies. Chemical, microbiological and physical analyzes technological peel blemish in flour and crackers cookie type , according to official methods . All regression models cookies with bark were significant blemish . The water activity of all the cookies found below the recommended limit for the growth of microorganisms . Cookies formulations with higher proportions of bark flour have minor blemish that cookies formulations with higher proportions of oatmeal and wheat flour thicknesses . Moreover , they absorb more water , show lower values of breaking strength and have a decrease in color parameters ( L *, a * and b *) . Cookies made with larger fractions of oatmeal have lower values of specific volume . Formulations with higher percentages of bark jabuticaba flour and wheat flour tend to be more soluble in water . Both the standard cookie and cookie with Jabuticaba bark , selected from the desirability test meal were accepted among children 9-14 years. Embed shell flour blemish on cookies cookie type , increases the health benefits , since this has a high antioxidant capacity and high content of dietary fiber . It is necessary to apply further studies shell flour blemish in food products , since high levels of dietary fiber and bioactive compounds were detected in the peel blemish flour and crackers which were developed in this study. / A jabuticaba (Myrciaria cauliflora Berg) é um fruto muito apreciado e de importância nutricional encontrado principalmente no centro-sul do Brasil. Sua casca é um subproduto descartado nas agroindústrias brasileiras. A grande preocupação com os impactos ambientais e o elevado índice de perdas e desperdícios leva à necessidade de se criarem meios para o aproveitamento desses resíduos. Este estudo teve como objetivo produzir farinha com casca de jabuticaba, desenvolver biscoito tipo cookie - enriquecido com a farinha de casca de jabuticaba em substituição da farinha de trigo e farinha de aveia, analisar os aspectos tecnológicos tanto da farinha quanto dos cookies elaborados a partir de modelos matemáticos e analisar os aspectos químicos e funcionais tanto da farinha quanto dos cookies elaborados. Foram realizadas análises físicas, químicas, microbiológicas e tecnológicas na farinha de casca de jabuticaba e nos cookies, conforme metodologias oficiais. Todos os modelos de regressão dos cookies com casca de jabuticaba foram significativos. A atividade de água de todas as formulações encontrou-se abaixo do limite recomendado para o desenvolvimento de microrganismos. Cookies com maiores proporções de farinha de casca de jabuticaba possuíram espessuras menores que formulações com maiores proporções de farinha de aveia e farinha de trigo. Além disso, absorveram mais água, apresentaram menores valores de força de ruptura e possuíam diminuição nos parâmetros de cor (L*, a* e b*). Cookies produzidos com frações de farinha de aveia maiores apresentaram menores valores de volume específico. Formulações com maiores percentuais de farinha de casca de jabuticaba e farinha de trigo foram mais solúveis em água. Tanto o cookie padrão e o cookie com farinha de casca de jabuticaba, selecionado a partir do teste de desejabilidade, foram aceitos entre crianças de 9 a 14 anos. Incorporar farinha de casca de jabuticaba em cookie, aumenta os benefícios à saúde, visto que, esta possui atividade antioxidante e fibras alimentars. Faz-se necessário mais estudos de aplicação de farinha de casca de jabuticaba em produtos alimentícios, uma vez que foram verificadas fibras alimentares e compostos bioativos na farinha de casca de jabuticaba e nos cookies que foram desenvolvidos neste estudo.

O morango representa boa fonte de vitamina C, flavonóides e derivados de ácido elágico e tem ampla aceitação pela população brasileira. Os objetivos deste trabalho foram: caracterizar diferentes cultivares quanto aos teores de compostos bioativos, otimizar a metodologia para quantificação do conteúdo de ácido elágico total, purificar e caracterizar estruturalmente os elagitaninos, avaliar sua potencial atividade antiproliferativa, anti-diabetes tipo 2 e anti-hipertensão, estudar a biodisponibilidade desses compostos in vivo. Os resultados demonstraram que existem diferenças significativas nos teores de compostos bioativos entre as cultivares. As melhores condições para determinação dos teores de ácido elágico total foram: extração em acetona 80% e posterior hidrólise com ácido trifluoracético (TFA) 2N a 120 °C por 60 minutos. A precipitação com acetato de itérbio foi o método mais eficiente para a purificação dos elagitaninos. Após administração por gavagem a ratos de um purificado de elagitaninos, nenhum composto foi detectado no plasma e tecidos analisados. / Strawberries are a good source of vitamin C, flavonoids and ellagic acid derivatives and are consumed in Brazil in large quantities. The objectives of the present work were: characterization of different cultivars with respect to the content of bioactive compounds, optimization of the method for quantification of total ellagic acid content, purification and structural identification of ellagitannins, evaluation of their antiproliferative, anti-type 2 diabetes and anti-hypertension potentials, investigation of their in vivo bioavailability. The results indicated that there were significant differences in bioactive compounds among the strawberry cultivars. The best conditions for the determination of the total ellagic acid content in strawberries were extraction with 80% acetone, and hydrolysis using 2N TFA at 120 °C for 60 minutes. The precipitation method using ytterbium acetate was the most efficient in purifying the ellagitannins. After administration of purified ellagitannins to rats by gavage, no compound was detected in plasma or tissue samples analyzed.

