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  • Development and validation method for simultaneous quantification of phenolic compounds in natural extracts and nanosystems
    Publication . Silva, Sara Baptista da; Oliveira, Ana; Ferreira, Domingos; Sarmento, Bruno; Pintado, Manuela
    IntroductionSage and savoury (Salvia sp. and Satureja montana, respectively) are plants used in traditional medicine. The quality control of their herbal formulations is of paramount concern to guarantee the expected biological activity of their anti-oxidant compounds. Objective To establish a simple and effective high-performance liquid chromatographic (HPLC) method to evaluate simultaneously quercetin and rosmarinic acid, in a pure form, in natural extracts (sage and savoury), and encapsulated into chitosan nanoparticles. Methods Chromatography was performed on an RP C-18-column, in a gradient mode with a mobile phase comprising methanol:formic acid:water 92.5:2.5:5 (v/v) at a flow rate of 0.75mL/min and at wavelength of 280nm. Results The method was specific, linear in the range of 0.05-1mg/mL (R-2=1.00), precise at the intraday and interday levels, accurate (recovery rate 90.50.6%), and robust to changes in equipment conditions. Conclusion The method established was effective for quercetin and rosmarinic acid characterisation in natural extracts and in chitosan nanoparticles, allowing the loading capacity determination, the association efficiency as well as the in vitro release. This study shows a new HPLC method validated to simultaneous quantification of rosmarinic acid and quercetin in natural extracts and chitosan nanoparticles. This method is also able for detection and quantification of other phenolic and flavanoid compounds in natural matrix.
  • Effect of modified atmosphere on phytochemical profile of pasteurized peach purées
    Publication . Oliveira, Ana; Coelho, Marta; Alexandre, Elisabete; Gomes, Maria Helena; Almeida, Domingos P. F.; Pintado, Manuela
    Peach 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.
  • The effect of sex, season and gametogenic cycle on gonad yield, biochemical composition and quality traits of Paracentrotus lividus along the North Atlantic coast of Portugal
    Publication . Rocha, Filipa; Baião, Luís F.; Moutinho, Sara; Reis, Bruno; Oliveira, Ana; Arenas, Francisco; Maia, Margarida R. G.; Fonseca, Antonio J. M.; Pintado, Manuela; Valente, Luisa M. P.
    Sea urchin population harvest in the North Atlantic coast of Portugal was characterized in terms of gonad yield, nutritional composition and important market-related traits, over one reproductive cycle (March 2016 to March 2017). Most of the quality attributes showed a seasonal variation strongly dependent on sea urchin sex. Maximum gonad yield (18%) was observed in March 2017. A single spawning event occurred between May and July. Gonads are rich sources of protein (12–18% WW) with low fat content (≤6% WW), that increase during the gametogenic stages of recovery and growing (November-December). Polyunsaturated fatty acids were the dominant class in both sexes (4.2–14.7 mg.g−1 WW), being preferentially accumulated in females. Total gonads carotenoid varied seasonally, with the highest level being observed in males during spawning season. Echinenone was the main pigment present in gonads, showing highest concentrations in males during spawning and gonad recovering. During the growing and early maturation period gonads were more reddish, yellowish and brighter, as well as more firm, irrespectively of the sex. Based on all seasonal changes affecting gonad yield and quality, the period between November and February seems the most suitable to harvest high quality gonads in the Atlantic coast of Portugal.
  • Long-term frozen storage and pasteurization effects on strawberry polyphenols content
    Publication . Oliveira, Ana; Coelho, Marta; Alexandre, Elisabete M. C.; Almeida, Domingos P. F.; Pintado, Manuela
    The 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.
