Elisa Rafaela Bonadio Bellucci Aplicação de extratos de pitaia e açaí em produtos cárneos: estudo da vida útil São José do Rio Preto 2022 Campus de São José do Rio Preto Elisa Rafaela Bonadio Bellucci Aplicação de extratos de pitaia e açaí em produtos cárneos: estudo da vida útil Tese apresentada como parte dos requisitos para obtenção do título de Doutor(a) em Engenharia e Ciência de Alimentos, junto ao Programa de Pós- graduação em Engenharia e Ciência de Alimentos (Área de concentração: Ciência e Tecnologia de Alimentos) do Instituto de Biociências, Letras e Ciências Exatas da Universidade Estadual Paulista “Júlio de Mesquita Filho”, Campus de São José do Rio Preto. Financiadora: CAPES Orientadora: Profa. Dra. Andrea Carla da Silva Barretto Coorientador: Prof. Dr. Jose Manuel Lorenzo Rodriguez São José do Rio Preto 2022 B449a Bellucci, Elisa Rafaela Bonadio Aplicação de extratos de pitaia e açaí em produtos cárneos: estudo da vida útil / Elisa Rafaela Bonadio Bellucci. -- São José do Rio Preto, 2022 141 p. Tese (doutorado) - Universidade Estadual Paulista (Unesp), Instituto de Biociências Letras e Ciências Exatas, São José do Rio Preto Orientadora: Andrea Carla da Silva Barretto Coorientador: José Manuel Lorenzo Rodriguez 1. Antioxidantes naturais. 2. Aditivos naturais. 3. Produtos cárneos saudáveis. 4. Oxidação lipídica. I. Título. Sistema de geração automática de fichas catalográficas da Unesp. Biblioteca do Instituto de Biociências Letras e Ciências Exatas, São José do Rio Preto. Dados fornecidos pelo autor(a). Essa ficha não pode ser modificada. Elisa Rafaela Bonadio Bellucci Aplicação de extratos de pitaia e açaí em produtos cárneos: estudo da vida útil Tese apresentada como parte dos requisitos para obtenção do título de Doutor(a) em Engenharia e Ciência de Alimentos, junto ao Programa de Pós- graduação em Engenharia e Ciência de Alimentos (Área de concentração: Ciência e Tecnologia de Alimentos) do Instituto de Biociências, Letras e Ciências Exatas da Universidade Estadual Paulista “Júlio de Mesquita Filho”, Campus de São José do Rio Preto. Comissão Examinadora Profa. Dra. Andrea Carla da Silva Barretto UNESP – Campus de São José do Rio Preto Orientadora Profa. Dra. Natália Soares Janzantti UNESP – Campus de São José do Rio Preto Profa. Dra. Marise A. Rodrigues Pollonio UNICAMP – Campus de Campinas Prof. Dr. Roger Darros Barbosa UNESP – Campus de São José do Rio Preto Profa Dra. Profa. Dra. Tayse Ferreira Ferreira da Silveira UNIFESP – Campus São Paulo São José do Rio Preto 18 de março de 2022 Dedico este trabalho a minha família! Aos meus pais, Edson e Dora, que nunca mediram esforços para me ajudar e estiveram ao meu lado em todos os momentos. Aos meus irmãos, Eder e Elisiane, por cada palavra de incentivo e por torcerem pelo meu sucesso. Dedico também aos meus avós (in memoriam), em especial ao meu avô Dino Orlando Bonadio, que me colocava em suas orações e acreditava na minha capacidade, sempre com muito orgulho. Sei que minhas ações ainda estão sob as bênçãos de Deus, devido a ele. AGRADECIMENTOS Iniciando os agradecimentos, gostaria de agradecer à minha orientadora, Profa. Dra. Andrea Carla da Silva Barretto, que não foi apenas uma ferramenta para meu aprendizado e uma guia nos meus conhecimentos, mas também foi uma amiga, na qual pude ter ao meu lado em muitas situações que se passaram nesses anos em que estive sob sua orientação. Meu eterno muito obrigada! Obrigada, Profa. Andrea, pelo apoio, incentivo, pelas “broncas” em momentos certeiros e por compartilhar todo seu conhecimento comigo. Agradeço ao meu coorientador, Dr. José Manuel Lorenzo Rodriguez, por todas as contribuições dadas para a realização desta tese de Doutorado, todas de grande importância para a conclusão deste trabalho. Agradeço também pela oportunidade a mim presenteada e por me receber no Centro Tecnológico da Carne, localizado na cidade de Ourense, Espanha. E, aqui, estendo meu agradecimento ao CTC e a todos os que estiveram comigo e puderam, de alguma forma, colaborar com a pesquisa que lá foi realizada. Agradeço ao Departamento de Engenharia e Tecnologia de Alimentos (DETA) da Universidade Estadual Paulista “Júlio de Mesquita Filho”, campus de São José do Rio Preto, pela infraestrutura oferecida para a execução da pesquisa aqui discutida. E, ao Programa de Pós-Graduação em Ciência e Tecnologia de Alimentos, bem como a todos os docentes pertencentes a ele, principalmente pelo apoio e convivência durante esses anos. Estendo o meu agradecimento aos Técnicos do DETA, Luiz, Tânia e Alana e aos demais funcionários que ali trabalham e que tive o prazer de conviver. Aos meus colegas de laboratório, que estiveram comigo nesses anos, meu muito obrigada! Em especial àqueles que não só foram meus colegas, mas se tornaram meus amigos, e que me ajudaram não somente durante análises e escritas, mas também no dia a dia. Agradeço aos amigos da vida, aqueles que estão comigo desde sempre. Agradeço a toda minha família, tios, tias, primos e primas, que estão sempre presentes em minha vida. Sinto o carinho e a preocupação de cada um e sei que estiveram ao meu lado em toda decisão que tomei, comemorando cada conquista obtida. Eu amo imensamente cada um! Aos meus pais, Edson e Dora, não há “obrigada” o suficiente para expressar o quanto sou grata por tudo que hão feito por mim. Sou muito abençoada por tê-los em minha vida e sei que não conseguiria sem o apoio e as orações de vocês. E aos meus irmãos, Eder e Elisiane, meu obrigada por estarem ao meu lado, por serem minha calmaria e meu porto seguro. E, em especial, agradeço a Deus por ter me proporcionado viver e concluir essa experiência. Por ter colocado em meu caminho somente pessoas maravilhosas e ter me colocado na família que tenho. Agradeço por ter me fortalecido nos momentos difíceis, por ter me dado discernimento nos momentos de dúvida e fé para seguir acreditando que tudo será conforme a vontade Dele. O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001, à qual agradeço, pelo suporte financeiro recebido e por contribuir para o desenvolvimento da pesquisa brasileira e para a capacitação de pesquisadores. RESUMO A mudança no hábito alimentar dos consumidores, que passaram a se preocupar mais com os alimentos que consomem, aliado aos riscos carcinogênico e tóxico associados aos aditivos sintéticos, levaram a um aumento na busca por alimentos com menores quantidades desses aditivos. Essa tendência refletiu no setor da carne impulsionando pesquisas para desenvolver produtos com adição de extratos naturais obtidos de diferentes fontes capazes de inibir ou retardar as reações de oxidação, a principal causa da deterioração desses produtos devido às alterações sensoriais e nutricionais. Extratos obtidos de frutas têm sido amplamente estudados. A pitaia vermelha e o açaí são frutos ricos em compostos bioativos com ação antioxidantes e com potencial para estender a vida útil de produtos cárneos, reduzindo as reações de oxidação lipídica e proteica, além de minimizar a descoloração durante o armazenamento. Assim, o objetivo deste trabalho foi obter extrato de pitaia vermelha para aplicação em hambúrguer suíno e avaliar os seus efeitos sobre a vida útil. Ainda neste contexto, obter extrato a partir da polpa de açaí e avaliar sua aplicação em hambúrguer suíno refrigerado e em salame tipo Italiano durante o tempo de maturação. O extrato de pitaia vermelha estudado foi adicionado em hambúrguer suíno com substituição de gordura suína por uma emulsão de óleo de chufa. Os tratamentos estudados foram: CON (sem adição de antioxidante), ERY (500 mg.kg-1 de eritorbato de sódio), PEL, PEM e PEH (250, 500 e 1000 mg.kg-1 de extrato de pitaia, respectivamente). O extrato de pitaia vermelha teve efeito sobre a coloração do produto, mantendo a coloração de hambúrguer suíno mais estável durante os 18 dias de armazenamento refrigerado sobre incidência de luz, além disso foi capaz de reduzir a oxidação lipídica e melhorar a aceitação sensorial, principalmente do atributo de cor. O extrato de açaí foi avaliado em ambos os produtos, hamburguer e salame, nas concentrações de 250, 500 e 750 mg.kg-1, e estas foram comparadas a um controle (sem adição de antioxidante) e um tratamento com eritorbato de sódio (500 mg.kg-1). Em hambúrguer, o extrato de açaí melhorou a estabilidade oxidativa, no entanto, as concentrações 500 e 750 mg.kg-1 (AEM e AEH) afetaram negativamente a coloração, o que não ocorreu na menor concentração estudada (AEL, 250 mg.kg-1), além disso, foi capaz de inibir as reações oxidativas tal como o eritorbato de sódio. Portanto, em hambúrguer suíno, o extrato de açaí adicionado a 250 mg.kg-1 pode ser usado como antioxidante natural em alternativa ao eritorbato de sódio. Já quando aplicado em salame, foi possível observar que o extrato de açaí aplicado nas concentrações de 500 e 750 mg.kg−1 melhoraram a estabilidade oxidativa, além de terem reduzido os valores de pH, e aumentado o crescimento de bactérias lácticas no tempo de fermentação. Diante disto, nestas concentrações, o extrato de açaí pode ser usado como antioxidante natural para melhorar a qualidade e prolongar a vida útil do salame do tipo italiano. Portanto, nas concentrações estudadas, tanto extrato de pitaia quanto o de açaí podem ser utilizados como antioxidantes para melhorar a vida útil de produtos cárneos. Palavras-chave: Antioxidantes naturais. Aditivos naturais. Produtos cárneos saudáveis. Oxidação lipídica. ABSTRACT The change in the eating habits of consumers, who have become more concerned with the food they consume, combined with the carcinogenic and toxic risks associated with synthetic additives, has driven an increase in the search for foods with lower amounts of these additives. This trend was reflected in the meat sector, boosting research to develop products with the addition of natural extracts obtained from different sources capable of inhibiting or delaying oxidation reactions, the main cause of deterioration of these products due to sensory and nutritional changes. Extracts obtained from fruits are widely studied. Red pitaya and açaí are fruits rich in bioactive antioxidant compounds that can act to extend the life of meat products, reducing the reactions of lipid and protein oxidation, in addition to minimizing discoloration during storage. Thus, the objective of this work was to obtain red pitaya extract for application in pork patties and to evaluate its effects on the stability of color and oxidation reactions, as well as on the sensory acceptance of this product. And, still in this context, to evaluate the effect of the extract obtained from the açaí pulp when applied in refrigerated pork patty and in Italian- type salami during the ripening time. The red pitaya extract studied was added to pork patties with pork fat replacement by an emulsion of chufa oil. The treatments studied were: CON (no antioxidant added), ERY (500 mg.kg-1 of sodium erythorbate), PEL, PEM and PEH (250, 500 and 1000 mg.kg-1 of pitaya extract, respectively). The red pitaya extract influenced the color of the product, keeping the pork patties color more stable during the 18 days of refrigerated storage under light incidence, in addition, it was able to reduce lipid oxidation and improve sensory acceptance, especially of the color attribute. The açaí extract was evaluated in both products, pork patties and Italian-type salami, at concentrations of 250, 500 and 750 mg.kg-1, and these were compared to a control (without the addition of antioxidant) and a treatment with sodium erythorbate (500 mg.kg-1). In pork patties, the açaí extract improved the oxidative stability, however, the concentrations of 500 and 750 mg.kg-1 (AEM and AEH) negatively affected the color, which did not occur at the lowest concentration studied (AEL, 250 mg.kg-1), in addition, it was able to inhibit oxidative reactions such as sodium erythorbate. Therefore, in pork patty, açaí extract added at 250 mg.kg-1 cans be used as a natural antioxidant as an alternative to sodium erythorbate. When applied in Italian-type salami, açaí extract applied at concentrations of 500 and 750 mg.kg−1 was able to improve oxidative stability, in addition to reducing the pH values and increasing the growth of lactic acid bacteria during the fermentation time. In this context, at these concentrations, the açaí extract can be used as a natural antioxidant to improve the quality and prolong the shelf life of Italian-style salami. Therefore, at the concentrations studied, both pitaya and açai extracts can be used as antioxidants to improve the shelf life of meat products. Keywords: Natural antioxidants. Natural additives. Healthy meat products. Lipid oxidation. LISTA DE FIGURAS CAPÍTULO 1 - ADDITION OF NATURAL EXTRACTS WITH ANTIOXIDANT FUNCTION TO PRESERVE THE QUALITY AND SHELF LIFE OF MEAT PRODUCTS Figure 1-1 Illustration representing the application of natural sources as an antioxidant in meat products as a means of preserving quality and shelf life. ......................................................... 