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Universidade Federal Do Rio De Janeiro André Mesquita Magalhães

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Universidade Federal Do Rio De Janeiro André Mesquita Magalhães Universidade Federal do Rio de Janeiro André Mesquita Magalhães Costa POMEGRANATE SEED OIL (PUNICA GRANATUM L.): CHEMICAL COMPOSITION AND PRODUCTION OF VALUE- ADDED MICROPARTICLES BY COMPLEX COACERVATION RIO DE JANEIRO 2018 André Mesquita Magalhães Costa POMEGRANATE SEED OIL (PUNICA GRANATUM L.): CHEMICAL COMPOSITION AND PRODUCTION OF VALUE- ADDED MICROPARTICLES BY COMPLEX COACERVATION Tese de Doutorado apresentada ao Programa de Pós-graduação em Ciência de Alimentos, Instituto de Química, Universidade Federal do Rio de Janeiro, como requisito parcial à obtenção do título de Doutor em Ciência de Alimentos. Orientadores: Prof. Dr. Alexandre Guedes Torres Drª Renata Valeriano Tonon Rio de Janeiro 2018 Dedication Aos meus pais, minha irmã e meus avós. Ao amor infinito, carinho e toda dedicação. Devo tudo a vocês. ACKNOWLEDGEMENTS Agradeço imensamente a todos que me ajudaram na realização desse trabalho, principalmente meus orientadores, Alexandre e Renata, minha família, meus companheiros de laboratório, meus amigos e acima de tudo DEUS. “Os nossos dias de luta São as nossas noites de glória.” Charlie Brown Jr. “Você não sabe o quanto caminhei Pra chegar até aqui.” Cidade Negra RESUMO ÓLEO DE SEMENTE DE ROMÃ (PUNICA GRANATUM L.): COMPOSIÇÃO QUÍMICA E PRODUÇÂO DE MICROPARTÍCULAS COM VALOR AGREGADO POR COACERVAÇÃO COMPLEXA André Mesquita Magalhães Costa Orientador: Prof. Dr. Alexandre Guedes Torres Co-orientadora: Drª Renata Valeriano Tonon O óleo de semente de romã (OSR) é um óleo funcional com uma composição singular em compostos bioativos, principalmente isômeros do ácido linolênico conjugado (cLnA). O encapsulamento é uma tecnologia capaz de aumentar a estabilidade do OSR e dessa forma viabilizar a sua adição em alimentos. Este trabalho apresentou 3 grandes objetivos: 1) determinar a composição físico-química detalhada de OSR comerciais provenientes da Turquia e Israel, e identificar possíveis tendências regionais relacionadas à composição em compostos bioativos (fenólicos totais, tocoferóis totais, cLnA total, β-caroteno); 2) encapsular o OSR por coacervação complexa utilizando o sistema whey protein-goma Arábica e investigar a influência da concentração de polímeros totais (Cp) e razão material de parede:óleo (WM:Oil) nas propriedades físicas, químicas e morfológicas das micropartículas; e 3) avaliar a estabilidade oxidativa em duas condições de estocagem (temperatura ambiente por 90 dias e 60 °C por 10 dias seladas com/sem vácuo) e as propriedades tecnológicas da melhor formulação de micropartícula. A aplicação das micropartículas em um potencial veículo (bebida instantânea de café com leite adicionada de micropartículas de OSR) para o consumo de OSR também foi avaliada. A caracterização dos OSR comerciais demonstrou similaridades entre as amostras quanto aos índices de qualidade, perfil de classes lipídicas e teor de ácido punícico (590 mg/g óleo). O conteúdo total de cLnA e teor total de tocoferóis foi influenciado pela origem das amostras. OSRs apresentaram um baixo teor de β-caroteno e perfis variados de compostos fenólicos e voláteis. Todas amostras apresentaram uma baixa estabilidade oxidativa, provavelmente pelo alto teor de cLnAs altamente oxidáveis. Segundo a análise de componente principal, a composição em bioativos das amostras promoveu uma discriminação preliminar baseada na origem. Em seguida, o OSR foi encapsulado, nove formulações foram testadas segundo um planejamento rotacional composto central, onde as variáveis independentes foram Cp (2,2-7,8%) e razão WM:Oil (0,5-5,0), e as variáveis dependentes, retenção de óleo, eficiência de microencapsulamento (EM), conteúdo de ácido punícico, índice de peróxido, conteúdo de umidade, atividade de água e tamanho de partícula. Cp e razão WM:Oil influenciaram a retenção de óleo, EM, conteúdo de ácido punícico, conteúdo de umidade e tamanho de partícula. Valores intermediários de Cp e WM:Oil produziram micropartículas com alta retenção de óleo e o maior conteúdo de ácido punícico, neste sentido a formulação do ponto central foi escolhida (Cp = 5%; WM:Oil = 2,75). Por fim, o processo de coacervação complexa foi comparado com um método de encapsulamento mais tradicional, spray drying. As micropartículas coacervadas apresentaram estabilidade oxidativa equivalente às micropartículas não coacervadas por 60 dias e a estocagem à vácuo reduziu a taxa de oxidação do OSR. A alta solubilidade e estabilidade térmica das micropartículas coacervadas possibilitam a sua aplicação no desenvolvimento de alimentos. A adição das micropartículas coacervadas no produto desenvolvido alterou apenas o conteúdo de umidade do mesmo. A composição físico-química detalhada do OSR foi determinada