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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.
- Nutritional value and chemical composition of three types of Petroselinum crispum leavesPublication . Ardohain, Elizandra; Fernandes, Ângela; Polyzos, Nikolaos; Petropoulos, Spyridon Α.; Pinela, José; Moreira, Glaucia; Ferreira, Isabel C.F.R.; Barros, LillianPetroselinum crispum Mill. Nym., commonly known as parsley, is an aromatic herb used to garnish and to give flavour and odour to dishes and salads [1]. The most common parsley types in the Mediterranean region are Petroselinum crispum ssp. neapolitanum (plain-leafed) and Petroselinum crispum ssp. crispum (curly- and bioactive properties, such as the genotype, the irrigation regime, the planting density, the sowing date and the climate conditions. The main objective of this study was to evaluate the crop diversification through the determination of the nutritional value and the chemical composition of leaves from twenty-five plain-leafed, curly-leafed and turnip-rooted parsley (Petroselinum crispum ssp. tuberosum) cultivars, cultivated in central Greece. The proximate composition was evaluated by AOAC official procedures, free sugars and tocopherols were determined using liquid chromatography coupled to a refraction index (HPLC-RI) and fluorescence (HPLC-FL) detectors, respectively. Fatty acids were determined by gas chromatography coupled to a flame ionization detector (GC-FID) and organic acids by ultra-fast liquid chromatography coupled to a diode detector (UPLC-DAD) [1]. The plain and curly-leafed type contains higher amount of fat and energy than the turnip-rooted one, protein content was higher in the curly-leafed and turnip-rooted type and no significant differences were observed between the three types in terms of the carbohydrates content. The total sugars content did not differ significantly among the tested -- linolenic and linoleic acid followed by palmitic acid, while only linoleic acid content differed among the cultivar types. The curly-leafed type contains the highest amount of malic, citric and total organic acids, whereas the highest oxalic acid content was recorded in both curly-leafed and turnip-rooted type. The results of our study showed a great variability in the nutritional value parameters and the chemical composition of twenty-five parsley cultivars from three distinct types, which indicates the great potential of the valorization of the existing genotypes to increase the agrobiodiversity and introduce the turnip-rooted type in the Mediterranean region.
- 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.
- Phytochemical composition and nutritional value of pot-grown turnip-rooted and plain and curly-leafed parsley cultivarsPublication . Fernandes, Ângela; Polyzos, Nikolaos; Petropoulos, Spyridon Α.; Pinela, José; Ardohain, Elizandra; Moreira, Glaucia; Ferreira, Isabel C.F.R.; Barros, LillianIn the present study plant growth, nutritional value and chemical composition of leaves from twenty-five plain-leafed, curly-leafed and turnip-rooted parsley cultivars were evaluated. Total fresh yield was higher for the plain-leafed cv. Rialto Bejo: 192 ± 11 g/pot, while significant differences were observed between the three types in the nutritional parameters, except for the carbohydrates content. The most abundant organic acid was malic acid (5.22–6.88 g/100 g dw), while the total sugars content did not differ significantly among the tested cultivar types. α-tocopherol was the major tocopherol detected in amount that ranged between 14.76–30.32 mg/100 g dw. The main fatty acids were α-linolenic and linoleic followed by palmitic acid, while only linoleic acid content being different among the cultivar types. In conclusion, the existing diversity in the parsley genotypes could be valorised to increase the agrobiodiversity in the broader Mediterranean region through the introduction of less cultivated curly-leafed and turnip-rooted types.
- Sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata) head oils recovered by microwave-assisted extraction: nutritional quality and biological propertiesPublication . Fuente, Beatriz de la; Pinela, José; Calhelha, Ricardo C.; Heleno, Sandrina A.; Ferreira, Isabel C.F.R.; Barba, Francisco Jose; Berrada, Houda; Caleja, Cristina; Barros, LillianMicrowave-assisted extraction (MAE) and Soxhlet extraction (SE) were used to obtain oil from European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) heads. The MAE technique allowed the recovery of more than 50% of the total lipid con- tent for both fish by-products in less than 11 min extraction. Based on their fatty acid composition, all fish head oils presented a healthy lipid profile and were found to be a good source of docosahexaenoic acid (DHA, 11–14%). Different lipid quality indices also revealed their cardiovascular protective potential. Oils obtained by MAE showed higher antibacterial and antifungal effects against food pathogens than those extracted by SE. Cellular antioxidant activity (29–35% oxidation inhibition) and anti-inflammatory poten- tial via NO production inhibition (IC50 = 14–21 μg/mL) were evaluated using murine macrophages cells (RAW 264.7). The highest cytotoxic effect (GI50 = 38–46 μg/mL) of fish head oils was observed against breast cancer cells (MCF-7). These results showed that sea bass and gilthead sea bream heads could be exploited for the production of oil with nu- tritional and bioactive properties in line with circular bioeconomy concepts.
- Valorization of Solanum melongena L. crop by-products: Phenolic composition and in vitro antioxidant, antidiabetic, anti-inflammatory, cytotoxic, and antimicrobial propertiesPublication . Añibarro-Ortega, Mikel; Dias, Maria Inês; Petrović, Jovana; Núñez, Sonia; Calhelha, Ricardo C.; Costa, Eduardo M.; Machado, M.; Pintado, Manuela; Soković, Marina; López, Víctor; Barros, Lillian; Pinela, JoséThis study explored the valorization of post-harvest eggplant aerial parts as a sustainable source of value-added ingredients by investigating their phenolic composition and in vitro bioactive properties. HPLC-DAD-ESI/MSn analysis identified chlorogenic acid derivatives as the predominant phenolic compounds (53 % of the phenolic fraction), followed by O-glycosylated kaempferol and quercetin. The extract displayed antioxidant activity in physiologically relevant cell-based assays and significant α-glucosidase inhibitory capacity that far exceeded that of the standard drug acarbose. It also inhibited the formation of advanced glycation end-products (AGEs), suggesting its potential to mitigate diabetes-related complications. Furthermore, the extract showed a modest pancreatic lipase inhibitory effect and capacity to suppress interleukin 6 production. Selective cytotoxicity against human gastric and colon adenocarcinoma cell lines and strong antimicrobial activity against foodborne pathogens were observed. Given the growing demand for natural alternatives to synthetic drugs, these findings position eggplant crop biomass as a promising, sustainable source of active compounds with potential applications in food, nutraceutical, and pharmaceutical formulations for managing type 2 diabetes and other oxidative stress-mediated conditions. This study not only contributes to the valorization of agricultural waste but also expands the research on by-products of Solanaceae crops, offering a pathway for sustainable resource utilization.
- 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.
- Thymus species from romanian spontaneous flora as promising source of phenolic secondary metabolites with health-related benefitsPublication . Babotă, Mihai; Frumuzachi, Oleg; Mafra, Isabel; Nicolescu, Alexandru; Dias, Maria Inês; Pinela, José; Barros, Lillian; Añibarro-Ortega, Mikel; Stojković, Dejan; Carevic, Tamara; Mocan, Andrei; López, Víctor; Crisan, GianinaWild thyme aerial parts (Serpylli herba) are recognized as a valuable herbal product with antioxidant, anti-inflammatory, and antibacterial effects. Although pharmacopoeial regulations allow its collection exclusively from Thymus serpyllum, substitution with other species is frequent in current practice. This study analyzed the phenolic composition, antioxidant, and enzyme-inhibitory and antimicrobial activity of the hydroethanolic extracts obtained from five Romanian wild thyme species (Thymus alpestris, T. glabrescens, T. panonicus, T. pulcherimus and T. pulegioides). The analysis of individual phenolic constituents was performed through LC-ESI-DAD/MS2, while for the in vitro evaluation of antioxidant potential, TEAC, FRAP, DPPH, TBARS and OxHLIA assays were employed. The anti-enzymatic potential was tested in vitro against tyrosinase, alpha-glucosidase and acetylcholinesterase. High rosmarinic acid contents were quantified in all species (20.06 +/- 0.32-80.49 +/- 0.001 mg/g dry extract); phenolic acids derivatives (including salvianolic acids) were confirmed as the principal metabolites of T. alpestris and T. glabrescens, while eriodictyol-O-di-hexoside was found exclusively in T. alpestris. All species showed strong antioxidant potential and moderate anti-enzymatic effect against alpha-glucosidase and acetylcholinesterase, showing no anti-tyrosinase activity. This is the first detailed report on the chemical and biological profile of T. alpestris collected from Romanian spontaneous flora.
