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- Blueberry juice as a nutritious and bioactive beverage to be included in novel food productsPublication . Backes, Emanueli; Saldanha, Ana Luísa; Molina, Adriana K.; Pereira, Carla; Dias, Maria Inês; Ferreira, Isabel C.F.R.; Barros, LillianBlueberry ( Vaccinium myrtillus L.) is a very popular fruit, native from the northern hemisphere and consumed worldwide. It has been widely studied for being a rich source of bioactive compounds with recognized beneficial properties for Human health [1]. Therefore, several industrialized products, such as juices and derivatives, have been developed from blueberry fruit, aiming at most practical forms of consumption. In this sense, the present work aimed to analyse the blueberry juice in terms of nutritional value, following AOAC procedures, and chemical compositions, namely in free sugars, by HPLC-RI, organic acids, by UFLC-PDA, and phenolic compounds, by HPLC-DAD/ESI-MS. Moreover, the antioxidant capacity, by TBARS and OxHLIA assays, was also assessed to validate its bioactive properties. Regarding the nutritional value, carbohydrates were the major macronutrients found in blueberry juice, followed by ash and protein. In terms of free sugars, fructose and glucose were detected, with a higher concentration of fructose. Four organic acids were also identified in the juice, namely oxalic, quinic, malic, and shikimic acids, being quinic acid the most abundant one. In what concerns the phenolic composition, four phenolic acids (quinic acid, caffeic acid hexoside, cis 5- O- caffeoylquinic acid, and trans 5- O -caffeoylquinic acid) and six anthocyanins (cyanidin-3- O -glucoside, cyanidin-3- O -pentoside, peonidin-3- O glucoside, peonidin-3- O -pentoside, malvidin-3- O -glucoside, and malvidin-3- O -pentoside) were detected. As expected, considering its chemical composition, the juice also presented a strong antioxidant capacity, being able of inhibiting the lipid peroxidation and the oxidative hemolysis. The results obtained in the present study validate the nutritional and bioactive quality of the juice obtained from Vaccinium myrtillus L., justifying its application in the development of novel foodstuff.
- Extraction of anthocyanins from red raspberry for natural food colorants development: Processes optimization and in vitro bioactivityPublication . Rocha, Rosiane; Pinela, José; Abreu, Rui M.V.; Añibarro Ortega, Mikel; Pires, Tânia C.S.; Saldanha, Ana Luísa; Alves, Maria José; Nogueira, António José M.; Ferreira, Isabel C.F.R.; Barros, LillianHeat (HAE)-and ultrasound (UAE)-assisted extraction methods were implemented to recover anthocyanins from red raspberry. Processing time, ethanol concentration, and temperature or ultrasonic power were the independent variables combined in five-level rotatable central composite designs coupled with response surface methodology (RSM) for processes optimization. The extraction yield and levels of cyanidin-3-O-sophoroside (C3S) and cyanidin-3-O-glucoside (C3G) were monitored by gravimetric and HPLC-DAD-ESI/MSn methods, respectively, and used as response criteria. The constructed theoretical models were successfully fitted to the experimental data and used to determine the optimal extraction conditions. When maximizing all responses simultaneously, HAE originated slightly higher response values (61% extract weight and 8.7 mg anthocyanins/g extract) but needed 76 min processing at 38â—¦ C, with 21% ethanol (v/v), while the UAE process required 16 min sonication at 466 W, using 38% ethanol (v/v). The predictive models were experimentally validated, and the purple-red extracts obtained under optimal condition showed antioxidant activity through lipid peroxidation and oxidative hemolysis inhibition, and antibacterial effects against food-related microorganisms, such as Escherichia coli and Enterococcus faecalis. These results highlight the potential of red raspberry extracts as natural food colorants with bioactive effects and could be exploited by industries interested in the production of anthocyanin-based products.
