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Abstract(s)
Este trabalho teve como objectivo o estudo do potencial antioxidante e anticancerígeno dos compostos presentes no efluente do processamento da cortiça. Recorrendo à tecnologia de membranas o efluente foi fraccionado em três fracções, enriquecidas em compostos com diferentes massas molares (designadas como fracção Pequena, Média e Grande.
Utilizaram-se dois lotes de membranas de ultrafiltração compostas por acetato de celulose, o CA1 e o CA4, com um limite de exclusão molecular de 3 e 74 kDa, respectivamente. Para a recuperação da fracção Média e Grande foi necessário recorrer a uma técnica de diafiltração. No final do processo de separação mediu-se a condutividade e a carga orgânica total das correntes de alimentação e permeado, onde se observou uma redução da carga orgânica comprovando a utilidade deste método no tratamento de efluentes industriais.
O efluente e as fracções recuperadas foram caracterizados em termos de Fenóis totais, Taninos e Açúcares redutores, tendo-se determinado que a fracção Grande é a mais rica nestes tipos de compostos, com 0,156 mg fenóis/mg extracto, 0,074 mg taninos/mg extracto e 0,278 mg glucose/mg extracto. Para além disso, foi utilizada a cromatografia líquida de fase reversa acoplada a espectrometria de massa na identificação dos compostos presentes no efluente e nas fracções, sendo que são maioritariamente compostos fenólicos, tais como, o ácido Quinico, Gálico, Protocatecuico, Elágico, entre outros. Para os compostos maioritários fez-se ainda o seu doseamento para cada fracção, tendo-se obtido para a fracção Grande 9 μg ácido Elágico/mg extracto.
Em termos de ensaios de actividade biológica foram realizados estudos de actividade antioxidante através do método do DPPH, em que o efluente e a fracção Grande demonstraram os melhores valores de EC50 de 23,48 e 23,79 μg/mL, respectivamente, e ensaios de citotoxicidade e inibição da proliferação celular em linhas celulares de cancro da mama MCF-7, onde mais uma vez a fracção Grande apresentou os valores mais promissores, tendo-se obtido um IC50 de 0,78 mg/mL para a citotoxicidade e 0,20 mg/mL para a inibição da proliferação celular. Os resultados obtidos permitiram concluir que nenhuma fracção é citotóxica, no entanto a fracção Grande apresenta potencial como agente citoestático das células de MCF-7.
Neste trabalho são ainda apresentados os resultados de um estudo preliminar sobre o efeito da fracção Grande sobre a linha celular tumoral em causa, nomeadamente nas suas proteínas e a permeação dos compostos.
This work aimed to study the antioxidant and anticancer potential of the compounds present in the cork processing wastewater. Using membrane technology the effluent was divided into three fractions, enriched in solutes of small, medium and large molecular weight. Two batches of cellulose acetate ultrafiltration membranes were used, the CA1 and CA4, with a molecular cut-off of 3 and 74 kDa, respectively. For the recovery of Medium and Large fractions it was used the diafiltration mode. At the end of the separation process, the conductivity and the total organic carbon were measured in the feed and in the permeate streams, where there was a reduction of the organic load demonstrating the utility of this method in the treatment of industrial effluents. The effluent and recovered fractions were characterized in terms of total phenols, tannins and reducing sugars, and it was found that the Large fraction is richer in these types of compounds with 0.156 mg phenol/mg extract, 0.074 mg tannin/mg extract and 0.278 mg glucose/mg extract. In addition, the liquid reverse phase chromatography coupled to mass spectrometry was used to identify the compounds present in the effluent and in the fractions, and they are mainly phenolic compounds, such as the Quinic acid, Gallic, Protocatechuic, Ellagic acid among others. The compounds present in higher quantity were quantified in the different fractions, and it was found that for the Large fraction there are 9 μg/mg extract of Ellagic acid. In terms of biological activity assays, the antioxidant activity was measured by DPPH method in which the effluent and the Large fraction showed the lowest EC50 values of 23.48 and 23.79 μg/mL, respectively. The cytotoxicity test and inhibition of cell proliferation in cell lines of breast cancer MCF-7 were also carried out, where again the Large fraction showed the most promising values, achieving an IC50 of 0.78 mg/ml for cytotoxicity and 0.20 mg/ml for inhibition of cell proliferation. The results showed that no fraction is cytotoxic, however the Large fraction has potential as a cytostatic agent of MCF-7 cells. In this work the results of a preliminary study on the effects of the Large fraction in the tumoral cell line are additionally presented, namely in the proteins and in the compounds permeation.
This work aimed to study the antioxidant and anticancer potential of the compounds present in the cork processing wastewater. Using membrane technology the effluent was divided into three fractions, enriched in solutes of small, medium and large molecular weight. Two batches of cellulose acetate ultrafiltration membranes were used, the CA1 and CA4, with a molecular cut-off of 3 and 74 kDa, respectively. For the recovery of Medium and Large fractions it was used the diafiltration mode. At the end of the separation process, the conductivity and the total organic carbon were measured in the feed and in the permeate streams, where there was a reduction of the organic load demonstrating the utility of this method in the treatment of industrial effluents. The effluent and recovered fractions were characterized in terms of total phenols, tannins and reducing sugars, and it was found that the Large fraction is richer in these types of compounds with 0.156 mg phenol/mg extract, 0.074 mg tannin/mg extract and 0.278 mg glucose/mg extract. In addition, the liquid reverse phase chromatography coupled to mass spectrometry was used to identify the compounds present in the effluent and in the fractions, and they are mainly phenolic compounds, such as the Quinic acid, Gallic, Protocatechuic, Ellagic acid among others. The compounds present in higher quantity were quantified in the different fractions, and it was found that for the Large fraction there are 9 μg/mg extract of Ellagic acid. In terms of biological activity assays, the antioxidant activity was measured by DPPH method in which the effluent and the Large fraction showed the lowest EC50 values of 23.48 and 23.79 μg/mL, respectively. The cytotoxicity test and inhibition of cell proliferation in cell lines of breast cancer MCF-7 were also carried out, where again the Large fraction showed the most promising values, achieving an IC50 of 0.78 mg/ml for cytotoxicity and 0.20 mg/ml for inhibition of cell proliferation. The results showed that no fraction is cytotoxic, however the Large fraction has potential as a cytostatic agent of MCF-7 cells. In this work the results of a preliminary study on the effects of the Large fraction in the tumoral cell line are additionally presented, namely in the proteins and in the compounds permeation.
Description
Trabalho final de mestrado para obtenção do grau de mestre em Engenharia Química e Biológica
Keywords
Efluente da cortiça Cork wastewater Membranas Membranes Ultrafiltração Ultrafiltration Actividades antioxidante e anticancerígena Antioxidant and anti-cancer activities Compostos fenólicos Phenolic compounds Linha celular do cancro da mama MCF-7 Breast cancer cell line MCF-7
Citation
GOMES, Luís Miguel Lopes - Estudo do efeito anti tumoral e antioxidante de diferentes fracções recuperadas do efluente da cortiça através do processo de membranas. Lisboa: Instituto Superior de Engenharia de Lisboa, 2016. Dissertação de mestrado.
Publisher
Instituto Superior de Engenharia de Lisboa