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- FIG (Ficus Carica L.) Bioresidues as sources of bioactive compounds and natural pigments for the food industryPublication . Shiraishi, Carlos S.H.; Zbiss, Yosra; Roriz, Custódio Lobo; Carocho, Márcio; Domingos, Sara; Calhelha, Ricardo C.; Alves, Maria José; Abreu, Rui M.V.; Prieto Lage, Miguel A.; Heleno, Sandrina A.; Barros, LillianThe 17 goals of sustainable development address several topics, such as: (2) Zero hunger and sustainable agriculture; (9) Industry, Innovation, and Infrastructure; (12) Responsible consumption and production; that are essential for the promotion of the circular economy, product development and conscious production [1]. Fig is a food matrix, cultivated in Portugal and valued by the Portuguese people for consumption in natura, being also used in wines, liqueurs, and jams. As this fruit is very appreciated and consumed, it´s cultivation leads to the production of tons of leaves, usually discarded [2]. Therefore, in the present work, the leaves of five fig varieties (Figure 1), namely Dauphine (Da), Longue d'Aout (La), Pasteliere (Pa), Marseille (Ma) and Bourjassote Noire (Bn), were nutritionally and chemically characterized to detect possible bioactive molecules. The antioxidant and antimicrobial, activities were also analyzed, to provide the food industry with natural additives in alternative to the artificial ones; and at the same time, promote the circular economy. Regarding the nutritional profile of the five leaves, La sample exhibited the highest amount in proteins (18.0±0.6 g/100g dw), while Pa revealed the highest content in fats (2.2±0.1 g/100g dw). The highest moisture content was presented by Da leaves (17.3±0.1 g/100g fw), and for the ashes, La sample was the one that presented the highest value (14.18±0.06 g/100g fw). Concerning the organic acids, these molecules were most abundant in Ma leaves, where it was possible to identify oxalic, malic and citric acids with a total of 139.6±0.4 mg/g dw. For the soluble sugars profiling, in all samples it was possible to identify five sugars, namely, fructose, glucose, sucrose, trehalose, and raffinose in different concentrations; however, Da leaves revealed the higher amount (17±1 g/100g dw). Tocopherols where also analyzed, and in all samples, three of the four isoforms where detected, being Pa sample standing for the predominance of these compounds (4.14±0.05 mg/100 g dw). For the bioactive analysis, different assays were performed, and Pa sample showed the strongest antioxidant potential for the TBARS assay, with an EC50 value of 105±5 mg/mL. For the antimicrobial activity assay, Da leaf extract was the one displaying the best results, by presenting Minimum Inhibitory Concentrations (MIC) ranging from 1.25 to 10 mg/mL against the tested bacterial strains. On the other hand, for the antifungal activity, the samples present very similar profiles, with the exception of the Pa sample, that present the lowest MIC of 5 mg/mL for Aspergillus fumigatus. In general, these leaf extracts can be used in the food industry namely in pastry and bakery products as promising sources of bioactive compounds, and at the same time, this reuse of biowaste promotes circular economy, and reduces the impact of biowaste resulting from the fig industry, thus meeting some of the goals of sustainable development.
- Virtual screening of a library of natural compounds against COX-2 proteinPublication . Shiraishi, Carlos S.H.; Souza, Sérgio F.; Prieto Lage, Miguel A.; Barros, Lillian; Abreu, Rui M.V.The use of nonsteroidal anti-inflammatory drugs (NSAIDs) in treating inflammatory diseases has been widespread, especially in treating rheumatoid arthritis. NSAIDs act mainly by promoting the inhibition of cyclooxygenase enzymes (COX-1 and COX-2), inducing significant anti-inflammatory, analgesic, and antipyretic activity. However, recent data show that prolonged use of NSAIDs can lead to cardiovascular side effects. Thus, the present work aims to identify COX-2 inhibitors alternatives from natural sources, specifically mushrooms, as an alternative to conventional inhibitors.
