Two Sides of the Same Coin: The Impact of Grain Legumes on Human Health: Common Bean (Phaseolus vulgaris L.) as a Case Study

Data from Food and Agriculture Organization indicate the worrying scenario of severe food insecurity in the world and the contrasting high prevalence of obesity (13% of the world adult population) in both developing and developed countries. Sustainable agriculture systems with increased inclusion of grain legume species and the boosting of public awareness about legume importance on diet should be a priority issue to eradicate malnutrition and promote public health. However, grain legume production and consumption are in constant state of decline, especially in the European Union. Assigned as the “poor man’s meat”, “promoters of flatulence”, or incorrectly classified as “starchy foods”, grain legumes have a negative image in modern societies. In fact, legumes represent an important source of protein, fiber, vitamins (e.g. folate) and minerals (e.g. magnesium). Moreover, legumes are rich in bioactive compounds (e.g. phenolic compounds, protease and α-amylase inhibitors) acting as a “double-edged sword” in human health. They may impair nutrients availability exerting at the same time beneficial biological activities in lipid profile, inflammation, glycaemia and weight. The present chapter is focused on the advantages of a legume-rich diet for health promotion at a global scale, reviewing legume nutritional and bioactive compounds, with particular emphasis on common bean

Process intensification by microwave pretreatment

Arbutus unedo L., commonly known as the strawberry-tree fruit, is an endemic species of the Mediterranean flora. Microwave extraction technology has been considered as a fast and "green" method for the production of extracts rich in bioactive compounds, although the energy consumption is high. To overcome this bottleneck, microwave was used as a pretreatment procedure in short time periods. This technique promotes the burst of intracellular vacuoles leading to an increase in the lixiviation of phenolic compounds. Different approaches were tested, namely a solvent-free irradiation (SFI), a solvent-assisted irradiation (SAI) and a pressurized solvent-assisted irradiation (PSAI). After irradiation, a solid-liquid extraction procedure was performed using a mixture of water and ethanol. A kinetic evaluation of the total phenolic content (TPC) was performed using the Folin-Ciocalteu method. For the total anthocyanin content, a UV-spectrophotometric method was used. HPLC-UV and LC-MS were used for TPC and identification of present compounds. Microwave irradiation led to an increase in TPC of extracts after SAI (52%) and PSAI (66%) along with a reduction in time of extraction from 30 min to less than 2 min. The anthocyanin content also increased by 66% for the SAI and PSAI extractions.publishersversionpublishe

Hydroxycinnamic acids and their derivatives in broa, a traditional ethnic maize bread

This research was funded by EUROPEAN UNION'S SEVENTH FRAMEWORK PROGRAMME for research, technological development and demonstration under grant agreement number 245058, by FUNDACAO PARA A CIENCIA E TECNOLOGIA and PORTUGAL 2020 to the Portuguese Mass Spectrometry Network, grant number LISBOA-01-0145-FEDER-402-022125 and by FUNDACAO PARA A CIENCIA E TECNOLOGIA through IF/01337/2014 FCT Investigator contract (MCVP) and research unit GREEN-IT (UID/Multi/04551/2020).Maize is one of the most interesting dietary sources of hydroxycinnamic acids, widely known for their beneficial health effects, namely antioxidant properties. This work aims to identify hydroxycinnamic acids and their derivatives in broa, a Portuguese traditional ethnic maize bread, and corresponding maize flours. Soluble and insoluble phenolic fractions of diverse maize flours and corresponding broas were prepared and analysed by HPLC‐DAD‐MS/MS (high‐performance liquid chromatography coupled with diode array detector and tandem mass spectrometry). Besides free hydroxycinnamic acids, mainly ferulic and p‐coumaric acids, several structural isomers and stereoisomers of insoluble ferulic acid dehydrodimers (n = 18) and trimers (n = 11), were also identified. Hydroxycinnamic acid amides consisting of coumaroyl and feruloyl conjugates (n = 22) were present in both soluble and insoluble fractions of maize flours and breads, in different isomeric forms. A new compound was putatively identified as bis‐N,N′‐diferuloyl putrescine. Additionally, more complex and insoluble hydroxycinnamic acid amides, derived from ferulic acid dehydrodimers (n = 47) and trimers (n = 18), were also putatively identified for the first time, suggesting that hydroxycinnamic acid amides are also linked to maize cell walls. Since hydroxycinnamic derivatives were not only identified in maize flours, but also in broas, they can contribute to the antioxidant properties and beneficial health effects of maize‐based foods.publishersversionpublishe

An improved HILIC HPLC-MS/MS method for the determination of β-ODAP and Its α isomer in lathyrus sativus

