Cistus, Acacia, and Lemon verbena Valorization through Response Surface Methodology: Optimization Studies and Potential Application in the Pharmaceutical and Nutraceutical Industries

Cistus ladanifer L., Acacia dealbata L., and Aloysia citrodora Paláu were subject to an optimization procedure for two extraction techniques (heat-assisted extraction (HAE) and ultrasound-assisted extraction (UAE)). The extracts were then analyzed by HPLC-DAD-ESI/MS for their phenolic profile (cistus—15 compounds, acacia—21 compounds, and lemon verbena—9 compounds). The response surface methodology was applied, considering four varying factors: ethanol percentage; extraction time; temperature/power; and S/L ratio, generating two responses (the major phenolic compound, or family of compounds, and the extraction yield). For cistus, both techniques optimized the extraction yield of punicalagins, with UAE proving to be the most efficient extraction method (3.22% ethanol, 22 min, 171 W, and 35 g/L). For acacia, HAE maximized the extraction of procyanidin (74% ethanol, 86 min, 24 °C, and 50 g/L), and UAE maximized the content of myricetin (65% ethanol, 8 min, 50 W, and 50 g/L). For lemon verbena, HAE favored the extraction of martynoside (13% ethanol, 96 min, 49 °C and 17 g/L) and forsythiaside UAE (94% ethanol, 25 min, 399 W, and 29 g/L). The optimal conditions for the extraction of compounds with high added value and potential for use in pharmaceuticals and nutraceuticals were defined.This research was funded by national funds through FCT/MCTES (PIDDAC): CIMO, UIDB/00690/2020 (https://doi.org/10.54499/UIDB/00690/2020) and UIDP/00690/2020 (https: //doi.org/10.54499/UIDP/00690/2020) and SusTEC, LA/P/0007/2020 (https://doi.org/10.544 99/LA/P/0007/2020), through the individual research grant (SFRH/BD/145467/2019) of Filipa Fernandes, through the individual scientific employment program-contract with Márcio Carocho (10.54499/CEECIND/00831/2018/CP1578/CT0001), and the institutional scientific employment program-contract with Sandrina A. Heleno and Lillian Barros, and the MICINN through the contract to researcher Miguel A. Prieto.info:eu-repo/semantics/publishedVersio

Differences in the phenolic composition and nutraceutical properties of freeze dried and oven-dried wild and domesticated samples of Sanguisorba minor Scop

Recent studies have dealt about the phenolic compounds and the nutraceutical properties of Sanguisorba minor Scop and about the possibility of their domestication to standardise the plant production. However, it is also known that the storage conditions can affect the bioactive compounds present in plants. Thus, wild (W) and domesticated (F1, F2, F3) S. minor samples were exposed to different drying methods (oven-drying at 60 °C until constant weight named OD or freeze-drying until constant weight named FD) and studied for their content in phenolic compounds, antioxidant, antimicrobial, cytotoxic and anti-inflammatory properties. In general, OD samples showed the highest nutraceutical properties and the highest content in phenolic compounds. The most abundant phenolic compounds in both drying methods were kaempferol-3-O-glucoside and caffeoyl ester, with some differences between wild and domesticated samples. Wild samples showed higher antimicrobial and cytotoxic activity than domesticated ones, except for the OD F3 and FD F2 samples that reported cytotoxicity against HeLa cells. This study provides important information to choose the most adequate methodology to retain phenolic compounds and nutraceutical properties of S. minor species. Further researches are necessary to standardise the domestication of the studied wild species and verify the highest efficiency of the OD method.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2020). L. Barros and C. Calhelha also thank the national funding by FCT – Foundation for Science and Technology, P.I., through the institutional scientific employment program-contract for their contracts, and Sandrina A. Heleno thanks to the national funding by FCT, P.I., through the individual scientific employment program-contract. The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_P and to European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project Norte-01- 0145-FEDER-000042: GreenHealth.info:eu-repo/semantics/publishedVersio

Novel incorporation of red-stage haematococcus pluvialis wet paste as a colourant and enhancer of the organoleptic and functional properties of filloas †

