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- Avaliação da capacidade funcional de extratos de Ruscus aculeatus LPublication . Rodrigues, Joana P.B.; Carvalho, Ana Maria; Barros, LillianAs plantas, utilizadas pelos seres humanos pelas suas capacidades aromáticas, alimentares e medicinais, são fonte de uma grande diversidade de compostos naturais. Deste modo, e também devido à intensificação da procura de substâncias naturais biologicamente ativas, o interesse da utilização de produtos naturais tem vindo a aumentar substancialmente. Contudo, as plantas têm na sua composição misturas complexas de componentes químicos que apresentam diversas ações biológicas e farmacológicas, pelo que a sua aplicação para fins medicinais carece ainda de muita investigação e experimentação, nomeadamente de ensaios clínicos, que aprovem a sua utilização terapêutica. Ruscus aculeatus L. (família botânica Asparagaceae), é uma espécie utilizada na medicina tradicional, em diversas partes do mundo. As partes aéreas são utilizadas principalmente como diuréticos e a parte subterrânea pode ser utilizada para vários fins, como por exemplo, no tratamento de distúrbios do sistema urinário e como laxante. Na presente dissertação, a parte aérea e, as raízes e rizomas de R. aculeatus L., foram caracterizados quimicamente no que se refere ao conteúdo em compostos fenólicos e propriedades bioativas. Para tal, foram preparados extratos distintos, aquosos (infusões e decocções) e hidroalcoólicos das 2 partes mencionadas da planta. O perfil fenólico de cada extrato foi avaliado por HPLC-DAD-ESI/MS e foram também analisados relativamente às suas bioatividades (atividade antioxidante, citotóxica, anti-inflamatória e antibacteriana). O potencial antioxidante foi avaliado através de dois ensaios celulares distintos: inibição da peroxidação lipídica em tecidos cerebrais de porco (TBARS) e inibição da hemólise oxidativa (OxHILA). As propriedades citotóxicas dos extratos foram avaliadas em quatro linhas celulares tumorais humanas: HeLa (carcionama cervical), NCI H460 (carcinoma de pulmão), MCF-7 (carcinoma da mama) e HepG2 (carcinoma hepatocelular); por sua vez, a citotoxicidade em células não tumorais foi avaliada numa cultura de células não tumorais: PLP2 (cultura primária de células de fígado de porco). A atividade anti-inflamatória foi avaliada utilizando uma linha celular de macrófagos de rato RAW 264.7. A atividade antibacteriana foi avaliada através da técnica de microdiluição, juntamente com o método colorimétrico de deteção rápida com cloreto de p-iodonitrotetrazólio (INT). Através da análise do perfil fenólico dos extratos de R. aculeatus, nove compostos fenólicos foram detetados nos diferentes extratos. O extrato hidroalcoólico da parte aérea apresentou os teores mais elevados em compostos fenólicos, seguido dos extratos aquosos. A apigenina-C-hexósido-C-pentósido isómero II foi o composto maioritário nos extratos aquosos e, no extrato hidroalcoólico foi a quercetina-O-desoxihexósido-hexósido seguida da apigenina-C-hexósido-C-pentósido isómero II. No que diz respeito às bioatividades dos extratos, os resultados apresentaram diferenças significativas consoante o tipo de extrato. No ensaio de TBARS, o extrato hidroalcoólico, foi o que apresentou melhor atividade tanto o da parte aérea como o das raízes e rizomas. No ensaio de OxHILA, os extratos que revelaram melhor atividade para a parte aérea e, para as raízes e rizomas foram a infusão e o hidroalcoólico, respetivamente. Os extratos aquosos da parte aérea e apenas a infusão do extrato de raízes e rizomas, não apresentaram toxicidade em células de fígado de porco saudáveis, enquanto os extratos hidroalcoólicos da parte aérea e raiz e rizoma, bem como a decocção da raiz e rizoma, apresentaram toxicidade contra esta linha celular. O extrato hidroalcoólico da parte aérea foi o que apresentou resultado efetivo na inibição de 50% da produção de óxido nítrico. Nos extratos aquosos, apenas a decocção das raízes e rizoma apresentou atividade anti-inflamatória. No ensaio da avaliação da atividade antibacteriana, a decocção da parte aérea foi efetiva contra a estirpe MRSA – Staphylococcus aureus resistente à meticilina. O presente trabalho permitiu comprovar o potencial biológico associado a esta espécie, em particular às suas bioatividades.