- Attalea speciosa mesocarp flour in-depth characterization and its application for the development of new bakery productsPublication . Souza, M.V.S.; Saldanha, Ana Luísa; Pereira, Carla; Ivanov, Marija; Soković, Marina; Steinmacher, Nádia Cristiane; Dias, Maria Inês; Barros, LillianNon-conventional food plants (NFPs) present themselves as a viable and efficient alternative for replacing the food products we consume today. Presenting a huge nutritional, chemical, physical, and biological potential, these plants are usually abundant and not competitors with other vegetable matrices used for human consumption.1 Attalea speciosa (Mart. Ex Spreng - Babassu) is a palm tree from the botanical family Arecaceae found in Brazil, and its mesocarp (BM) represents about 20.4% of the fruit and most of its application it’s in the manufacture of animal feed.2 Thus, the present work aimed to deepen the study the nutritional characterization of BM by AOAC methods; the determination, by chromatographic methods, of free sugars (HPLC-RI), fatty acids (GC-FID), and organic acids content (UPLC-DAD); and the study of the phenolic profile (HPLC-DAD/ESI(MS) and bioactivities (antioxidant, antimicrobial, anti-inflammatory and cytotoxicity activities) of the hydroethanolic extracts. Furthermore, given the possible industrial application, bakery products (bread) were developed with 12, 18, and 24% substitutions of wheat flour by BM flour. BM flour presented low moisture, fat, ash levels, being carbohydrates the main macronutrients, and palmitic (C16:0) and stearic (C18:0) acids were the main fatty acids found in the sample. Regarding phenolic composition, nine phenolic compounds were tentatively identified, six flavan-3-ols (catechin and epicatechin derivatives) and three Oglycosylated flavonoids (quercetin derivatives). Specifically, the flavan-3-ols group represented 99% of total amount of phenolic compounds mainly due to the presence of β-type (Epi)catechin dimer. The hydroethanolic extract of BM showed a high antioxidant capacity to inhibit lipid peroxidation and high anti-hemolytic capacity, showing an IC50 99% and 75% more effective than the positive control used (Trolox) in these assays, respectively. Moreover, it was able to inhibit all the tumor cell lines tested (MCF-7, NCIH460, CaCo, and AGS), however, it showed some toxicity towards healthy cells of PLP2 and VERO lines. The results obtained for the antibacterial activity of the BM extract were lower when compared to the positive controls used (E211 and E224). Comparing the results obtained with 100% wheat flour products, it was noted that the formulations with substitutions by BM presented remarkable results, providing a reasonable increase in the PUFAs content (despite decreasing protein content), maintaining the antioxidant capacity with lower IC50 values than the positive control used, and not presenting hepatotoxic activity (first validation of this flour for the incorporation in food matrices). In general, the formulation B24 was the one that presented greater similarity to the control bread (B0), relatively to the nutritional aspects, however, concerning the physical parameters it was the one that presented bigger discrepancy, especially regarding the specific volume, texture, and the color parameter L* (luminosity), tending for a darker and opaque coloration. All the formulations presented higher percentages of loss of rheological characteristics in the first three days of elaboration, however, formulation B24 was the one that presented the lowest percentage. This study presented innovative results regarding the nutritional, chemical and bioactive characterization of both the babassu mesocarp itself and the bread made from it, showing great potential to be applied in the food industry, however, there is still much to explore, especially in preparations that do not yet use non-conventional ingredients.