- Red seaweeds as a source of nutrients and bioactive compounds: Optimization of the extractionPublication . Carpena Rodríguez, María; Caleja, Cristina; Pereira, Eliana; Pereira, Carla; Ćirić, Ana; Soković, Marina; Soria-Lopez, Anton; Fraga-Corral, Maria; Simal-Gandara, Jesus; Ferreira, Isabel C.F.R.; Barros, Lillian; Prieto Lage, Miguel A.The present work aimed to determine the nutritional composition (ash, protein, fat, carbohydrate content and energy value), phenolic compounds, pigments and organic acids content of three typical red algae from the Northwest of Spain: Chondrus crispus, Mastocarpus stellatus, and Gigartina pistillata; as well as their antioxidant and antimicrobial activities. Furthermore, the present work compared two extraction techniques: conventional heat assisted extraction (HAE) and high pressure assisted extraction (HPAE) to maximize the yield and the concentration of target compounds. Different independent variables were considered for the response study. Time (t) and percentage of ethanol of the solvent (S) were chosen for both techniques and temperature (T) and pressure (P) were used for HAE and HPAE, respectively. The experiments were designed following a response surface methodology (RSM) approach. The obtained results showed a similar nutritional composition between algae samples: low-fat content and high content of proteins, carbohydrates and energy. All tested algae showed good antioxidant and antimicrobial properties. Finally, HEA demonstrated to be the most efficient extraction technique. This study confirms the potential of red algae to be part of the human diet as a source of non-animal protein, due to its nutritional content, phenolic profile, pigments concentration and bioactive properties, which proves that HAE is the optimum technique for the extraction maximization.
- Optimization of heat- and ultrasound-assisted extraction of anthocyanins from Hibiscus sabdariffa calyces for natural food colorantsPublication . Pinela, José; Prieto Lage, Miguel A.; Pereira, Eliana; Jabeur, Inès; Barreiro, M.F.; Barros, Lillian; Ferreira, Isabel C.F.R.calyces. The extraction variables, time (t), ethanol proportion (S), and temperature (T) or ultrasonic power (P), were combined in a 5-level experimental design and analysed by response surface methodology for process optimization. The delphinidin-3-O-sambubioside (C1) and cyanidin-3-O-sambubioside (C2) levels were monitored by LC-DAD-ESI/MSn and used as response criteria. The developed models were successfully fitted to the experimental data and used to determine optimal extraction conditions. UAE was the most efficient method yielding 51.76 mg C1+C2/g R under optimal conditions (t=26.1 min, P=296.6W and S=39.1% ethanol, v/ v). The dose-response effects of the solid/liquid ratio on the extraction rate were also determined. The anthocyanin levels herein reported are higher than those found in the literature, which support the potential use of H. sabdariffa as a sustainable source of natural colorants with application in different industrial sectors.
- Extraction of essential oils from the residues of two shrub species aiming for their revalorization: chemical characterization and antioxidant, antimicrobial and cytotoxic activitiesPublication . Xavier, Virginie; Heleno, Sandrina A.; Prieto Lage, Miguel A.; Amaral, Joana S.; Mediavilla Ruiz, Irene; Esteban Pascual, Luis Saul; Ferreira, Isabel C.F.R.; Barros, LillianIn recent years, essential oils have been subject of research for their bioactive properties, such as antimicrobial, fungitoxic, anti-viral, anti-inflammatory and antioxidant activities. Owing to these properties they are potentially interesting for diverse industries including the food industry since one of its main problems concerns microbes and associated toxins that are responsible for food spoilage. Although the application of essential oils in the food industry may have some limitations, such as impact on the organoleptic properties and low solubility, different delivery strategies such as nanoencapsulation, active packaging and coatings are promising technologies that may overcome these issues without compromising nutritional properties in food systems [1]. In this view, increased knowledge on the composition and activity of different essential oils is needed, particularly regarding novel potential sources of essential oils such as agricultural wastes or species grown in marginal lands, on a perspective of circular economy. Therefore, in the scope of the BeonNAT project, biomass from different tree and shrub species are being screened as possible sources of essential oils and respective bioactivity evaluated. In this work, the essential oil extracted by steam distillation from the branches (>20mm) of two shrub species grown in Spain, Juniperus communis L. and Cistus ladanifer L., was characterized for their chemical composition by gas chromatography coupled with mass spectrometry (GC-MS) as well as for their antioxidant, antimicrobial, antiinflammatory and cytotoxic activities. GC-MS