- Chemical and bioactive screening of green polyphenol-rich extracts from chestnut by-products: an approach to guide the sustainable production of high-added value ingredientsPublication . Rodrigues, Daniele Bobrowski; Verissimo, Lavinia; Finimundy, Tiane C.; Rodrigues, Joana P.B.; Oliveira, Izamara; Fernandes, Isabel P.; Barros, Lillian; Heleno, Sandrina A.; Calhelha, Ricardo C.Opportunities for the valorisation of agro-industrial residues of the chestnut (Castanea sativa Mill.) production chain have been fostered with the production of multifunctional polyphenol-rich extracts with the potential to be introduced as natural additives or active components in several products. Nonetheless, it is crucial to explore the feasibility of different extracts from the various by-products for these applications through the exhaustive study of their composition and bioactivities without losing sight of the sustainable character of the process. This work aimed at the screening of the phenolic compound composition and bioactivities of different green extracts of chestnut burs, shells and leaves, as the first step to establish their potential application as natural ingredients, primarily as food preservatives. To this end, maceration (MAC) as a conventional extraction method besides ultrasound and microwave-assisted extractions (UAE and MAE) was employed to obtain the extracts from chestnut by-products using water (W) and hydroethanolic solution (HE) as solvents. Phenolic compounds were analysed by HPLC-DAD-(ESI-)MS/MS; the antioxidant capacity was assessed by colourimetric assays, and the antimicrobial activity was evaluated against several strains of food-borne bacteria and fungi. The leaf extracts obtained by MAC-HE and UAE-HE presented the highest concentration of phenolic compounds (70.92 & PLUSMN; 2.72 and 53.97 & PLUSMN; 2.41 mg.g(-1) extract dw, respectively), whereas, for burs and shells, the highest recovery of total phenolic compounds was achieved by using UAE-HE and UAE-W (36.87 & PLUSMN; 1.09 and 23.03 & PLUSMN; 0.26 mg.g(-1) extract dw, respectively). Bis-HHDP-glucose isomers, chestanin and gallic acid were among the most abundant compounds. Bur extracts (MAC-HE and UAE-HE) generally presented the highest antioxidant capacity as measured by TBARS, while the best results in DPPH and reducing power assays were found for shell extracts (MAE-W and MAC-HE). Promising antibacterial activity was noticed for the aqueous extracts of burs, leaves and hydroethanolic extracts of shells, with emphasis on the MAE-W extract of burs that showed bactericidal activity against E. cloacae, P. aeruginosa and S. aureus (MBC 5 mg.mL(-1)). Overall, it can be concluded that chestnut by-products, including burs, shells and leaves, are sources of polyphenolic compounds with significant antioxidant and antimicrobial activities. The choice of extraction method and solvent greatly influenced the composition and bioactivity of the extracts. These findings highlight the potential of chestnut by-products for the development of natural additives, particularly for food preservation, while also emphasizing the importance of sustainable utilization of agricultural waste materials. Further research is warranted to optimize extraction techniques and explore additional applications for these valuable bioactive compounds.