27 Figure 1-2 Mechanisms of the oxidative processes that occur in the meat system (Ribeiro et al., 2019). ........................................................................................................................................ 29 Figure 1-3 Mechanism of antioxidant in lipid oxidation at propagation step (Falowo at al., 2014). ........................................................................................................................................ 31 CAPÍTULO 2 - RED PITAYA EXTRACT AS NATURAL ANTIOXIDANT IN PORK PATTIES WITH TOTAL REPLACEMENT OF ANIMAL FAT Figure 2-1. Evolution of TBARs values (A) and total carbonyl content (B) in pork patties during refrigerated storage........................................................................................................ 84 CAPÍTULO 3 - AÇAÍ EXTRACT POWDER AS NATURAL ANTIOXIDANT ON PORK PATTIES DURING THE REFRIGERATED STORAGE Figure 3-1 Photographs of the A) raw pork patties, B) cooked pork patties. ........................ 111 Figure 3-2 Antioxidant activity (%DPPH) of pork patties with erythorbate and different levels of açai extract (AE) during 10 days of refrigerate storage. .................................................... 114 LISTA DE TABELAS CAPÍTULO 1 - ADDITION OF NATURAL EXTRACTS WITH ANTIOXIDANT FUNCTION TO PRESERVE THE QUALITY AND SHELF LIFE OF MEAT PRODUCTS Table 1-1 Fruit (pulp, pigment, peel and seeds) as a source of antioxidants applied in meat products. ................................................................................................................................... 35 Table 1-2 Spices, products derivates from them and essential oil as naturals source of antioxidants applied in meat products ...................................................................................... 41 Table 1-3 Flowers, leaves and vegetable residues as naturals source of antioxidants applied in meat products. ........................................................................................................................... 51 CAPÍTULO 2 - RED PITAYA EXTRACT AS NATURAL ANTIOXIDANT IN PORK PATTIES WITH TOTAL REPLACEMENT OF ANIMAL FAT Table 2-1. Proximate composition, cooking loss and texture parameters of pork patties ....... 79 Table 2-2. Evaluation of pH values of pork patties during refrigerated storage ...................... 81 Table 2-3. Colour parameters of pork patties during refrigerated storage. .............................. 82 Table 2-4. In vitro antioxidant activity (μg Trolox/g) of pork patties during refrigerated storage. ...................................................................................................................................... 86 Table 2-5. Fatty acid profile of pork patties (g/100 g of total fatty acids) ............................... 87 Table 2-6. Acceptance scores and sum of orders according to the preference of consumers on day 0 of cooked pork patties ..................................................................................................... 89 CAPÍTULO 3 - AÇAÍ EXTRACT POWDER AS NATURAL ANTIOXIDANT ON PORK PATTIES DURING THE REFRIGERATED STORAGE Table 3-1 Proximate composition of pork patties with sodium erythorbate and different levels of açai extract (AE) during refrigerated storage. .................................................................... 106 Table 3-2 Evaluation of pH values of pork patties with sodium erythorbate and different levels of açai extract (AE) during refrigerated storage........................................................... 107 Table 3-3 Cooking loss and shrinkage of pork patties with sodium erythorbate and different levels of açai extract (AE) during refrigerated storage........................................................... 108 Table 3-4 Color parameters of pork patties with erythorbate and different levels of açai extract (AE) during refrigerated storage. ................................................................................ 110 Table 3-5 Evolution of TBARs values of pork patties with sodium erythorbate and different levels of açai extract (AE) during refrigerated storage........................................................... 112 CAPÍTULO 4 - APPLICATION OF AÇAÍ EXTRACT POWDER AS NATURAL ANTIOXIDANT IN ITALIAN-TYPE SALAMI UNDER RIPENING CONDITIONS: EVALUATION ON PHYSICOCHEMICAL AND MICROBIOLOGICAL PROPERTIES Table 4-1 Weight loss, pH, and water activity of Italian-type salami with sodium erythorbate and açaí extract during ripening time. .................................................................................... 128 Table 4-2 Lactic acid bacterial (LAB) count of Italian-type salami with sodium erythorbate and açaí extract during ripening time. .................................................................................... 130 Table 4-3 Color parameters of Italian-type salami with sodium erythorbate and distinct levels of açaí extract (EA) during processing. .................................................................................. 132 Table 4-4 Lipid oxidation values of Italian-type salami with sodium erythorbate and distinct levels of açaí extract (EA) during processing (mg MDA/kg of sample). ............................... 134 Table 4-5 Centesimal composition (g/100g) and instrumental texture profile of Italian-type salami with sodium erythorbate and different levels of açaí extract (EA). ............................ 135 SUMÁRIO RESUMO .............................................................................................................................. 6 ABSTRACT .......................................................................................................................... 8 LISTA DE FIGURAS ........................................................................................................ 10 LISTA DE TABELAS ....................................................................................................... 11 INTRODUÇÃO GERAL .................................................................................................. 17 APRESENTAÇÃO DO TRABALHO ............................................................................. 18 REFERÊNCIAS BIBLIOGRÁFICA ............................................................................... 19 OBJETIVOS ....................................................................................................................... 23 a) Objetivo Geral ............................................................................................................. 23 b) Objetivos Específicos ................................................................................................... 23 Capítulo 1 ............................................................................................................................. 24 ADDITION OF NATURAL EXTRACTS WITH ANTIOXIDANT FUNCTION TO PRESERVE THE QUALITY AND SHELF LIFE OF MEAT PRODUCTS .............. 25 Introduction ........................................................................................................................ 26 Oxidation reactions in meat and meat products ............................................................. 27 Antioxidants used in meat products ................................................................................. 30 Antioxidant substances derived from fruits (pulp, pigment, peel and seeds) applied in meat products ..................................................................................................................... 33 Spices and essential oils as naturals source of antioxidants applied in meat products40 Flowers, leaves and vegetable residues as naturals source of antioxidants applied in meat products ..................................................................................................................... 49 Other sources of natural antioxidants applied in meat products .................................. 53 Final considerations ........................................................................................................... 54 Acknowledgements ............................................................................................................ 55 References ........................................................................................................................... 55 Capítulo 2 ............................................................................................................................. 68 RED PITAYA EXTRACT AS NATURAL ANTIOXIDANT IN PORK PATTIES WITH TOTAL REPLACEMENT OF ANIMAL FAT .................................................. 69 1. Introduction .................................................................................................................... 70 2. Materials and Methods .................................................................................................. 71 2.1. Red pitaya extract (PE) .................................................................................................. 71 2.2. Total phenolic content and antioxidant activity of the PE ............................................. 71 2.3. Manufacture of pork patties ........................................................................................... 72 2.4. Chemical composition of pork patties ........................................................................... 73 2.5. Colour parameters and pH of pork patties .................................................................... 73 2.6. Cooking loss and texture profile analysis of pork patties .............................................. 74 2.7. Lipid oxidation and protein oxidation of pork patties ................................................... 74 2.8. In vitro antioxidant activity of pork patties ................................................................... 75 2.9. Fatty acid profile of pork patties ................................................................................... 76 2.10. Sensory analysis of raw and cooked pork patties ........................................................ 77 2.10. Statistical analyses ....................................................................................................... 77 3. Results and discussion ................................................................................................... 78 3.1. Total phenolic content and antioxidant capacity of PE ................................................. 78 3.2. Chemical composition of pork patties ........................................................................... 79 3.3. Cooking loss and texture profile analysis of pork patties .............................................. 80 3.4. pH and colour parameters of pork patties ..................................................................... 80 3.5. Lipid and protein oxidation of pork patties ................................................................... 