Thesis (M. Sc. (Zoology)) -- University of Limpopo, 2015 / The unprecedented expansion in human population and industry, since the industrial revolution in the late 1700s, has led to increased anthropogenic activities which have indisputably impacted freshwater ecosystems and biological communities therein, including fish. Although this has understandably been the focus, under natural aquatic conditions, no organism is only affected by pollution. Parasites have also been shown in a number of interdisciplinary studies to affect the health of aquatic hosts (amphibians, crustaceans, fish, and mammals). This is illustrated in a number of comprehensive studies the detrimental effects parasites exacerbate when their hosts (fish) are stressed. Therefore, the ability of parasites to interact with anthropogenic stressors, as well as effects they have on the genetic, cellular or tissue level of their host is crucial in conservation and sustaining aquatic biodiversity. As such, the present study examined the combined effects of pollution and metazoan parasites on the health and oxidative stress biomarkers, evaluated for the first time for silver carp, Hypophthalmichthys molitrix (Valenciennes, 1844) and rednose mudfish, Labeo rosae Steindachner, 1894, in one of South Africa’s impacted freshwater ecosystems, Flag Boshielo Dam, Olifants River System, Limpopo Province. Seasonal surveys were conducted from February 2012–January 2013. A total of 111 H. molitrix and 116 L. rosae fish specimens were collected using conventional angling gear, scoop and gill nets with stretched mesh sizes of 30–110 mm. The two selected cyprinid fish species were assessed for oxidative stress biomarkers [Glutathione S-transferase (GST), lipid peroxidation (MDA) and Total Antioxidant Capacity (TAC)] and parasitism of metazoan parasites. Concentrations of biomarkers of oxidative damage and antioxidant defense in the gill and liver tissue were measured to assess how these major organs of the immune system responded to oxidative stress associated with parasitic infections. In addition, water quality analyses were carried out by testing an assay of physico-chemical parameters to establish the level of contamination. Fish health was assessed using the Health Assessment Index (HAI), refined Parasite Index (PI), Inverted Parasite Index (IPI) and Condition Factor (K) protocols. Relative to previous studies at Flag Boshielo Dam, water quality results showed an increase of nutrients, major ions and several metals which may have adverse effects that may comprise fish health; however, this dam remains moderately polluted in a mesotrophic state. The fish health assessment results indicated that H. molitrix was more affected in terms of the necropsy and parasite based assessments (HAI, IPI and K) with mean±SD of 65.68±35.51; 68.29±25; 0.82±0.20, respectively, as compared to 39.14±22.44; 28.79±18.33; 1.17±0.21 for L. rosae during the study. In addition, significantly higher parasitic infections (mean prevalence of infection with any species of parasite = 45.3±0.13) were observed for H. molitrix than L. rosae (12.0±0.05). Furthermore, there was considerable variation in biomarker concentration between highly infected and non-infected fish, for and between each species and tissues with regard to parasite infection, suggesting that the specific functions of each tissue are associated with their susceptibility to oxidative stress, as well as their ability to defend against oxidative damage. These results illustrate that although fish are affected by aquatic contaminants they are to an extent affected by parasites, which may act synergistically on the health of the two fish species. Most importantly, it was suggested that knowledge on the parasites of alien H. molitrix when compared to indigenous L. rosae may give an indication of how adaptive this fish are to new localities as well as expands the information on the rarely studied biology, epizootiology and ecological interactions of these two cyprinid species. Keywords: Health Assessment Index, refined Parasite Index, Inverted Parasite Index, Condition Factor, water quality, lipid peroxidation, Glutathione S-transferase, Total Antioxidant Capacity, Hypophthalmichthys molitrix, Labeo rosae, Flag Boshielo Dam.