- Red raspberry waste as a source of anthocyanin-rich food colorants: extraction process optimization and functionality assessmentPublication . Añibarro-Ortega, Mikel; Rocha, Rosiane; Pinela, José; Pires, Tânia C.S.; Abreu, Rui M.V.; Saldanha, Ana Luísa; Alves, Maria José; Nogueira, António José M.; Ferreira, Isabel C.F.R.; Barros, LillianFood colorants are increasingly used in the food industry to preserve, improve or change the food color. While the quite controversial artificial colorants are widely used in this sector, the natural counterparts have been less selected in part due to the limited availability of options and stability issues [1]. Within this class, anthocyanins are naturally occurring colorants that can be found in different plant matrices, including berries such as red raspberry (Rubus idaeus L.). These water-soluble pigments show attractive colors ranging from red to purple and present health-promoting effects [2,3]. Therefore, this work aimed to develop a novel anthocyanin-rich food colorant from red raspberry waste through the optimization of a sustainable extraction methodology and to characterize this ingredient for its functionality. Heat (HAE)- and ultrasound (UAE)- assisted extraction methods were implemented to recover the anthocyanins from red raspberry. Processing time, ethanol concentration, and temperature or ultrasonic power were the independent variables analyzed in a central composite design coupled with response surface methodology for processes optimization. The extraction yield and levels of anthocyanins (cyanidin-3-O-sophoroside and cyanidin-3-O-glucoside) were monitored gravimetrically and by HPLC-DAD-ESI/MSn , respectively, and used as response criteria. The constructed theoretical models were successfully fitted to the experimental data and used to determine the optimal extraction conditions. Overall, HAE originated slightly higher response values (61% extract weight and 8.7 mg anthocyanins/g extract) but needed 76 min processing at 38 °C, with 21% ethanol, while the UAE process required 16 min sonication at 466 W, using 38% ethanol, to obtain 58% extract weightand 8.3mganthocyanins/gextract.Then,thepredictivemodelswereexperimentallyvalidatedand the purple red extracts obtained under optimal condition showed antioxidant activity through lipid peroxidation and oxidative hemolysis inhibition, and antibacterial effects against food-related bacteria, such as Escherichia coli and Enterococcus faecalis [4]. These results highlight the potential of red raspberry extracts as natural food colorants with bioactive effects. In future studies, it will be interesting to investigate the stability of the developed anthocyanin-rich extracts when exposed to different stability factors and in real food matrices.
- Caracterización detallada de la harina del mesocarpio de Attalea speciosa y su aplicación para el desarrollo de nuevos productos de panaderíaPublication . Souza, M.V.S.; Saldanha, Ana Luísa; Pereira, Carla; Ivanov, Marija; Soković, Marina; Steinmacher, Nádia Cristiane; Dias, Maria Inês; Barros, Lillian; Morais, CarlosLas plantas alimenticias no convencionales se presentan como una alternativa viable y eficiente para reemplazar los productos alimenticios que consumimos hoy. Presentando un enorme potencial nutricional, químico, físico y biológico, estas plantas suelen ser abundantes y no competidoras de otras matrices vegetales [1]. Attalea speciosa (Mart. Ex Spreng - Babassu) es una palmera que se encuentra en Brasil, y su mesocarpio (MB) representa cerca del 20,4% del fruto, presentando aplicabilidad en la fabricación de alimentos para animales [2]. Así, el presente trabajo tuvo como objetivo profundizar el estudio de la caracterización nutricional de MB por métodos de la AOAC; la determinación del contenido de azúcares libres (HPLC-RI), ácidos grasos (GC-FID), ácidos orgánicos (UPLC-DAD) y compuestos fenólicos (HPLCDAD/ ESI(MS); y propiedades bioactivas (antioxidante, antimicrobiana, antiinflamatoria, citotóxica y nefrotoxicidad) de los extractos hidroetanólicos. Además, se desarrollaron productos de panadería (pan) con 12, 18 y 24% de sustitución de harina de trigo por harina MB. La harina presentó bajos niveles de humedad, grasa y cenizas, siendo los carbohidratos los principales macronutrientes, y los ácidos palmítico y esteárico los principales ácidos grasos. Se identificó tentativamente nueve compuestos fenólicos, seis flavan-3-oles (99% de la cantidad total, sobre todo por la presencia del dímero de (epi)catequina tipo β) y tres flavonoides O-glicosilados. El extracto hidroetanólico mostró una alta capacidad antioxidante y antihemolítica, mostrando una IC50 de 99% y 75% más efectiva que el control positivo (Trolox), respectivamente. Además, fue capaz de inhibir todas las líneas celulares tumorales, per mostró cierta toxicidad hacia las células sanas. Para la actividad antibacteriana fueron obtenidas concentraciones mínimas inhibitorias inferiores a la de los controles positivos (E211 y E224). Finalmente, se observó que las formulaciones de pan con sustituciones presentaron un aumento razonable en el contenido de PUFAs, manteniendo la capacidad antioxidante con valores de IC50 más bajos que los del control positivo, y no presentando actividad hepatotóxica (primera validación de esta harina para la incorporación en matrices alimentarias). En general, la formulación B24 fue la que presentó mayor similitud con el pan control, en relación a los aspectos nutricionales, sin embargo, en cuanto a los parámetros físicos fue la que presentó mayor discrepancia (en volumen específico, textura, y luminosidad). Todas las formulaciones presentaron mayores porcentajes de pérdida de características reológicas en los primeros tres días de elaboración. Este estudio presentó resultados innovadores en cuanto a la caracterización nutricional, química y bioactiva tanto del propio mesocarpio de babasú como del pan elaborado a partir de él, mostrando un gran potencial para ser aplicado en la industria alimentaria, sin embargo, aún queda mucho por explorar, especialmente en preparaciones que todavía no use ingredientes no convencionales
