Desenvolvimento de um aditivo natural à base de catequina: otimização da extração e estabilização a partir de frutos de Arbutus unedo L.

The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources.The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources. This study compares and optimizes the maceration, microwave and ultrasound extraction techniques in the recovery of a catechin extract from Arbutus unedo L. fruits and evaluate the stability of flavan-3-ols during storage and application processes. To obtain conditions that maximize the catechin extraction yield, a response surface methodology was used. Maceration and microwave extractions were found to be the most effective methods, capable of yielding 1.38±0.1 and 1.70±0.3 mg of catechin/g dry weight (dw) in the corresponding optimal extraction conditions. The optimal conditions for maceration were 93.2±3.7 min, 79.6±5.2 ºC and 23.1±3.7 % of ethanol, while for the microwave extraction were 42.2±4.1 min, 137.1±8.1 ºC and 12.1±1.1 % of ethanol. The microwave system was a quicker solution, conducting to slightly higher yields of catechin than maceration, but this one needed lower temperatures to reach similar yields. The ultrasound method was the least effective solution in terms of catechin yield extraction (0.71±0.1 mg/g at 42.4±3.6 min, 314.9±21.2 W and 40.3±3.8 %. ethanol). The stability was tested with of the catechin-enriched extract (60% flavan-3-ols and 22% catechin), obtained under the best maceration conditions, was tested. Therefore, catechin-enriched extracts were submitted to physical and chemical stability studies, considering the main affecting variables (time, temperature and pH): i) a stability study of the extracts during storage as powder system; and ii) a stability study of the extracts in simulated food environment (aqueous solution system). The measured responses were the flavan-3-ols and catechin contents, determined by HPLC-DAD, and the antioxidant activity of the extracts evaluated by hydrophilic assays. Mechanistic and phenomenological equations were used to describe the responses, and the optimal conditions for flavan-3-ols (including catechin) stability as powder extract during a month were pH= 5.4 and T= -20ºC; while its stability in aqueous solution remained during the 24 h of application at pH<4 and T<30ºC. This study compares and optimizes the maceration, microwave and ultrasound extraction techniques in the recovery of a catechin extract from Arbutus unedo L. fruits and evaluate the stability of flavan-3-ols during storage and application processes. To obtain conditions that maximize the catechin extraction yield, a response surface methodology was used. Maceration and microwave extractions were found to be the most effective methods, capable of yielding 1.38±0.1 and 1.70±0.3 mg of catechin/g dry weight (dw) in the corresponding optimal extraction conditions. The optimal conditions for maceration were 93.2±3.7 min, 79.6±5.2 ºC and 23.1±3.7 % of ethanol, while for the microwave extraction were 42.2±4.1 min, 137.1±8.1 ºC and 12.1±1.1 % of ethanol. The microwave system was a quicker solution, conducting to slightly higher yields of catechin than maceration, but this one needed lower temperatures to reach similar yields. The ultrasound method was the least effective solution in terms of catechin yield extraction (0.71±0.1 mg/g at 42.4±3.6 min, 314.9±21.2 W and 40.3±3.8 %. ethanol). The stability was tested with of the catechin-enriched extract (60% flavan-3-ols and 22% catechin), obtained under the best maceration conditions, was tested. Therefore, catechin-enriched extracts were submitted to physical and chemical stability studies, considering the main affecting variables (time, temperature and pH): i) a stability study of the extracts during storage as powder system; and ii) a stability study of the extracts in simulated food environment (aqueous solution system). The measured responses were the flavan-3-ols and catechin contents, determined by HPLC-DAD, and the antioxidant activity of the extracts evaluated by hydrophilic assays. Mechanistic and phenomenological equations were used to describe the responses, and the optimal conditions for flavan-3-ols (including catechin) stability as powder extract during a month were pH= 5.4 and T= -20ºC; while its stability in aqueous solution remained during the 24 h of application at pH<4 and T<30ºC.

Otimização da extração de ergosterol assistida por microondas a partir de Agaricus bisporus L., aplicando a técnica estatística de superfície e resposta combinada com a técnica de HPLC-UV