Haematococcus pluvialis Flotow is a microalga used as a nutraceutical, due to its high content in bioactive compounds, mainly carotenoids, in which astaxanthin stands out [1]. Furthermore, H. plu- vialis has shown a high antioxidant potential, and combined with its intense red colour, this microalga could have dual functionality: as a colourant and a bioactive ingredient [ 2]. The process to obtain this ingredient involves several transformation steps—namely, lyophilisation and saponification—raising the costs to develop and obtain free astaxanthin, which paradoxically presents greater instability and solubility than its esterified counterpart [ 3]. Thus, this study provides an alternative approach for the application of red, astaxanthin-rich, H. pluvialis wet paste as a partial substitute for wheat flour (7% and 13% w/w) in the preparation of filloas (Galician pancakes), a typical dessert from the northwestern region of the Iberian Peninsula. To evaluate its power as a natural pigment, the stability of colour over time (3, 6, and 9 days) was measured, and the results were compared with those of a commercial colourant. At the same time, its physicochemical properties such as the microbiological profile were measured to determine its functionality as a food preservative. As a result, redness stability (a*) of 8% higher than that of the commercial colourant was obtained for the maximum concentration of H. pluvialis analysed. The texture showed a significant response (p < 0.02), improving its properties as the concentration of the microalga increased, which showed a tenacity of 3.23 N and extensibility of 15.10 mm during the first 6 days, i.e., a 52% and 19% improvement, respectively, in relation to the control group. In turn, an enrichment of carotenoids, fatty acids, and phenolic com- pounds, in combination with a potential moderator of microbiological degradation by this unicellular organism, gives added value to this food matrix.This research was funded by project ED431 2020/06 (Galician Competitive Research Groups Xunta de Galicia). This study was supported by project EQC2018-005011-P (Ministry of Science, Innovation and Universities, Spain). All these programmes are co-funded by FEDER (EU). The authors are also grateful to Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); to FCT for S. Heleno (CEECIND/03040/2017) and L. Barros contracts through the individual and institutional scientific employment programme contract, respectively. This article is based upon work from the Sample Preparation Study Group and Network, supported by the Division of Analytical Chemistry of the European Chemical Societyinfo:eu-repo/semantics/publishedVersio

Magnesium and manganese induced changes on chemical, nutritional, antioxidant and antimicrobial properties of the pansy and Viola edible flowers

composition was determined using standard methods. Free sugars, fatty acids, organic acids, tocopherols, and phenolic compounds were analyzed using various HPLC and GC devises. The extract’s antimicrobial, antioxidant, cytotoxicity, and anti-inflammatory activity were assessed. The results indicated that Mg enrichment negatively affected plant growth and mineral accumulation but improved photosynthetic performance. The edible flowers contained significant amounts of protein, low levels of fat, and varying sugar contents, such as glucose and fructose. Various fatty acids and phenolic compounds were identified, with different concentrations depending on the treatment. The flowers exhibited antioxidant potential, antimicrobial activity, cytotoxic effects, and antiinflammatory properties. The correlations between the investigated parameters not only expand knowledge on Mg and Mn interaction but also catalyze significant advancements in sustainable agriculture and food health, fostering a healthier and more conscious future.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES (PIDDAC) to CIMO (UIDB/00690/2020 and UIDP/00690/ 2020) and SusTEC (LA/P/0007/2020). L. Barros, Ricardo C. Calhelha, S. A. Heleno and T. C. Finimundy thank the national funding by FCT through the institutional scientific employment program-contract for her contract, while M. Carocho thanks FCT through the individual scientific employment program-contract (CEECIND/00831/2018). I. Oliveira thanks FCT for her PhD grant (BD/06017/2020).info:eu-repo/semantics/publishedVersio

Exploring the bioactive compounds from berry biowaste: Eugenia involucrata DC. as a study case

Aiming at valorising the Eugenia involucrata residue, the present work aimed in investigating the chemical profile and in vitro bioactivities of the residue (EIR) and its individual parts, i.e., seeds (EIS) and peels (EIP). Chromatographic analyses showed that EIP had the highest content of organic acids, tocopherols, and anthocyanins, whereas EIS was the richest in hydrolysable tannins. bis-HHDP-glucoside isomers and cyanidin-3-O-galactoside were the main detected phenolic compounds. By the cellular antioxidant activity (CAA) assay, the samples showed similar. antioxidant potential with 71%, 77%, and 80% inhibition for EIP, EIR, and EIS, respectively, while EIS demonstrated the best antioxidant activity by thiobarbituric acid reactive substances (TBARS, IC50 = 4.96 mu g/mL) and oxidative inhibition assay (OxHLIA, IC50 = 40 mu g/mL). The samples were able to inhibit the growth of seven bacteria and two fungi. Between the microorganisms tested, all samples showed a strong antibacterial activity (MIC = 0.156 mg/mL) against Yersinia enterocolitica and Pseudomonas aeruginosa (MIC = 0.156-0.625 mg/mL). Moreover, they also display a high antifungal activity against Arpergillus brasiliensis (MIC = 0.625 mg/mL). Furthermore, only EIR and EIS had anti-inflammatory effect via NO production inhibition (IC50 = 168-259 mu g/mL, respectively) and anti-proliferative activity towards four tumour and one non-tumour cell lines. These findings suggest that the biowaste generated during the processing of E. involucrata fruit has potential as a source of bioactive compounds.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES (PIDDAC) to CIMO (UIDB/00690/2020 and UIDP/00690/2020) and SusTEC (LA/P/0007/2020) . To the "La Caixa" Foundation and to FCT for the financial support through the mobilizing Project "Aquae Vitae - Thermal water as a source of life and health", "PROMOVE - O futuro do Interior" call 2020. To FCT for the contracts of J. Pinela (CEECIND/01011/2018) , R.C. Calhelha and L. Barros (CEEC Institutional) , and the PhD grants of B.R. Albuquerque (SFRH/BD/136370/2018 and COVID/BD/152908/2022) . To the MICINN for the Juan de la Cierva Formacion contract for T.C.S.P. Pires (FJC20120- 045405-I) . R.C.G. Correa is a research grant recipient of Cesumar Institute of Science, Technology, and Innovation (ICETI) .info:eu-repo/semantics/publishedVersio

