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

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

Frutos biológicos de Vaccinum myrtillus L. na base de novos produtos enriquecidos: caracterização química e nutricional

Vaccinum myrtillus L. pertence à família Ericaceae, sendo comumente conhecida pelos seus pequenos frutos e doces: os mirtilos. Largamente consumidos em fresco, estes frutos também são usados em compotas e marmeladas, devido à suas propriedades digestivas e hipoglicémicas bem como, devido à presença de vários compostos bioativos [1]. Portanto, tornou-se uma matriz muito apelativa para o desenvolvimento de novos produtos funcionais que, para além das suas propriedades nutricionais, adicionam um efeito benéfico fisiológico e para a saúde de longo prazo [2]. No presente trabalho, três novos produtos desenvolvidos pela RBR Foods Company (Portugal) tendo como base o mirtilo, foram caracterizados pelas suas propriedades nutricionais e químicas: hidratos de carbono, cinzas, proteínas, gordura e valor energético (seguindo métodos oficiais de análise de alimentos AOAC), perfil de ácidos gordos (por GC-FID), açúcares solúveis (por HPLC-RI), ácidos orgânicos (por HPLC-DAD) e tocoferóis (por HPLC de fluorescência). Os produtos resultam de uma mistura dos frutos com pétalas de rosa (P1), pétalas de calêndula (P2) e bagas goji e maçã (P3). Os frutos de mirtilo foram utilizados como amostra de controlo. O perfil nutricional dos novos produtos mostrou-se muito semelhante ao da amostra controlo: os hidratos de carbono foram os macronutrientes mais abundantes, seguido de proteínas e de gordura total. Em relação aos açúcares, frutose, glucose e sacarose foram identificados em todas as amostras. P1 e P2 não apresentaram diferenças significativas em relação ao controlo, no entanto, P3 revelou uma menor concentração de açúcares. Em termos de composição de ácidos gordos, todas as amostras estudadas apresentaram maiores teores em ácidos gordos polinsaturados, especialmente devido à contribuição dos ácidos linoléico e alfa-linolénico. Os resultados de tocoferóis revelaram que a amostra controlo apresentou apenas duas isoformas de tocoferóis, α- e γ-tocoferol, sendo a mesmo observado em P3. No entanto, P1 revelou a presença de todas as isoformas de tocoferóis, enquanto P2 não apresentaou δ-tocoferol; Estes resultados estão relacionados com a contribuição das pétalas de rosa e calêndula, respetivamente. A isoforma α-tocoferol foi a mais abundante em todas as amostras estudadas. Em geral, este trabalho contribuiu para a caracterização de novos produtos nutricionais com base de mirtilo sendo parte de um projecto mais abrangente que tem por objetivo o estudo detalhado destes produtos, para serem utilizados como alimentos funcionais.