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- Nutritional and bioactive oils from salmon (Salmo salar) side streams obtained by Soxhlet and optimized microwave-assisted extractionPublication . Pinela, José; Mandim, Filipa; Heleno, Sandrina A.; Ferreira, Isabel C.F.R.; Barba, Francisco Jose; Berrada, Houda; Caleja, Cristina; Barros, Lillian; Fuente, Beatriz de laThe efficiency of the microwave-assisted extraction (MAE) technique on recovering nutritional and bioactive oils from salmon (Salmo salar) side streams was evaluated and compared to Soxhlet extraction. The response surface methodology (RSM) coupled with a central composite rotatable design was used to optimize time, microwave power, and solid/liquid ratio of the MAE process in terms of oil yield. The optimal MAE conditions were 14.6 min, 291.9 W, 80.1 g/L for backbones, 10.8 min, 50.0 W, 80.0 g/L for heads, and 14.3 min, 960.6 W, 99.5 g/L for viscera, which resulted in a recovery of 69% of the total lipid content for backbones and heads and 92% for viscera. The oils obtained under optimal MAE conditions showed a healthy lipid profile as well as cytotoxic, antioxidant, anti-inflammatory, or antimicrobial properties. These results highlight that oils from underutilized salmon by-products could be exploited by different industrial sectors under the circular economy approach.
- Upcycling fish by-products into bioactive fish oil: the suitability of microwave-assisted extractionPublication . Pinela, José; Rodrigues, Matilde; Pires, Tânia C.S.P.; Mandim, Filipa; Almeida, André; Dias, Maria Inês; Caleja, Cristina; Barros, LillianThe seafood industry is often left out of the food waste discussion, but this sector is no exception, as it generates large amounts of various by-products. This study aimed to explore the potential of the microwave-assisted extraction (MAE) technique to obtain high-quality oil from fish by-products. The independent variables, which were time (1-30 min), microwave power (50-1000 W), and solid/liquid ratio (70-120 g/L) were combined in a 20-run experimental design coupled with the response surface methodology (RSM) for process optimization. The obtained oil yield values were fitted to a quadratic equation to build the theoretical models, which were statistically validated based on statistical criteria and used to predict the optimal MAE condition. The oil yields were significantly affected by the three independent variables through linear, quadratic, and/or interactive effects. Compared to a conventional Soxhlet extraction (SE), the optimal MAE conditions allowed between 60 and 100% of oil to be recovered in less than 19 min and with less solvent consumption. The fatty acid profiles of the oils obtained through SE and optimized MAE were characterized by gas chromatography with flame ionizing detection (GC-FID) after a derivatization process. These oils were constituted mainly of health, beneficial unsaturated fatty acids, such as oleic, docosahexaenoic (DHA), linoleic, and eicosapentaenoic (EPA) acids, which were not affected (p > 0.05) by the extraction methods. Interestingly, the oils obtained through MAE showed the best microbial growth inhibition results may have been due to thermolabile compounds, preserved via this unconventional non-thermal method. The oils also exhibited anti-inflammatory effects via nitric oxide production inhibition and cytotoxic potential especially, against breast and gastric adenocarcinoma cells. However, the threshold of toxicity should be further investigated. Overall, this work emerges as a future-oriented approach to upcycling fish by-products into high-quality oils that can be used in the formulation of pet food and other products.
