Integral valorization of pineapple (Ananas comosus l.) by-products through a green chemistry approach towards added value ingredients

We would like to thank the scientific collaboration of Escola Superior de Biotecnologia of Universidade Católica Portuguesa through CBQF under FCT–Fundação para a Ciência e a Tecnologia, to accept Débora Campos through a Ph.D. grant SFRH/BD/104074/2014.Industrial by-products are produced every day through fruit processing industries. Pineapple is not an exception; when processed, around 60% (w/w) of its weight are peels, stem, trimmings, and crown, the only used fruit part for human consumption. Due to high concerns of sustainability in the food system and negative high impact of human practice in the environment, a strategy has to be developed. Therefore, a green chemistry approach was applied to pineapple by-products to make an integrated valorization by the extraction of bioactive molecules. Two pineapple by-products (peels and stems) were studied, applying a green chemistry approach, which means the non-use of organic solvents or extreme methodologies. A subdivision of each by-product was done by the application of a juice machine. The peels and stems in the fresh state were ground separately, creating two fractions for each by-product—a juice and a wet pulp (press cake). The press cake was characterized, dried, and ground to create a fine powder flour. To the juice, a precipitation methodology with polysaccharides was applied, which allowed the bromelain separation (developing of an enzymatic fraction) from the fruit juice. The enzymatic extract was freeze-dried, and the juice was spray-dried, developing two more fine powders. Thus, three new ingredients were produced from each by-product, creating a total of six new ingredients. Overall, the enzymatic fractions represented around 0.26% (w/w) of pineapple weight. Pineapple stem juice represented 4.8% (w/w), and peel juice represented 17.3% (w/w). Pineapple stem flour represented 3.1% (w/w), and peel flour represented 11.4% (w/w) of the total pineapple weight. To valorize the by-products juices, a full characterization was performed of bioactive molecules and biological activities. When comparing the two juices, the peel juice showed lower content of total phenolic compounds, lower antioxidant capacity, and lower content of vitamin C. The different phenolic compounds were identified by HPLC analysis in the two pineapple by-products juices. However, the same compounds in both juices were quantified (chlorogenic, caffeic, and ferulic acids). On the other hand, the by-products flours had a high content of insoluble dietary fiber (IDF), mainly cellulose and hemicellulose. Therefore, the approach applied in this work opens the door to the production of green products, as a result of by-products valorization. This could be applied not only in the food industry but also in the nutraceutical and cosmetic industries.This research was funded by BiValBi–Biotechnologies to Valorize the regional Biodiversity in Latin America, in the funding project FP/-PEOPLE-2013-IRSES PEOPLE Marie Curie Program through project reference PIRSES-GA-2013-611493 and by FCT through project Multibiorefinery–Multi-purpose strategies for the creation of an integrated biorefinery (POCI-01-0145-FEDER-0066) and the project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

Study of olive pomace antioxidant dietary fibre powder throughout gastrointestinal tract as multisource of phenolics, fatty acids and dietary fibre

Supplementary data to this article can be found online at https://doi. org/10.1016/j.foodres.2020.110032.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.Tˆania B. Ribeiro thanks the Fundação para a Ciência e Tecnologia (FCT), Portugal and Association BLC3 – Technology and Innovation Campus, Centre Bio R&D Unit for the PhD Grant SFRH/BDE/108271/2015. This work was supported by the project “Biological tools for adding and defending value in key agro-food chains (bio – n2 – value)”, n◦NORTE-01-0145-FEDER-000030, funded by Fundo Europeu de Desenvolvimento Regional (FEDER) and TransfereEmpreende, n◦NORTE-02-0651-FEDER-000081 under Programa Operacional Regional do Norte - Norte2020. We would also like to thank the scientific collaboration of the project UID/Multi/50016/2019 and local olive pomace producers from Tr ́as-os-Montes region of Portugalinfo:eu-repo/semantics/publishedVersio

Impact of extraction process in non-compliant ‘Bravo de Esmolfe’ apples towards the development of natural antioxidant extracts

