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Nephelium lappaceum L., popularmente conhecido como rambutan, é um fruto tropical nativo da Ásia que, atualmente, se encontra amplamente distribuído em outras regiões tropicais, como a América Latina, a Austrália e alguns países africanos [1,2], sendo muito apreciado pela sua aparência exótica e sabor agradável, o que tem aumentado a sua comercialização e processamento em todo o mundo [1- 3]. No entanto, apenas uma pequena porção deste fruto é comestível/processado, sendo que até 48% do seu peso corresponde ao seu epicarpo não comestível, o que pode gerar um alto volume de bioresíduos e perdas económicas [2,3]. No sentido de propor uma valorização para o epicarpo de rambutan como fonte de moléculas bioativas, o presente estudo teve como objetivo elucidar a sua composição química em termos de tocoferóis (HPLC-FD), ácidos orgânicos (UFLC-PDA) e compostos fenólicos antociânicos e não antociânicos (HPLC-DAD/ESI-MSn) e determinar as bioatividades, nomeadamente atividade antioxidante (AA) e atividade antimicrobiana (AM), do seu extrato hidroetanólico por ensaios in vitro. A AA foi avaliada através do ensaios de substâncias reativas ao ácido tiobarbitúrico (TBARS) e de inibição da hemólise oxidativa (OxHLIA); a AM foi avaliada pelo método de microdiluição usando seis bactérias (Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium e Enterobacter cloacae) e seis fungos (Aspergillus fumigatus, Aspergillus versicolor, Aspergillus niger, Penicillium funiculosum, Penicillium verrucosum var. cyclopium e Trichoderma víride). O epicarpo de rambutan apresentou três isómeros de tocoferol, sendo o γ-tocoferol o mais representativo; cinco ácidos orgânicos, com destaque para a concentração de ácido chiquímico; cinco compostos fenólicos derivados de elagitaninos e duas antocianinas O-glicosiladas derivadas de delfinidina. Em relação às bioatividades, o seu extrato hidroetanólico foi capaz de inibir a oxidação lipídica e proteger os eritrócitos da hemólise oxidativa em baixas concentrações (EC50 de 2,79 ± 0,03 e 72 ± 2 μg/mL). Além disso, as atividades antibacteriana e antifúngica do extrato foram alcançadas em concentrações semelhantes ou inferiores aos controlos utilizados (benzoato de sódio (E211) e metabissulfito de potássio (E224)) e as amostras também apresentaram atividade bactericida e fungicida em todos os microrganismos avaliados. Em conclusão, o epicarpo de N. lappaceum possui uma composição química rica em compostos bioativos, principalmente derivados de elagitaninos, além de apresentar boa atividade antioxidante e potencial para inibir o crescimento de microrganismos. Os resultados encontrados revelam que este subproduto pode ser uma interessante fonte de ingredientes bioativos que podem ser aplicados em diversos setores da indústria alimentar e farmacêutica. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e aos fundos nacionais FCT/MCTES (PIDDAC) pelo apoio financeiro ao CIMO (UIDB/00690/2020 e UIDP/00690/2020) e SusTEC (LA/P/0007/2020), pela bolsa de doutoramento de B.R.A (SFRH/BD/136370/2018), e pelos contratos de M.I.D., C.P. e L.B. no âmbito da celebração do contrato-programa de emprego científico info:eu-repo/semantics/publishedVersion

Eugenia brasilensis Lam (EB) and Eugenia invoucrata DC (EI are Brazilian fruits that are still little known; however, some local food insustries use these fruits in the confection of frozen pulps and ice creams, processing that can generate bioresidues with no commercial value, but that can have interesting nurtrition value and couls be a source of biocative compounds. info:eu-repo/semantics/publishedVersion

As frutas tropicais são amplamente apreciadas pelo seu sabor adocicado e geralmente conhecidas pela sua composição rica em compostos bioativos info:eu-repo/semantics/publishedVersion