- The Effect of Cropping System and Irrigation Regime on the Plant Growth and Biochemical Profile of Cichorium spinosumPublication . Paschoalinotto, Beatriz H.; Polyzos, Nikolaos; Liava, Vasiliki; Mandim, Filipa; Pires, Tânia C.S.P.; Añibarro-Ortega, Mikel; Dias, Maria Inês; Barros, Lillian; Petropoulos, Spyridon A.This study evaluated the effects of three irrigation treatments (control (rain-fed plants), deficit irrigation (DI: 50% of maximum field capacity), full irrigation (FI: 100% of maximum field capacity)), and two crop-management treatments (with or without crop rotation with bean, CR, and NCR, respectively) on the plant growth and chemical composition of C. spinosum. The results indicated that deficit irrigation combined with crop rotation increased the weight of leaves per plant, followed by rain-fed plants for the same crop-management treatment. Additionally, these two factors significantly influenced the nutritional profile, free sugars, and organic acid content in a variable manner. Moreover, the control treatment and deficit irrigation increased the content of K, Na, and Mg, which are highly mobile nutrients, whereas the levels of moderately mobile nutrients such as Fe, Mn, Cu, and Zn decreased. Deficit irrigation without crop rotation significantly increased the content of total tocopherols, followed by deficit irrigation with crop rotation and full irrigation without crop rotation. The main fatty acids were α-linolenic acid (C18:3n3), followed by palmitic acid (C16:0) and linoleic acid (C18:2n6), while the control and deficit irrigation treatments combined with crop rotation increased PUFA and decreased SFA content. Furthermore, deficit irrigation and crop rotation induced the accumulation of phenolic compounds, flavonoids, and phenolic acids, especially the content of the major compounds (e.g., chicoric acid, quercetin-O-hexurunoside, and luteolin-O-hexurunoside). The leaf extracts exhibited varied antioxidant activity (assessed by TBARS and OxHLIA assays), and antimicrobial activity. On the other hand, no antifungal, antiproliferative (except for AGS cell line), hepatotoxic, or anti-inflammatory effects were recorded. In conclusion, the combination of deficit irrigation and crop rotation with bean positively affected the quality traits and the fresh weight of leaves, thus suggesting that such eco-friendly practices could have beneficial effects in the cultivation of C. spinosum plants within the context of climate-change mitigation strategies.
- Plantas de pimento dulce (Capsicum annuum L.) em poscesacha como fuente de ingredientes antioxidantes y antidiabéticosPublication . Añibarro-Ortega, Mikel; López, Víctor; Dias, Maria Inês; Mandim, Filipa; Barros, Lillian; Pinela, JoséEl estrés oxidativo desempeña un papel fundamental en el desarrollo de enfermedades crónicas y metabólicas como la diabetes, enfermedades cardiovasculares y neurodegenerativas, obesidad y cáncer. Esto se debe a la producción excesiva de especies reactivas de oxígeno que dañan los lípidos, proteínas y ADN de los organismos, comprometiendo procesos celulares vitales [1]. Los antioxidantes naturales, como los polifenoles, pueden prevenir estos daños y hoy en día existe un creciente interés por el uso de extractos ricos en estos compuestos como ingredientes funcionales naturales. En el caso del cultivo de pimiento dulce (Capsicum annuum L.), tras la cosecha se genera una grande cantidad de biomasa vegetal infravalorada, que podría contener metabolitos secundarios de alto valor comercial. Así, el objetivo de este estudio fue valorizar estos subproductos agrícolas en línea con los objetivos de la Agenda 2030 de Desarrollo Sostenible de las Naciones Unidas. Para ello, los subproductos de la planta fueron empleados para preparar extractos hidroetanólicos [2], los cuales se caracterizaron con respecto a su perfil de compuestos fenólicos mediante HPLC-DAD-ESI/MSn y sus propiedades antioxidante, antidiabética, anti-obesidad, antiinflamatoria y citotóxica con base en ensayos celulares y enzimáticos in vitro. El análisis cromatográfico reveló un perfil fenólico con predominio de ácidos fenólicos, como el ácido clorogénico, y flavonas O-glicosiladas, concretamente luteolina y apigenina. Aunque los extractos presentaron cierta actividad en todos los ensayos realizados, donde más se destacaron fue en la actividad antioxidante, antidiabética y citotóxica en ciertas líneas celulares tumorales. Como conclusión, se puede afirmar que los subproductos de plantas de pimiento dulce son una prometedora fuente de compuestos bioactivos de alto valor añadido. Estos pueden tener propiedades beneficiosas para la salud del consumidor al ser incorporados en productos alimentarios. Además, fomentar esta práctica de circularidad será importante para la producción sostenible de ingredientes alimentarios naturales.