O aumento de consumo de cogumelos tem-se verificado em todo o mundo, não só pelo seu valor nutricional, sabor apurado e textura, mas também pelas suas propriedades medicinais. Existem vários estudos científicos que descrevem os benefícios do consumo de cogumelos, que advêm da sua riqueza em compostos bioativos, tais como micosteróis, em particular, ergosterol. Agaricus bisporus L. é o cogumelo mais consumido em todo o mundo, sendo a sua fração de esteróis constituída essencialmente por ergosterol (90%) [1], tornando a sua extração um tópico de elevado interesse já que esta molécula apresenta elevado valor comercial e inúmeras aplicações nas indústrias alimentar, farmacêutica e cosmética. Segundo a literatura, o teor de ergosterol pode variar entre 3 e 9 mg por g de cogumelo seco. Atualmente, os métodos tradicionais tais como a maceração e a extração em Soxhlet estão a ser substituídos por metodologias emergentes, nomeadamente a extração assistida por microondas, visando diminuir a quantidade de solvente utilizado e o tempo de extração e, naturalmente, aumentar o rendimento da mesma. No presente trabalho, utilizou-se A. bisporus como fonte de ergosterol, tendo-se otimizado as seguintes variáveis relevantes para a sua extração pela tecnologia de microondas (MAE): tempo (0-20 min), temperatura (60-210 ºC) e razão sólido-líquido (1-20 g/L). O solvente utilizado foi o etanol tendo-se aplicado a técnica estatística de superfície de resposta por forma a gerar modelos matemáticos que permitissem maximizar a resposta e otimizar as variáveis que afetam a extração de ergosterol. O conteúdo em ergosterol foi monitorizado por HPLC-UV. Os resultados demonstraram que a técnica MAE é promissora para a extração de ergosterol, tendo-se obtido, para as condições ótimas (20,4 min, 121,5ºC e 1,6 g/L), 569,4 mg ergosterol/100 g de massa seca, valor similar ao obtido com extração convencional por Soxhlet (671,5±0,5 mg/100 g de massa seca). Em síntese, a extração assistida por microondas demonstrou ser uma tecnologia eficiente para maximizar o rendimento de extração em ergosterol.

A importância dos ácidos fenólicos na atividade anti-inflamatória de cogumelos comestíveis

Os cogumelos comestíveis são uma fonte rica de moléculas bioativas que lhes conferem importantes atividades biológicas. Moléculas como os Polissacáridos, terpenóides e os compostos fenólicos têm sido descritos como os componentes mais importantes no que respeita á atividade anti-inflamatória dos cogumelos (1). No presente trabalho, os extratos etanólicos de cogumelos comestíveis foram obtidos por maceração e caracterizados quimicamente em termos de ácidos fenólicos por técnicas de HPLC-DAD. Além disso, derivados metilados e glucuronados dos ácidos fenólicos identificados foram também sintetizados com o objetivo de mimetizar reações de metabolização no organismo e estudar a capacidade destas moléculas de manter a bioatividade exibida inicialmente. Os extratos obtidos, os ácidos fenólicos e compostos sintetizados foram avaliados pela sua atividade anti-inflamatória. De entre as amostras analisadas, B. impolitus revelou o mais elevado conteúdo em ácidos fenólicos (675 ± 23 μg/g), seguido de C. cibarius > A. caesaria > L. deliciosus > B. aereus > M. esculenta > B. edulis; devido à contribuição do ácido cinâmico que foi encontrada em maior quantidade nesta amostra (505 ± 12 μg/g). Mais ainda, B. impolitus apresentou também maior inibição da produção de NO (EC50=166 ± 10 μg/mL) seguido das amostras A. caesaria > C. cibarius > L. deliciosus > M. esculenta > B. aereus > B. edulis. No que respeita aos compostos individuais, o ácido cinâmico (CA) revelou a atividade mais forte (EC50 = 182 ± 16 μM), seguido pelos ácidos p-hidroxibenzóico (HA) (239 ± 29 μM) e p-Cumárico (CoA) (442± 33 μM), o que realça a importância destas moléculas para a atividade anti-inflamatória dos cogumelos. Comparando a atividade exibida pelos ácidos fenólicos com os respetivos derivados, é possível verificar a seguinte ordem de atividades: ácido p-hidroxibenzóico: HA > HA-M3 > HA-M2 > HA-M1 > HA-G; ácido p-cumárico : CoA-M1 > CoA-G > CoA-M2 > CoA-M3 > CoA e ácido cinâmico: CA-G > CA > CA-M1. Perante os resultados obtidos é de realçar a importância dos ácidos fenólicos na contribuição para a bioatividade exibida pelos cogumelos em estudo. Mais ainda, foi possível concluir que as alterações das moléculas pelas reações de conjugação no organismo têm influência na bioativade das moléculas iniciais, sendo que muitas vezes esta atividade é aumentada.

Preenchimento sedimentar de duas zonas de dinâmica diferenciada no sapal de Castro Marim

Dissertação de mest. em Estudos Marinhos e Costeiros Ramo Gestão Costeira, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2002

Avaliação da biodessulfurização de 4-metildibenzotiofeno por Rhodococus rhodochrous (NRRL B-2149)