Garcinia mangostana L., known as mangosteen, is a tropical fruit belonging to the Clusiaceae family, native from South Asia but can also be found in other tropical territories, such as South America [1,2,3]. The fruit comprises an inedible dark purple epicarp (> 60%) that encases an edible succulent pulp [2]. Nowadays, mangosteen pulp and pericarp have been used in beverages as food supplement by virtue of the traditional knowledge about its health benefits. However, correlation studies between the folk medicine usage and its chemical composition are scarce in the literature [2,3]. Aiming to elucidate part of the chemical composition, the present study carried out the determination of the main flavonoids, including anthocyanin compounds, present in mangosteen pericarp by High Performance Liquid Chromatography coupled to a diode array detector and mass spectrometry by electrospray ionization (HPLC-DAD-ESI/MSn). Furthermore, the cytotoxicity effects of its hydroethanolic extracts were evaluated on four human tumor cell lines (NCI-H460 - lung carcinoma, MCF-7 - breast carcinoma, HepG2 - hepatocellular carcinoma, and HeLa - cervical carcinoma) by the Sulforhodamine B (SRB) assay. Mangosteen pericarp presented nine non-anthocyanin flavonoid compounds, most of which belonging to the procyanidin class (seven compounds), one taxifolin derivative (taxifolin-O-rhamnoside, found in low concentrations), and one quercetin derivative (quercetin-3-O-rutinose, found in trace amounts). Regarding the anthocyanin flavonoids compounds group, two were found and tentatively identified as cyanidin-O- dihexoside and delphinidin-O-dihexoside. Regarding the total amount of flavonoids, the extracts presented 53 ± 1 mg of non-anthocyanin flavonoids/g of extract, 3.66 ± 0.02 mg of anthocyanins/g of extract. Concerning the cytotoxic activity, the hydroethanolic extracts presented activity against all tumor cell lines studied (GI50 < 75 μg/mL). The results obtained from the present study showed that mangosteen pericarp could be an interesting natural source of high added value and bioactive compounds, with the potential to the applied in several industrial fields including pharmaceutical, nutraceutical, among others. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). National funding by FCT, P.I., through the individual scientific employment program- contract for M.I. Dias, R.C. Calhelha, and L. Barros contracts and B.R. Albuquerque research grant (SFRH/BD/136370/2018). This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER- 024479: ValorNatural®.The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E and the project TRANSCoLAB 0612_TRANS_CO_LAB_2_P. info:eu-repo/semantics/publishedVersion

Tropical fruits are widely appreciated for their sweet taste and generally known for their rich composition in bioactive compounds. However, the processing of this type of fruits can generate a large amount of non-edible and non-tradable by-products, consisting mainly in peels and seeds. These by- products have already been described as having a greater amount of bioactive compounds than the edible parts; this fact has been increasing the interest of the scientific community as also of several industrial fields for the application of these by-products. 1 In this context, the rigid non-edible epicarp of Sicana odorifera (vell.) Naudin, a purple-black tropical fruit from Brazil, 2 was evaluated for the anthocyanin and non-anthocyanin phenolic composition by High-Performance Liquid Chromatography coupled to a diode-array detector and a mass spectrometer functioning by electrospray ionization (HPLC-DAD/ESI-MS). The bioactive potential was assessed thought several in vitro assays: the antioxidant activity was determined by thiobarbituric acid reactive substances assay (TBARS) and by the oxidative hemolysis inhibition assay (OxHLIA) and the antimicrobial activity was tested in four bacteria and four fungi strains using the microdilution method. S. odorifera epicarp hydroethanolic extract presented four phenolic compounds, namely two O- glycosylated quercetin and kaempherol derivatives and two O-glycosylated anthocyanins, being the latest group the most abundant one with a total amount of 24±1 mg of anthocyanins/g of dry epicarp. Concerning its bioactive potential, S. odorifera hydroethanolic extract revealed considerable antioxidant activity (EC50 values of 48.2±0.5 and 27±1 μg/mL for TBARS and OxHLIA assays, respectively) and antimicrobial action against all bacteria and fungi strains evaluated (minimal inhibitory concentrations (MICs) ≤ 2.2 mg/mL). The results obtained allow to classify this tropical fruit epicarp as a potential source of bioactive compounds with great market value, since they can be applied in several industrial sectors, including the food and pharmaceutical industries. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020) and B.R. Albuquerque research grant (SFRH/BD/136370/2018). National funding by FCT, P.I., through the institutional scientific employment program-contract for M.I.D., C.P., and L.B. contracts. The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_PT and to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth: Norte-01-0145-FEDER-000042 and the Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. This work has also been supported by the Ministry of Education, Science and Technological Development of Republic of Serbia (451-03-68/2020-14/200007). info:eu-repo/semantics/publishedVersion