- Extraction of chlorophylls from natural sourcesPublication . Gomes, Leonardo Corrêa; Pereira, Carla; Dias, Maria Inês; Molina, Adriana K.; Saldanha, Ana Luísa; Ferreira, Isabel C.F.R.; Barros, LillianThe growing consumers’ concern for possible long-term adverse effects of artificial molecules commonly used in food industry has led to an increased interest in natural products. At the same time, there is a demand for a more eco-sustainable use of natural matrices, which justifies the search for byproducts that have no other application to be explored in the development of novel food products [1,2]. In this context, the present study was designed to exploit natural pigments, more specifically chlorophylls, from bioresidues (aerial parts of carrot and tomato) for the development of food colorants. These are the most abundant pigments in plants and present, beyond their great coloring capacity, several bioactive properties, which corroborates the importance of their application in foodstuff. In this work, different extraction methodologies and techniques (maceration, ME, and ultrasound-assisted, USE) were applied to the lyophilized aerial parts of carrot and tomato to maximize the chlorophyll extraction yield. For the extraction, green solvents were prioritized, namely water, ethanol (90%), and hexane. The parameters affecting the pigments recovery were varied for each technique, namely the time, power, and solvent for USE, and the time and solvent for ME. The extractions were performed protecting the samples from light and the results were monitored through the implementation of a new chromatographic method, HPLC coupled to a diode array detector (DAD) and mass spectrometry (MS), to determine the concentration of chlorophylls and the best procedure to be performed. Both aerial parts presented chlorophylls and derivatives in significant concentrations and extraction yields up to 88% for the ethanolic extracts. The applied chromatographic method revealed to be appropriate for the analysis of this class of pigments, allowing a good peak resolution and separation, but also characteristic TIC spectrum for the tentative identification of the compounds. Therefore, the results of the present study can be explored for the development of chlorophyl-based colorants from these bioresidues, but also from similar byproducts.
- Carotenoid-based solutions for the replacement of artificial colorants in pastry productsPublication . Martins, Pedro Miguel Mota; Dias, Maria Inês; Pereira, Carla; Molina, Adriana K.; Saldanha, Ana Luísa; Ferreira, Isabel C.F.R.; Barros, LillianColour has a great importance in the first consumers’ impression, allowing to infer about the overall quality, the taste, the smell, the texture, and even the safety of foodstuff [1]. For these reasons, there is a massive use of colorants in food products. Nevertheless, the most applied compounds are of artificial origin and some of them have been increasingly associated to health issues, with allergic reactions, children attention deficit, and cancer pointed out as the most common consequences [2]. These facts have been driving new research in this field, through the exploitation of natural sources of colouring molecules to be applied in detriment of artificial colorants. Among the numerous natural matrices potentially used for the extraction of colouring compounds, the fruits from the genus Solanum represent promising sources of pigments, namely carotenoids [3]. Together with the fact that large amounts of fresh tomato wastes (resulting from crop growing, packaging, processing, storage, and sale) are discarded worldwide, the recovery of valuable colorant biomolecules from these agri-food wastes represents a crucial step of the circular economy by re introducing them into the food chain as ingredients [3]. The need to process these bio-wastes for the recovery of coloring molecules, has led to the use of more eco-sustainable extraction methodologies in detriment of more conventional techniques, such as maceration. Ultrasound-assisted extraction methodology arises as one of the most promising alternatives, with lower extraction times, use of greener solvents, and higher recovery yields, but also with the possibility to be scaled-up to respond to the high demands of the industrialized world [3]. Carotenoid compounds are lipophilic pigments responsible for the yellow, orange, and red colours of certain plant matrices, with a vast structural diversity, but prone to isomerization and oxidation [4]. However, the colouring capacity of these molecules overcomes any instability problem (that can be solved with stabilization strategies) and, therefore, carotenoid-based colorants appear as a valid solution for application in the pastry sector, that greatly relies on yellow/orange artificial colorants.