  • Non-compliant fruit as new functional food ingredients
    Publication . Vilas-Boas, Ana A.; Gómez-García, Ricardo; Campos, Débora A.; Oliveira, Ana; Pintado, Manuela
    Over the last decade, the high amount of food waste has received increased attention towards a more sustainable future, such as reducing food losses and valorize food by-products. Non-compliant fruit represents a significant part of fruit losses during production as a result of aesthetic requirements (color, low size, and deformations in the shape of the products). These fruits do not match commercialization standards when sold as fresh fruit. Selling this kind of fruit at a cheap price might have a great impact on the reduction of fruit loss, nevertheless, this option does not offer a long-time solution, being required an alternative and sustainable approach. These fruits represent a rich source of bioactive compounds (phenolic compounds and dietary fiber) that could be used as new functional ingredients/products. The production of new functional ingredients/products from non-compliant fruit consists of a valuable and sustainable approach for taking advantage of these food losses. Therefore, the purpose of this chapter is to provide an overview of new strategies to recover from non-compliant fruit value-added ingredients, impacting society, environment, and economy.
  • Phytochemicals preservation in strawberry as affected by pH modulation
    Publication . Oliveira, Ana; Gomes, Maria Helena; Alexandre, Elisabete M. C.; Poças, Fátima; Almeida, Domingos P. F.; Pintado, Manuela
    Strawberries purees are incorporated in foods and subjected to pH modulation according to the expected final food matrix. The effect of pH on strawberry polyphenols stored at 4 and 23 degrees C for 90 days was evaluated. Total antioxidant activity and total phenolics content were only affected by time according to a first order model. The pH 4.5 induced higher decrease in (-)-epigallocatechin gallate (71% and 79%) and quercetin-3-glucoside (29% and 36%), for both storage temperatures. For pH 2.5 and 3.0, ellagic acid increased 84% for 4 degrees C. and 185% for 23 degrees C. Anthocyanins concentration changes along storage were well described by first order model. The pH value of 2.5 presented the lower kinetic constant rate where cyanidin-3-glucoside, pelargonidin-3-glucoside and pelargonidin-3-rutinoside had a k = 0.04, 0.05 and 0.03 day(-1). Lower storage temperature (4 degrees C) and lower pH (2.5) were the best condition for the preservation of polyphenols in pasteurized strawberry during a 90-day storage period.
  • Polyphenol extraction by different techniques for valorisation of non-compliant portuguese sweet cherries towards a novel antioxidant extract
    Publication . Vilas-Boas, Ana A.; Campos, Débora A.; Nunes, Catarina; Ribeiro, Sónia; Nunes, João; Oliveira, Ana; Pintado, Manuela
    Currently, there is special interest in the recovery of polyphenols from non-compliant fruits that have no market value; e orts to find value-added solutions for these food areas are a key option for a sustainable bio-economy. Saco cherries are a traditional Portuguese cherry variety, and although they are a nutritionally important food, rich in powerful dietary polyphenols, significant amounts of these cherries are not sold due to their small size. In this context, this work aimed to select the best method to produce novel antioxidant polyphenol-rich extracts from low calibre and non-compliant Saco cherries. Based on the results, microwaves-assisted extraction (MAE) allowed us to obtain a polyphenol-rich extract with a high antioxidant capacity (50.46 + 1.58 mg Trolox equivalent (TE)/g dry extract (DE) by oxygen radical absorbance capacity (ORAC), 10.88 + 0.38 mg ascorbic acid equivalent (AA)/g DE by 2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and 9.58 + 0.42 mg TE/g DE by 2,2-diphenyl-1-picrylhydrazyl (DPPH)) and a high content of polyphenols, namely, hydroxycinnamic acids (neochlorogenic and p-coumaric acids) and anthocyanins (cyanidin-3-rutinoside and cyanidin-3-glucoside), compared with those of conventional extractions with low and high temperature and ultrasound-assisted extraction. The antioxidant extract produced from MAE could be a new alternative for the valorisation of non-compliant cherries since these extracts proved to be a functional ingredient due to the high content of antioxidants, which are linked to the prevention of diseases.