The generally recognized as safe (GRAS) microalgae Haematococcus pluvialis (wet) as a multifunctional additive for coloring and improving the organoleptic and functional properties of foods

This work proposes the application of astaxanthin-rich H. pluvialis wet paste (HPW) as a partial substitute for wheat flour in the preparation of filloas, a dish that combines the basic ingredients of industrial bakery. The nutritional and color profile of HPW-enriched samples was evaluated by comparative analysis with a mixture of synthetic food dyes. The highest content of carotenoids (798 ± 12 µg g−1) and fatty acids (76 ± 2mgg−1) was obtained for a filloa fortified with H. pluvialis in contrast to a non-significant dye response. Subsequently, the color stability of the fortified filloa was evaluated over time (3, 6 and 9 days), as well as its physicochemical properties and microbiological profile. As a result, HPW provided filloas with a longer shelf life, brightness (*L), and texture, in comparison with a mixture of synthetic dyes. In addition, an inhibitory effect of HPW towards mesophilic aerobic microorganisms in the food was obtained.This research was funded by project ED431 2020/06 (Galician Competitive Research Groups Xunta de Galicia). All these pro- grammes are co-funded by FEDER (EU). The authors are also grateful to the Foundation for Science and Technology (FCT; Portugal) for financial support through national funds FCT/ MCTES (PIDDAC) to CIMO (UIDB/00690/2020 and UIDP/ 00690/2020) and SusTEC (LA/P/0007/2021). Castillo A. acknowl- edges the support of the European Grouping of Territorial Cooperation Galicia-Norte Portugal (GNP, EGTC) under the IACOBUS Program and the MCI of Spain for his contract part of the grant DIN2021-011976 funded by the MCIN/AEI/ 10.13039/501100011033. L. Barros and S. Heleno acknowledge the national funding by FCT, P. I., through the institutional scientific employment program-contract for their contracts, and Filipa A. Fernandes for the PhD grant (SFRH/BD/145467/ 2019).info:eu-repo/semantics/publishedVersio

Chemical and Bioactive Evaluation of Essential Oils from Edible and Aromatic Mediterranean Lamiaceae Plants

The Lamiaceae family, which includes several well-known aromatic plants, is scientifically relevant due to its essential oils (EOs). In this work, four EOs from Mediterranean species, namely Origanum vulgare L., Rosmarinus officinalis L., Salvia officinalis L., and Thymus vulgaris L., were evaluated for their volatile profiles and the biological activity in vitro to assess their potential use in the food and cosmetic sector. GC/MS analysis revealed dominant compounds, such as carvacrol, thymol, and eucalyptol. Regarding biological action, the samples exhibited antioxidant, cytotoxic, anti-inflammatory, antimicrobial, and antifungal activities, with O. vulgare and T. officinalis standing out. T. vulgaris showed the lowest EC50 in the reducing power assay, and O. vulgare had the lowest EC50 in the DPPH assay. Most EOs also displayed excellent anti-inflammatory responses and antifungal properties, with O. vulgare and T. vulgaris also demonstrating antibacterial activity. All EOs from Mediterranean species showed cytotoxicity against tumoral cell lines. Overall, the selected EOs stood out for their interesting bioactivities, with the obtained results underscoring their potential as natural preservatives and bioactive agents in various industrial applications, including food, pharmaceuticals, and cosmetics.This work was supported by national funding by FCT/MCTES (PIDDAC): CIMO, UIDB/00690/2020 (DOI: 10.54499/UIDB/00690/2020) and UIDP/00690/2020 (DOI: 10.54499/UIDP/00690/2020); national funding by SusTEC, LA/P/0007/2020 (DOI: 10.54499/LA/P/0007/2020); national funding by FCT, P.I., through the institutional and individual scientific employment program-contracts for L. Barros, R. Calhelha, and E. Pereira (2021.03908.CEECIND); and research contracts ofC. Caleja, T. Finimundy, and T.C.S.P. Pires (VIIAFOOD project no. C644929456-00000040) and RafaelMascoloti Sprea (2020.08092.BD)info:eu-repo/semantics/publishedVersio

