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- Effect of modified atmosphere on phytochemical profile of pasteurized peach puréesPublication . Oliveira, Ana; Coelho, Marta; Alexandre, Elisabete; Gomes, Maria Helena; Almeida, Domingos P. F.; Pintado, ManuelaPeach phytochemical profile was analysed during 90 days storage under three different gas mixtures: 10 kPa O2+90 kPa N2, 100 kPa N2 and air (78 kPa N2+21 kPa O2+0.03 kPa CO2) for both 4 and 23 °C. The antioxidant activity increased when peach purée was stored under air at 4 °C and 100 kPa N2 at 23 °C, while total phenolic content was not affected by atmospheres. Total carotenoids were more stable for 100 kPa N2, where it decreases 48 and 58% respectively for 4 and 23 °C. The content of (+)-catechin decreased 42% for 10 kPa O2 at 4°C, remaining constant in other atmospheres. Chlorogenic and neochlorogenic acids were well preserved in atmospheres containing oxygen, presenting an increase of 14 and 24% for 10 kPa O2 at 4°C. The higher content of carotenoids was obtained for 100 kPa N2 at 23 °C. Principal component analysis shows that atmospheres variance was associated with carotenoids while storage time was more related with total phenolics and total antioxidant capacity.
- Long-term frozen storage and pasteurization effects on strawberry polyphenols contentPublication . Oliveira, Ana; Coelho, Marta; Alexandre, Elisabete M. C.; Almeida, Domingos P. F.; Pintado, ManuelaThe profile of the strawberry polyphenols was analysed on fresh strawberry, immediately after freezing or after pasteurization and during frozen storage of strawberry at -20 A degrees C for 360 days. Freezing increased the extractability of (-)-epigallocatechin gallate (60 %), (-)-epicatechin (48 %) and ellagic acid (51 %) and decreased the pelargonidin-3-glucoside (18 %). Direct pasteurization of fresh strawberry increased the extractability for (-)-epigallocatechin gallate (73 %), (-)-epicatechin (45 %) and ellagic acid (143 %) and decreased for pelargonidin-3-glucoside (16 %). During storage at -20 A degrees C for 360 days the extractable levels of pelargonidin-3-rutinoside, (-)-epigallocatechin gallate, (+)-catechin, (-)-epicatechin and ellagic acid decreased by 17, 20, 37, 65 and 80 %, respectively, while those of pelargonidin-3-glucoside increased 13 %. Changes in polyphenols content, produced by pasteurization after 360 days frozen storage, were minimal. Polyphenols content decreased during storage of strawberry pieces at -20 A degrees C for 360 days, while the thermal treatment applied after freezing storage did not induce any significant change.
- 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.
- Variation of anthocyanins and other major phenolic compounds throughout the ripening of four Portuguese blueberry (Vaccinium corymbosum L) cultivarsPublication . Silva, Sara; Costa, Eduardo M.; Coelho, Marta C.; Morais, Rui M.; Pintado, Manuela E.Blueberries are widely recognised as one of the richest sources of bioactive compounds, among which are anthocyanins, though the ripeness of berries has been reported as affecting the phytochemical composition of fruits. Therefore, the present work aimed to evaluate the variation of anthocyanins, and other major phenolics, throughout five ripening stages in four blueberry cultivars. The results showed that the antioxidant capacity and anthocyanin content increased during ripening, reaching the highest values when the blueberries are collected from bunches comprised of 75% ripe blueberries. Antagonistically, the amount of phenolic acid decreases, while the quercetin-3-glucoside levels remain stable. Furthermore, Goldtraube blueberries appear to possess, systematically, higher amounts of phenolic compounds than the other cultivars studied. Thus, when seeking the highest yield of anthocyanins, the preferred harvest should occur in bunches that contain ca 75% of ripe blueberries and, considering the cultivars assayed, the Goldtraube cultivar appears to bethe richest in phenolic compounds.
