Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds

Detalhes bibliográficos
Autor(a) principal: Rosales, Thiécla Katiane Osvaldt
Data de Publicação: 2022
Tipo de documento: Tese
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/9/9131/tde-14102022-160404/
Resumo: Anthocyanins are sensitive flavonoid pigments and are widely distributed in nature. Currently, these phenolic compounds are being extensively researched due to intense interest in their beneficial properties. Due to their functionalities, they can be added to food products for natural coloring, functional ingredients, and in the development of nutritional supplements. The consumption of these antioxidant pigments can be a strategy for reducing the risk of developing chronic non-communicable diseases in the population. However, the molecular instability of these compounds limits their technological use. Nanoencapsulation based on pectin and lysozyme is considered a viable option to stabilize it and enable this inclusion. Thus, the present thesis had as primary objectives: 1) to develop a new methodology for nanoencapsulation of anthocyanins based on pectin and lysozyme; 2) to characterize physical and chemically the nanostructures with encapsulated anthocyanins; 3) to evaluate stability in different pH and temperature ranges; 4) to observe the stability of nanostructures and the preservation of intact anthocyanins in a simulated digestion system; 5) to analyze the effect on cell viability, cytotoxicity and in vitro uptake in human cells; 6) to discuss possible biological effects and feasibility for inclusion in food products. Mathematic models were used for optimization parameters. To characterize the nanostructures, analysis of Dynamic Light Scattering, Scanning Microscopy, Differential Scanning Calorimetry, Fourier Transforms Infrared Spectral, and Encapsulation Efficiency were performed. To analyze the stability different pH titrations and temperatures of 4 °C, 25 °C, and 40 °C were observed, simulated in vitro digestion (INFOGEST) was evaluated, and the effect in cells two- and three-dimensional cell models were used to determine viability, toxicity, and cellular incorporation. The nanostructures were formed by a complex network of molecular self-organization. The results showed that the formed nanostructures have adequate physicochemical characteristics, an average size of 190 nm, spherical and homogeneous morphology, a Zeta Potential of about -30, and an efficiency of encapsulation of 79%. The nanostructures were stable in different pH and temperature ranges, simulating storage. In the gastrointestinal system, anthocyanins were gradually released, and they maintained stable colloid characteristics in the intestine. The viability of cells treated with nanoparticles was maintained, they had no cellular toxicity, and the nanostructures successfully absorbed the nanostructures in cells, as observed by microscopy. The pectin covered the nanostructures, while lysozyme provide internal stability, and anthocyanin was effectively encapsulated. Future in vivo experiments are indicated to analyze the bioavailability. Pectin-based nanostructures were effective in maintaining stability and for prospecting inclusion in food products and supplements.
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spelling Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compoundsNanoencapsulação de antocianinas à base de pectina e lisozima: uma nova abordagem tecnológica para aumento da estabilidade físicoquímica de compostos fenólicosCianidina-3-O-glucosideoColloidal StabilityCyanidin-3-O-glucosideDigestão SimuladaEstabilidade ColoidalFood SupplementsNanoestruturasNanostructuresPolissacarídeosPolysaccharidesProteinProteínaSimulated digestionSuplemento alimentarAnthocyanins are sensitive flavonoid pigments and are widely distributed in nature. Currently, these phenolic compounds are being extensively researched due to intense interest in their beneficial properties. Due to their functionalities, they can be added to food products for natural coloring, functional ingredients, and in the development of nutritional supplements. The consumption of these antioxidant pigments can be a strategy for reducing the risk of developing chronic non-communicable diseases in the population. However, the molecular instability of these compounds limits their technological use. Nanoencapsulation based on pectin and lysozyme is considered a viable option to stabilize it and enable this inclusion. Thus, the present thesis had as primary objectives: 1) to develop a new methodology for nanoencapsulation of anthocyanins based on pectin and lysozyme; 2) to characterize physical and chemically the nanostructures with encapsulated anthocyanins; 3) to evaluate stability in different pH and temperature ranges; 4) to observe the stability of nanostructures and the preservation of intact anthocyanins in a simulated digestion system; 5) to analyze the effect on cell viability, cytotoxicity and in vitro uptake in human cells; 6) to discuss possible biological effects and feasibility for inclusion in food products. Mathematic models were used for optimization parameters. To characterize the nanostructures, analysis of Dynamic Light Scattering, Scanning Microscopy, Differential Scanning Calorimetry, Fourier Transforms Infrared Spectral, and Encapsulation Efficiency were performed. To analyze the stability different pH titrations and temperatures of 4 °C, 25 °C, and 40 °C were observed, simulated in vitro digestion (INFOGEST) was evaluated, and the effect in cells two- and three-dimensional cell models were used to determine viability, toxicity, and cellular incorporation. The nanostructures were formed by a complex network of molecular self-organization. The results showed that the formed nanostructures have adequate physicochemical characteristics, an average size of 190 nm, spherical and homogeneous morphology, a Zeta Potential of about -30, and an efficiency of encapsulation of 79%. The nanostructures were stable in different pH and temperature ranges, simulating storage. In the gastrointestinal system, anthocyanins were gradually released, and they maintained stable colloid characteristics in the intestine. The viability of cells treated with nanoparticles was maintained, they had no cellular toxicity, and the nanostructures successfully absorbed the nanostructures in cells, as observed by microscopy. The pectin covered the nanostructures, while lysozyme provide internal stability, and anthocyanin was effectively encapsulated. Future in vivo experiments are indicated to analyze the bioavailability. Pectin-based nanostructures were effective in maintaining stability and for prospecting inclusion in food products and supplements.As antocianinas são pigmentos flavonoides sensíveis e amplamente distribuídos na natureza. Atualmente, esses compostos fenólicos estão sendo extensivamente pesquisados devido ao intenso interesse em suas propriedades benéficas. Devido às suas funcionalidades, podem ser adicionados a produtos alimentícios como corante natural, ingredientes funcionais e no desenvolvimento de suplementos nutricionais. O consumo desses pigmentos antioxidantes pode ser uma estratégia para reduzir o risco de desenvolvimento de doenças crônicas não transmissíveis na população. No entanto, a instabilidade molecular desses compostos limita seu uso tecnológico. A nanoencapsulação à base de pectina e lisozima é considerada uma opção viável para estabilizá-las e possibilitar essa inclusão. Assim, a presente tese teve como objetivos principais: 1) desenvolver uma nova metodologia à base de pectina e lisozima para nanoencapsulação de antocianinas; 2) caracterizar física e quimicamente as nanoestruturas com antocianinas encapsuladas; 3) avaliar a estabilidade em diferentes faixas de pH e temperatura; 4) observar a estabilidade das nanoestruturas e a preservação de antocianinas intactas em um sistema de digestão simulada; 5) analisar o efeito na viabilidade celular, citotoxicidade e internalização in vitro em células humanas; 6) discutir possíveis efeitos biológicos e viabilidade para inclusão em produtos alimentícios. Para isso, modelos matemáticos foram usados para parâmetros de otimização. Para caracterizar as nanoestruturas, foram realizadas análises de Espalhamento Dinâmico de Luz, Microscopia de Varredura, Calorimetria de Varredura Diferencial, Espectro de Infravermelho por Transformada de Fourier e Eficiência de Encapsulamento. Para analisar a estabilidade, a titulação em diferentes pH e temperaturas de 4 °C, 25 °C e 40 °C foram avaliadas, digestão in vitro simulada (INFOGEST) foi analisada e o efeito em células modelos bi e tridimensionais foram usados para determinar a viabilidade, toxicidade e incorporação celular. As nanoestruturas foram formadas por uma complexa rede de auto-organização molecular. Os resultados indicam que as nanoestruturas possuem características físico-químicas adequadas, tamanho médio de 190 nm, morfologia esférica e homogênea, Potencial Zeta em torno de -30, e eficiência de encapsulamento de 79%. As nanoestruturas foram estáveis em diferentes faixas de pH e temperatura, simulando o armazenamento. No sistema gastrointestinal simulado, as antocianinas foram liberadas gradativamente e as estruturas mantiveram características de coloides estáveis no intestino. A viabilidade de células tratadas com as nanopartículas foi mantida, não apresentaram toxicidade celular, e as nanoestruturas foram absorvidas com sucesso nas células, conforme observado por microscopia. A pectina recobriu as nanoestruturas, enquanto a lisozima forneceu estabilidade interna e a antocianina foi encapsulada com eficácia. Futuros experimentos in vivo são indicados para analisar a biodisponibilidade. As nanoestruturas à base de pectina foram eficazes na manutenção da estabilidade e na prospecção para inclusão em produtos alimentícios e suplementos.Biblioteca Digitais de Teses e Dissertações da USPFabi, João PauloRosales, Thiécla Katiane Osvaldt2022-09-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/9/9131/tde-14102022-160404/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPReter o conteúdo por motivos de patente, publicação e/ou direitos autoriais.info:eu-repo/semantics/openAccesseng2024-08-13T18:36:48Zoai:teses.usp.br:tde-14102022-160404Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-13T18:36:48Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
Nanoencapsulação de antocianinas à base de pectina e lisozima: uma nova abordagem tecnológica para aumento da estabilidade físicoquímica de compostos fenólicos
title Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
spellingShingle Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
Rosales, Thiécla Katiane Osvaldt
Cianidina-3-O-glucosideo
Colloidal Stability
Cyanidin-3-O-glucoside
Digestão Simulada
Estabilidade Coloidal
Food Supplements
Nanoestruturas
Nanostructures