Nephelium lappaceum L., popularly known as rambutan, is a tropical fruit belonging to the Sapidaceae family. It is native to Asia, but widely distributed in other tropical regions, such as Latin America, Australia, and some African countries [1,2]. This fruit is very appreciated for its exotic appearance and pleasant taste, and its commercialization and processing has been growing all over the world [1-3]. However, only a small portion of this fruit is edible/processed, with up to 67% corresponding to its inedible epicarp, which can generate a high volume of bioresidues and economic losses [2,3]. In order to propose a valorisation of rambutan epicarp as a source of bioactive molecules, the present study aimed to identify its anthocyanin and non-anthocyanin phenolic compounds by High-Performance Liquid Chromatography coupled to a diode-array detector and a mass spectrometer functioning by electrospray ionization (HPLC-DAD/ESI-MS), and determine the antioxidant activity of its hydroethanolic extract by two in vitro assays: thiobarbituric acid reactive substances assay (TBARS) and oxidative haemolysis inhibition assay (OxHLIA). The rambutan epicarp extract presented seven phenolic compounds, among which two anthocyanin compounds (O-glycosylated delphinidin derivatives) and five non-anthocyanin compounds (ellagitannin derivatives), in a total concentration of 11.57±0.08 and 31.6±0.5 mg/g of extract, respectively. Delphinidin isomers, geraniin isomers, and ellagic acid were the compounds detected in higher concentrations. In terms of antioxidant activity, rambutan extract was able to inhibit the lipid peroxidation in a low concentration (EC50 value of 2.79 ± 0.03 μg/mL), and a moderate amount of extract was required to exert oxidative haemolysis inhibition (EC50 value of 72 ± 2 μg/mL). The results obtained allow to conclude that rambutan epicarp could be an interesting matrix to be used as source of bioactive compounds for further application in food/pharmaceutical fields. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020) and B.R. Albuquerque research grant (SFRH/BD/136370/2018). National funding by FCT, P.I., through the institutional scientific employment program-contract for M.I.D., C.P., and L.B. contracts. To FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_PT and to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth: Norte-01-0145-FEDER-000042 and the Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

Tropical fruits are widely appreciated for their sweet taste and generally known for their rich composition in bioactive compounds. However, the processing of this type of fruits can generate a large amount of non-edible and non-tradable by-products, consisting mainly in peels and seeds. These by-products have already been described as having a greater amount of bioactive compounds than the edible parts; this fact has been increasing the interest of the scientific community as also of several industrial fields for the application of these by-products. 1 In this context, the rigid non-edible epicarp of Sicana odorifera (vell.) Naudin, a purple-black tropical fruit from Brazil, 2 was evaluated for the anthocyanin and non-anthocyanin phenolic composition by High-Performance Liquid Chromatography coupled to a diode-array detector and a mass spectrometer functioning by electrospray ionization (HPLC-DAD/ESI-MS). The bioactive potential was assessed thought several in vitro assays: the antioxidant activity was determined by thiobarbituric acid reactive substances assay (TBARS) and by the oxidative hemolysis inhibition assay (OxHLIA) and the antimicrobial activity was tested in four bacteria and four fungi strains using the microdilution method. S. odorifera epicarp hydroethanolic extract presented four phenolic compounds, namely two O-glycosylated quercetin and kaempherol derivatives and two O-glycosylated anthocyanins, being the latest group the most abundant one with a total amount of 24±1 mg of anthocyanins/g of dry epicarp. Concerning its bioactive potential, S. odorifera hydroethanolic extract revealed considerable antioxidant activity (EC50 values of 48.2±0.5 and 27±1 μg/mL for TBARS and OxHLIA assays, respectively) and antimicrobial action against all bacteria and fungi strains evaluated (minimal inhibitory concentrations (MICs) ≤ 2.2 mg/mL). The results obtained allow to classify this tropical fruit epicarp as a potential source of bioactive compounds with great market value, since they can be applied in several industrial sectors, including the food and pharmaceutical industries. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020) and B.R. Albuquerque research grant (SFRH/BD/136370/2018). National funding by FCT, P.I., through the institutional scientific employment program-contract for M.I.D., C.P., and L.B. contracts. The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_PT and to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth: Norte-01-0145-FEDER-000042 and the Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. This work has also been supported by the Ministry of Education, Science and Technological Development of Republic of Serbia (451-03-68/2020-14/200007). info:eu-repo/semantics/publishedVersion