- Incorporation of strawberries preparation in yoghurt: impact on phytochemicals and milk proteinsPublication . Oliveira, Ana; Alexandre, Elisabete M. C.; Coelho, Marta; Lopes, Cláudia; Almeida, Domingos P. F.; Pintado, Manuela E.An immediate decrease in the total antioxidant activity (23%) and total phenolic content (14%) was observed after addition of strawberry preparations to yoghurt. The total anthocyanin content did not change immediately, but decreased 24% throughout the yoghurt shelf-life. The individual compounds, (+)-catechin (60%), (-)-epicatechin (60%), kaempferol (33%) and quercetin-3-rutinoside (29%) decreased after 24 h in the yoghurt made with the strawberry preparation. During the remaining period of storage these compounds increased by 47%, 6%, 4% and 18%, respectively. Pelargonidin-3-glucoside decreased 49% after 28 d. Immediately after the addition of the strawberry preparation to yoghurt, beta-lactoglobulin decreased to values lower than the limit of detection and alpha-lactalbumin by approximately 34%, and was reduced further slowly throughout yoghurt self-life. An immediate interaction between the carrageenan present in the strawberry preparation and beta-LG was observed. The variations of both polyphenols and protein in the presence of carrageenan and the potential interactions were discussed.
- Ohmic-heating treatment as a new process for development of functional ingredients through valorisation of tomato and grape by-productsPublication . Coelho, Marta Isabel Correia; Pintado, Maria Manuela Estevez; Teixeira, José António Couto; Rodrigues, António SebastiãoTomato and wine production are the main agro-industrial sectors in Portugal. These activities produce and accumulate large amounts of waste, especially seeds, peels and pulp residues, with high economic and environmental concerns worldwide. These residues are typically rich in bioactive compounds (BC), such as proteins, sugars and lipids, and phenolic compounds (PC) and carotenoids. Therefore, they can be an economical source of high added value, with potential food, cosmetic, or pharmaceutical industry applications. However, current extraction methods (e.g. extraction with chemical solvents) and environmental hazards can also degrade BC and promote toxicity, reducing biological properties and health benefits, making it difficult to use as value-added products. The main objective of this thesis was to explore the value of grape and tomato bagasse to promote the principle of sustainable development, with the achievement of ingredients of added value and zero waste. Thus, an optimisation study of BC extraction from these by-products was carried out, using ohmic heating (OH) method and solvents used in the food industry, to ensure the maintenance of compositional integrity, functionality and safety of bioactive extracts obtained and compared with the traditional method in which it is used organic solvents (CONV). Initially, a group of grape by-products was used, namely stalk and bagasse samples from white and red wine and tomato bagasse, obtained from the processing industry. The compositional characterisation and phytochemical properties of the byproducts were carried out to verify the potential of the matrix to be used and to define the recovery strategy. Grape bagasse presented higher protein, carbohydrate, and BC content than stalks, mainly characterised by high fibre content. Concerning tomato bagasse, samples of the same Heinz tomato cultivar from two different industries were used, and the results were compared. Both samples presented protein contents between 16.3 and 19.4 g / 100 g DW; fibre content between 57.8 and 59.0 g / 100 g DW, mainly polyunsaturated, linoleic acid, oleic and palmitic acid. After complete characterisation, the grape and tomato bagasses were selected as the most promising byproducts and OH was applied to extract the BC and reduce the environmental impacts based on a circular economy strategy. As solvents, water was used alone or combined with ethanol to promote the extraction of more lipophilic compounds, such as carotenoids. In parallel, a CONV method was used to compare the results. After extraction, two different fractions were obtained: the liquid fraction (LF) rich in BC solubilised and the remnant, the solid fraction (SF) with relevant nutritional and functional potential. In the case of bagasse, LF did not present significant differences (p>0.05) between CONV and OH for total phenolic compounds (TPC), 2.84 ± 0.037 and 3.28 ± 0.46 mg / g DW equivalent of gallic acid, respectively. The same trend was found for antioxidant activity (AA), 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 main anthocyanins identified were malvidin-3-O-acetylglucoside, delphinidin-3-Oglucoside, petunidin-3-O-glucoside. These extracts exhibited antimicrobial potential against the microorganisms Yersinia enterocolitica, Pseudomonas aeruginosa, Salmonella Enteritidis, Methicillin-sensitive Staphylococcus aureus, methicillin-resistant S. aureus (MRSA) and Bacillus cereus. SFOH showed a composition with potential functional ingredients, showing higher proteins and BC bound to fibre than CONV. Regarding tomato bagasse, a LF rich in carotenoids, polyphenols, and sugars and a SF rich in fibres linked to