- Chemical composition and biological activity of Commelina erecta: an edible wild plant consumed in BrazilPublication . Cavichi, Lucas V.; Liberal, Ângela; Dias, Maria Inês; Mandim, Filipa; Pinela, José; Kostić, Marina; Soković, Marina; Kalschne, Daneysa Lahis; Fernandes, Ângela; Canan, Cristiane; Barros, Lillian; Amaral, Joana S.In recent years, the interest in products of natural origin has boosted the exploitation and use of plants as food and sources of bioactive compounds, especially wild plants widely used in different cultures for several purposes. Commelina erecta is a wild edible plant (WEP) traditionally used as food and medicine, about which few studies exist. Thus, this study aimed at enhancing the knowledge about its nutritional, chemical and bioactive profile, considering different plant parts and development stages, in order to increase its inclusion in the diet of South American communities. The nutritional profile was found to be similar to other WEP frequently consumed in Brazil. Thirteen phenolic compounds (HPLC-DAD-ESI/MS) were tentatively identified, with apigenin, luteolin and quercetin derivatives being the most abundant. Fructose and oxalic acid were the major sugar and organic acid, respectively, in the aerial parts of C. erecta, and four isoforms of tocopherols were also identified. Regarding the plant’s antioxidant activity, the EC50 values varied between 18.4 and 1060 µg/mL in the inhibition of lipid peroxidation assay (TBARS) and between 53 and 115 µg/mL in the oxidative haemolysis inhibition (OxHLIA) assay. The hydroethanolic extract obtained from stems at the flowering stage also presented anti-inflammatory activity. In general, all the extracts evidenced promising antimicrobial activity. Altogether, these results reinforce the traditional use of this plant species as food and medicine to support the diet of needier populations and also promote food sovereignty and sustainability.
- Phenolic profile and bioactive potential of cardoon (Cynara cardunculus L.) inflorescences: selection of the best genotype for food applicationPublication . Mandim, Filipa; Pinela, José; Dias, Maria Inês; Barracosa, Paulo; Santos-Buelga, Celestino; Ferreira, Isabel C.F.R.; Barros, LillianCynara cardunculus L. (syn. cardoon) belongs to the Asteraceae family and is native to the Mediterranean basin. This species is widely used for milk coagulation of PDO cheeses and consumed due to its rich nutritional value and related medicinal properties (e.g. choleretic, cardiotonic, antidiabetic, liver disease treatment). In the last decade, the interest in this crop increases due to its multifaceted industrial applications. It has been explored for animal feed and lightwood panels, biodiesel, paper pulp, and food oil production. Their production countries estimate a generation of approximatly 15 to 30 t/ha of dry biomass. The adequate exploitation and characterization of the species are essential for circular economic stimulation and environmental impact reduction. The hydroethanolic extracts of four cardoon genotypes and their vegetable tissues (stigma, corolla, bracts, pappus, receptacle) were studied. The polyphenolic profile was analyzed by HPLC-DAD-ESI/MS. The antioxidant activity was evaluated TBARS and OxHLIA cell-based assays.The anti-inflammatory activity was assessed through the extracts’ capacity to inhibit the pro-inflammatory mediator nitric oxide. Finally, the cytotoxic potential was evaluated against four tumor and a non-tumor cell line (PLP2) using the sulforhodamine B assay. Fourteen phenolic compounds were tentatively identified. The corolla presented the higher variety and bracts the higher content of phenolic compounds. In terms of bioactivity, none of the tested extracts exhibit anti-inflammatory potential. Genotype F1-1-1 exhibits a higher cytotoxic potential. All the four genotypes and vegetable tissues tested exhibited antioxidant capacity, especially corolla of F4-1-4 genotype for TBARS assay (IC50 = 38 μg/mL) and its receptacle for OxHLIA (IC50 = 71 μg/mL). In conclusion, this study showed that the phenolic composition and biological activities of cardoon are influenced by both genotype and plant tissue. Further studies are needed to determine the genetic information for obtaining the highest bioactivity.