analysis allowed the identification of 98.1% of compounds in J. communis EO, corresponding to a total of 63 identified compounds, alfa-pinene being the major compound (35.1%), followed by limonene (15.0%), sabinene (6.7%), cis-tujopsene (8.0%), beta-myrcene (3.2%) and beta-caryophyllene (3.5%). In general, the chemical composition is in agreement with that of juniper berry EO, defined in the European Pharmacopoeia and ISO 8897 standard, except for limonene (15.0%) which was slightly higher than the defined range (Eur. Ph of 2-12% and ISO standard of 2-8%). For C. ladanifer EO, 61 compounds were identified corresponding to 92.8% of the total compounds, with viridiflorol being the main compound (24.0%), followed by alfa-pinene (19.3%), ledol (6.9%), camphene (6.7%) and bornyl acetate (5.0%), which is in good agreement with previous data [2]. Both oils showed potential against the panel of bacteria selected according to their importance in public health and foodborne diseases, highlighting the rock-rose EO that showed interesting activity against Escherichia coli, Morganella morganii, Pseudomonas aeruginosa, Enterococcus faecalis, Listeria monocytogenes and methicillin resistant Staphylococcus aureus in a concentration range of 0.039-2.5%(v/v). Regarding the antioxidant activity, both oils showed promising results, with EC50 values of 1.35 ± 0.19 mg/mL and 1.30 ± 0.07 mg/mL in the reducing power assay and 68% and 83% inhibition of oxidation according to the cellular antioxidant activity assay, for J. communis and C. ladanifer, respectively. The essential oils showed antiinflammatory (IC50 of 24±1 µg/mL and 21±2 µg/mL for juniper and rock-rose-rose, respectively) and cytotoxic activity, with the best results obtained with the rock-rose EO in the inhibition of stomach-AGS, colon-CaCo, breast-MCF-7 and lung-NCI-H460 cancer cell lines (GI50 between 47±5 µg/mL and 58±1 µg/mL). Juniper EO did not evidenced cytotoxicity in non-tumoral Vero cells at the highest tested concentration (400 µg/mL) wich can be an indicator of its safety. Overall, the results demonstrated that shrubs biomass can be a source of EO with similar composition to that reported for respective berries and leaves. The EOs showed interesting antibacterial and antioxidant activity thus being potential candidates for further studies on their safety and potential application in food systems.
- Crop rotation and irrigation regime affects the nutritional and chemical profile of Cichorium spinosumPublication . Paschoalinotto, B.H.; Prieto Lage, Miguel A.; Compocholi, Maria; Polyzos, Nikolaos; Petropoulos, Spyridon Α.; Ferreira, Isabel C.F.R.; Dias, Maria Inês; Barros, LillianThe sustainable management of agricultural systems offers synergistic opportunities for the co-production of agricultural and natural capital outcomes.1 A properly sized agricultural system is essential for the sustainable and ecological maintenance of crop productivity. Irrigation management is an important adaptation strategy to improve crop resilience to global climate change while crop rotation brings benefits such as increased crop yields through high soil fertility and reduced fertilizer inputs.2 Cichorium spinosum L. (spiny chicory) is a wild edible plant that has received very recent attention as a potential alternative/complementary crop. It is a plant traditionally consumed in the so-called Mediterranean diet due to its high nutritional value and various beneficial health effects.3 The study aims to improve and integrate the cultivation of this species in farming systems of the Mediterranean region. Thus, a combination of full or deficit irrigation with or without crop rotation with maize was established in an attempt to establish the commercial cultivation of spiny chicory. Two control samples were cultivated: C0 (rain-feed with crop rotation with maize) and C00 (rain-feed without crop rotation). The nutritional profile was evaluated using AOAC methods. Energy was calculated according to the equation: energy (kcal per 100 g) = 4 x (g protein + g carbohydrate) + 2 x (g total dietary fiber) + 9 x (g fat). The profile of organic acids, minerals, fatty acids and sugars were performed using UFLC-PDA, atomic absorption spectrophotometry, GC-FID and HPLC-RI, respectively. Although the impacts that a sustainable farming system generates on the crop involved is a long-term assessment and after the system has been repeated for several growing periods, however some changes are already noticeable in the first growing period. In the nutritional profile, there were no differences between the six experimental treatments, with the exception of the total dietary fiber content which samples C0 (control) and CFIC (full irrigation with crop rotation with maize) showed the highest levels. The samples presented low values of total fat, being the sample C0 the one that presented the highest value (3.5 g/100g dry weight). Promising levels of crude protein were indicated by all samples, however once again the control sample had the highest content (C00). The