- Phenolic compounds and bioactive properties of Ruscus aculeatus L.: na underexploited subshrubPublication . Rodrigues, Joana P.B.; Fernandes, Ângela; Dias, Maria Inês; Pereira, Carla; Pires, Tânia C.S.; Calhelha, Ricardo C.; Carvalho, Ana Maria; Ferreira, Isabel C.F.R.; Barros, LillianRuscus aculeatus L. is a subshrub used in traditional medicine in different parts of the world, namely in Europe and Iberian Peninsula [1]. According to reported folk knowledge, the aerial parts are mainly used as diuretics and the underground parts are used for the treatment of disorders of the urinary system and as laxative [2]. The present work reports a comprehensive study on R. aculeatus phenolic compounds and bioactivities, intending to contribute to the knowledge of the chemical composition of this unexplored species. The plant material of R. aculeatus was harvested in April 2019 inside woodlands and hedgerows, in Valpaços, Portugal. Two distinct parts were gathered, the aerial part (cladodes or laminar stems and lateral branches) and the underground organs (rhizomes with roots). After lyophilization, the distinct parts of the plant were further analyzed. Hydroethanolic extracts and aqueous (infusions and decoctions) preparations from the two mentioned parts of the plant were prepared. Phenolic compounds were determined by high performance liquid chromatography (HPLC) coupled to a diode array detector (DAD) and mass spectrometry (MS) using the electrospray ionization interface (ESI). Multi-resistant clinical bacterial strains were used to screen the antimicrobial activity and the antioxidant activity was evaluated through two different in vitro assays: inhibition of lipid peroxidation in brain homogenates by thiobarbituric acid reactive substances (TBARS) and oxidative hemolysis inhibition assay (OxHLIA). The cytotoxic activity was assayed using MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung carcinoma), HeLa (cervical carcinoma) and HepG2 (hepatocellular carcinoma) human tumor cell lines, and also a non-tumor cell line (porcine liver primary cells, PLP2). Nine phenolic compounds were detected in all extracts. Apigenin-C-hexoside-Cpentoside isomer II was the major compound in aqueous extracts and, in the hydroethanolic extract, the most abundant one was quercetin-O-deoxyhexoside-hexoside, followed by apigenin-C-hexoside-C-pentoside isomer II. All extracts revealed antioxidant activity and potential to inhibit some of the assessed bacteria; the aqueous extracts of the aerial part and infusions of roots and rhizomes did not show cytotoxic effects in a non-tumour primary cell culture. This preliminary study provides suggestions of the biological potential associated with the empirical uses and knowledge of this species, in particular its bioactivities.
- Nutritional profile of Brassica rapa L. (rapini) from different comercial genotypesPublication . Rodrigues, Joana P.B.; Almeida, Daiana; Gioia, Francesco di; Petropoulos, Spyridon Α.; Ferreira, Isabel C.F.R.; Fernandes, Ângela; Barros, LillianThe Brassica genus comprises a large and diverse group of important vegetables widely consumed throughout the world. Part of this group, Brassica rapa L., also known as Rapini or Broccoli Rabe, is a green leafy vegetable appreciated in the cuisine of countries such as China, Greece, Italy, Portugal, and Spain. The consumption of the edible parts of Rapini (young leaves, stems, and flower buds) is associated with various health benefits, due to their composition in macronutrients and bioactive compounds. The aim of the present work was to evaluate the nutritional value of 45 commercially available Rapini varieties. The nutritional profile was assessed following AOAC procedures. Carbohydrates were the major macronutrients found (48.28–64.1 g/100 g dry weight (dw)), followed by proteins (23.7–39.2 g/100 g dw) and ash (7.52– 16.9 g/100 g dw), with samples Leccese centoventina, Novantina leccese a cima grande and Quarantino extra presenting the highest contents, respectively. Lipids (2.68–5.09 g/100 g dw) were less abundant in all samples, highlighting Quarantino extra for the smallest amount. Finally, the energy values found were between 350.4 and 390.4 Kcal/100g dw, with the highest content obtained by sample Novantina riccia San Marzano. In conclusion, the results show that the varieties tested present good nutritional value, which indicate that Rapini can be considered an interesting option of food, as it is a good source of important macronutrients.