83 3.6. In vitro antioxidant activity of pork patties ................................................................... 85 3.7. Fatty acid profile of pork patties ................................................................................... 86 3.8. Sensory analysis of pork patties ..................................................................................... 88 4. Conclusion ...................................................................................................................... 89 References ........................................................................................................................... 90 Capítulo 3 ............................................................................................................................. 98 AÇAÍ EXTRACT POWDER AS NATURAL ANTIOXIDANT ON PORK PATTIES DURING THE REFRIGERATED STORAGE .............................................................. 99 1. Introduction ............................................................................................................... 100 2. Materials and Methods ............................................................................................. 101 2.1. Preparation of Açaí extract (AE) powder and determination of antioxidant activity . 101 2.1.1. Preparation of AE ................................................................................................ 101 2.1.2. Total phenolic compounds (TPC) and antioxidant activity of AE ....................... 102 2.2. Manufacture of pork patties ........................................................................................ 102 2.3. Proximate composition and pH ................................................................................... 103 2.4. Instrumental color parameters .................................................................................... 103 2.5. Cooking proprieties ..................................................................................................... 104 2.6. Lipid oxidation ............................................................................................................. 104 2.7. Radical scavenging activity assay (in vitro) ................................................................ 105 2.8. Statistical analyses ...................................................................................................... 105 3. Results and discussion ............................................................................................... 105 3.1. Total phenolic compounds (TPC) and antioxidant activity of AE ............................... 105 3.2. Proximate composition and pH of pork patties ........................................................... 106 3.3. Cooking parameters .................................................................................................... 108 3.4. Instrumental color parameters .................................................................................... 109 3.5. Lipid oxidation ............................................................................................................. 112 3.6. In vitro antioxidant activity of pork patties ................................................................. 113 4. Conclusion .................................................................................................................. 113 Acknowledgements .......................................................................................................... 114 References ......................................................................................................................... 114 Capítulo 4 ........................................................................................................................... 121 APPLICATION OF AÇAÍ EXTRACT POWDER AS NATURAL ANTIOXIDANT IN ITALIAN-TYPE SALAMI UNDER RIPENING CONDITIONS: EVALUATION ON PHYSICOCHEMICAL AND MICROBIOLOGICAL PROPERTIES .............. 122 1. Introduction ............................................................................................................... 123 2. Materials and Methods ............................................................................................. 124 2.1. Obtaining and characterization of açaí extract powder extract ................................. 124 2.2. Manufacturing of Italian-type salami added with açaí extract powder ...................... 124 2.3. Water activity (Aw), Weight loss and Microbiological analysis (LAB) ...................... 125 2.4. Instrumental color Italian-type salami and pH determination .................................... 126 2.5. Lipid oxidation (TBARS assay) ................................................................................... 126 2.6. Chemical composition ................................................................................................. 127 2.7. Texture Profile Analysis (TPA) of Italian-type salami ................................................ 127 2.8. Statistical analysis ....................................................................................................... 127 3. Results and Discussion .............................................................................................. 128 3.1. Weight loss, pH, Water activity (Aw) and LAB count ................................................. 128 3.2. Color Parameters ........................................................................................................ 131 3.3. Lipid oxidation during ripening .................................................................................. 133 3.4. Chemical Composition and Texture Analyses Profile at the end of ripening ............. 134 4. Conclusion .................................................................................................................. 135 Acknowledgements .......................................................................................................... 135 References ......................................................................................................................... 136 CONCLUSÃO GERAL ................................................................................................... 141 17 INTRODUÇÃO GERAL Há um aumento do consumo de carne a nível mundial e apesar de ter sido correlacionado com algumas implicações a saúde, a carne ainda é considerada a mais importante fonte de proteínas de alta qualidade, de vitaminas do grupo B e minerais essenciais (BURRI et al., 2020; JIANG; XIONG, 2016). No entanto, carnes e produtos cárneos são acometidos de reações de oxidação devido principalmente a alta proporção de ácidos graxos insaturados e também a composição dos fosfolipídios, presença de íons metálicos, ferro presente na porção heme da mioglobina, presença de oxigênio e, devido a etapas de processamento como moagem e cozimento (BISWAS; CHATLI; SAHOO, 2012; BURRI et al., 2020; DOMÍNGUEZ et al., 2019; LORENZO et al., 2018a). As reações de oxidação reduzem a qualidade de produtos cárneos, tendo como consequência a formação de odor de ranço, redução das características nutricionais e formação de compostos tóxicos (FAN et al., 2019). Contra esse fenômeno são adicionados aditivos a produtos cárneos com a finalidade de estender a vida útil e principalmente evitar a descoloração. Entre os antioxidantes utilizados estão o eritorbato de sódio, ascorbato de sódio, hidroxianisol butilado (BHA), butilado hidroxitolueno (BHT), terc-butilhidroquinona (TBHQ), galato de propila e sais de nitrito e nitrato de sódio e/ou potássio que se encontram associados a efeitos toxicológicos e carcinogênicos (BALDIN et al., 2018; ECHEGARAY et al., 2018; KARRE; LOPEZ; GETTY, 2013; MERCADANTE et al., 2010; RIBEIRO et al., 2019). O risco associado aos antioxidantes sintéticos somado a mudança no habito alimentar da população mundial, que passou a se preocupar em consumir alimentos com características mais saudáveis, impulsionou a indústria a desenvolver produtos com rótulos limpos, ou seja, rótulos com ingredientes reconhecidos pelos consumidores, estimulando a pesquisa por ingredientes naturais que possam substituir aditivos químicos comumente utilizados em alimentos (BREWER, 2011; KUMAR et al., 2015). Estudos estão sendo conduzidos para investigar o efeito antioxidante de compostos extraídos de fontes naturais, avaliando sua ação na garantia da qualidade sensorial e nutricional em produtos cárneos, mostrando serem boas alternativas aos antioxidantes sintéticos devido aos altos níveis de compostos fenólicos presentes e outras substâncias antioxidantes (BALDIN et al., 2018; FAN et al., 2019; LORENZO et al., 2018a, 2018b; MUNEKATA et al., 2017; POGORZELSKA et al., 2018; RIBEIRO et al., 2019). 18 Neste contexto, as frutas têm se destacado como fontes de compostos bioativos com propriedades antioxidantes para aplicação em produtos cárneos. A pitaia vermelha (Hylocereus polyrhizus) é um fruto exótico com propriedades interessantes envolvendo sua coloração e valor nutricional, pois é rica em compostos fenólicos, vitamina C e açucares, além de possuir pigmentos como betalaínas e flavonoides, ambos com comprovada capacidade antioxidante (FERRERES et al., 2017; HUA et al., 2018; OTÁLORA et al., 2019). As betalaínas são responsáveis pela cor rosa da polpa e da casca desses frutos, são pigmentos solúveis em água que possuem nitrogênio em sua estrutura (GARCÍA-CRUZ et al., 2017). Alguns estudos mostram efeito da betalaína como corante em produtos cárneos com reduzido teor de nitrito (BELLUCCI et al., 2021; BLOUKAS; ARVANITOYANNIS; SIOPI, 1999; MARTÍNEZ et al., 2006). A Euterpe oleracea é uma palmeira nativa da floresta Amazônica e é mais conhecido como açaizeiro e seu fruto, o açaí, possui alto potencial econômico devido a comercialização de seu suco e polpa, os quais são reconhecidas fontes de energia (BRUNSCHWIG et al., 2016). No Brasil, os maiores produtores de açaí são os estados do Pará, com produção anual de mais de 1,3 milhão toneladas, seguido do Amazonas e de Roraima, que juntos somam mais de 55 mil toneladas (ABRAFRUTAS, 2019). Antocianina é o principal bioativo presente no açaí, sendo associada com inativação de radicais livres e inibição da oxidação, podendo ser utilizado como antioxidante (CARVALHO et al., 2017; KANG et al., 2011). Ainda não há estudos sobre a aplicação de extrato de polpa de pitaia vermelha e de polpa de açaí em produtos cárneos como potencial antioxidante natural. APRESENTAÇÃO DO TRABALHO Este trabalho foi organizado em quatro capítulos para a melhor distribuição e entendimento dos assuntos abordados. O capítulo 1 consiste em uma revisão bibliográfica sobre o tema abordado na tese. A revisão intitulada de “Addition of natural extracts with antioxidant function to preserve the quality and shelf life of meat products” trata-se do levantamento bibliográfico sobre o uso de aditivos naturais em produtos cárneos como alternativa a antioxidantes químicos. Este capítulo foi submetido à Revista Food Reviews International. 