Multifunctions of Pleurotus sajor-caju (Fr.) Singer: a highly nutritious food and a source for bioactive compounds

A study with Pleurotus sajor-caju was conducted to: evaluate the nutritional and chemical composition of the fruiting bodies; optimize the preparation of bioactive phenolic extracts; and characterize the optimized extract in terms of bioactive compounds and properties. P. sajor-caju revealed an equilibrated nutritional composition with the presence of hydrophilic (sugars and organic acids) and lipophilic (tocopherols and PUFA) compounds. p-Hydroxybenzoic, p-coumaric and cinnamic acids were identified in the extract obtained with ethanol (30 g/l ratio) at 55 °C for 85 min. This extract showed antioxidant properties (mainly reducing power and lipid peroxidation inhibition), antibacterial activity against MRSA and MSSA and cytotoxicity against NCI-H460, MCF-7 and HeLa. Furthermore, as the extract showed capacity to inhibit NO production in Raw 264.7 macrophages, molecular docking studies were performed to provide insights into the anti-inflammatory mechanism of action, through COX-2 inhibition by the phenolic acids identified.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Program PT2020 for financial support to CIMO (UID/AGR/00690/2013) and for L. Barros and R. Calhelha contracts. T.C. Finimundy thanks CAPES Foundation, Ministry of Education of Brazil (CAPES fellow, process number 88881.134581/2016-01). To Xunta de Galicia for financial support for the post-doctoral researcher of M.A. Prieto.info:eu-repo/semantics/publishedVersio

Phytochemical composition and bioactive potential of Melissa officinalis L., Salvia officinalis L. and Mentha spicata L. extracts

Plants are rich in bioactive phytochemicals that often display medicinal properties. These can play an important role in the production of health-promoting food additives and the replacement of artificial ones. In this sense, this study aimed to characterise the polyphenolic profile and bioactive properties of the decoctions, infusions and hydroethanolic extracts of three plants: lemon balm (Melissa officinalis L.), sage (Salvia officinalis L.) and spearmint (Mentha spicata L.). Total phenolic content ranged from 38.79 mg/g extract to 84.51 mg/g extract, depending on the extract. The main phenolic compound detected in all cases was rosmarinic acid. The results highlighted that some of these extracts may have the ability to prevent food spoilage (due to antibacterial and antifungal effects) and promote health benefits (due to anti-inflammatory and antioxidant capacities) while not displaying toxicity against healthy cells. Furthermore, although no anti-inflammatory capacity was observed from sage extracts, these stood out for often displaying the best outcomes in terms of other bioactivities. Overall, the results of our research provide insight into the potential of plant extracts as a source of active phytochemicals and as natural food additives. They also support the current trends in the food industry of replacing synthetic additives and developing foods with added beneficial health effects beyond basic nutrition.info:eu-repo/semantics/publishedVersio

Recovery of high valuable bioactive molecules from Vaccinium myrtillus L. bioresidues

Purpose The interest in using bioresidues produced by the agri-food industry is increasingly intrinsic to the world economy. As such, researchers started to look for new ways to enhance its use, developing innovations to transform these residues into high-value raw materials with industrial application, promoting the circular economy concept. Methods Therefore, pruning residues from the production of Vaccinium myrtillus L. could be an interesting exploitation feld, given its position as a superfood due to its well-known antioxidant properties. The antioxidant, antibacterial, anti infammatory, anti-tyrosinase, and cytotoxicity potentials of aqueous and hydroethanolic extracts of V. myrtillus aerial parts were evaluated. Results From the obtained results, ultrasound-assisted and maceration extracts were found to be as efective as, if not more efective than, conventional antibiotics against methicillin-resistant Staphylococcus aureus. Aside from antibacterial activ ity, the extracts also showed antioxidant and anti-tyrosinase efects, which were found to be favourably related to the level of cafeoylquinic acid derivatives. Conclusion The obtained results highlight the bioactive potential and the importance of exploiting this bio residue as a novel candidate for industrial application, taking advantage of their biological properties. Graphical AbstractFoundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); the contract of L. Barros through the institutional scientific employment program-contract, the contracts of M. Carocho and S.A. Heleno (CEEC-IND/00831/2018 and CEECIND/ 03040/2017) through the individual scientific employment program- contract, and the PhD studentship granted to M. Añibarro-Ortega (2020.06297.BD). To the “La Caixa” Foundation and to FCT for the financial support through Project “Aquae Vitae — Thermal water as a source of life and health”, Promove Mobilizer programme. This work is also supported by MICINN for the Juan de la Cierva Formación contract for T. Oludemi (FJC2019-042549-I).info:eu-repo/semantics/publishedVersio