  • Nanoencapsulation of polyphenols towards dairy beverage incorporation
    Publication . Silva, Sara; Veiga, Mariana; Costa, Eduardo M.; Oliveira, Ana L.; Madureira, Ana Raquel; Pintado, Manuela
    Phenolic compounds, while widely recognized for their biological potential, when added into food matrixes may interact with food constituents. One example of this is the interaction between phenolic compounds and proteins, that may result in the formation of complexes and alter the bioavailability of both phenolic compounds and the nutrient availability. Moreover, when adding compounds to improve the functionality of a food matrix, these interactions may compromise the perceived benefits of the additions. Nanoencapsulation has been considered one of the means to circumvent these interactions, as they may function as a physical barrier between the phenolic compounds and the matrix (preventing not only the loss of bioactivity, but eventual sensorial alterations of the foods), protect phenolic compounds through the gastrointestinal tract, and may enhance phenolic absorption through cellular endocytosis. However, despite these advantages the food industry is still limited in its nanotechnological solutions, as special care must be taken to use food-grade encapsulants which will not pose any deleterious effect towards human health. Therefore, this review aims to provide an encompassing view of the existing advantages and limitations of nanotechnology, associated with the inclusion of phenolic compounds in dairy beverages
  • Total and sustainable valorisation of olive pomace using a fractionation approach
    Publication . Ribeiro, Tânia B.; Oliveira, Ana L.; Costa, Cristina; Nunes, João; Vicente, António A.; Pintado, Manuela
    Olive pomace management represents a great concern to the olive oil industry. This work focused on the development of a “zero waste” strategy for olive pomace based on a fractionation approach resulting in the obtention of different value-added fractions. The physicochemical composition of edible fractions obtained (liquid and pulp) was analysed. The potential use as a solid biofuel of the non-edible fraction (stones) was evaluated. High amounts of hydroxytyrosol (513.61–625.76 mg/100 g dry weight) were present in the liquid fraction. Pulp fraction was demonstrated to be a good source of fibre (53–59% dry weight) with considerable antioxidant activity both from free and bound phenolics. The stones fraction exhibited substantial high heating values (18.65–18.94 megajoule (MJ/kg). All these results support the added value of the olive pomace fractions combining the biofuel potential from the stones fraction and the functional food ingredients’ potential both from liquid and pulp fractions. The present methodology seems to be a feasible whole valorisation approach to achieve the circularity in the olive oil sector, prioritising obtaining high over low added-value products.
  • Study of olive pomace antioxidant dietary fibre powder throughout gastrointestinal tract as multisource of phenolics, fatty acids and dietary fibre
    Publication . Ribeiro, Tânia; Campos, Débora; Oliveira, Ana; Nunes, João; Vicente, António A.; Pintado, Manuela
    Pulp-enriched powder (POPP) was obtained from olive pomace solid fraction, a derived from the new value chain established for olive by-products. As a multifunctional powder, POPP retains several bioactive compounds (fatty acids, dietary fibre and phenolics) under potential synergic interaction, even more, reactive throughout the digestion. So, in this study, the potential multifunctionality of POPP was evaluated after the gastrointestinal tract. A significant loss of phenolics occurred during oral digestion (62.48 %). However, the potential role of dietary fibre as phenolics’ carrier and its possible liberation in the stomach allowed recovering a significant amount of phenolics (77.11 %) and a bioaccessibility index of at least 50 % (mainly for tyrosol and its glucoside). POPP also provides high content of dietary fibre mainly insoluble fibre (69.68 g/100 g dry weight) linked to a substantial amount of bound phenolics (7.63 mg of gallic acid equivalents/g fibre dry weight), with a positive effect on the fatty acids bioaccessibility [decreased the saturated (5-6 %) and facilitated the unsaturated fatty acids bioaccessibility (4-11%)]. PCA analysis became evident the negative effect of simulated gastrointestinal digestion upon POPP as mainly linked to phenolics’ loss. Despite all negative effects of the simulated digestion on POPP bioactive composition, phenolics and unsaturated fatty acids showed to be bioaccessible in significant amount, and the amount of bound phenolics associated to fibre retained in the colon have the potential to exert gut health benefits.