19 O capítulo 2 trata-se do artigo científico “Red pitaya extract as natural antioxidant in pork patties with total replacement of animal fat” publicado na Revista Meat Science de autoria de Elisa Rafaela Bonadio Bellucci, Paulo E. S. Munekata, Mirian Pateiro, José M. Lorenzo e Andrea Carla da Silva Barretto. Este capítulo aborda a obtenção de extrato a partir da polpa de pitaia vermelha e os efeitos da sua aplicação em hambúrguer suíno com substituição da gordura animal por uma emulsão de óleo de chufa sobre a estabilidade oxidativa e de cor e sobre a aceitação sensorial. O capítulo 3 trata-se do artigo “Açaí extract powder as natural antioxidant on pork patties during the refrigerated storage” publicado na Revista Meat Science, de autoria de Elisa Rafaela Bonadio Bellucci, João Marcos dos Santos, Larissa Tátero Carvalho, Taís Fernanda Borgonovi, José M. Lorenzo, Andrea Carla da Silva Barretto. Este capítulo aborda a obtenção de extrato a partir da polpa açai e os efeitos da sua aplicação em hambúrguer suíno sobre a estabilidade oxidativa e de cor durante armazenamento refrigerado. O capítulo 4 trata-se de um artigo elaborado para ser submetido na Revista Meat Science e aborda os efeitos da aplicação do extrato obtido a partir da polpa açaí em salame tipo Italiano sobre a estabilidade oxidativa e de cor durante a etapa de processamento (maturação). Este artigo foi intitulado “Application of açaí extract powder as natural antioxidant in Italian-type salami under ripening conditions: evaluation on physicochemical and microbiological properties” e é de autoria de Elisa Rafaela Bonadio Bellucci, Larissa Tátero Carvalho, João Marcos do Santos, José M. Lorenzo e Andrea Carla da Silva Barretto. REFERÊNCIAS BIBLIOGRÁFICA Associação Brasileira dos Produtores Exportadores de Frutas e Derivados (ABRAFRUTAS). Açaí: A pequena fruta que movimenta milhões na economia paraense. Publicado em: 13 de agosto de 2019. Disponível em: < https://abrafrutas.org/2019/08/13/acai-a-pequena-fruta-que- movimenta-milhoes-na-economia-paraense/>. Acessado em: 28 de julho de 2020. BALDIN, J. C. et al. Effect of microencapsulated Jabuticaba (Myrciaria cauliflora) extract on quality and storage stability of mortadella sausage. Food Research International, v. 108, p. 551–557, 2018. BELLUCCI, E. R. B. et al. Natural colorants improved the physicochemical and sensorial 20 properties of frozen Brazilian sausage (linguiça) with reduced nitrite. Scientia Agricola, v. 78, n. 3, 2021. BISWAS, A. K.; CHATLI, M. K.; SAHOO, J. Antioxidant potential of curry (Murraya koenigii L.) and mint (Mentha spicata) leaf extracts and their effect on colour and oxidative stability of raw ground pork meat during refrigeration storage. Food Chemistry, v. 133, n. 2, p. 467–472, 2012. BLOUKAS, J. G.; ARVANITOYANNIS, I. S.; SIOPI, A. A. Effect of natural colourants and nitrites on colour attributes of frankfurters. Meat Science, v. 52, n. 3, p. 257–265, 1999. BREWER, M. S. Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Comprehensive Reviews in Food Science and Food Safety, v. 10, n. 4, p. 221–247, 2011. BURRI, S. C. M. et al. Lipid oxidation inhibition capacity of plant extracts and powders in a processed meat model system. Meat Science, v. 162, p. 108033, abr. 2020. CARVALHO, A. V. et al. Chemical composition and antioxidant capacity of açaí (Euterpe oleracea) genotypes and commercial pulps. Journal of the Science of Food and Agriculture, 2017. DOMÍNGUEZ, R. et al. A comprehensive review on lipid oxidation in meat and meat products. Antioxidants, v. 8, n. 10, p. 429, 25 set. 2019. ECHEGARAY, N. et al. Chestnuts and by-products as source of natural antioxidants in meat and meat products: A review. Trends in Food Science & Technology, v. 82, n. 3, p. 110–121, dez. 2018. FAN, X.-J. et al. Effects of Portulaca oleracea L. extract on lipid oxidation and color of pork meat during refrigerated storage. Meat Science, v. 147, p. 82–90, jan. 2019. FERRERES, F. et al. Optimization of the recovery of high-value compounds from pitaya fruit by-products using microwave-assisted extraction. Food Chemistry, v. 230, p. 463–474, set. 2017. GARCÍA-CRUZ, L. et al. Betalains and phenolic compounds profiling and antioxidant capacity of pitaya (Stenocereus spp.) fruit from two species (S. Pruinosus and S. stellatus). Food Chemistry, v. 234, p. 111–118, nov. 2017. 21 HUA, Q. et al. Metabolomic characterization of pitaya fruit from three red-skinned cultivars with different pulp colors. Plant Physiology and Biochemistry, v. 126, p. 117–125, maio 2018. JIANG, J.; XIONG, Y. L. Natural antioxidants as food and feed additives to promote health benefits and quality of meat products: A review. Meat Science, v. 120, p. 107–117, out. 2016. KANG, J. et al. Flavonoids from acai (Euterpe oleracea Mart.) pulp and their antioxidant and anti-inflammatory activities. Food Chemistry, 2011. KARRE, L.; LOPEZ, K.; GETTY, K. J. K. Natural antioxidants in meat and poultry products. Meat Science, v. 94, n. 2, p. 220–227, jun. 2013. KUMAR, Y. et al. Recent trends in the use of natural antioxidants for meat and meat products. Comprehensive Reviews in Food Science and Food Safety, v. 14, n. 6, p. 796–812, nov. 2015. LORENZO, J. M. et al. Berries extracts as natural antioxidants in meat products: A review. Food Research International, v. 106, p. 1095–1104, 2018a. LORENZO, J. M. et al. Influence of pitanga leaf extracts on lipid and protein oxidation of pork burger during shelf-life. Food Research International, v. 114, p. 47–54, dez. 2018b. MARTÍNEZ, L. et al. Comparative effect of red yeast rice (Monascus purpureus), red beet root (Beta vulgaris) and betanin (E-162) on colour and consumer acceptability of fresh pork sausages packaged in a modified atmosphere. Journal of the Science of Food and Agriculture, v. 86, n. 4, p. 500–508, mar. 2006. MERCADANTE, A. Z. et al. Effect of natural pigments on the oxidative stability of sausages stored under refrigeration. Meat Science, v. 84, n. 4, p. 718–726, 2010. MUNEKATA, P. E. S. et al. Effect of natural antioxidants in Spanish salchichón elaborated with encapsulated n-3 long chain fatty acids in konjac glucomannan matrix. Meat Science, v. 124, p. 54–60, fev. 2017. OTÁLORA, M. C. et al. Encapsulated betalains (Opuntia ficus-indica) as natural colorants. Case study: Gummy candies. LWT - Food Science and Technology, v. 103, p. 222–227, abr. 2019. POGORZELSKA, E. et al. Antioxidant potential of Haematococcus pluvialis extract rich in 22 astaxanthin on colour and oxidative stability of raw ground pork meat during refrigerated storage. Meat Science, v. 135, p. 54–61, 2018. RIBEIRO, J. S. et al. Natural antioxidants used in meat products: A brief review. Meat Science, v. 148, p. 181–188, 2019. 23 OBJETIVOS a) Objetivo Geral Este estudo teve por objetivo avaliar o efeito da adição de extratos naturais obtidos da polpa da pitaia vermelha e da polpa do açaí sobre as propriedade físico-químicas como cor, oxidação lipídica e as propriedades sensoriais em hambúrguer suíno durante o armazenamento refrigerado em salame tipo Italiano durante o processamento (maturação). b) Objetivos Específicos  Produzir extratos aquosos a partir da polpa de pitaia e do açaí.  Determinar a quantidade de compostos fenólicos totais e a atividade antioxidantes dos extratos obtidos através das análises FRAP e DPPH.  Aplicar o extrato de pitaia em hambúrguer suíno com substituição total de gordura suína, e adição de emulsão de óleo de chufa (Cyperus esculentus) embalado em atmosfera modificada armazenado sob refrigeração e avaliar seu efeito sob a estabilidade de cor, oxidação e a aceitação sensorial  Aplicar o extrato de açaí em hambúrguer suíno armazenado sob refrigeração e avaliar seu efeito sob a estabilidade de cor e oxidação lipídica.  Aplicar o extrato de açaí em salame tipo Italiano e avaliar seu efeito durante o processamento (maturação) sob a estabilidade de cor e oxidação lipídica. 24 Capítulo 1 ADDITION OF NATURAL EXTRACTS WITH ANTIOXIDANT FUNCTION TO PRESERVE THE QUALITY AND SHELF LIFE OF MEAT PRODUCTS Elisa Rafaela Bonadio Bellucc, Camila Vespúcio Bis-Souza, José M. Lorenzo, Andrea Carla da Silva Barretto Este capítulo será submetido no periódico Food Reviews International 25 ADDITION OF NATURAL EXTRACTS WITH ANTIOXIDANT FUNCTION TO PRESERVE THE QUALITY AND SHELF LIFE OF MEAT PRODUCTS Elisa Rafaela Bonadio Belluccia, Camila Vespúcio Bis-Souza, José M. Lorenzob,c, Andrea Carla da Silva Barrettoa a Department of Food Technology and Engineering, UNESP – São Paulo State University, Street Cristóvão Colombo, 2265, Zip Code 15054-000, São José do Rio Preto, SP, Brazil. b Centro Tecnológico de la Carne de Galicia, Avda. Galicia nº 4, Parque Tecnológico de Galicia, San Cibrao das Viñas, 32900 Ourense, Spain. c Universidade de Vigo, Area de Tecnología de los Alimentos, Facultad de Ciencias de Ourense, 32004 Ourense, Spain *Corresponding author: Email: andrea.carla@unesp.br Abstract - Antioxidants are used to prevent oxidation reactions and inhibit the development of unwanted sensory characteristics that decrease nutritional quality, acceptance and shelf life of processed meat products, improving their stability. Synthetic antioxidants, although efficient, are related to the development of diseases because they present toxic and carcinogenic effects. Thus, researchers and industry study natural alternatives to synthetic antioxidants to be used in meat products, thus meeting the demand of consumers who seek foods without additives in their composition. These natural extracts have compounds that exert antioxidant activity in different meat products by different mechanisms faction. Thus, this review work aimed to gather studies that applied natural extracts derived from different plant sources or not as possible antioxidants in meat products and their action in preserving the quality of these products. 26 Keywords: natural antioxidants, synthetic antioxidants, bioactive compounds, lipid oxidation, meat discoloration, shelf life. Introduction Meat and meat products are affected by several factors during the shelf life that compromise sensory acceptance due to changes in color, odor, flavor and nutritional quality (de Carvalho et al., 2019). Changes in sensory characteristics can result in the rejection of the product and, due to the association with the freshness and safety of the product, discoloration negatively affect the purchase intention by consumers (Agregán et al., 2019). The main cause of deterioration of products of animal origin is oxidation reactions, thus, the use of antioxidants is a way found by the industry to maintain the characteristics of fresh foods for longer by inhibiting these oxidation reactions (Domínguez et al., 2019). Chemical additives with antioxidant action are used to extend the shelf life and specially to avoid discoloration in meat products. Among the most used antioxidants are sodium erythorbate, sodium ascorbate, propyl gallate, butylated hydroxyanisol (BHA), tert-butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), and curing salts as nitrite and nitrate and all of them are associated with toxicological and carcinogenic effects (Baldin et al., 2018; Echegaray et al., 2018; Ribeiro et al., 2019). It is evident that consumers are concerned about the use of chemical additives manly due its carcinogenicity, which results in an increased demand for clean label foods using only natural ingredients. In this way, the risk associated with synthetic antioxidants added to the change in the food habit of the world population, who started to worry about consuming foods with healthier characteristics, prompted the industry to look for natural additives which have greater potential for application due the consumer’s acceptability in relation to synthetics (Nikmaram et al., 2018). In response to this new trend, studies are being carried out in order to find natural alternatives for synthetic additives and among the possibilities found is the use of plants extracts in meat matrices evaluating their action in maintaining sensory and nutritional quality in meat products. As these extracts contain high levels of phenolic compounds present and other antioxidant substances effective in delaying oxidative reactions (Baldin et al., 2018; Fan et al., 2019; Lorenzo, Pateiro, et al., 2018; Lorenzo, Vargas, et al., 2018; Munekata et al., 27 2017; Pogorzelska et al., 2018; Ribeiro et al., 2019). Thus, this review work aimed to gather studies that applied natural extracts derived from different matrices such as fruits, spices in addition to essential oils and residues and by-products of the industry as possible antioxidants in meat products and their action in preserving quality during shelf life (Figure 1-1). Figure 1-1 Illustration representing the application of natural sources as an antioxidant in meat products as a means of preserving quality and shelf life. Oxidation reactions in meat and meat products Oxidation reactions are the main non-microbial causes of meat deterioration during the storage period (Agregán et al., 2019; Cunha, Monteiro, Costa-Lima, et al., 2018) reducing its quality. The presence of unsaturated fatty acids and an increase in oxygen exposure through process steps such as milling, slicing and cooking intensifies these reactions (Lorenzo, Pateiro, et al., 2018). Oxidation reactions initially compromise the sensory quality of meat and its derivatives due the changes in texture, formation of a characteristic flavor of a rancid product and manly discoloration, that influences the acceptance by consumers. In addition, these reactions decrease the nutritional due to the loss of essential fatty acids and vitamins. However, the most impacting consequence refers to the formation of toxic products during lipid oxidation that are considered harmful to health because they are associated with some pathologies such as atherosclerosis, cancer, inflammation and aging processes, among others (Domínguez et al., 2019; Fan et al., 2019; Ribeiro et al., 2019). 