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- Chitosan mouthwash: toxicity and in vivo validationPublication . Costa, E. M.; Silva, S.; Costa, M. R.; Pereira, M.; Campos, D. A.; Odila, J.; Madureira, A. R.; Cardelle-Cobas, A.; Tavaria, F. K.; Rodrigues, A. S.; Pintado, M. M.A previous study showed that a chitosan mouthwash would be a valid alternative to current mouthwashesas it demonstrated, in vitro, significantly higher antibiofilm activity than two commercial mouthwashes.As such, the aim of this work was to verify the safety of the developed product and to validate, in vivo, thebiological activity ascertained in vitro. Chitosan mouthwash safety was evaluated through Ames, MTT andV79 chromosomal aberration assay while antimicrobial activity was evaluated through in vivo assays. Theresults showed that the chitosan mouthwash was safe, presenting lower cytotoxicity than a commercialmouthwash, and that it effectively reduced viable counts of Streptococcus spp. and Enterococcus spp. by ca.5.5 log of CFU. Furthermore, in direct comparison with a commercial mouthwash the chitosan mouthwashpossessed significantly higher antimicrobial activity. The conjunction of these results proves that thechitosan mouthwash is a safe, effective, natural alternative to the existent chemical mouthwashes.
- Anthocyanin recovery from grape by-products by combining ohmic heating with food-grade solvents: phenolic composition, antioxidant, and antimicrobial propertiesPublication . Coelho, Marta; Silva, Sara; Costa, Eduardo; Pereira, Ricardo N.; Rodrigues, António Sebastião; Teixeira, José António; Pintado, ManuelaUsually, wine-making by-products are discarded, presenting a significant environmental impact. However, they can be used as a source of bioactive compounds. Moreover, consumers’ increasing demand for naturally nutritious and healthy products requires new formulations and food product improvement, together with sustainable, environmentally friendly extraction methods. Thus, this work aimed to compare ohmic heating (OH) with conventional methodology (CONV), using food-grade solvents, mainly water, compared to standard methanol extraction of anthocyanins. No significant differences were found between the CONV and OH for total phenolic compounds, which were 2.84 ± 0.037 and 3.28 ± 0.46 mg/g DW gallic acid equivalent, respectively. The same tendency was found for antioxidant capacity, where CONV and OH presented values of 2.02 ± 0.007 g/100 g and 2.34 ± 0.066 g/100 g ascorbic acid equivalent, respectively. The major anthocyanins identified were malvidin-3-O-acetylglucoside, delphinidin-3-O-glucoside, petunidine-3-O-glucoside, cyanidin-3-O-glucoside, and peonidine-3-O-glucoside. These extracts displayed antimicrobial potential against microorganisms such as Yersinia enterocolitica, Pseudomonas aeruginosa, Salmonella enteritidis, methicillin-sensitive Staphylococcus aureus, a methicillin-resistant Staph. aureus (MRSA), and Bacillus cereus. In conclusion, OH provides similar recovery yields with reduced treatment times, less energy consumption, and no need for organic solvents (green extraction routes). Thus, OH combined with water and citric acid allows a safe anthocyanin extraction from grape by-products, thus avoiding the use of toxic solvents such as methanol, and with high biological potential, including antimicrobial and antioxidant activity.