- Phenolic compounds and bioactive properties of Ruscus aculeatus L. (asparagaceae): the pharmacological potential of an underexploited subshrubPublication . Rodrigues, Joana P.B.; Fernandes, Ângela; Dias, Maria Inês; Pereira, Carla; Pires, Tânia C.S.; Calhelha, Ricardo C.; Carvalho, Ana Maria; Ferreira, Isabel C.F.R.; Barros, LillianRuscus aculeatus L. is a subshrub used in traditional medicine in different parts of the world, namely in Europe and the Iberian Peninsula. According to reported folk knowledge, the aerial parts are mainly used as diuretics and the underground organs are used for the treatment of disorders of the urinary system and as a laxative. In this work, the aerial part and the roots and rhizomes of R. aculeatus were chemically characterized with regard to the content of phenolic compounds and bioactive properties. Aqueous (infusions and decoctions) preparations and hydroethanolic extracts from the two mentioned parts of the plant were prepared. Nine phenolic compounds were detected in all the extracts. Apigenin-C-hexoside-C-pentoside isomer II was the major compound in aqueous extracts and, in the hydroethanolic extract was quercetin-O-deoxyhexoside-hexoside followed by apigenin-C-hexoside-C-pentoside isomer II. All extracts revealed antioxidant activity and potential to inhibit some of the assayed bacteria; aqueous extracts of the aerial part and infusions of roots and rhizomes did not show cytotoxic effects on a non-tumor primary cell culture. This preliminary study provides suggestions of the biological potential associated with the empirical uses and knowledge of this species, in particular its bioactivities.
- Agri-food surplus, waste and loss as sustainable biobased ingredients: a reviewPublication . Rodrigues, Joana P.B.; Liberal, Ângela; Petropoulos, Spyridon Α.; Ferreira, Isabel C.F.R.; Oliveira, Beatriz; Fernandes, Ângela; Barros, LillianEnsuring a sustainable supply of food for the world’s fast growing population is a major challenge in today’s economy, as modern lifestyle and increasing consumer concern with maintaining a balanced and nutritious diet is an important challenge for the agricultural sector worldwide. This market niche for healthier products, especially fruits and vegetables, has increased their production, consequently resulting in increased amounts of agri-food surplus, waste, and loss (SWL) generated during crop production, transportation, storage, and processing. Although many of these materials are not utilized, negatively affecting the environmental, economic, and social segments, they are a rich source of valuable compounds that could be used for different purposes, thus preventing the losses of natural resources and boosting a circular economy. This review aimed to give insights on the efficient management of agri-food SWL, considering conventional and emerging recovery and reuse techniques. Particularly, we explored and summarized the chemical composition of three worldwide cultivated and consumed vegetables (carrots, broccoli and lettuce) and evaluate the potential of their residues as a sustainable alternative for extracting value-added ingredients for the development of new biodynamic products
- Bioactive properties of Ruscus aculeatus L.: na underexploited subshrubPublication . Rodrigues, Joana P.B.; Fernandes, Ângela; Dias, Maria Inês; Pereira, Carla; Pires, Tânia C.S.; Calhelha, Ricardo C.; Carvalho, Ana Maria; Ferreira, Isabel C.F.R.; Barros, LillianRuscus aculeatus L. is a sub-shrub used in traditional medicine in different parts of the world, namely in Europe and the Iberian Peninsula[1]. Traditionally, the aerial parts are mainly used as diuretics and the underground parts are used for the treatment of urinary tract diseases and as a laxative[2]. In the present work, the aerial part, roots, and rhizomes of R. aculeatus were characterized regarding their bioactive properties in order to contribute to the knowledge of the chemical composition of this unexplored species. The plant material of R. aculeatus was harvested in April 2019 inside woodlands