28 The lipid oxidation process is complex and has many mechanisms that interact with each other (Domínguez et al., 2019). Figure 1-2 illustrate the mechanism of the oxidative processes that occur in the meat system. In simple terms, oxidation consists of the loss of one or more electrons by the molecule, most characterized by the loss of hydrogen and oxygen gain (Oswell et al., 2018). According to Lorenzo, Pateiro, et al. (2018) oxygen is the main factor that affects lipid oxidation, reacting with unsaturated fatty acids and producing peroxides. The oxidation process occurs in three stapes called initiation, propagation and termination (Lorenzo, Pateiro, et al., 2018; Ribeiro et al., 2019) and result in the modification of fatty acid through a self- catalytic mechanism called self-oxidation (Kumar et al., 2015). Initiation occurs through the action of pro-oxidizing agents or reactive oxygen species (ROS) and conditions favorable to the reaction such as thermal process, presence of light, forming a fatty acid radical (R •) through the removal of the hydrogen radical. In the second stage, the propagation phase, there is the formation of peroxide radical (ROO •) through the reaction between the radical previously formed in the initiation step and the molecular oxygen (O2). In a next reaction, primary products are formed, the so-called hydroperoxides, which are considered relatively stable products (Kumar et al., 2015). Peroxide and hydroperoxide radicals have no taste and odor. However, metal ions, light and heat cause isomerization and decomposition reactions of hydroperoxides giving rise to secondary products such as hexanal, pentanal, 4-hydroxynonenal and malonaldehyde (MDA), among others, responsible for the odor, flavor and texture characteristic of rancidity. Finally, stable or non-reactive products are formed in the termination phase from various reaction combinations that occur between free radicals or from the reaction of free radicals with non-radical compounds such as antioxidants (Domínguez et al., 2019; Ribeiro et al., 2019). One of the most important aldehydes produced during lipid oxidation is malonaldehyde or MDA (1,3-propanedial) because it can produce rancid aroma even in small quantities and it is used to quantify lipid oxidation through analytical methods in meat and meat products. The main methods of analysis used to quantify MDA is the thiobarbituric acid (TBA) test which consists in the colorimetric measurement of the complex formed between TBA and MDA. However, there are other oxidation products that are reactive to thiobarbituric acid with color formation and, therefore, the method determines substances reactive to thiobarbituric acid (TBARs) (Domínguez et al., 2019). There are studies that relate the values of lipid oxidation between 2 and 2.5 mg MDA/kg are considers as accepted limit and still no affect the sensory acceptability of the product (Zhang et al., 2019). Other studies point out different acceptable 29 limits for lipid oxidation that ranged from 0.5 to 2.0 mg MDA/kg (Campo et al., 2006b; Šojić et al., 2020; Tarladgis et al., 1960). Figure 1-2 Mechanisms of the oxidative processes that occur in the meat system (Ribeiro et al., 2019). Many components of the meat can be affected by oxidation reactions, among them proteins, which being "attacked" by reactive oxygen species lose the sulfhydryl groups, generating carbonyl compounds. The protein oxidation mechanism is considered complex, and the formation of carbonyl compounds is considered the main change in oxidized protein (Domínguez et al., 2022). There are four ways that carbonyls can form from protein oxidation, the first is when the amino acid side chain is directly oxidized, the second way is the peptide structure fragmentation, the third is when protein and reducing sugars react to each other and, at least, when a bond between protein and non-protein carbonyl compounds occur (Domínguez et al., 2022). In meat and meat products, the analytical method most used to measure protein oxidation is based on the spectrophotometric quantification of carbonyl compounds during the conversion of 2,4 dinitrophenylhydrazine to hydrazones (Lorenzo, Pateiro, et al., 2018). Protein contributes to important technological, nutritional and sensory properties, so it is considered the main constituent of meat. Modification in its structures results in protein denaturation or proteolysis, affecting such meat properties (Domínguez et al., 2022). Due to protein oxidation, meat systems become more susceptible to the action of proteolytic enzymes, in addition to undergoing polymerization of proteins producing soluble aggregates that can cause gelation and emulsification, or insoluble aggregates that prevent binding with water 30 affecting its solubility. Protein oxidation can also alter its digestibility, affecting nutritional quality (Jiang & Xiong, 2016). Finally, the impact caused by protein oxidation that most affect the quality of meat and meat products is the color changes, more specifically called pigment oxidation (Domínguez et al., 2022). Myoglobin, a protein and a pigment of the meat, is formed by a protein (globin) and a non-protein prosthetic group (heme) that contains an iron molecule. and the color variation of this pigment depends on the oxidative state that the iron. Myoglobin is purplish red in color, however it becomes bright red in the presence of high oxygen concentration, as it has become oxymyoglobin and the iron ion has become reduced (Fe++). It is concluded that the partial pressure of O2 is responsible for the oxidation states of myoglobin and that a low partial pressure favors the formation of deoxymyoglobin, an unstable pigment, which in the presence of oxygen and through the action of free radicals, becomes metmyoglobin, brown in color and iron is in oxidized form (Fe+++) (Lorenzo, Pateiro, et al., 2018; Ribeiro et al., 2019). Therefore, from a commercial point of view, with color being the sensory attribute that most influences the consumer's purchase intention (Bellucci, Barretto, et al., 2021; Ruiz- Capillas et al., 2015), oxidation reactions increase the possibility of product rejection. Therefore, the shelf life of meat and meat products consists of the moment when the consumer can detect the oxidation products that confer the rancid characteristic (mainly volatile) or observe changes in the color of the meat (Domínguez et al., 2019). Antioxidants used in meat products Antioxidants are used by the industry as the main strategy to decrease the intensity of oxidation reactions in meats and their processed derivatives, increasing the useful life of these products (Cunha, Monteiro, Lorenzo, et al., 2018; Domínguez et al., 2019). Synthetic compounds with phenolic structure capable of acting through the elimination of peroxyl radicals or canceling the formation of free radicals are used for this purpose (Lorenzo, Pateiro, et al., 2018). The reaction of antioxidants with oxidation is believed to occur by donating electrons to break and terminate the oxidation at the propagation step and thereby preventing additional lipid and protein radicals from forming (Falowo at al., 2014) as showing in Figure 1-3. 31 Figure 1-3 Mechanism of antioxidant in lipid oxidation at propagation step (Falowo at al., 2014). In Brazil, the list of allowed antioxidants is different depending on the meat product that will be prepared, however, in general, the use of ascorbic acid and its derivatives is allowed (sodium, calcium and potassium ascorbate, erythorbate and sodium isoascorbate, among others) in the amount necessary to obtain the desired technological effect (quantum sadis) (Brasil, 2019). For specific products such as fresh, cooked and dried processed meat products and for canned meat and mixed, the use of propyl gallate (GP), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) in a maximum quantity between 0.01 and 0.02 g/100g on the fat content. According to Ribeiro et al. (2019), it is important to know the maximum allowed limits of synthetic antioxidants that will be added to meat products because their consumption can have a toxic effect. Antioxidants such as BHA and BHT can cause DNA mutations and increase blood cholesterol levels, in defense the body produces enzymes that can destroy important compounds such as vitamin D, causing hives and dermatitis. Also, propyl gallate cause gastric irritations and allergic reactions, change in vision and respiratory problems when consumed in large concentrations. Ascorbic acid, on the other hand, must be used according to Good Manufacturing Practices (GMP) to be considered a safe additive. 32 Nitrite and nitrate salts can also be used to reduce oxidation reactions in meat products, as these salts, in addition to their preservative and curing action, also have an antioxidant action (Bellucci, Barretto, et al., 2021). However, these salts are widely associated with an increased risk of cancer due to the formation of harmful compounds called nitrosamines through the reaction of nitrite with components present in meat such as secondary amines (Gómez et al., 2015). Although these compounds have high efficiency, even at lower concentrations, consumers and the food industry started to consider the relationship between food ingredients and health, which increased the demand and interest in natural ingredients (Bernardo et al., 2021; Cunha, Monteiro, Lorenzo, et al., 2018). In parallel, there has been a change in the habits of the population in recent years, as they have become more food conscious, concerned with consuming healthy, affordable and good-looking foods (Rodrigues et al., 2020). Teixeira & Rodrigues (2021) also state that the way meat products are produced is a concern of consumers, so the industry started to reformulate its meat products to meet this new demand for cleaner and healthier labels by reducing fat, salt, curing salts (nitrite and nitrate) and replacing synthetic antioxidants. In this regard, synthetic antioxidants are being rejected by consumers due its potential toxicity use of natural extracts has also been of great interest to the meat industry since they started to be considered safe for consumers (Munekata et al., 2020). So, studies are being conducted in order to find compounds obtained from plants capable of reducing oxidative reactions in meat products and replacing synthetic antioxidants (Pateiro, Gómez-Salazar, et al., 2021). In addition, such replacement has been widely accepted by consumers (Munekata et al., 2017). To be considered as antioxidants, natural extracts cannot negatively interfere with sensory characteristics, act at low concentrations, must be cheap and stable over time and storage conditions and, most importantly, cannot be harmful or toxic in the concentrations used (Munekata et al., 2020). Different plants are being tested in order to investigate their action as