- Gastrointestinal digestion of yerba mate, rosemary and green tea extracts and their subsequent colonic fermentation by human, pig or rat inoculaPublication . Correa, Vanesa G.; Garcia-Manieri, Jessica Amanda Andrade; Dias, Maria Inês; Pereira, Carla; Mandim, Filipa; Barros, Lillian; Ferreira, Isabel C.F.R.; Peralta, Rosane M.; Bracht, AdelarPolyphenolic compounds are common constituents of human and animal diets and undergo extensive metabolism by the gut microbiota before entering circulation. In order to compare the transformations of polyphenols from yerba mate, rosemary, and green tea extracts in the gastrointestinal tract, simulated gastrointestinal digestion coupled with colonic fermentation were used. For enhancing the comparative character of the investigation, colonic fermentation was performed with human, pig and rat intestinal microbiota. Chemical analysis was performed using a HPLC system coupled to a diode-array detector and mass spectrometer. Gastrointestinal digestion diminished the total amount of phenolics in the rosemary and green tea extracts by 27.5 and 59.2 %, respectively. These reductions occurred mainly at the expense of the major constituents of these extracts, namely rosmarinic acid (-45.7 %) and epigalocatechin gallate (-60.6 %). The yerba mate extract was practically not affected in terms of total phenolics, but several conversions and isomerizations occurred (e.g., 30 % of trans-3-Ocaffeoylquinic acid was converted into the cis form). The polyphenolics of the yerba mate extract were also the least decomposed by the microbiota of all three species, especially in the case of the human one (-10.8 %). In contrast, the human microbiota transformed the polyphenolics of the rosemary and green extracts by 95.9 and 88.2 %, respectively. The yerba mate-extract had its contents in cis 3-O-caffeoylquinic acid diminished by 78 % by the human microbiota relative to the gastrointestinal digestion, but the content of 5-O-caffeoylquinic acid (also a chlorogenic acid), was increased by 22.2 %. The latter phenomenon did not occur with the rat and pig microbiota. The pronounced interspecies differences indicate the need for considerable caution when translating the results of experiments on the effects of polyphenolics performed in rats, or even pigs, to humans.
- Influence of the growth cycle on the bioactive properties and phenolic composition of Cynara cardunculus L. var altilisPublication . Mandim, Filipa; Petropoulos, Spyridon Α.; Dias, Maria Inês; Fernandes, Ângela; Pinela, José; Kostić, Marina; Soković, Marina; Barros, Lillian; Santos-Buelga, Celestino; Ferreira, Isabel C.F.R.Cynara cardunculus L. (syn. Cardoon), is an erect perennial herbaceous plant, native to the Mediterranean basin. It is characterized by a high variety and richness in compounds with health benefits. Also, its cultivation and economic impact are highly important due to the diverse industrial applications (i.e. vegetable rennet, biomass, bioenergy, etc.) [1, 2]. Due to its increasing consumption and commercial interest, this study purposes the analysis of phenolic compounds and bioactive properties of different cardoon parts (heads, bracts, and seeds) collected in Greece during different maturation stages.The phenolic composition was determined by HPLC-DAD-ESI/MS. The antibacterial and antifungal activities were evaluated by applying the microdilution method. The cytotoxic effects were evaluated in four human tumor cell lines using the sulforhodamine B assay, while the anti- inflammatory activity was evaluated through the inhibition of NO production in macrophage cells. Finally, the antioxidant activity was measured through the TBARS and OxHLIA assays. The content in phenolic compounds decreased with the maturation process, except in the seed extract. All the tested samples exhibited antibacterial and antifungal activity, where lower MICs were observed when the plant reached maturity. Regarding the cytotoxic and anti-inflammatory activities, the earliest harvest stages revealed the highest activity, except for seeds.Moreover, with the maturation process, extracts presented higher capacity to OxHLIA and TBARS inhibition. The heterogeneity of the biological results reveals that other compounds than phenolic ones may be correlated with these bioactivities. This study proved the high biological potential of cardoon parts as also its possible use as a source of important bioactive compounds. Nevertheless, further studies are needed to understand which compounds are responsible for the observed bioactivities, as well as to find the stage of maturity that provides the best bioactive properties.