Sicana odorifera (Vell.) Naudin, belonging to the Cucurbitaceae family, is a fruit probably native to Brazil, also found in other Latin America countries [1,2]. This fruit presents a rigid purple- black inedible epicarp, considered as a bioresidue after the fruit processing, but which could be an interesting source of bioactive compounds, mainly anthocyanins [1,2]. In order to provide a valorization of this bioresidue, the present work sought to determine part of the chemical composition, namely tocopherols, organic acids and phenolic compounds, including anthocyanins, present in the epicarp of S. odorífera. Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020) and B.R. Albuquerque research grant (SFRH/BD/136370/2018). National funding by FCT, P.I., throug the institutional scientific employment program-contract for M.I.D., and L.B. contracts. FEDER-Interreg España-Portug programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_PT and to the Europea Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Projec GreenHealth: Norte-01-0145-FEDER-000042 and the Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

Garcinia mangostana L., commonly known as mangosteen or queen of fruit, is a tropical fruit belonging to the Clusiaceae family. This shrub is native from South Asia, but can also be found in other tropical territories, such as South America [1,2,3]. The fruit comprises an inedible dark purple or red pericarp (> 60%) that encases the edible succulent pulp [2]. Nowadays, mangosteen pulp and pericarp have been used in beverages as a food supplement by virtue of the traditional knowledge about its benefits. However, a poor correlation between the medicinal use of mangosteen and its chemical composition has been described [2,3]. Aiming to contribute to the elucidation of the phytochemical composition of this fruit pericarp, the present study was carried out to characterize anthocyanins by HPLC-DAD-ESI/MS. In addition, the bioactivities of the hydroethanolic extract obtained by solid-liquid extraction were evaluated. The extract cytotoxicity to four human tumour cell lines (NCI-H460 - lung carcinoma, MCF-7 - breast carcinoma, HepG2 - hepatocellular carcinoma, and HeLa - cervical carcinoma) was tested by the sulforhodamine B assay. The anti-inflammatory activity was evaluated by the ability to inhibit the lipopolysaccharide-induced nitric oxide (NO) production by murine macrophage RAW 264.7 cells. Lastly, the antioxidant activity was measured by the extract capacity to inhibit the oxidative haemolysis (OxHLIA) and the formation of thiobarbituric acid reactive species (TBARS), using sheep erythrocytes and porcine brain cells as oxidizable substrates. The chromatographic analysis allowed the identification of two anthocyanins in the mangosteen pericarp, namely cyanidin-O-dihexoside, which was the most abundant, and delphinidin-O-pentoside. Regarding the in vitro evaluated bioactive properties, the extract showed cytotoxic activity against the four human tumour cell lines, with GI50 values lower than 75 μg/mL. It also exhibited antioxidant and anti-inflammatory activities at low concentrations, with IC50 values of 8.7 and 17.3 μg/mL for the TBARS and OxHLIA assays, and an EC50 value of 85 μg/mL for the NO production inhibition assay, respectively. The results obtained in this study allow to conclude that G. mangostana pericarp contains an interesting amount of anthocyanins and also exhibits strong bioactivities, which makes it an interesting source of bioactive ingredients. To the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020). M.I. Dias, J. Pinela, R.C. Calhelha and L. Barros thank their research contracts (national funding by FCT, P.I., through the institutional scientific employment program-contract) and B.R. Albuquerque thanks her PhD grant (SFRH/BD/136370/2018). This work was funded by the FEDER through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. To FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

Jabuticaba (Myrciaria jaboticaba (Vall.) O. Berg) is a Brazilian berry very appreciated for in natura consumption. However, its epicarp is not normally consumed, and in manufacture of products from jabuticaba fruit, it is responsible for the generation of large amounts of residues [1,2]. The exploration of by-products is becoming important for the obtainment of valuable bioactive compounds for food and pharmaceutical industries [2]. In this context, the present work aimed in the characterization of the main bioactive compounds present in jabuticaba epicarps and its potential antioxidant. Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020); national funding by FCT, P.I., through the institutional scientific employment program-contract for M.I. Dias, L. Barros, and C. Pereira contract; and B. Albuquerque (SFRH/BD/136370/2018) PhD grant; ERDF through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®; FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