antioxidants in meat and meat products and extracts are obtained from different sources such as pulp, peel and seeds of fruits, spices and essential oils, as well as other sources such as flowers, nuts, leaves, vegetable residues, etc. (Basanta et al., 2018; Bellucci et al., 2022; Bellucci, Munekata, et al., 2021; Burri et al., 2020; Dadi et al., 2019; Khemakhem et al., 2019). The mechanism of action of natural antioxidants is different depending on the compound. In general, they antioxidant effect is associated with their eliminating reactive 33 species capability, curbing autoxidation by donating a hydrogen atom to stabilize the first oxidation products, their potential chelation of metal ions (pro-oxidants) and their role as inhibitory of the hydro peroxide’s degradation (Nikmaram et al., 2018). Phenolic compounds are the main natural antioxidants, among them the most important are tocopherols, flavonoids and phenolic acids. These compounds have a strong ability to donate hydrogen and a free radical scavenging capability (Kumar et al., 2015; Munekata et al., 2021; Nikmaram et al., 2018). According to Oswell et al. (2018), the compounds that have antioxidant activity in meat products are carotenoids, hydroxycinnamic acid, flavonoids, terpenes and antioxidant vitamins. Antioxidant substances derived from fruits (pulp, pigment, peel and seeds) applied in meat products Studies using fruit or extracts derived from fruits as a source of antioxidant compounds have been widely carried out in meat products, as many works have been published in recent years (Baldin et al., 2016, 2018; Basanta et al., 2018; Bellucci, Barretto, et al., 2021; Bellucci et al., 2022; Cunha, Monteiro, Costa-Lima, et al., 2018). Fruits are rich sources of bioactive compounds and pigments such as phenolic compounds, including anthocyanins and catechins, in addition to carotenes, betacyanin that have high antioxidant capacity. Table 1-1 shows studies and results about the application of substances derived from fruits (pulp, pigment, peel and seeds) applied in meat products Some works evaluated pink guava fruit as natural antioxidant in meat products. In this regard, Chang et al. (2020) observed that flesh of this fruit presented an important antioxidant activity for containing large amount of vitamin C and phytochemicals as lycopene, carotenoid, and flavonoids. Also, the phenolic compounds like apigenin, myricetin, anthocyanins, and ellagic acid contribute with the ascorbic acid to its antioxidant activity. Therefore, the antioxidant action of pink guava pulp was evaluated by Joseph et al. (2014a) in raw pork emulsion for nine days in refrigerated storage and aerobic packaging. The authors found that guava pulp improved the color stability during storage and stated that the increase in added concentration resulted in an increase in a* values and correlated this effect to the lycopene present in the emulsion (1.9 vs. 4 .4 mg.kg-1 in 0 day and 1.7 vs. 3.5 mg.kg-1 in 9 storages for 5 vs. 10%). Regarding the antioxidant effect, they observed that the pink guava pulp was able to minimize lipid oxidation, showing higher values of TBARS for the control without addition of pulp and the values were reduced according to the increase in the added concentration of the 34 pink guava pulp. Pink guava pulp was also studied, in addition to tomato-derived products (tomato puree, freeze-dried tomato peel and tomato pulp), by Joseph et al. (2014b) in pork emulsified products (Table 1-1). In this study, the tested antioxidants improved the color, reduced the formation of metmyoglobin and minimized lipid oxidation in raw meat emulsion, at the concentrations studied. According to Domínguez et al. (2020), tomato by-products such as seed, skin, pulp residues are rich in a variety of compounds with antioxidant and coloring properties, such as carotene (lycopene, beta-carotene, phytoene, phyto fluorine and lutein), phenolic compounds (phenolic acids and flavonoids) and vitamins A and C. In addition, García-Cruz et al. (2017) affirmed that phenolic compounds are photochemical associated with beneficial effects to the body, which may have a cardio protective, antioxidant, antimicrobial, ant allergic action and act against thrombosis. Moreover, Mercadante et al. (2010) investigated the effect of natural pigments (norbixin, lycopene, zeaxanthin and β-carotene) in refrigerated sausage (Table 1-1) and reported that the dyes improved the oxidative stability of the product, wherein zeaxanthin and norbixin were the most efficient. Basanta et al. (2018) prepared and extracted fiber microparticles from the pulp and skin of Japanese plum by extraction with ethanol and then freeze dried to be added to breast chicken patties (Table 1-1). According to the authors, the microparticles retain carotenoids, tocopherol and phenolic compounds (pentameric proanthocyanidins) and the cyanidin present in high amounts is responsible for the intense red color of the fiber. The microfiber obtained from the pulp was able to improve the color within 10 days of refrigerated storage and both (added at 2.0%) reduced lipid oxidation around 50%. Pomegranate peel as phenolic compounds source: Advanced analytical strategies and practical use in meat products, the pomegranate peel represents 40 to 50% of the total weight of the fruit and, being rich in phenolic compounds, it is a valuable residue of the industry. Pomegranate peel has considerable amounts of phenolic compounds, including flavonoids (anthocyanins, catechins and other complex flavonoids) and hydrolysable tannins, as well as phenolic and organic acids, which gives them an important antioxidant, antimicrobial, antifungal and coloring action (Smaoui et al., 2019). Turgut et al. (2017) investigated the antioxidant action of an aqueous and freeze-dried extract obtained from pomegranate peel (0.5 and 1%) in frozen beef meatballs (Table 1-1). The extract added in concentration not only kept the color stable for 6 months of storage, but also delayed lipid and protein oxidation without impairing sensory acceptance. 35 Table 1-1 Fruit (pulp, pigment, peel and seeds) as a source of antioxidants applied in meat products. Source or active compound Levels added Sample Storage conditions Effect Reference Pink guava pulp (PGP) (ß-carotene and lycopene) 5%, 7.5% and 10% Raw pork emulsion Nine days in refrigerated storage and aerobic packaging Increase redness (improves color) Reduces metmyoglobin formation High inhibition of lipid oxidation at 10% Joseph et al., 2014a Tomato products and PGP Tomato puree - 10%, tomato pulp - 12.5%, lyophilized tomato peel -6 %, and PGP - 10 %) Pork emulsion 9 days at 4±1 °C in darkness and aerobic conditions Improves color and reduce lipid oxidation Joseph et al., 2014b Natural pigments: Norbixin, lycopene, zeaxanthin, ß-carotene 0.05 g/100 g Sausage Vacuum package and under refrigeration (4 °C) for 45 days Norbixin and zeaxanthin presented the lower lipid oxidation values after 45 days of refrigerated storage. Mercadante et al., 2010 Plum (Prunus salicina) peel and pulp micro particles 2.0 % w/w level Breast chicken patties Polyethylene films (oxygen permeability= 5.20×10−12 m3·m−2·s−1·Pa−1); dark, at 4.0 ± 0.5 °C for 10 days Improve color end reduced lipid oxidation around 50% during 10-days storage Basanta et al., 2018 36 Source or active compound Levels added Sample Storage conditions Effect Reference Red pitaya extract 500, 750 and 1000 mg.kg-1 Pork patties with replacement of animal fat Modified atmosphere (80% O2 and 20% CO2); fluorescent light during 18 days at 2 ± 1 °C Did not affect cooking loss and texture profile; increased redness and improve color stability; Reduce lipid oxidation and improve sensorial scores Bellucci et al., 2021 Açaí extract powder 250, 500 and 750 mg.kg-1 Pork patty Packed in nylon- polyethylene bags without vacuum during 10 days in the dark at 2 °C Reduced lipid oxidation; not affect pH, cooking parameters (weight loss and shrinkage); 500 and 750 mg.kg-1 compromised color parameters; 250 mg.kg-1 was effective as natural antioxidant Bellucci et al., 2022 Microencapsulated extract of pitaya (Hylocereus c.) peel 100 and 1000 ppm Pork patty submitted to high pressure processing 9 days of storage at 4 °C under aerobic packaging Able to reduce protein oxidation after 9 days of storage; not affect lipid oxidation Cunha et al., 2018 Microencapsulated jaboticaba (Myrciaria cauliflora) extract 2 and 4% Fresh pork sausage Dark and aerobic condition for 15 days under refrigeration (1 ± 1°C) Reduce lipid oxidation; 4% affect color, texture and overall acceptance of the pork sausages; 2% not affect global acceptance and color Baldin et al., 2016 Lyophilized powder of pomegranate peel extract 0.5 and 1.0% PE Beef meatballs Oxygen-permeable bags; frozen storage at−18±1 °C for 6 months Increase L* values; reduce discoloration (higher values of a* at the end of storage); reduce protein and lipid oxidation Turgut et al., 2017 37 Source or active compound Levels added Sample Storage conditions Effect Reference Dry red grape pomace 1 and 2 % Meat emulsion system/cooked sausage Vacuum package and storage at 4 °C Affected color; not compromise sensory acceptance; increase the oxidative stability Riazi et al., 2016 Grape seed extract 100, 300 and 500 ppm Beef sausage model system Package in PVC; frozen at - 18 °C for 4 months All concentrations protected the meat product model system against lipid oxidation Kulkarni et al., 2011 1000 mg.kg-1 Pig liver pâté Packed in glass containers (150 g); cooked; stored in the dark at 4°C for 24 weeks Reduced lipid oxidation better than BHT Pateiro et al., 2014 Guarana seeds extract 250, 500 and 1000 mgkg Pork patty Modified atmosphere (80% O2 and 20% CO2); under light during 18 days at 2 ± 1 °C Shown higher inhibitory effect on oxidation reactions (lipidic and protein) than BHT at 250 and 500 mg.kg-1; improvement of product color Pateiro et al., 2018 Rosemary, acerola and citrus (lemon, orange and buffered vinegar) extracts Rosemary extract -0,3%; acerola extract – 0,25%; citrus extract – 0,6% Beef patty Packaged in polystyrene trays lined with an oxygen permeable PVC film and displayed simulating retail conditions during 6 days at 4 ± 1 °C Citrus extract inhibited the lipid oxidation during all display period and reduced the aerobic plate count; citrus and acerola improved patty color Fruet et al., 2019 38 In a study, Riazi et al. (2016) showed that the addition of red grape pomace powder in beef sausage emulsion reduced the lipid oxidation in the product during refrigerated storage compared to the control, and that, despite making the samples darker, sensory acceptance was not compromised. Also in this study, the authors stated that the grape pomace powder had the potential to be used as a healthy ingredient and to act in the oxidative stability of meat products. Grape pomace is a residue from the production of grape juice consisting of skin, seed and stem that contains high amounts of polysaccharides and phenolic compounds, which are mostly anthocyanins, catechins, procyanidins and phenolic acids. Grape seed can be considered an alternative to preserve oxidative stability in meat products because it has a high content of polyphenolic compounds and proanthocyanidins and mainly because it presents benzoic acids (gallic acid and protocatechuic acid), flavan-3-ols monomer and oligomeric procyanidins as main compounds (Pateiro et al., 2014). Kulkarni et al. (2011), for example, studied the effect of grape seed extract on pre-cooked, frozen, and re- heated beef sausage model system. In this study, the extract was added at concentrations of 100, 300 and 500 mg.kg-1 and compared to a control, ascorbic acid, and propyl gallate, both added at a concentration of 100 mg.kg-1. The authors found that the extract used at the lowest concentrations showed similar results to propyl gallate for oxidative, color and sensory stability in the 4 months of storage. In pork