- Integrated approaches for socio-economic boosting of the sustainable production and consumption of Montesinho mushroomsPublication . Saldanha, Ana Luísa; Gomes, Leonardo Corrêa; Pinela, José; Fernandes, Ângela; Martins, Anabela; Carvalho, Ana Maria; Nobre, Sílvia; Coimbra, Manuel A.; Barros, Lillian; Dias, Maria Inês; Pereira, CarlaMontesinho Natural Park (MNP) represents a mountain area with a unique mycological heritage. Among the approximately two hundred edible mushrooms found in this protected area, most of them have been consumed since ancient times for their exquisite taste and nutritional value. 1 Nevertheless, the availability of these mountain products is limited by their seasonality and weather conditions, an issue intensified by climate change. On the other hand, unsustainable mushroom collection practices and illegal trade of high-value species have also been common practices with negative impacts in the ecosystem and regional and national economy. All these issues, together with the incapacity of local collectors to guarantee the authenticity/safety of the collected mushrooms, have led many restaurants to avoid their inclusion in their menus, safeguarding their business and consumer’s health. Thus, it is proposed the production of appreciated edible mushrooms in controlled ex-situ environment. An extensive nutritional/chemical/biological characterization are being performed to ensure the high quality of the produced species and the preservation of their original characteristics. The development of a quality and safety seal, “Safe2Taste”, that guarantee the traceability of the entire production chain, aims to increase consumers’ confidence/loyalty on the products.
- Enfoques integrados para el impulso socioeconómico de la producción y consumo sostenible de hongos de MontesinhoPublication . Saldanha, Ana Luísa; Gomes, Leonardo Corrêa; Pinela, José; Coimbra, Manuel A.; Barros, Lillian; Dias, Maria Inês; Pereira, CarlaEl Parque Natural de Montesinho representa un área de montaña con un patrimonio micológico único. Entre las aproximadamente doscientas setas comestibles que se encuentran en este espacio protegido, la mayoría se consumen desde la antigüedad por su exquisito sabor y valor nutritivo [1]. Sin embargo, la disponibilidad de estos productos de montaña está limitada por su estacionalidad y condiciones climáticas, cuestión que se intensifica con el cambio climático. Por otro lado, las prácticas insostenibles de recolección de hongos y el comercio ilegal de especies de alto valor también han sido prácticas comunes con impactos negativos en el ecosistema y la economía regional y nacional. Todas estas cuestiones, junto con la incapacidad de los recolectores locales para garantizar la autenticidad/inocuidad de las setas recolectadas, han llevado a muchos restaurantes a evitar su inclusión en sus menús, salvaguardando su negocio y la salud de los consumidores. Así, se propone la producción de apreciados hongos comestibles en ambiente controlado ex-situ. Se está realizando una extensa caracterización nutricional, química y biológica para asegurar la alta calidad de las especies producidas y la preservación de sus características originales. El desarrollo de un sello de calidad y seguridad, “Safe2Taste”, que garantiza la trazabilidad de toda la cadena de producción, tiene como objetivo aumentar la confianza/fidelidad de los consumidores en los productos
- Red raspberry waste as a source of anthocyanin-rich food colorants: extraction process optimization and functionality assessmentPublication . Añibarro-Ortega, Mikel; Rocha, Rosiane; Pinela, José; Pires, Tânia C.S.; Abreu, Rui M.V.; Saldanha, Ana Luísa; Alves, Maria José; Nogueira, António José M.; Ferreira, Isabel C.F.R.; Barros, LillianFood colorants are increasingly used in the food industry to preserve, improve or change the food color. While the quite controversial artificial colorants are widely used in this sector, the natural counterparts have been less selected in part due to the limited availability of options and stability issues