As incertezas a respeito do consumo seguro de alguns aditivos artificiais e a procura por alimentos mais saudáveis têm promovido a utilização de corantes naturais na indústria alimentar [1]. Nos últimos anos, a obtenção deste tipo de aditivos a partir de fontes naturais tornou-se numa área de investigação ativa e em crescimento [2]. Neste contexto, o epicarpo de jabuticaba, que normalmente não é consumido e muitas vezes descartado durante o processamento industrial, apresenta uma interessante composição em antocianinas, compostos com grande potencial para produção de agentes corantes com propriedades bioativas [3]. Por esta razão, a presente investigação teve como objetivos: i) otimizar a extração de antocianinas por maceração (ME) e ultrassons (UE) utilizando a metodologia de superfície de resposta; e ii) validar a capacidade corante do extrato obtido nas condições previamente otimizadas através da sua incorporação numa matriz alimentar, nomeadamente macarrones. Para otimização por RSM, foi um desenho composto central de 5 níveis combinando as variáveis tempo, percentagem de etanol e temperatura ou potência. Os teores de delfinidina-3-O-glucósido e cianidina-3-O-glucósido, determinados por HPLC-DAD-ESI/MS, foram usados como variáveis de resposta. O software Design-Expert foi usado para construir e validar os modelos e obter as condições que maximizam a extração. O extrato obtido nas melhores condições foi posteriormente incorporado em macarrones a fim de se avaliar o potencial corante. Para isso, o produto formulado foi analisado quanto aos parâmetros de cor e valor nutricional ao longo do tempo de prateleira e os resultados foram comparados com os obtidos com um corante comercial à base de antocianinas. Dentre as técnicas testadas, a ME foi a mais eficaz em extrair ambas as antocianinas identificadas no epicarpo de jabuticaba; este método permitiu extrair mais do dobro do conteúdo de antocianinas obtido com UE. Para maiores rendimentos, a extração ME necessitou de uma percentagem de etanol mais baixa e uma temperatura mais elevada, enquanto o processo UE foi caracterizado por um tempo de extração mais curto e uma elevada potência de ultrassons. Quanto aos macarrones formulados com o extrato antociânico, estes exibiram uma coloração estável e não apresentaram nenhuma diferença no valor nutricional quando comparados com macarrones formulados como o corante comercial. Conclui-se que o epicarpo de jabuticaba é uma fonte interessante de antocianinas e que o método ME foi mais indicado para extração destes compostos. O extrato obtido nas condições ótimas incorporado em macarrones proporcionou uma coloração estável e não interferiu na composição nutricional do produto. O biorresíduo estudado apresentou, assim, potencial para produção de corantes naturais. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER, no âmbito do programa PT2020, pelo apoio financeiro ao CIMO (UID/AGR/00690/2019); à FCT pela bolsa de doutoramento de B. Albuquerque (SFRH/BD/136370/2018) e pelos contratos-programa de emprego científico institucional de J. Pinela e L. Barros. Ao FEDER pelo financiamento através do Programa Operacional Regional Norte 2020 no âmbito do Projeto NORTE-01-0145-FEDER-023289 (DeCodE) e projeto Mobilizador Norte-01-0247-FEDER- 024479 (ValorNatural); ao FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E e ao projeto TRANSCoLAB (0612_TRANS_CO_LAB_2_P). info:eu-repo/semantics/publishedVersion

Myrciaria cauliflora (Mart.) O. Berg, known as jabuticaba, is a native species to Brazil, more specifically to Atlantic Rainforest biome. Its fruits are small berries with a diameter of 2.0 – 3.5 cm that contain between one and four seeds, with a sweet gelatinous pulp and thick dark purple epicarp when mature, which is not commonly consumed. Due to the properties of its pulp, the consumption and production of jellies and liqueurs from jabuticaba has increased. However, the epicarp that corresponds to about 50% of the fruit is not used, being converted into a bioresidue [1]. The intense colour of this part of the fruit is due to the pigments, namely anthocyanins, present in its composition. In addition to attractive staining, these molecules have been associated with bioactive properties, such as antioxidant activity, which makes their recovery interesting for application in various industrial segments, for instance, food and pharmaceutical [2,3]. With the objective of valorising this bioresidue, the present work aimed the determination of the main anthocyanins present in jabuticaba epicarp (by HPLC-DAD-ESI/MS) and the evaluation of the bioactivity of its ethanolic extract (through in vitro assessment of cytotoxic, antimicrobial, and antioxidant activities). Thus, cytotoxicity was evaluated in four tumour cell lines (NCI-H460 - lung carcinoma, MCF-7 - breast carcinoma, HepG2 - hepatocellular carcinoma, and HeLa - cervical carcinoma) and in a primary culture of non-tumour liver cells (PLP2), by the Sulforodamine B (SRB) assay. The antimicrobial activity was evaluated on five Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Morganella morgani, Proteus mirabilis, and Pseudomonas aeruginosa) and three Gram-positive bacteria (Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis). Lastly, the antioxidant activity was tested through the oxidative haemolysis inhibition assay (OxHLIA). The bioresidue of jabuticaba fruits presented two anthocyanins, identified as cyanidin 3-O-glucoside and delphinidin 3-O-glucoside, being the first one the most abundant. Regarding bioactivity, the ethanolic extract revealed antiproliferative activity in all tumour cell lines evaluated (GI50 < 300 μg/mL), except for NCI-H460, and did not show toxicity for PLP2 at the maximal tested concentration (400 μg/mL). It also exhibited bacteriostatic properties in all the analysed bacterial strains (ranging from 20 to 10 mg/mL). Regarding haemolysis inhibition, the extract was able to protect 50% of the erythrocyte population for 120 minutes in a lower concentration than the positive control (Trolox). The results obtained in this study allow to conclude that jabuticaba epicarp is a rich source of anthocyanins and also exhibits strong bioactivity, which makes it suitable for use as colorant. 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/00690/2019), L. Barros, R.C. Calhelha, and C. Pereira contracts and B. Albuquerque research grant (SFRH/BD/136370/2018). This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®.The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