The ‘Bravo de Esmolfe’ apple (BE) is rich in antioxidants and represents one of the most important traditional cultivars in Portugal. A large amount of non-compliant BE, with no market value, it is generated every year. Thus, an effort to find value-added solutions is of utmost importance. For the first time, green extraction (microwave-assisted-extraction (MAE) and ultrasound-assisted-extraction (UAE)) were compared against conventional extraction (with (CET) and without temperature (CE)) to select the most efficient process to produce a natural antioxidant extract from non-compliant BE. The results showed that MAE and CET are good methodologies for developing a BE antioxidant extract. However, MAE is more sustainable than CET, requiring less time to reach high temperatures. The MAE-extract showed great antioxidant activity (14.80 ± 0.70 mg TE/g DE by ORAC, 3.92 ± 0.25 mg AAE/g DE by ABTS, and 5.11 ± 0.13 mg TE/g DE by DPPH). This extract revealed high amount of chlorogenic acid (0.48 ± 0.07 mg/g DE), (-)-epicatechin (0.30 ± 0.02 mg/g DE) and phloridzin (0.13 ± 0.01 mg/g DE). This study shows that non-compliant BE is a useful source of antioxidants, being a sustainable way for the recovery of value-added compounds from the rejected fruit in line with sustainable and circular bioeconomy principles.info:eu-repo/semantics/publishedVersio

Total and sustainable valorisation of olive pomace using a fractionation approach

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.This research was funded by National Funds from FCT—Fundação para a Ciência e a Tecnologia, through the project MULTIBIOREFINERY-SAICTPAC/0040/2015 (POCI-01-0145-FEDER-016403) and the project “Biological tools for adding and defending value in key agro-food chains (bio-n2-value)”, nº NORTE-01-0145-FEDER-000030, funded by Fundo Europeu de Desenvolvimento Regional (FEDER), under Programa Operacional Regional do Norte-Norte2020. We would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

Serra da Estrela PDO cheese authentication: RAPD and SCAR approaches for identification of adulterant breed’s milk

Serra da Estrela cheese is a Protected Denomination of Origin (PDO) product and is perceived as a unique high-quality food, being the most famous Portuguese cheese and presenting high commercial value. It is legitimately manufactured from raw milk of the autochthonous sheep breed Serra da Estrela; however, the adulteration of production with cheaper and/or lowerquality milks from non-autochthones ovine breeds compromises the quality of the final product and undervalues the original PDO cheese. Considering that these fraudulent productions may lead to serious problems at both social and economic levels, it is urgent to develop low-cost, sensitive, fast and reliable analytical techniques that efficiently allow traceability of the breed origin of milk in Serra da Estrela PDO cheese. Here we describe a Randomly Amplified Polymorphic DNA (RAPD) method capable of efficient detection of adulterant breeds in milk mixtures, containing Serra da Estrela milk, used for fraudulent production of this cheese. Taking this into account, we suggest RAPD to be a valuable tool for identification of sheep breed in a first stage of milk authentication in dairy industry. Furthermore, considering the possible degradation of DNA during milk processing, Sequence Characterized Amplified Region (SCAR) markers were designed envisioning the detection of milk adulteration in processed dairy foods. RAPD-SCAR techniques have been here used, for the first time, to identify breed origin in milk samples, establishing its applicability for quality control on dairy industry, being capable of milk authentication in the final products. In this sense, our findings will play an important role on the valorization of not only the Serra da Estrela PDO cheese, but also on other high-quality dairy products prone to adulteration, contributing to the further development of the dairy industry.Project “Valor Queijo” (CENTRO-07-0202-FEDER-030372

Incorporation of olive pomace ingredients into yoghurts as a source of fibre and hydroxytyrosol: Antioxidant activity and stability throughout gastrointestinal digestion