liver pate, the addition of grape seed extract (1000 mg.kg-1) had a better effect on lipid oxidation than the treatment added with BHT (200 mg.kg-1) during storage at 4 °C for 24 weeks, confirming its antioxidant capacity in meat products (Pateiro et al., 2014). In pork patties, the effect of adding extract of guarana seeds was evaluated by (Pateiro, Vargas, et al., 2018). In this work, the patties were evaluated for 18 days of storage at 2 ± 1 °C (Table 1-1) and it was observed that the guarana seed extract at both concentrations (250 and 500 mg.kg-1) not only reduced lipid oxidation when compared to BHT (200 mg.kg-1) as well as improved color, lipid, and protein stability. It should also be noted that the guarana seed extract has many phenolic compounds, with tyrosol and other low molecular weight phenolics appearing in greater quantities. An interesting approach was tested to improve the stability of beef patty during storage (6 days at 4 °C under retail conditions). With the purpose of evaluating the effect of a new combination of extracts, Fruet et al. (2019) developed an antioxidant with lemon, orange and buffered vinegar and compared it with acerola and rosemary extracts in beef patties. Although rosemary and acerola extract (0.30 and 0.25 %) were similar in relation to antioxidant activity, 39 there was an increase in lipid oxidation products during storage. However, the proposed extract avoided the formation of these lipid oxidation products in the concentration used (0.6%), inhibited the increase in TBARS values during storage. Moreover, it was efficient inhibited de growing of aerobic bacteria during storage due the buffered vinegar in this extract. Betacyanin as betanin are present in pitaya or dragon fruit, and they are known not only for their color, but also for their antioxidant activity that record up to double of some anthocyanins (Shaaruddin et al., 2017). According to Cunha, Monteiro, Costa-Lima, et al. (2018), pitaya has a large amount of beta-carotene, betacyanin, lycopene, vitamin E, vitamin C and polyphenols. In this context, a study was developed to investigate the antioxidant effect of red pitaya extract in pork patties with animal fat replacement (Bellucci, Munekata, et al., 2021). In this work, an aqueous extract and lyophilized of red pitaya pulp (250, 500 and 1000 mg.kg- 1) were added to the patties as a natural antioxidant. A control (without antioxidant) and a treatment added of sodium erythorbate (500 mg.kg-1) was elaborated. The authors observed a reduction in discoloration over time due to pitaya extract. Regarding oxidative stability, the extract was effective in reducing lipid oxidation reactions and carbonyl formation compared to the control without the addition of antioxidants. Another extract that has been studied for application in meat products was obtained from açai palm fruit (Euterpe oleracea Mart.). This fruit is natural from Amazon region and is widely known for having high antioxidant activity in vitro due its rich composition in phenolic compounds such as different anthocyanins, flavones, and phenolic acids (Gordon et al., 2012). Therefore, Bellucci et al. (2022) applied an extract obtained from the açaí pulp in pork patty to evaluate its antioxidant activity (Table 1-1). The results of this study showed that açaí extract reduced lipid oxidation in pork hamburger as much as sodium erythorbate. However, bioactive compounds derivate of plant are instable chemically and can undergo to oxidative degradation by exposure to oxygen, light, metal ions, pH and other parameters, and their direct incorporation in food is very restricted (Baldin et al., 2016; Smaoui et al., 2021). In this regard, microencapsulation is an innovative technique that can be used to preserve the bioactive present in plant extracts, enabling greater stability and preserving their functional quality (Cunha, Monteiro, Costa-Lima, et al., 2018; Munekata et al., 2020; Shaaruddin et al., 2017; Smaoui et al., 2021). Studies were performed to investigate the preservation of bioactive by a microencapsulation (Dadi et al., 2019; do Valle Calomeni et al., 2017; Ghaderi-Ghahfarokhi et al., 2017; Leyva-Porras et al., 2021; Sharayei et al., 2020; Taofiq et al., 2019). On the other hand, some authors investigated the use of microcapsules containing 40 bioactive compounds in meat products as alternative to synthetic antioxidants (Baldin et al., 2018; Cunha, Monteiro, Costa-Lima, et al., 2018; Surendhiran et al., 2020; Tometri et al., 2020). In this regard, Cunha, Monteiro, Costa-Lima, et al. (2018) evaluated the action of microencapsulated extract from pitaya peels on pork hamburger submitted to a high-pressure process. The process causes protein denaturation and membrane damage, releasing non-heme iron, and consequently increasing lipid oxidation. The authors reported that the microencapsulated extract from pitaya peels (100 and 1000 mg.kg-1), despite not having reduced lipid oxidation as much as BHT (100 mg.kg-1), was able to minimize it and was also able to reduce protein oxidation after 9 days of refrigerated storage in an emulsified pork product subjected to a high-pressure process. Also, the extract rich in anthocyanin (cyanidin-3- o-glucoside) obtained from Jaboticaba peel was microencapsulated by spray-dryer into maltodextrin to be applied in fresh pork sausage as natural antioxidant (Baldin et al., 2016, 2018). In this work, jaboticaba extract microencapsulated were added at the concentrations of 2 and 4% reduced lipid oxidation during 15 days of refrigerated storage, presenting TBARS values lower than the other treatments (cochineal carmine and control) (Baldin et al., 2016). Still on this study, the extract (2%) did not affect the global acceptance of fresh sausage, showing itself to be an alternative as a natural coloring agent. Spices and essential oils as naturals source of antioxidants applied in meat products Herbs and spices or products derivates from them have been used to preserve food to be considered as natural antioxidant, mainly in meat products and its use is beneficial to prevent the oxidation reactions and develop clean label products, desirable for consumers (Khanam & Prakash, 2021). Table 1-2 shows studies and results about the application spices and essential oils as naturals source of antioxidants in meat products. Many studies have been conducted in recent years to investigate its effect on the stability of lipid oxidation, protein and color in meat products. For example, the aqueous extract of Dog rose (Rosa canina L., RC) was tested by Vossen et al. (2012) in porcine sausage (frankfurter) for presenting relevant amounts of phenolic compounds, such as procyanidins and catechins, and ascorbic acid. The concentrations of 5 and 30 g/kg of extract were compared to both positive control treatment added to ascorbic acid (0.5 g/kg) and sodium nitrite (0.1 g/kg) and a negative control without the addition of any antioxidant. In this study, dog rose extract decreased the formation of lipid oxidation products as much as the combination of sodium nitrite and ascorbic 41 Table 1-2 Spices, products derivates from them and essential oil as naturals source of antioxidants applied in meat products Source or active compound Levels added Sample Storage conditions Effect Reference Dog rose (R. canina L.) extract 5 and 30 g.kg-1 Porcine sausage (frankfurter) Oxygen permeable poly-vinyl chloride film; dispensed in polypropylene trays; stored for 60 days at 2 °C in the dark Both concentrations inhibited lipid and protein oxidation Vossen et al., 2012 Rosemary extract 0, 250, 500 and 750 mg.kg-1 Reduced nitrite cooked liver pattè Seven days in the dark at 4 °C; Significantly reduced the lipid oxidation. However, it shows no effect on color stability. Doolaege et al., 2012 1500 and 2500 ppm Raw frozen and cooked- frozen pork sausage Packed in high-barrier film pouches; frozen in an air-blast freezer at -30 °C; stored at -20 °C. In raw frozen: Inhibited the lipid oxidation, shown lower values than samples with BHA/BHT; improve and preserved the color. Cooked-frozen: oxidation results were similar to BHA/BHT treatments. Sebranek et al., 2005 Ranged from 500 to 3000 ppm Fresh- refrigerated pork sausage Placed on foam-board trays, overwrapped with oxygen- permeable film and stored on shelves under 7.0 lux florescent lighting at 2–4°C 2500 and 3000 was effective in inhibiting lipid oxidation such as the use of BHA/BHT; Sebranek et al., 2005 42 Source or active compound Levels added Sample Storage conditions Effect Reference Turmeric (Curcuma longa L.) extract 250, 500 and 750 mg.kg-1 Fresh lamb sausage with fat replacement by Tiger nut (Cyperus esculentus L.) oil Package under modified atmosphere (80% O2 and 20% CO2) in 300 mm thick PET- EVOH-PE trays, sealed with multilayer PE-EVOH-PE film, stored at 2 ± 1 °C under light for 18 days. 750 mg.kg-1 shown the highest antioxidant activity throughout the storage period; reduced lipid oxidation; de Carvalho et al., 2020 Turmeric and black pepper spices For 250 g of raw patty: turmeric – ranged from 0 to 6 g; black pepper – ranged from 0 to 1,1 g. Cooked meat patties Stored at -20ºC until tested. A decrease in lipid peroxidation. Zhang et al., 2015 Pink pepper extract Pink pepper extract concentration equivalent to 90 mg GAE/kg meat Chicken burger Two different conditions: aerobic packing (PVC film) and vacuum; stored at 2 °C with white light incidence; evaluated for consecutive 7 days Pink pepper extract reduced oxidative products as BHT synthetic antioxidant and improved color (presented higher redness). Regarding to packing condition, no effect was observed to pink pepper and BHT treatments on lipid oxidation. Menegali et al., 2020 43 Source or active compound Levels added Sample Storage conditions Effect Reference Clove extract 0.1 % Cooked beef patties Packaged in a polyethylene pack and storage under refrigerated conditions for 10 days The clove extract was more efficient to reduce lipid oxidation in cooked beef patties than BHT (0,02%) and ascorbic acid (0,05%); more efficient in preserve protein oxidation than BHT and similar to ascorbic acid at 10 days Zahid et al., 2020 Rosemary essential oil 150, 300 and 600 ppm Different types of frankfurters: produced with tissues from Iberian pigs (IF) or white pigs (WF) Storage in the dark for 60 days at 4 °C. IF: 300 and 600 ppm was effective to reduce lipid oxidation and 600 ppm reduce hexanal formation. WF: 300 and 600 ppm increased lipid and protein oxidation Estévez & Cava, 2006 0.2 % Poultry fillets Two different conditions: air- packaging and modified atmosphere The combination of rosemary essential oil and modified- atmosphere packaging reduced the level of lipid oxidation. Kahraman et al., 2015 Blend of essential oils (garlic, cinnamon, cloves and rosemary); Rose hips extract 1 g/kg of meat; 300 mL/kg of meat Iberian cooked hams Vacuum-packed and storage for five months under refrigeration conditions (4 °C) The essential oils blend was more effective against lipid oxidation than rose hips extract and commercial antioxidant. Whereas rose hips presented better results regard the protein oxidation. Armenteros et al., 2016 44 Source or active compound Levels added Sample Storage conditions Effect Reference Essential oils: clove; holy basil; cassia and thyme oil clove oil (0.25%), holy basil oil (0.125%), cassia oil (0.25%) and thyme oil (0.125%) Fresh chicken sausage Packed in vacuum and stored at 18 ± 2 ° C for 45 days Clove oil was the most effective regarding reduce lipid oxidation. All essential oil presented antioxidant action and reduced discoloration during storage. Sharma et al., 2017 Satureja montana L. essential oil 7.80, 15.60 and 31.25 µl/g. Mortadella- type sausage with different concentration of sodium nitrite Storage at room temperature (25 °C) for 30 days. Color was negatively affected by addition of high concentrations of essential oil; essential oil at 15.60 μl/g with