This research was funded by EUROPEAN UNION'S SEVENTH FRAMEWORK PROGRAMME for research, technological development and demonstration under grant agreement number 613551 and by FUNDACAO PARA A CIENCIA E TECNOLOGIA (IF/01337/2014, PTDC/AGR-TEC/0992/2014) and PORTUGAL 2020 to the Portuguese Mass Spectrometry Network, grant number LISBOA-01-0145-FEDER-402-022125.β-N-Oxalyl-l-α,β-diaminopropionic acid (β-ODAP) is a non-protein amino acid present in Lathyrus sativus (grass pea) and other Lathyrus species, in parallel with its nontoxic isomer, α-ODAP. When consuming grass pea for several months as staple food, β-ODAP may cause neurolathyrism, a motor neuron degeneration syndrome. Therefore, the independent quantification of both ODAP isomers instead of only the total amount in grass pea allows the identification of less toxic varieties and the development of tools to support breeding for improving grass pea quality. In this work, a simple and fast HPLC-MS/MS method was developed without sample derivatization, using a hydrophilic interaction chromatography (HILIC) column and an isocratic gradient of eluents for 18 min, which allowed the determination of both α- and β-ODAP. The proposed method was fully validated and applied to the determination of α- and β-ODAP contents in a diverse collection of 107 grass pea accessions representative of the main grass pea-growing geographical regions in the world, with the prompt identification of contrasting accessions. β-ODAP content in the analyzed grass pea samples ranged from 0.45 ± 0.02 to 6.04 ± 0.45 mg g-1. The moderate correlation found between α- and β-ODAP contents (0.65) in this collection reinforces the importance of the independent quantification of both ODAP isomers.publishersversionpublishe

Relating Bioactivities with the Phenolic Compounds

Funding Information: V.W. acknowledges the Fundação para a Ciência e Tecnologia for PhD fellowship 2021.08511.BD. This work was supported by the Associate Laboratory for Green Chemistry—LAQV, which is financed by national funds from FCT/MCTES (UIDB/50006/2020). The authors acknowledge the financial support from the Fundação para a Ciência e Tecnologia and Portugal 2020 to the Portuguese Mass Spectrometry Network (LISBOA-01-0145-FEDER-402-022125). This work was funded by the Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). A.T.S. also thanks FCT/MCTES for the individual grant CEECIND/04801/2017. This research was also funded by the Sociedade Ponto Verde—Sociedade de Resíduos de Embalagens, S.A., through the project entitled “Bioplastics and Edible, Vegan Films”. Publisher Copyright: © 2023 by the authors.Arabinoxylan has prebiotic properties, as it is able to resist digestion in the small intestine and undergoes fermentation in the large intestine. In this work, arabinoxylan was extracted from corn fiber using an alkaline solution and further purified with membrane processing. It was found that the extracts were mainly composed of xylose (50–52%), arabinose (37–39%), galactose (9%) and glucose (1–4%), with an A/X ratio of 0.72–0.77. All the extracts were composed of phenolic compounds, including ferulic acid derivatives such as dimers, trimers and tetramers. The purified extract had a lower concentration of ferulic and p-coumaric acid (0.004 and 0.02 mg/mgdry_weight, respectively) when compared to raw extract (19.30 and 2.74 mg/mgdry_weight, respectively). The same effect was observed for the antioxidant activity, with purified extracts having a lower value (0.17 ± 0.02 µmol TEAC/mg) when compared to the raw extract (2.20 ± 0.35 µmol TEAC/mg). The purified extract showed a greater antiproliferative effect against the HT29 cell line with EC50 = 0.12 ± 0.02 mg/mL when compared to the raw extract (EC50 = 5.60 ± 1.6 mg/mL). Both raw and purified extracts did not show any cytotoxicity to the Caco-2 cell line in the maximum concentration tested (10 mg/mL).publishersversionpublishe

Shedding light on the volatile composition of broa, a traditional portuguese maize bread