- Untargeted phytochemical profiling and biological activity of small yellow onion (Allium flavum L.) from different regions of RomaniaPublication . Moldovan, Cadmiel; Frumuzachi, Oleg; Babotă, Mihai; Pinela, José; Barros, Lillian; Rocchetti, Gabriele; López, Víctor; Lucini, Luigi; Crisan, Gianina; Mocan, AndreiThis study examined the phytochemical profiles (mainly phenolics, carotenoids, and organosulfur compounds) and biological effects of hydroalcoholic extracts of Allium flavum (AF), a species of the Allium genus commonly known as small yellow onion. Unsupervised and supervised statistical approaches revealed clear differences between extracts prepared with samples collected from different areas of Romania. Overall, the AFFF (AF flowers collected from Faget) extract was the best source of polyphenols, also showing the highest antioxidant capacity evaluated through both in vitro DPPH, FRAP, and TEAC anti-radical scavenging assays and cell-based OxHLIA and TBARS assays. All the tested extracts exhibited & alpha;-glucosidase inhibition potential, while only the AFFF extract exhibited anti-lipase inhibitory activity. The phenolic subclasses annotated were positively correlated with the assessed antioxidant and enzyme inhibitory activities. Our findings suggested that A. flavum has bioactive properties worth exploring further, being a potential edible flower with health-promoting implications.
- Mycorrhization and micropropation of chestnut (Castanea sativa Mill.) seedlings as tools to obtain high added-value phenolic compoundsPublication . Dias, Maria Inês; Pinela, José; Pereira, Carla; Ferreira, Patrícia; Oliveira, Maria de Fátima; Martins, Anabela; Afonso, Andreia; Barros, LillianMiChestnut3 is a project of the company DEIFIL whose main objective is to produce more resistant and productive hybrid chestnut (Castanea sativa Mill.) seedlings through micropropagation and mycorrhization techniques. In this work, in addition to the agronomic traits of the micropropagated mycorrhizal chestnut seedlings, it was also important to evaluate the changes induced by mycorrhization in the phenolic profile of these plants. Phenolic compounds are plant secondary metabolites involved in plant-microbe interactions/symbiosis and act as signaling molecules in the establishment of arbuscular mycorrhizal symbioses, as well as in plant defense mechanisms [1]. According to the literature, considerable increases in phenolic compounds in host plants as a result of arbuscular mycorrhizal fungus inoculation have been reported during the progression of the infection [1]. Therefore, this work was carried out to study the impact of the type of fungal inoculum and the period of mycorrhization (before or after potting) on the qualitative and quantitative profile of phenolic compounds of the roots and leaves of the chestnut seedlings produced by DEIFIL. After collection and lyophilization of the plant material, hydroethanolic extracts were prepared and the phenolic compounds were characterized by HPLC-DAD-ESI/MS [2]. Ellagic acid derivatives and Oglycosylated flavonoids were the major phenolic compounds in both plant roots and leaves, which agreed with previous reports [2,3]. A statistical analysis showed that the type of inoculum and period of mycorrhization significantly (p<0.05) affected the phenolic profile of the chestnut hybrids. In general, the mycorrhizal seedlings with the fungi Amanita caesarea and Boletus edulis were those that presented the highest levels of phenolic compounds. Relationships between the levels of these signaling compounds and the agronomic performance of chestnut seedlings were also found. The results bring new perspectives into the future production of a hybrid chestnut tree resistant to ink disease in the main traditional Portuguese varieties