CFIC and CFIN samples (full irrigation with and without crop rotation, respectively) showed the lowest values of carbohydrates. The sample CDIC (deficit irrigation with crop rotation with maize) showed the highest energy (276.3 kcal/100g dry weight) probably due to the low fiber content and consequently the high carbohydrate content. Five organic acids were identified in the spiny chicory samples, mostly quinic acid, except in the CFIN sample in which oxalic acid had the highest concentration. In terms of minerals, the samples with full irrigation showed higher concentrations of iron, manganese and copper and lower calcium, while the samples without crop rotation showed lower concentrations of potassium. The predominant fatty acids identified and quantified were linolenic, linoleic, and palmitic acids, the sum of which represented 82 to 86% in the studied samples, while the sample with deficit irrigation and without crop rotation (CDIN) presented the lowest percentage. Finally, the sugars identified in higher concentrations were sucrose, glucose, fructose, and trehalose, respectively, however, it is suggested that crop rotation with maize altered the profile of sugars by increasing their concentrations. Considering that these are preliminary results, it was possible to point out positive impacts of the tested agronomic practices on nutritional parameters of the species that could be commercially applied aiming to integrate wild edible species in sustainable and low inputs farming systems.
- The nutritional and bio-active constituents, functional activities, and industrial applications of cashew (Anacardium occidentale): a reviewPublication . Chen, Ying-Ying; Li, Ningyang; Guo, Xu; Huang, Hai-jie; Garcia-Oliveira, Paula; Sun, Jin-yue; Zhang, Jian-dong; Prieto Lage, Miguel A.; Guo, Zhi-qing; Liu, ChaoCashew nut is a globally popular nut not only for its delicious and crunchy flavor but also for bioactive compounds, which present positive benefits to human health. Different parts of cashew have characteristic compounds. The kernel of cashew nut is rich in fatty acids, the testa is rich in polyphenols, cashew apple is rich in polyphenols and carotenoids, and phenols with aliphatic long chain are present in the cashew nutshell liquid. Therefore, the edible part of cashew possesses a wide range of bioactive compounds, with reported biological properties, including anti-tumor, brain health, cardiovascular and cerebrovascular protection, anti-diabetic, gastroprotection, pathophysiological disorders protection, and antioxidant. On the other hand, the inedible part of cashew, the cashew nutshell liquid, could play an important role in the industry as surface coatings, insulating formulations and so on. Hence, there are considerable demands to increase the added value of cashew, thereby exploring greater economic value. Thus, this review summarizes nutrients, bioactive compounds, biological functions, and applications of Anacardium occidentale to provide a theoretical basis and inspiration for further in-depth studies and utilization.
- Valorisation of tomato wastes for development of nutrient-rich antioxidant ingredients: A sustainable approach towards the needs of the today's societyPublication . Pinela, José; Prieto Lage, Miguel A.; Barreiro, M.F.; Carvalho, Ana Maria; Oliveira, Beatriz; Curran, Thomas P.; Ferreira, Isabel C.F.R.Nutrient-rich antioxidant ingredients were produced from tomato fruit wastes using a microwave-assisted extraction (MAE) process. Different conditions of extraction time (t), temperature (T), ethanol concentration (Et) and solid/liquid ratio (S/L) were combined in a circumscribed central composite design and optimized by response surface methodology. The model was statistically validated and used for prediction in the experimental range. Under the global optimal MAE conditions (t = 20 min, T = 180 °C, Et = 47.4% and S/L = 45 g/L), it was possible to obtain an extraction yield of 75.5% and ingredients with high levels of sugars, proteins, phenolics, and flavonoids, and interesting antioxidant properties measured via ABTS[rad] + scavenging activity and oxidative haemolysis inhibition assay (OxHLIA). The antioxidant capacity of the extracts was lower compared to the one of commercial food additives. However, the sustainably developed ingredients may be used in the fortification and functionalisation of food, as well as for incorporation in feed products. Industrial relevance This study addresses current needs of the agri-food sector, namely the recycling of plant wastes and production of valuable extracts for the food/feed industry. A MAE process was developed and optimized to maximize the recovery of nutrients and antioxidants from tomato fruit wastes. The optimum processing conditions established in this study allowed a high extraction yield and reduced solvent consumption. MAE can be considered as a sustainable alternative to conventional extraction methods. These findings will contribute to promote a more sustainable bioeconomy in the agro-food sector.