Funding Information: Funding: This research was funded by the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement number 245058, and by Fundação para a Ciência e Tecnologia through research unit GREEN-IT (UID/Multi/04551/2020). Funding Information: This research was funded by the European Union?s Seventh Framework Programme for research, technological development and demonstration under grant agreement number 245058, and by Funda??o para a Ci?ncia e Tecnologia through research unit GREEN-IT (UID/Multi/04551/2020). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.In Portugal, maize has been used for centuries to produce an ethnic bread called broa, em-ploying traditional maize varieties, which are preferred by the consumers in detriment of commercial hybrids. In order to evaluate the maize volatiles that can influence consumers’ acceptance of broas, twelve broas were prepared from twelve maize varieties (eleven traditional and one commercial hybrid), following a traditional recipe. All maize flours and broas were analyzed by HS-SPME-GC-MS (headspace solid-phase microextraction) and broas were appraised by a consumer sensory panel. In addition, the major soluble phenolics and total carotenoids contents were quantitated in order to evaluate their influence as precursors or inhibitors of volatile compounds. Results showed that the major volatiles detected in maize flours and broas were aldehydes and alcohols, derived from lipid oxidation, and some ketones derived from carotenoids’ oxidation. Both lipid and carotenoids’ oxidation reactions appeared to be inhibited by soluble phenolics. In contrast, phenolic compounds appeared to increase browning reactions during bread making and, consequently, the production of pyranones. Traditional samples, especially those with higher contents in pyranones and lower contents in aldehydes, were preferred by the consumer sensory panel. These findings suggest that, without awareness, consumers prefer broas prepared from traditional maize flours with higher contents in health-promoting phenolic compounds, reinforcing the importance of preserving these valuable genetic resources.publishersversionpublishe

Extraction of Biocompatible Collagen From Blue Shark Skins Through the Conventional Extraction Process Intensification Using Natural Deep Eutectic Solvents

ERC-2016-CoG 725034The disposal of large amounts of skin waste resulting from the blue shark fishing industry presents several industrial and environmental waste management concerns. In addition, these marine subproducts are interesting sources of collagen, a fibrous protein that shows high social and economic interest in a broad range of biomedical, pharmaceutical, and cosmetic applications. However, blue shark wasted skins are a poorly explored matrix for this purpose, and conventional collagen recovery methodologies involve several pre-treatment steps, long extraction times and low temperatures. This work presents a new green and sustainable collagen extraction approach using a natural deep eutectic solvent composed of citric acid:xylitol:water at a 1:1:10 molar ratio, and the chemical characterization of the extracted collagen by discontinuous electrophoresis, thermogravimetric analysis, Fourier transformed infrared spectroscopy and circular dichroism. The extracted material was a pure type I collagen, and the novel approach presented an extraction yield 2.5 times higher than the conventional one, without pre-treatment of raw material and reducing the procedure time from 96 to 1 h. Furthermore, the in vitro cytotoxicity evaluation, performed with a mouse fibroblasts cell line, has proven the biocompatibility of the extracted material. Overall, the obtained results demonstrate a simple, quick, cheap and environmentally sustainable process to obtain marine collagen with promising properties for biomedical and cosmetic applications.publishersversionpublishe

Olive pomace phenolic compounds stability and safety evaluation: From raw material to future ophthalmic applications

Funding Information: This project has received funding from the European Union?s Horizon 2020 research and innovation program under the Marie Sk?odowska-Curie Initial Training Network (ITN) ?IT-DED3? (H2020-MSCA-ITN-2017) grant agreement No. 765608. The authors would like to acknowledge the financial support from Fundac?a?o para a Cie?ncia e Tecnologia (FCT) and Portugal 2020 to the Portu-guese Mass Spectrometry Network (Rede Nacional de Espectrometria de Massa RNEM; LISBOA 01 0145 FEDER 402 022125). Soraya Rodriguez Rojo acknowledges the University of Valladolid for her postdoctoral contract. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Nowadays, increasing interest in olive pomace (OP) valorization aims to improve olive’s industry sustainability. Interestingly, several studies propose a high-value application for OP extracts containing its main phenolic compounds, hydroxytyrosol and oleuropein, as therapy for ocular surface diseases. In this work, the stability and accessibility of OP total phenolic and flavonoid content, main representative compounds, and antioxidant activity were assessed under different pretreatment conditions. Among them, lyophilization and supercritical CO2 extraction were found to increase significantly most responses measured in the produced extracts. Two selected extracts (CONV and OPT3) were obtained by different techniques (conventional and pressurized liquid ex-traction); Their aqueous solutions were characterized by HPLC-DAD-MS/MS. Additionally, their safety and stability were evaluated according to EMA requirements towards their approval as ophthalmic products: their genotoxic effect on ocular surface cells and their 6-months storage stability at 4 different temperature/moisture conditions (CPMP/ICH/2736/99), together with pure hydroxy-tyrosol and oleuropein solutions. The concentration of hydroxytyrosol and oleuropein in pure or extract solutions was tracked, and possible degradation products were putatively identified by HPLC-DAD-MS/MS. Hydroxytyrosol and oleuropein had different stability as standard or extract solutions, with oleuropein also showing different degradation profile. All compounds/extracts were safe for ophthalmic use at the concentrations tested.publishersversionpublishe