and hedgerows, in Valpaços, Portugal. Two distinct parts were gathered: the aerial part (cladodes or laminar stems and lateral branches) and the underground organs (rhizomes with roots). Hydroethanolic extracts and aqueous (infusions and decoctions) preparations from the two mentioned parts of the plant were prepared. Multi-resistant clinical bacterial strains (Enterococcus faecalis, Listeria monocytogenes, methicillin-resistant Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Morganela morganii, Proteus mirabilis and Pseudomonas aeruginosa) were used to screen the antimicrobial activity and the antioxidant activity was evaluated through two different in vitro assays: inhibition of lipid peroxidation in brain homogenates by thiobarbituric acid reactive substances (TBARS) and oxidative hemolysis inhibition assay (OxHLIA). The cytotoxic activity was assayed using MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung carcinoma), HeLa (cervical carcinoma) and HepG2 (hepatocellular carcinoma) human tumor cell lines, and also a non-tumor cell line (porcine liver primary cells, PLP2). All extracts revealed antioxidant activity and potential to inhibit some of the assessed bacteria; except for the infusion of the aerial part (for the lines MCF7 and HepG2) and decoction extracts (line MCF7), all the remaining extracts presented effective results in the inhibition of the tested cell lines; the aqueous extracts of the aerial part and infusions of roots and rhizomes did not show cytotoxic effects in a non-tumor primary cell culture. This preliminary study provided innovative and interesting results in relation to the bioactive properties of this little-studied and explored wild plant.
- Propriedades bioativas de Ruscus aculeatus L.: um subarbusto inexploradoPublication . Rodrigues, Joana P.B.; Fernandes, Ângela; Pereira, Carla; Pires, Tânia C.S.; Calhelha, Ricardo C.; Carvalho, Ana Maria; Ferreira, Isabel C.F.R.; Barros, LillianRuscus aculeatus L. é um subarbusto utilizado na medicina tradicional em várias partes do Mundo, nomeadamente na Europa e na Península Ibérica 1' 1. Tradicionalmente, as partes aéreas são utilizadas como diuréticos e as subterrâneas no tratamento de doenças do trato urinário e como laxante 1' 1. No presente trabalho, a parte aérea, raízes e rizomas de R. aculeatus foram caracterizados quanto às suas propriedades bioativas, nomeadamente: atividade antioxidante, antimicrobiana, anti-inflamatória e citotóxica. O material vegetal de R. aculeatus foi colhido em abril de 2019 em parcelas florestais, em Valpaços, Portugal. Foram preparados extratos hidroetanólicos e aquosos (infusões e decocções) das partes aéreas (caules laminares e ramificações) e das partes subterrâneas (rizomas e raízes). A atividade antioxidante dos extratos foi avaliada mediante a inibição da peroxidação lipídica (TBARS) e da hemólise oxidativa (OxHLIA). A atividade antibacteriana foi avaliada através da técnica de microdiluição, juntamente com o método colorimétrico de deteção rápida com o cloreto de piodonitrotetrazólio (INT). A atividade anti-inflamatória foi avaliada através da inibição da produção de NO utilizando uma linha celular de macrófagos de rato RAW 264.7. As propriedades citotóxicas foram avaliadas utilizando linhas celulares tumorais humanas e numa linha celular não tumoral (células primárias de fígado de porco, PLP2). Todos os extratos revelaram atividade antioxidante e potencial para inibir algumas das bactérias utilizadas. O extrato hidroalcoólico da parte aérea foi o que apresentou melhores resultados na inibição da produção de óxido nítrico seguido pela decocção das raízes e rizoma. Todos os extratos inibiram o crescimento das linhas celulares tumorais, exceto a infusão da parte aérea e os extratos de decocção. Os extratos aquosos da parte aérea e a infusão do extrato de raízes e rizomas, não apresentaram toxicidade para a linha celular PLP2. Este estudo preliminar forneceu resultados inovadores e interessantes em relação às propriedades bioativas desta planta silvestre pouco estudada e explorada.