- Nutrients, Phytochemicals, and In Vitro Biological Activities of Goldenberry (Physalis peruviana L.) Fruit and CalyxPublication . Añibarro-Ortega, Mikel; Dias, Maria Inês; Petrović, Jovana; Mandim, Filipa; Núñez, Sonia; Soković, Marina; López, Víctor; Barros, Lillian; Pinela, JoséThis study provides a comprehensive characterization of Physalis peruviana L., covering the nutritional composition of the fruit and the phytochemical profiles and in vitro bioactive properties of berry and calyx extracts. The fresh fruit stood out as a source of dietary fiber (5.16 g/100 g) and is low in fat (0.49 g/100 g). A 100-g serving also contained notable amounts of ascorbic acid (32.0 mg), tocopherols (2.34 mg), potassium (253 mg), phosphorus (45 mg), and magnesium (20 mg). HPLC-DAD-ESI/MS analysis tentatively identified five physalin derivatives and one withanolide in the fruit extract, which showed significant antiproliferative activity against human colorectal adenocarcinoma (Caco-2) and non-small-cell lung carcinoma (NCI-H460) cells. The calyx extracts contained three phenolic acids and four flavonoids, demonstrating high antioxidant activity through physiologically relevant cell-based assays, the ability to inhibit advanced glycation end products (AGEs) formation and nitric oxide production, and also antiproliferative properties. These findings highlight goldenberry as a nutrient-dense fruit rich in vitamins and functional compounds with potential health benefits, supporting its recognition as a “superfruit”. Furthermore, the fruit calyx emerged as a valuable source of bioactive secondary metabolites with potential applications in food and pharmaceutical industries and related sectors.
- Green and Innovative Extraction: Phenolic Profiles and Biological Activities of Underutilized Plant Extracts Using Pulsed Electric Fields and MacerationPublication . Pallarés, Noelia; Berrada, Houda; Ferrer, Emilia; Rached, Wahiba; Pinela, José; Mandim, Filipa; Pires, Tânia C.S.P.; Finimundy, Tiane C.; Barba, Francisco J.; Barros, LillianUnderutilized plant species such as Asteriscus graveolens (Forssk.) Less., Haloxylon scoparium Pomel, and Ruta chalepensis L. have been historically valued in traditional medicine for their potential health benefits. These species present an untapped source of bioactive compounds with significant applications in the food and pharmaceutical industries, including the development of functional foods and additives. Recent advances in food processing have introduced innovative methods, such as pulsed electric fields (PEFs), to enhance the extraction of valuable compounds without compromising their integrity or quality. This study investigates the impact of PEF technology on the recovery of bioactive compounds from these plants, comparing it with conventional maceration (MAC) techniques. Phenolic compound profiles and biological activities, including antioxidant, antimicrobial, anti-inflammatory, and cytotoxic effects, were evaluated. The results demonstrated that for R. chalepensis, PEF extraction achieved comparable phenolic content (58 mg/g) to MAC (72 mg/g). However, MAC generally provided higher phenolic yields for other plants. A. graveolens extracts exhibited significant antitumoral and anti-inflammatory potentials. The antimicrobial results indicated that MAC extracts were more effective against bacterial growth, while PEF extracts outperformed MAC against A. brasiliensis (MIC: 10 mg/mL). Antioxidant potential was observed in both methods, with TBARS IC50 values ranging from 17 to 79.5 µg/mL. While MAC generally yielded superior results, PEF extraction showed great promise as an environmentally sustainable alternative, eliminating the need for organic solvents and aligning with green extraction principles.