polyphenols and carotenoids were obtained. Also, new molecules were identified by UPLC-qTOF-MS analysis, such as phene-di-hexane and N-acetyl-Dtryptophan. After first evaluating the feasibility of applying OH, an experimental design was made to obtain better yields. Different moderate electric fields (MEF) of different intensities (i.e., 4, 6 and 11 V.cm-1) were also used to identify non-thermal effects in the extraction process and their influence on the bioactive properties of the extracted compounds. BC extraction using OH was successfully optimised with the best extraction conditions corresponding to 70 ºC for 15 min using 70% ethanol as a solvent, which allowed a more significant recovery of rutin in 77% than the control samples. It allowed to recover up to 4.93 μg / g lycopene of tomato by-products without resorting to organic solvents, showing itself as a selective extraction method, depending on the compounds of interest. Since there are no studies on the impact of the extraction methodologies and the action of the gastrointestinal tract (GIT) on the bioaccessibility of BC obtained from tomato by-products, the GIT was simulated for each of the LF and SF obtained by the OH and CONV methods. In LF, the results showed that extraction significantly influences the bioaccessibility of the BC present, with OH demonstrating a positive impact on the preservation of BC and consequently on the associated biological properties such as antioxidant, antihypertensive, prebiotic and anti-inflammatory. The main BC identified by UPLC-qTOF-MS were p-cumáric acid (163 m/z), naringenin (271 m/z) and luteolin (285 m/z). In addition, extract obtained by OH after GIT simulation exhibited a prebiotic effect on different strains of Bifidobacterium and Lactobacillus. Regarding FS, the results showed that the OH treatment originated a flour (SFOH) with higher total fibre than the flours obtained with CONV (SFCONV), 62.47 ± 1.24 and 59.06 ± 0.67 g / 100 g DW, respectively. Both flours presented high resistant protein content, representing between 11 and 16% of the insoluble dietary fibre. The main carotenoids identified were lycopene, phytofluene and lutein, all known as health promoters. Despite the higher initial polyphenols and carotenoids in SFOH, BC were more bioaccessible and presented higher antioxidant capacity than those present in SFCONV, throughout the simulated GIT. Finally, and because there is scientific support on the positive effects of prebiotics on the intestinal microbiota, the fermentability of SFOH and SFCONV of tomato were evaluated. The results showed a higher growth of Bacteroidetes with SFOH and the highest values of Bacteroides with SFCONV. A correlation between the growth of microorganisms and short-chain fatty acids was also found. Therefore, the OH treatment allowed similar recovery yields with reduced treatment times and without the need for organic solvents (green extraction routes). In a sense, this work will contribute to the sustainability of the wine and tomato processing industries in the context of circular economy, as it presents an environmentally friendly, fast and economical process capable of recovering BC with high application potentials, such as functional ingredients for food, nutraceutical or cosmetic application.
- Are olive pomace powders a safe source of bioactives and nutrients?Publication . Ribeiro, Tânia B.; Oliveira, Ana; Coelho, Marta; Veiga, Mariana; Costa, Eduardo M.; Silva, Sara; Nunes, João; Vicente, António A.; Pintado, ManuelaBACKGROUND Olive oil industry generates significant amounts of semi-solid wastes, namely the olive pomace. Olive pomace is a by-product rich in high-value compounds (e.g. dietary fibre, unsaturated fatty acids, polyphenols) widely explored to obtain new food ingredients. However, conventional extraction methods frequently use organic solvents, while novel eco-friendly techniques have high operational costs. The development of powdered products without any extraction step has been proposed as a more feasible and sustainable approach. RESULTS The present study fractionated and valorised the liquid and pulp fraction of olive pomace obtaining two stable and safe powdered ingredients, namely a liquid-enriched powder (LOPP) and a pulpenriched powder (POPP). These powders were characterized chemically, and their bioactivity was assessed. LOPP exhibited a significant amount of mannitol (141 g/ kg), potassium (54 g/ kg) and hydroxytyrosol/ derivatives (5 mg/g). POPP exhibited high amount of dietary fibre (620 g/ kg) associated to significant amount of bound phenolics (7.41 mg GAE/ g fibre DW) with substantial antioxidant activity. POPP also contained an unsaturated fatty acids composition similar to olive oil (76% of total fatty acids) and showed potential as a reasonable source of protein (12 %). Their functional properties (solubility, water‐holding and oil‐holding capacity), antioxidant capacity and antimicrobial activity were also assessed, and their biological safety was verified. CONCLUSION The development of olive pomace powders to apply in the food industry could be a suitable strategy to add-value to olive pomace and obtain safe multifunctional ingredients with higher health-promoting effects than dietary fibre and polyphenols itself.