- In vitro gastrointestinal digestion impact on the bioaccessibility and antioxidant capacity of bioactive compounds from tomato flours obtained after conventional and ohmic heating extractionPublication . Coelho, Marta C.; Ribeiro, Tânia B.; Oliveira, Carla; Batista, Patrícia; Castro, Pedro; Monforte, Ana Rita; Rodrigues, António Sebastião; Teixeira, José; Pintado, ManuelaIn times of pandemic and when sustainability is in vogue, the use of byproducts, such as fiber-rich tomato byproducts, can be an asset. There are still no studies on the impact of extraction methodologies and the gastrointestinal tract action on bioactive properties. Thus, this study used a solid fraction obtained after the conventional method (SFCONV) and a solid fraction after the ohmic method (SFOH) to analyze the effect of the gastrointestinal tract on bioactive compounds (BC) and bioactivities. Results showed that the SFOH presents higher total fiber than SFCONV samples, 62.47 ± 1.24–59.06 ± 0.67 g/100 g DW, respectively. Both flours present high amounts of resistant protein, representing between 11 and 16% of insoluble dietary fiber. Furthermore, concerning the total and bound phenolic compounds, the related antioxidant activity measured by 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical cation decolorization assay presented significantly higher values for SFCONV than SFOH samples (p < 0.05). The main phenolic compounds identified in the two flours were gallic acid, rutin, and p-coumaric acid, and carotenoids were lycopene, phytofluene, and lutein, all known as health promoters. Despite the higher initial values of SFCONV polyphenols and carotenoids, these BCs’ OH flours were more bioaccessible and presented more antioxidant capacity than SFCONV flours, throughout the simulated gastrointestinal tract. These results confirm the potential of ohmic heating to modify the bioaccessibility of tomato BC, enhancing their concentrations and improving their antioxidant capacity.
- Quercus based coffee-like beverage: effect of roasting process and functional characterizationPublication . Coelho, Marta; Silva, Sara; Rodríguez-Alcalá, Luis Miguel; Oliveira, Ana M.; Costa, Eduardo; Borges, André; Martins, Célia; Rodrigues, António S.; Pintado, Maria Manuela E.Coffee is one of the world’s most widely consumed beverages but intake it is not encouraged in consumers with some health conditions. In this way, with the growing interest in developing healthier substitutes, a coffee-like beverage obtained from Quercus ilex and Quercus suber acorn´s species, was elaborates maintaining the flavor. Although, this beverage is a promising coffee alternative, little is known about effects of roasting process in its composition. To that end the antioxidant capacity and toxicity of the developed coffee were analyzed and phenolic compounds and fatty acids (esterified and free forms) were characterized through HPLC-DAD and GC-FID, respectively. The results showed that Quercus based beverages presented antioxidant capacity related to their phenolic content, mainly to ellagic acid as the primary phenolic compound identified. Due to this composition, the beverage also presented antimutagenic activity. The main fatty acids in the esterified lipids were mainly oleic, linoleic, palmitic, stearic and cis vaccenic. In the free fatty acids fraction (FFA) they were oleic, linoleic and palmitic acids. Heat processing produced a reduction in total fatty acid concentration in TG and FFA fraction of Q. ilex. Nevertheless, for Q. suber alterations were only found for FFA. Coffee production did not form genotoxic or cytotoxic compounds. Overall, these results show the feasibility of Quercus acorn-based foodstuffs and its potential to produce a functional coffee-like beverage.
- Extraction of tomato by-products’ bioactive compounds using ohmic technologyPublication . Coelho, Marta; Pereira, Ricardo; Rodrigues, António S.; Teixeira, José A.; Pintado, Manuela E.Tomato peels and seeds are the main by-products of the tomato industry and represent an interesting source of bioactive compounds (BC) including carotenoids, which can be then used as colorant to commercial aquaculture. The aim of the present work was to optimize the extraction of BC from tomato by-products using Ohmic heating (OH) technology. OH extraction experiments were done in the presence of moderate electric fields (MEF) of different intensity (i.e. 4, 6 and 11 V cm−1) to identify the presence of non-thermal effects on the extraction process and its influence on bioactive properties of the extracted compounds. Polyphenol extraction using OH was successfully optimized with the best extraction conditions being 70 °C for 15 min using 70% ethanol as a solvent, which exhibited rutin recovers of 77% higher than control samples. It allowed to recover up to 4.93 μg/gFW lycopene from tomato by-products without resorting to organic solvents. OH can be used as an environmental-friendly, fast and economic process to polyphenols recover from industrial tomato by-products. In addition, the use of different MEF during extraction shows to have a high potential to cause different levels of permeabilization and cell stress that can help to define a selective extraction process of valuable components from tomato by-products.