sodium nitrite (100 mg.kg-1) presented the best results regarding reduce lipid oxidation Coutinho de Oliveira et al., 2012 45 acid, however, it was shown to be more efficient in inhibiting protein oxidation during the 60 days of refrigerated storage (2 °C). In addition, the authors report a possible contribution of the extract in the pink sausage coloration. The antioxidant effect of rosemary extract has been demonstrated in many studies in different products such as hamburgers, sausages, fresh and cooked sausages (Fruet et al., 2019; Gao et al., 2019; Heck et al., 2018; Schilling et al., 2018). Doolaege et al. (2012), for example, investigated the action of different concentrations of rosemary extract (0, 250, 500 and 750 mg.kg-1) on lipid oxidation and color stability in liver pâté added from different concentrations of sodium nitrite (40, 80 and 120 mg.kg-1). Rosemary extract contains phenolic diterpenes, such as carnosic acid and carnosol that act as hydrogenic donors in the chain reaction of free radicals, having its antioxidant action well known. What was proven in this study, since the addition of the extract provided lower values of TBARS, therefore, decreased lipid oxidation in raw and cooked liver pâté, however, no concentration effect was observed between 250 and 750 mg.kg- 1. At the same time maintained high concentrations of antioxidants ascorbic acid, α-tocooferol and carnoic acid. Regarding sodium nitrite, it was concluded that the addition of 80 mg.kg-1 of nitrite would be sufficient when associated with the addition of the extract without impairing the oxidative and color stability of the pâté. On the other hand, Sebranek et al. (2005) carried out two experiments under study for antioxidant efficacy of rosemary extract, the first concentrations of 1500 and 2500 mg.kg-1 were added in frozen and precooked sausages and, the second, from 500 to 3000 mg.kg-1 in refrigerated sausages and the addition of BHA and BHT in the proportion of 1:1 (200 mg.kg-1) was compared. In the first experiment, rosemary extract was more efficient leaf than synthetic antioxidants, presenting better results for oxidation stability and instrumental and sensory color in raw sausage stored under freezing for 112 days. The same did not occur in cooked-frozen sausage, because the concentrations of the extract presented similar results to each other and did not differ from the treatment with BHA/BHT in relation to the antioxidant potential. Regarding the second experiment, 2500 and 3000 mg.kg-1 of rosemary extract had a similar effect to BHA/BHT in fresh chilled sausage. In this work it was interesting to realize that the extract at 2500 mg.kg-1 was as effective as the maximum permitted concentrations of BHA/BHT in fresh and chilled pork sausage and in the boiled frozen sausage but was superior to the BHA /BHT in raw frozen sausage, showing that applications and conditions should always be considered. Either way, rosemary extract can be used with a substitute for synthetic antioxidants. 46 Turmeric (Curcuma longa L.) is widely used as seasoning, preservative and dye in food in Southeast Asian countries, India and China. Several sesquiterpenes and curcuminoids were isolated from the turmeric rhizome, and curcuminoids were responsible by their yellow color action and equivalent between 2 and 7% of their composition (de Carvalho et al., 2020; Gul & Bakht, 2015). The condiment is obtained by boiling the roots, followed by drying, but for the study conducted by de Carvalho et al. (2020), the extract was obtained from the extraction by supercritical fluid and presented high antioxidant capacity for the extract from the DPPH and ABTS assays (42.92 mg Trolox/g and 1490.53 mg ascorbic acid/100g, respectively). When applied in lamb sausage with fat substitution by vegetable oil emulsion, the extract at the concentrations used (250, 500 and 750 mg.kg-1) had better results in preventing lipid oxidation than sodium erythrobate (500 mg.kg-1) during the 18 days of refrigerated storage. Another study was conducted with the addition not only of turmeric, but also of black pepper (Piper nigrium) in cooked meat patties (Zhang et al., 2015). For the authors, curcuminoids exhibit high antioxidant activity due to their cityhood of trapping oxygen radicals and breaking the oxidation chain. In this study, black pepper did not present antioxidant effect, but there was a synergistic effect with turmeric, with its antioxidant activity increased when associated with aromatic constituents and terpenes present in black pepper. In a different study, pink pepper extract (Schinus terebinthifolius Raddi) was added to chicken burgers with antioxidant effect as much as BHT during refrigerated storage for 7 days (Menegali et al., 2020). The authors also state that pink pepper extract, which contains bioactive compounds such as ascorbic acid, phenolic and carotenoid compounds, can be used as an antioxidant in a meat product, reducing lipid oxidation, without affecting sensory attributes. Clove is another widely studied spice as it is an aromatic condiment widely used in food and are added to increase shelf life and reduce food deterioration. It also has a variety of bioactive compounds, such as sesquiterpenes and triterpenoids and eugenol (4-alil-2- methoxyphenol), its main bioactive compound that has insecticide and antioxidant properties and is classified as a safe additive (El-Maati et al., 2016). After studying on different solvents to obtain clove extract, El-Maati et al. (2016) concluded that ethanol is more appropriate for the extraction of phenolic and flavonoid stations and that the antioxidant capacity was directly proportional to the amount of total phenolic compounds. Zahid et al. (2020), however, used water as a solvent to obtain clove extract and applied it in cooked beef patties stored under refrigeration. Still, in 10 days of storage, the aqueous extract of clove (0.1%) showed the best antioxidant activity, as it had lower values of lipid and 47 protein oxidation compared to BHT (0.02%). Regarding ascorbic acid (0.05%), the extract had higher values a* and color notes in the sensory analysis preserving the color of hamburgers, in addition to lower TBARS values. All antioxidants used reduced lipid and protein oxidation, maintained better a* values and preserved the stability of sensory attributes when compared to control without added antioxidants. The incorporation of essential oils in meat products is an alternative to preserve oxidation reactions because they are rich in bioactive compounds, such as terpenes, terpenoids and phenolics that have proven antimicrobial, antifungal, antioxidant, antiviral, antimycotic, antiparasitic and insecticide properties (Falleh et al., 2020; Pateiro, Munekata, et al., 2021). In addition, essential oils are classified General Recognized as Safe (GRAS), allowing their use in food products as safe additives, besides being widely accepted by consumers, thus generating greater interest in their application in meat products (Pateiro, Barba, et al., 2018). Ginger, oregano, rosemary, sage, thyme, mint and many other aromatic plants are the main sources of essential oils. Conventional distillation methods using steam and hydro distillation are employed, while innovative techniques are being proposed to obtain essential oil such as hydro distillation with the aid of microwaves or extraction with supercritical fluids and assisted ohmic distillation (Pateiro, Barba, et al., 2018). Many studies were carried out in order to investigate the antioxidant and antimicrobial potential of several essential oils in meat products (Sharma et al., 2017; Smeti et al., 2018; Vieira et al., 2019). According to Pateiro, Barba, et al. (2018), the bioactive present in the essential oils responsible for antioxidant activity are alcohols, aldehydes, phenylpropanoids, terpenes and keones, while compounds such as thymol, cymenone p, γ-terpineno and carvacrol are related to their antimicrobial activity and may act against various bacteria including Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Clostridium perfringens, Clostridium sporogeneses in various meat products. Estévez & Cava (2006) studied the effect of rosemary essential oil on lipid and protein oxidation in several types of frankfurters, produced with Iberian pig and white pig. In white pig frankfurters, 300 and 600 ppm reduced TBARS values, and the highest level also reduce hexanal formation. Whereas levels of 150 ppm of rosemary essential oil were sufficient to inhibit lipid and protein oxidation, while higher concentrations (300 and 600 ppm) increase oxidation reactions in frankfurters formulated with Iberian pig. Therefore, the authors reported that the oxidative stability of the product depends on the concentration of the natural antioxidant used and on the components of the meat. In this sense, Pateiro, Barba, et al. (2018) claim that 48 some essential oils can act as pro-oxidants when added in high concentrations. Another study was carried out by applying rosemary essential oil to chicken fillets packed different conditions: air-packing and modified atmosphere (Kahraman et al., 2015). At concentrations of 0.2%, the rosemary essential oil inhibited the oxidation reaction, when in combinate with modified atmosphere packaging showed grater antioxidant activity and resulting in a reduction of lipid oxidation. All treatments added essential oil presented lower TBARS values than those no added. Although it had no effect on the growth rate of the bacteria tested, reduced the L* values and increased the values of a* in 5 and 7 days of storage at 4° C. Armenteros et al. (2016) evaluated a blend of essential oils (garlic, cinnamon, clove and rosemary) and rose hip (R. canina L.) extract as natural antioxidant compared to a commercial antioxidant (citrate and sodium erythrobate, 1:1; w/w) in Iberian cooked ham via injected brine. In this study, after150 days of refrigerated storage, the mixture of essential oils (1 g/kg of meat) was more effective in controlling lipid oxidation on cooked ham than the other antioxidants studied. Regarding this, the authors affirmed that there was an abundant amount of antioxidant compounds in the spices studied, including carnosol and rosmarinic acid that was present in clove and rosemary. Whereas, for protein oxidation, rose hip and commercial antioxidant presented the lowest values after 150 days of storage. Rosehip was attributed to a high ability to prevent protein oxidation and carbonyl formation from proteins due to the high amount of ascorbic acid and other redox-active phenolic compounds. In this study, all antioxidants were effective against oxidation reactions compare to control (without antioxidants). Spice essential oils were study as preservative in meat product by Sharma et al. (2017). Four different essential oils, clove oil (0.25%), holy basil oil (0.125%), cassia oil (0.25%) and thyme oil (0.125%), were incorporated into fresh chicken sausages packed in vacuum and stored at 18 ± 2 °C for 45 days. According to the authors, clove oil presented the lowest values of TBARS followed by cassia oil, however, all essential oils were effective against lipidic oxidation when compared to control. Regarding antimicrobial activity, cassia oil showed the lowest values in the total count of microorganisms, psychotropic and molds and yeasts. Sensorially, the essential oils added in chicken fresh sausage protected against discoloration during the storage. The essential oil of Satureja montana L. was studied to evaluate its effects on TBARS and color in mortadella-type sausages with different concentrations of sodium nitrite (0, 100 and 200 mg.kg-1) (Coutinho de Oliveira et al., 2012). Satureja montana is a plant native to the Mediterranean is widely used as a spice in food or as tea, and in its essential oil there is the 49 presence of compound such as thymol and carvacrol (terpenes) and some phenolic compounds, responsible for their