Polissacarídeos
Polysaccharides
Protein
Proteína
Simulated digestion
Suplemento alimentar
title_short Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
title_full Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
title_fullStr Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
title_full_unstemmed Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
title_sort Nanoencapsulation of anthocyanins based on pectin and lysozyme: a new technological approach to increase the physicochemical stability of phenolic compounds
author Rosales, Thiécla Katiane Osvaldt
author_facet Rosales, Thiécla Katiane Osvaldt
author_role author
dc.contributor.none.fl_str_mv Fabi, João Paulo
dc.contributor.author.fl_str_mv Rosales, Thiécla Katiane Osvaldt
dc.subject.por.fl_str_mv Cianidina-3-O-glucosideo
Colloidal Stability
Cyanidin-3-O-glucoside
Digestão Simulada
Estabilidade Coloidal
Food Supplements
Nanoestruturas
Nanostructures
Polissacarídeos
Polysaccharides
Protein
Proteína
Simulated digestion
Suplemento alimentar
topic Cianidina-3-O-glucosideo
Colloidal Stability
Cyanidin-3-O-glucoside
Digestão Simulada
Estabilidade Coloidal
Food Supplements
Nanoestruturas
Nanostructures
Polissacarídeos
Polysaccharides
Protein
Proteína
Simulated digestion
Suplemento alimentar
description Anthocyanins are sensitive flavonoid pigments and are widely distributed in nature. Currently, these phenolic compounds are being extensively researched due to intense interest in their beneficial properties. Due to their functionalities, they can be added to food products for natural coloring, functional ingredients, and in the development of nutritional supplements. The consumption of these antioxidant pigments can be a strategy for reducing the risk of developing chronic non-communicable diseases in the population. However, the molecular instability of these compounds limits their technological use. Nanoencapsulation based on pectin and lysozyme is considered a viable option to stabilize it and enable this inclusion. Thus, the present thesis had as primary objectives: 1) to develop a new methodology for nanoencapsulation of anthocyanins based on pectin and lysozyme; 2) to characterize physical and chemically the nanostructures with encapsulated anthocyanins; 3) to evaluate stability in different pH and temperature ranges; 4) to observe the stability of nanostructures and the preservation of intact anthocyanins in a simulated digestion system; 5) to analyze the effect on cell viability, cytotoxicity and in vitro uptake in human cells; 6) to discuss possible biological effects and feasibility for inclusion in food products. Mathematic models were used for optimization parameters. To characterize the nanostructures, analysis of Dynamic Light Scattering, Scanning Microscopy, Differential Scanning Calorimetry, Fourier Transforms Infrared Spectral, and Encapsulation Efficiency were performed. To analyze the stability different pH titrations and temperatures of 4 °C, 25 °C, and 40 °C were observed, simulated in vitro digestion (INFOGEST) was evaluated, and the effect in cells two- and three-dimensional cell models were used to determine viability, toxicity, and cellular incorporation. The nanostructures were formed by a complex network of molecular self-organization. The results showed that the formed nanostructures have adequate physicochemical characteristics, an average size of 190 nm, spherical and homogeneous morphology, a Zeta Potential of about -30, and an efficiency of encapsulation of 79%. The nanostructures were stable in different pH and temperature ranges, simulating storage. In the gastrointestinal system, anthocyanins were gradually released, and they maintained stable colloid characteristics in the intestine. The viability of cells treated with nanoparticles was maintained, they had no cellular toxicity, and the nanostructures successfully absorbed the nanostructures in cells, as observed by microscopy. The pectin covered the nanostructures, while lysozyme provide internal stability, and anthocyanin was effectively encapsulated. Future in vivo experiments are indicated to analyze the bioavailability. Pectin-based nanostructures were effective in maintaining stability and for prospecting inclusion in food products and supplements.
publishDate 2022
dc.date.none.fl_str_mv 2022-09-12
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://www.teses.usp.br/teses/disponiveis/9/9131/tde-14102022-160404/
url https://www.teses.usp.br/teses/disponiveis/9/9131/tde-14102022-160404/
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv
dc.rights.driver.fl_str_mv Reter o conteúdo por motivos de patente, publicação e/ou direitos autoriais.
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Reter o conteúdo por motivos de patente, publicação e/ou direitos autoriais.
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.coverage.none.fl_str_mv
dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
dc.source.none.fl_str_mv
reponame:Biblioteca Digital de Teses e Dissertações da USP
instname:Universidade de São Paulo (USP)
instacron:USP
instname_str Universidade de São Paulo (USP)
instacron_str USP
institution USP
reponame_str Biblioteca Digital de Teses e Dissertações da USP
collection Biblioteca Digital de Teses e Dissertações da USP
repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)
repository.mail.fl_str_mv virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br
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