The thesis deals with one group of small fruits, the genus Ribes, ie currants, and gooseberries. It assesses their physical and chemical parameters with respect to the possibility of use in these properties for the design of a new type of product – beverages. Titration, spectrometric and other methods were used for the evaluation of these properties. A total of fifteen of these methods were available (total juice yield, juice pH, total dry matter, soluble fruit dry matter, titratable acidity, formolic number, reducing carbohydrate content, D-glucose, D-fructose and sucrose content, total polyphenolic substance content, anthocyanins, vitamin C and total antioxidant capacity) and a total of nine analytes (copper, iron, magnesium, phosphorus, zinc, manganese, potassium, calcium and sodium) were selected for the determination of mineral elements. For two years (harvest 2014 and 2015), these parameters were monitored for color variants of currants (white, red and black) and gooseberries (green-fruited, yellow-fruited and red-fruited). A total of six white varieties were tested in the group of currants - Olin, Jantar, Primus, Blanka, Viktoria and Orion, eleven varieties of red currants - Junnifer, Jesan, Detvan, Rovada, Rubigo, J.V.Tets, Tatran, Losan, Kozolupský raný, Stanca, NŠLS 11/6 and eleven varieties of black currant varieties - Ometa, Démon, Triton, Ben Hope, Ruben, Ben Gairm, Ben Lomond, Moravia, Ben Conan, Fokus and Ceres. A total of five varieties of green-fruited gooseberries were tested - Zebín, Mucurines, Rixanta, Rodnik and Prima. Yellow-fruited gooseberries were available in four varieties - Lemon Giant, Invicta, Golden Fig and Darek. The red-fruited gooseberries included varieties - Rolonda, Alan, Karat, Karmen, Krasnoslawjanskij, Himnomacki Rot, Remarka, Tamara and Black Neguš. Using Tukey's test, an analysis was performed and by scoring individual varieties based on the results of individual analyzes, a list of recommended varieties was compiled for further work on the development of a new beverage. Unfortunately, in 2015 there were very few fruits of green-fruited and yellow-fruited gooseberries, so it was decided that the volume of fruit will fall on the testing of bioactive substances. Therefore, at the end of the work, it is not possible to objectively evaluate these varieties of gooseberries in the whole range of analyzes. Therefore, the point evaluation of varieties was based on the averages of individual varieties. The red-fruited varieties were available in full, and therefore a full evaluation could take place. The most significant differences between red gooseberry varieties were in the content of bioactive substances (the content of polyphenolic substances, anthocyanin dyes, and vitamin C) and antioxidant capacity. These results were provided to the food subject, which continued to work with these values. Based on this project, utility model No. PUV 2016-33171 and the resulting combined drink based on wine and fruit juice were developed.

Philosophiae Doctor - PhD / The aim of this thesis is first, to evaluate various culture conditions to improve embryo development, and secondly, to analyze spent culture media for any biomarkers that may be predictive of embryo health. Single-step and sequential culture media were compared in both Planer and EmbryoScope™ incubators. Single-step media resulted in better blastocyst development compared to sequential media and the EmbryoScope™ incubation system showed slight improvements in embryo development than the Planer system. The benefits of supplementing the culture medium with either insulin or insulin-like growth factor 1 (IGF-1) or culturing in a 2% O2 environment, using two different strains of mice (hybrid and C57), as well as the suitability of these strains for quality control were compared. In insulin, hybrid embryos were slower to blastulate and had a lower blastocyst rate, whereas C57 embryos were slower to the morula and faster to blastocyst stages, and lower blastocyst rate than the controls. IGF-1 showed no difference in time-lapse morphokinetics (TLM) or blastocyst rates compared to controls in both hybrid and C57 embryos. Under 2% O2, hybrid embryos showed no significant difference in TLM up to the 8-cell stage, but slowed down afterwards, resulting in blastocysts with significantly lower cell counts than the 6% O2 group. The C57 embryos were slower to reach morula and expanded blastocyst, and had lower blastocyst rates in 2%O2 vs 6%O2. The C57 strain had significant slower overall embryo development for all time points than hybrid embryos in insulin, IGF-1 and ultra-low O2, as well as lower blastocyst rates. Measurement of growth differentiation factor 9 (GDF-9) and oxidation-reduction potential (ORP) in spent media as markers for embryo health were evaluated. Day 5 human blastocysts yielded higher pregnancy rates and GDF-9 levels in spent media compared to Day 6 blastocysts, but TLM parameters showed no impact on pregnancy outcome. In Day 6 blastocysts, the non-pregnant group showed significantly faster embryo development compared to the clinically pregnant group up to the 8-cell stage and start of blastulation. GDF-9 did not show any significant differences between non-pregnant and pregnant groups of Day 5 or Day 6 embryo transfers. ORP in spent media from good quality Day 3 embryos that developed into blastocysts were significantly higher than from those that did not, with no difference in control medium ORP. Spent media from arrested embryos showed lower ORP than their corresponding controls. Arrested embryos had slower development at syngamy, morula, blastulation and blastocyst stages. The single step medium in the EmbryoScope™ is the preferred choice for embryo culture. Insulin or IGF-1 media supplementation or 2% O2 culture did not provide any benefit to embryo development. The C57 mouse strain is more sensitive and may be better to detect changes in culture conditions, and therefore better model for quality control assays. GDF-9 values decrease from Day 5 to Day 6 which gives new insight to understanding the role of GDF-9 during embryogenesis. ORP in spent media indicate that embryos that developed into blastocysts did not contribute to ROS, but maintained ORP balance.