- Thermochemical characterization of eight seaweed species and evaluation of their potential use as an alternative for biofuel production and source of bioactive compoundsPublication . Cassani, Lucia; Lourenço-Lopes, Catarina; Barral Martínez, Marta; Chamorro Rivo, Franklin Noel; García-Pérez, Pascual; Simal-Gandara, Jesus; Prieto Lage, Miguel A.Algae are underexplored resources in Western countries and novel approaches are needed to boost their industrial exploitation. In this work, eight edible seaweeds were subjected to their valorization in terms of nutritional characterization, thermochemical properties, and bioactive profile. Our results suggest that seaweeds present a rich nutritional profile, in which carbohydrates are present in high proportions, followed by a moderate protein composition and a valuable content of !-3 polyunsaturated fatty acids. The thermochemical characterization of seaweeds showed that some macroalgae present a low ash content and high volatile matter and carbon fixation rates, being promising sources for alternative biofuel production. The bioactive profile of seaweeds was obtained from their phenolic and carotenoid content, together with the evaluation of their associated bioactivities. Among all the species analyzed, Porphyra purpurea presented a balanced composition in terms of carbohydrates and proteins and the best thermochemical profile. This species also showed moderate anti-inflammatory activity. Additionally, Himanthalia elongata extracts showed the highest contents of total phenolics and a moderate carotenoid content, which led to the highest rates of antioxidant activity. Overall, these results suggest that seaweeds can be used as food or functional ingredient to increase the nutritional quality of food formulations.
- Extraction optimization of bioactive compounds from Thymus ulgaris L.: comparison of heat-assisted and ultrasound-assisted extractionsPublication . Caleja, Cristina; Pereira, Eliana; Sprea, Rafael M.; Finimundy, Tiane C.; Carocho, Márcio; Amaral, Joana S.; Prieto Lage, Miguel A.; Barros, LillianPlants have been used since ancient times due to their biological action associated mainly with several phytochemicals, including phenolic compounds1. Thymus vulgaris L. is a species described as a potential source of natural molecules with applications in the food industry, but also in the cosmetic and pharmaceutical industry due to its antibacterial, antifungal, and anti-inflammatory effects2. In this sense, the present study aimed to compare extraction systems in terms of phenolic compounds from T. vulgaris through the heat-assisted (HAE) and ultrasound-assisted extraction (UAE) techniques, aiming to improve its potential for industrial applications. For this propose, an extraction optimization was studied using three independent variables, time (t, minutes), solvent (S, % of ethanol), and temperature (T, ºC) or power (P, W), in a three-level Box-Behnken design, analyzing the surface methodology (RSM). The extraction yield and the content in phenolic compounds identified by HPLC-DAD-ESI/MS were the experimental responses applied. The polynomial models were successfully fitted to the experimental data and used to determine the optimal HAE and UAE conditions. Nineteen phenolic compounds were identified, nine of which were phenolic acids (mainly caffeic acid derivatives) and ten flavonoids: flavan-3-ols (catechin derivatives), flavones (quercetin derivatives), flavonols (apigenin and luteolin derivatives) and flavanones (eriodictyol derivatives). The conditions obtained for the optimal extraction of phenolic compounds, that allowed the experimental validation of the predictive model are 3% ethanol (v/v) for 89 minutes at a temperature of 98º C for HAE and 77% ethanol (v/v) for 17 minutes at 207 W for UAE. Extraction optimization is important from an industrial point of view, as it allows obtaining extracts with a high content of bioactive compounds. Considering that the phenolic composition appears in literature as a direct influence on bioactive properties, the optimization processes will allow a better exploitation of extracts, ensuring the greatest cost reduction for the industry.