A jabuticabeira é uma árvore frutífera nativa do Brasil, sendo encontrada principalmente na Mata Atlântica. Os seus frutos, denominados por jabuticabas, são pequenas bagas de aproximadamente 2 – 3,5 cm que possuem uma coloração preta a roxa, quando maduros. São muito apreciados para consumo in natura devido ao agradável sabor da sua polpa; no entanto, o epicarpo, que é rígido e possui um sabor adstringente, não é comumente consumido, sendo responsável por grande parte do resíduo gerado no processamento de produtos derivados de jabuticaba [1]. A intensa coloração desta parte dos frutos é devida à concentração de pigmentos presentes na sua composição, nomeadamente antocianinas, que são compostos de grande interesse devido à sua coloração atrativa e às suas propriedades benéficas para a saúde [2]. Além disso, a presença de outros compostos fenólicos também tem sido relatada no epicarpo de jabuticaba [1,3]. Com o objetivo de valorizar este biorresíduo, o presente estudo teve como foco a determinação do seu perfil fenólico e a avaliação das suas propriedades bioativas, nomeadamente a atividade antioxidante, antimicrobiana e antiproliferativa. Os compostos fenólicos presentes no extrato do epicarpo de jabuticaba (EEJ) foram detetados por HPLC-DAD- ESI/MS. Os ensaios de bioatividade foram realizados por métodos in vitro: a atividade antioxidante foi determinada através da inibição da formação de substâncias reativas do ácido tiobarbitúrico (TBARS) e da inibição da hemólise oxidativa (OxLHIA); a atividade antimicrobiana foi avaliada pelo método da microdiluição contra oito bactérias patogénicas; e a citotoxicidade foi analisada pelo ensaio da sulforrodamina B com quatro linhas celulares tumorais e uma cultura primária de células não-tumorais. O epicarpo de jabuticaba apresentou um perfil fenólico constituído por catorze compostos fenólicos não antociânicos e dois antociânicos, sendo a cianidina-3-O-glucósido o composto mais abundante. Em relação às propriedades bioativas, o EEJ apresentou uma elevada atividade antioxidante, atividade antiproliferativa contra três linhas celulares tumorais e atividade antimicrobiana contra todas as bactérias avaliadas. Não foi observada hepatotoxicidade em células não-tumorais na maior concentração de extrato avaliada. Conclui-se, assim, que o EEJ apresenta uma composição rica em compostos fenólicos, principalmente em antocianinas, e que possui consideráveis propriedades bioativas, o que potencia o interesse da exploração deste biorresíduo para a obtenção de compostos de interesse para a indústria alimentar e farmacêutica. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2019); financiamento nacional da FCT, PI, através do contrato de programa institucional de emprego científico para o contrato de L. Barros e R.C. Calhelha; o contrato de C. Pereira através da celebração contrato de programa previso no no 4, 5 e 6 do artigo 23o do Decreto-Lei no 57/2016, de 29 de agosto, alterado pela Lei no 57/2017, de 19 de julho; e bolsas de doutoramento de B. Albuquerque (SFRH/BD/136370/2018) e T.C.S.P. Pires (SFRH/BD/129551/2017); FEDER através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289 (DeCodE) e projeto Mobilizador Norte-01- 0247-FEDER-024479: ValorNatural®; FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E e TRANSCoLAB 0612_TRANS_CO_LAB_2_P. info:eu-repo/semantics/publishedVersion