e o mesmo foi encapsulado com sucesso por coacervação complexa, se mantendo estável por 60 dias, possibilitando assim sua aplicação no desenvolvimento de produtos alimentícios com potencial funcionalidade. Palavras chaves: análise de alimentos, composição de alimentos, óleo de semente de romã, ácido punícico, ácidos fenólicos, gama-tocoferol, compostos voláteis, origem geográfica, variação na composição química, planejamento experimental, coacervação complexa, whey protein, goma Arábica, spray drying, isômeros de cLnA, estabilidade oxidativa, estocagem, hidroperóxidos, propriedades tecnológicas e alimento funcional. ABSTRACT POMEGRANATE SEED OIL (PUNICA GRANATUM L.): CHEMICAL COMPOSITION AND PRODUCTION OF VALUE-ADDED MICROPARTICLES BY COMPLEX COACERVATION André Mesquita Magalhães Costa Supervision: Prof. Dr. Alexandre Guedes Torres Co-supervision: Drª Renata Valeriano Tonon Pomegranate seed oil (PSO) is a functional edible oil with a unique composition of bioactive compounds, especially conjugated linolenic acids (cLnA) isomers. The microencapsulation technique could be applied to increase PSO’s stability, enabling food addition. This work had three major objectives: 1) perform a detailed physical-chemical characterization of commercial PSOs from Turkey and Israel, and identify possible regional trends related with bioactive compounds (total phenolics, total cLnAs, total tocopherols and β-carotene) composition; 2) encapsulate PSO by complex coacervation utilizing the whey protein-gum Arabic system and investigate the influence of total polymer concentration (Cp) and wall material:oil ratio (WM:Oil) on microparticles’ physical, chemical and morphological characteristics; and 3) evaluate the oxidative stability in two storage conditions (ambient temperature during 90 days and 60 °C during 10 days with/without vacuum) and technological properties of the best PSO microparticle’s formulation. Microparticles’ application on a potentional vehicle (instant coffee latte drink added with PSO’s microparticles) for PSO consumption, was also investigated. Commercial PSOs characterization indicated that samples were similar concerning quality indexes, lipid classes and punicic acid content (ca. 590 mg/g oil). Total cLnA content and total tocopherols were influenced by samples origin. PSOs showed low contents of β-carotene and a very distinct profiles of phenolics and volatile compounds. Samples showed low oxidative stability, possibly because of cLnA’s oxidizability. Bioactive compounds data provided a marginal discrimination of PSO’s geographical origin, by principal component analysis. Next step was PSO encapsulation, nine formulations were tested according to a rotatable central composite design. Independent variables were: Cp (2.2-7.8%) and WM:Oil ratio (0.5-5.0). Oil retention, microencapsulation efficiency (ME), punicic acid content, peroxide value, moisture content, water activity and particle size were analyzed as responses. Cp and WM:Oil ratio affected oil retention, ME, punicic acid content, moisture content and particle size. Intermediate values of Cp and WM:Oil ratio produced microparticles with the highest punicic acid content and 100% oil retention, therefore central point formulation (Cp = 5%; WM:Oil = 2.75) was selected to continue in the study. In the third phase of the study, complex coacervation process was compared throughout the study with spray drying, a more traditional encapsulation method. At ambient temperature, coacervate microparticles showed similar oxidative stability as non- coacervate microparticles up to day 60. These results might be associated with coacervate microparticles’ more porous structure demonstrated by morphological analysis. At 60 °C, storage under reduced O2 concentration diminished microparticles’ oxidation ratio. Coacervate microparticles were applicable to wide range of products, due to its high solubility and thermal stability. Microparticles addition only affected product’s moisture. PSO detailed physical-chemical composition was determined and successfully stabilized by complex coacervation, showing an overall 60 days stability, enabling it application in the development of potential functional food products. Keywords: Food analysis, food composition, pomegranate seed oil, punicic acid, phenolic acids, tocopherol, volatile compounds, geographical origin, chemical composition variation, experimental design, complex coacervation, whey protein, gum Arabic, spray drying, cLnA isomers, oxidative stability, storage, hydroperoxides, technological properties and functional food. LIST OF FIGURES Section Page Chapter 1 Figure 1. Punica granatum cv ‘Bhagua’ structures. A) Pomegranate plant; B) Flowering stem; C) Developing fruit; D) Matured fruit; E) Fruit split open; white arrow: outside peel; black arrow: white septal membranes; F) Arils. ------------------- 26 Figure 2.
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