- Influence of harvesting factors on sensory attributes and phenolic and aroma compounds composition of Cymbopogon citratus leaves infusionsPublication . Coelho, Marta; Rocha, C.; Cunha, L.M.; Cardoso, L.; Alves, L.; Lima, R.C.; Pereira, M. J.; Campos, Francisco M.; Pintado, M. E.Herbal infusions are receiving increasing attention for the number of physiological benefits that can bring to human health. Cymbopogon citratus is one the most used plants in traditionalmedicine besides its characteristics and unique aroma appreciated by the consumers; however, little is known about the effects of harvesting on functional and sensory properties of this plant. The present work aimed to assess the lemongrass infusions, which were harvested following a factorial plan according to the type of cut (manual and mechanical) and part of the plant (tips and 2nd half leaves). Total phenolic content, antioxidant capacity, aroma compounds composition (terpenoids) and sensory profiles and consumer perception of different samples were assessed. The quantitative descriptive analysis (QDA) and Check-All-That-Apply (CATA) methods were used to describe the lemongrass infusion, complemented with the overall liking evaluation, using a 9-point scale. Results showed that there were significant differences between type of cut and part of the plant concerning phenolic compounds: samples obtained by mechanical cut showed in general higher content of individual phenolic compounds (in particular for chlorogenic acid) and tips showed in general the highest contents for both types of cut. Consumers panel did not found significant differences between samples. Generally, consumers indicate a bitter taste in all infusions when the content of the phenolic compounds was higher, in particular for p-coumaric acid. Concerning the aroma compounds no significant differences were observed between type of cut and part of the plant, and citral was the terpenoid present in higher quantity. Regarding the sensory profile methods, it was found that QDA and CATA were both good methods to describe this infusion. Considering the role of consumers' evaluation throughout the value chain, the infusion preparedwith plant tips of lemongrass was selected as the premium herbal tea.
- Increasing phenolic and aromatic compounds extraction and maximizing liking of lemon verbena (Aloysia triphylla) infusions through the optimization of steeping temperature and timePublication . Rocha, C.; Coelho, M.; Lima, R. C.; Campos, F. M.; Pintado, M.; Cunha, L. M.Herbal infusions are a new emerging trend among consumers, appearing in the market in many different forms. Due to intense competition, the commercial success of herbal infusions largely depends on flavour. The identification of the best combinations of steeping time and water temperature for the preparation of Aloysia triphylla infusions using a central composite design and response surface methodology was performed. The procedure aimed to maximize its antioxidant activity, phenolic and aromatic compounds, and consumers’ liking of the infusions. The antioxidant activity and phenolic content were evaluated using the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid radical cation) method and the Folin–Ciocalteu procedure. The analysis of aromatic compounds was performed based on the method of terpenic compounds. Sensory evaluation encompassed overall liking using a 9-point hedonic scale. Antioxidant activity and extraction of most phenolic compounds was higher for longer steeping times and higher temperatures, with some presenting a contrasting effect due to degradation at higher temperatures for longer steeping times. Results for terpenic compounds showed similar contrasting patterns. Based on the quadratic response surface, it was possible to predict the maximum overall liking for lemon verbena infusions prepared by brewing for 6 min at 96 ℃. Under such conditions, extraction of bioactive compounds was kept at a high level, close to the maximum attainable, while reducing the extraction of bitter compounds. These results are considered of great importance for the development of premium infusions from organic lemon verbena leaves.
- Impact of storage on phytochemicals and milk proteins in peach yoghurtPublication . Oliveira, Ana; Coelho, Marta; Alexandre, Elisabete M. C.; Pintado, ManuelaYoghurt supplemented with peach is a dairy product consumed around the world with a shelf-life of ca. 30 days. During this period protein–polyphenol complexes can be formed and have a detrimental effect on the in vivo solubility of both phenolic and proteins. Yoghurt enriched with peach preparate were produced and stored up to 28 days at 2 °C. Total antioxidant activity decreased 24%, total carotenoids increased 66%, while total phenolics remained unchanged during the yoghurt shelf-life. Carotenoids presented significant differences between peach enriched yoghurt and peach preparate stored under the same conditions. After 28 days the zeaxantin + lutein, β-cryptoxanthin and β-carotene increased 133, 122, and 84% respectively, for peach preparate, while in peach yoghurt they increased 68, 40, and 15%, indicating interactions of carotenoids with matrix components mainly proteins and fat present in yoghurt. Polyphenols presented higher stability when added to yoghurt with slight variations in their content of ca. 10% for (+)-catechin and 12% for p-coumaric acid, and no variations were recorded in neochlorogenic and chlorogenic acids and in rutin. The α-lactalbumin content decreased 22% after 24 h of peach addition while β-lactoglobulin content did not change. Peach polyphenols were very stable during yoghurt shelf-life while carotenoids were the compounds that presented higher interactions with the matrix.