produz frutos ricos em compostos fenólicos, em particular as catequinas [l]. Catequinas são flavan-3-óis que apresentam alta capacidade antioxidante [2]. No entanto, os flavan-3-óis estão susceptíveis a degradação por diversos fatores ambientais [3,4] e a estabilidade destes compostos durante o processamento e armazenamento é crucial para a preservação das suas propriedades bioativas. O presente trabalho teve como objetivo otimizar a extração de catequina a partir de frutos de A. unedo por maceração (ME), micro-ondas (MÃE) e ultrassons (UAE), utilizando a metodologia de superfície de resposta para avaliar os efeitos das principais variáveis de extração de cada técnica (tempo, temperatura ou potência e mistura hidroetanólica) e determinar a estabilidade do extraio rico em catequina sob influência da temperatura, do tempo e do pH, durante o armazenamento (sistema em pó) e aplicação numa matriz alimentar (sistema aquoso). As respostas medidas foram o conteúdo de flavan-3-óis e catequina, determinados por HPLC-DAD. Foram utüizadas equações mecanicistas e fenomenológicas para descrever as respostas para a estabilidade dos compostos. Dentre as técnicas de extração avaliadas, a ME e a MÃE foram mais eficientes, capazes de produzir 1,38±1 e 1,70±0,3 mg catequina/g matéria seca, respetivamente. As condições ótimas para a ME foram 93,2±3,7 min, 79,6±5,2 °C e 23, 1±3,7% de etanol, enquanto para a MÃE foram 42,2±4,1 min, 137, 1±8,1 °C e 12, 1±1,1% de etanol. A UAE apresentou menor rendimento nas condições otimizadas (0,71±0,1 mg/g em 42,4 min, 314,9±21,2 W e 40,3±3,8% de etanol). O extrato rico em catequina (60% flavan-3-óis e 22% catequina) obtido sob as condições ótimas para a ME apresentou maior estabilidade no armazenamento a -20°C e pH 5,4 durante o período avaliado (30 dias), enquanto a estabilidade dos compostos em sistema aquoso permaneceu estável durante as 24h de aplicação a um pH

Mestrado com dupla diplomação com a Universidade Tecnológica Federal do Paraná The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources.The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources. This study compares and optimizes the maceration, microwave and ultrasound extraction techniques in the recovery of a catechin extract from Arbutus unedo L. fruits and evaluate the stability of flavan-3-ols during storage and application processes. To obtain conditions that maximize the catechin extraction yield, a response surface methodology was used. Maceration and microwave extractions were found to be the most effective methods, capable of yielding 1.38±0.1 and 1.70±0.3 mg of catechin/g dry weight (dw) in the corresponding optimal extraction conditions. The optimal conditions for maceration were 93.2±3.7 min, 79.6±5.2 ºC and 23.1±3.7 % of ethanol, while for the microwave extraction were 42.2±4.1 min, 137.1±8.1 ºC and 12.1±1.1 % of ethanol. The microwave system was a quicker solution, conducting to slightly higher yields of catechin than maceration, but this one needed lower temperatures to reach similar yields. The ultrasound method was the least effective solution in terms of catechin yield extraction (0.71±0.1 mg/g at 42.4±3.6 min, 314.9±21.2 W and 40.3±3.8 %. ethanol). The stability was tested with of the catechin-enriched extract (60% flavan-3-ols and 22% catechin), obtained under the best maceration conditions, was tested. Therefore, catechin-enriched extracts were submitted to physical and chemical stability studies, considering the main affecting variables (time, temperature and pH): i) a stability study of the extracts during storage as powder system; and ii) a stability study of the extracts in simulated food environment (aqueous solution system). The measured responses were the flavan-3-ols and catechin contents, determined by HPLC-DAD, and the antioxidant activity of the extracts evaluated by hydrophilic assays. Mechanistic and phenomenological equations were used to describe the responses, and the optimal conditions for flavan-3-ols (including catechin) stability as powder extract during a month were pH= 5.4 and T= -20ºC; while its stability in aqueous solution remained during the 24 h of application at pH