Liquid-enriched powder (LOPP) and pulp-enriched powder (POPP) obtained from olive pomace were incorporated into yoghurt, not only, to increase its content in dietary fibre, hydroxytyrosol and unsaturated fatty acids, but also to understand the lipids-phenolics interaction by simultaneous incorporation of olive oil. POPP (2%) and LOPP (1%) addition to yoghurt allowed fulfilling the condition of being a source of fibre and provided 5 mg of hydroxytyrosol and derivatives in a standard yoghurt (120 g), respectively. Yoghurts unsaturated fatty acids profile was positively influenced by the addition of only POPP and olive oil + LOPP or + POPP. All OP powder-fortified yoghurts exhibited higher total phenolic content and antioxidant activity than the control (p<0.05). After in vitro digestion the bioaccessibility of total phenolics (more 25.58%) and hydroxytyrosol (more 68.71%) in LOPP-yoghurts was improved by the addition of olive oil. In conclusion, OP powders´ incorporation gave additional and essential healthy properties to yoghurt.Tânia I. B. Ribeiro thanks the FCT - Fundação para a Ciência e Tecnologia (FCT - Fundação, Portugal and Association BLC3 – Technology and Innovation Campus, Centre Bio R&D Unit for the PhD Grant SFRH/BDE/108271/2015. This work was supported by National Funds from Fundo Europeu de Desenvolvimento Regional (FEDER), under Programa Operacional Temático Competitivdade e Internacionalização – COMPETE 2020, through the project 3i Bioeconomy project POCI-01-0246-FEDER-026758, and FCT - Fundação para a Ciência e a Tecnologia through the project MULTIBIOREFINERY - SAICTPAC/0040/2015 (POCI-01-0145-FEDER-016403). We would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

In vitro gastrointestinal digestion impact on the bioaccessibility and antioxidant capacity of bioactive compounds from tomato flours obtained after conventional and ohmic heating extraction

In 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 ± 1.2459.06 ± 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.The authors would like to thank to “MultiBiorefinery: Estratégias multiuso para a valorização de uma gama alargada de subprodutos agroflorestais e das pescas: Um passo em frente na criação de uma biorrefinaria” financiado pelo Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-016403) e pelo Programa Operacional Regional de Lisboa(LISBOA-01-0145-FEDER-016403), na sua componente FEDER e pela Fundação para a Ciência e Tecnologia, I.P. na componente nacional (SAICTPAC/0040/2015). The authors would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019, UID-BIM-00009- 2020, and UID/BIO/04469/2020. The author Marta Coelho would like to acknowledge FCT for your grant with the reference (grant number SFRH/BD/111884/2015).info:eu-repo/semantics/publishedVersio

Evaluation of minimum inhibitory concentrations of plant extracts against environmental fungi and dermatophytes

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Antioxidant activity, phenolic profile, cytotoxicity and genotoxicity of plant extracts

The outbreak of COVID-19 disease caused by SARS-CoV-2 forced the scientific world to search for new alternatives to help control the virus. Plant extracts have natural compounds that might provide a starting point for the research on the use of plants as an excellent source of new antiviral agents against viruses, including COVID-19 to be included in disinfectants, fabrics or other materials. In this study, the polyphenols content (Folin-Ciocalteu), antioxidant capacity (DPPH, ABTS and ORAC) and the phenolic profile (HPLC) of different hydroethanolic (ethanol:H2O 50:50 v/v) extracts of medicinal plants cultivated under controlled conditions in Portugal (echinacea, rosemary, laurel, thyme and rock rose) were determined, as well as the cytotoxicity effect against a keratinocyte cell line using cell viability assay by PrestoBlue and genotoxicity effect using the AMES test. According to the results, total phenolic content ranged from 204.54 ± 1.78 / 274.20 ± 3.14 (mg EAG/g extract) with the rock rose extract presenting the highest content (p < 0.05). The extracts showed a good antioxidant capacity demonstrated by the high values found for ORAC, which ranged 2855.03 ± 9.75 / 5285.35 ± 60.04 µMol Trolox/mg extract. HPLC analysis revealed the presence of different compounds in the extracts such as the kaempferol-O-glucuronide, catechin, protocatechuic acid and galloyl glucoside, representing a potential source of bioactive components with antioxidant capacity. No toxicity was observed towards the keratinocyte cells and none of them showed mutagenic effects. Based on the results of safety and high polyphenols content of the extracts they demonstrate a great potential as antimicrobial agents. This will allow the design of new experiments aimed at evaluating the antiviral activity of these extracts, especially against SARS-CoV-2.info:eu-repo/semantics/publishedVersio

Antioxidant activity, phenolic profile, cytotoxicity and genotoxicity of plant extracts

info:eu-repo/semantics/publishedVersio