Crude oil has between 0.04 up to 5% of sulphur, the higher the oil the higher the sulphur levels. Sulphur usually gives problems such as corrosion in refinery, and once burnt produces SO2 that goes to atmosphere. This work aim to investigate the capacity of Rhodococcus rhodochrous (NRRL B-2149) to metabolize the model compound 4-methyldibenzotiophene (4-MDBT), to remove the sulphur and transform it in 2-hydroxybiphenyl (2-HBF) and sulphite using the 4S pathway. Kynetic runs were carried out in shaker at 120 rpm and 32°C. Samples were taken every 12h to assay substrate consume as well as cells production using HPLC. Results showed that R. rhodochrous NRRL B-2149 can use the 4S pathway in order to remove sulphur without change the carbon chain of the molecule as well as that cells and 4-MDBT affects the product formation. The production of 2-hydroxybiphenyl has interest for industry once it is a potent biocide. However, evaluation is necessary in order to obtain better results compatible with industry needs

Potencial larvicida de extratos de plantas regionais no controle de larvas de Aedes Aegypti (Diptera:Culicidae)

Dengue, amongst the virus illnesses one can get by vectorial transmission, is the one that causes more impact in the morbidity and mortality of world s population. The resistance to the insecticides has caused difficulties to control of vector insect (Aedes aegypti) and has stimulated a search for vegetables with larvicidal activity. The biodiversity of Caatinga is barely known and it is potential of use even less. Some plants of this biome are commercialized in free fairs northeast of Brazil, because of its phytotherapics properties. The vegetables in this study had been selected by means of a questionnaire applied between grass salesmen and natives of the Serido region from Rio Grande do Norte state; culicids eggs had been acquired with traps and placed in container with water for the larva birth. Thirty larvae had been used in each group (a group control and five experimental groups), with four repetitions four times. The vegetables had been submitted to the processes of decoction, infusion and maceration in the standard concentration of 100g of the vegetable of study in 1l of H2O and analyzed after ½, 1, 2, 4, 8, 12, 24 and 48 hours for verification of the average lethal dose (LD50) from the groups with thirty larva. The LD50 was analyzed in different concentrations (50g/l, 100g/l, 150g/l, 200g/l e 300g/l) of Aspidosperma pyrifolium Mart. 48 extracts of rind, leaf and stem of the seven vegetal species: Aspidosperma pyrifolium Mart., Mimosa verrucosa Benth, Mimosa hostilis (Mart.) Benth., Myracrodruon urundeuva Allemão, Ximenia americana L, Bumelia sartorum Mart Zizyphus joazeiro Mart, had been analyzed. The extracts proceeding from the three methods were submitted to the freezedrying, to evaluate and to quantify substances extracted in each process. The results had shown that Aspidosperma pyrifolium Mart. and Myracrodruon urundeuva Allemão are the species that are more distinguished as larvicidal after 24 hours of experiment, in all used processes of extraction in the assays. The Zizyphus joazeiro Mart species has not shown larvicidal activity in none of the assays. In relation to the extraction method, the decoction was the most efficient method in the mortality tax of the A. aegypti larvae

Contents and bioconversion of B-glycoside isoflavones to aglycones in the processing conditions of soybean tempeh.

The objective of this work was to evaluate the effect of the processing conditions of soybean tempeh on the contents of ??glycoside isoflavones and on their bioconversion into aglycones. Different times of soaking (6, 12, and 18 hours), cooking (15, 30, and 45 minutes), and fermentation (18, 24, and 30 hours) with Rhizopus oligosporus at 37°C were evaluated for tempeh preparation. Grains from the cultivar 'BRS 267' were used, and the experiment was carried out according to a central composite design (23). The response functions comprised the contents of genistin, malonyldaidzin, malonylgenistin, daidzein, and genistein, quantified by ultraperformance liquid chromatography (UPLC). Soaking, cooking, and fermentation times change the content, profile, and distribution of the different forms of isoflavones in tempeh. The highest bioconversion of glycoside isoflavones into aglycones occurred in 6?hour soaked soybean grains, whose cotyledons were cooked for 15 minutes and subjected to 18?hour fermentation. RESUMO:O objetivo deste trabalho foi avaliar o efeito das condições de processamento do tempeh de soja sobre o conteúdo de isoflavonas ??glicosídeos e sobre sua bioconversão em agliconas. Diferentes tempos de maceração (6, 12 e 18 horas), cozimento (15, 30 e 45 minutos) e fermentação (18, 24 e 30 horas) com Rhizopus oligosporus a 37°C foram avaliados na preparação do tempeh. Foram utilizados grãos da cultivar 'BRS 267', e o experimento foi realizado de acordo com um delineamento composto central (23). As funções?respostas compreenderam o teor de genistina, malonildaidzina, malonilgenistina, daidzeína e genisteína, quantificadas por cromatografia líquida de ultraeficiência (CLUE). Os tempos de maceração, cozimento e fermentação alteraram o conteúdo, o perfil e a distribuição das diferentes formas de isoflavonas no tempeh. A maior bioconversão de ??glicosídeos em agliconas ocorreu em grãos de soja macerados por 6 horas, cujos cotilédones foram cozidos por 15 minutos e submetidos à fermentação por 18 horas.