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Solar drying is affordable, requiring low energy and an eco-friendly method. Thus, the present paper studies the efficiency and characteristics of the indirect solar convective drying in the fruits of Arbutus unedo L. as well as its effects on the fruit phenolic compounds. The fruit samples were dried at 60 °C, 70 °C, and 80 °C. Phenolic compounds were investigated using a Liquid Chromatography platform. Experimental results revealed that the effective moisture diffusivity determined by Fick’s second law varied from 1.51 × 10−9 to 4.68 × 10−9 m2/s, and the activation energy recorded was 2203.62 kJ/kg. Both the total energy consumption and the specific electrical energy of the dried fruits decreased as temperature increased. The Midilli–Kucuk model was selected as the best-fitted model for drying Arbutus unedo L. Significant effect of temperature on phenolics was observed. The concentration of the phenolic compounds decreased by 15.54, 39, and 40.63% at 60, 70, and 80 °C, respectively.

This work focused on the extraction, quantification, and characterization of bioactive compounds of Arbutus unedo L. fruits, comparing the results obtained from the different ripening states. Extractions were performed by different methods (such as maceration extraction and ultrasonic extraction) and food grade solvents (aqueous and hydroalcoholic solvents) in each of the all ripening states (four states considered, associated with four different colors, i.e., green, yellow, orange, and red). The presence of (poly)phenols was quantified and characterized, and scavenging activity was determined by the Folin–Ciocâlteu reagent and the DPPH method, respectively. The content of bioactive compounds was characterized by LC-MS/MS, such as multiple reaction monitoring (MRM) mass spectrometry. The results showed that ultrasound-assisted extraction (UAE) performed better than maceration extraction; ethanol–water mixture extracts showed a more positive effect than the use of aqueous extracts regarding the content of total phenolic compounds. Overall, the total phenolic compounds in the EtOH:H2O mixture at a ratio of 7:3 (v:v) were higher than that of the other solvents for both extraction methods. Some bioactive molecules were characterized for the first time in the extracts of A. unedo. The chemical profile of the strawberry tree extracts depended on the degree of fruit ripeness. The results suggest that A. unedo fruits may be of great interest for food and nutraceutical applications.

The objective of this study was to identify, quantify, and elucidate the polyphenols, flavonoids, and anthocyanins, and their antioxidant activities (via 13 2,2-diphenyl-1-picrylhydrazyl) (DPPH) radical and 2,2ʹ-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation scavenging abilities and bleaching β-carotene tested in vitro in the whole fruit, fruit skin of strawberry tree fruits of 5 genotypes and to find out the most valuable fruit for disease preventing diets. Total phenols, total flavonoids, total anthocyanins, antioxidant activity (DPPH, ABTS and β-carotene bleaching assay), pH, titrable acidity, soluble solids, and organic acids were investigated in five strawberry tree genotypes belonging to several areas in Morocco. Qualitative and quantitative analyses of individual phenolic compounds by high-performance liquid chromatography (HPLC) were also carried out. Significant differences (p˂0.05) were observed across the five genotypes in total phenols (25.37–39.06 mg GAE/g DW), total flavonoids (3.30–7.07 mg RE/g DW), total anthocyanins (0.15–0.64 mg cya-3-glu/100 g DW), pH (2.44–3.92), titrable acidity (0.65–1.01 g malic acid/100 g fw), and soluble solids (14.83–18.53%). The antioxidant activity was evaluated by three assays. The values were 3.33–21.08, 2.25–19.58, and 1.08–13 mg AAE/g DW for DPPH scavenging test, ABTS, and β-carotene bleaching, respectively. Gallocatechol and catechin were the most abundant phenolic compounds. Principal component analysis showed that the first three components formed 90.25% of the total inertia. Chlorogenic acid, ellagic acid derivative I, ellagic acid, rutin, and cyanidin-3,5-diglucoside were the most involved variables in the total variance explained.

The Republic of Croatia has a long tradition in fruit growing due to its geographical position, climatic conditions and high quality of fruit crops. The autochthonous fruit varieties are characterized by good morphological/pomological properties, but their nutritional and biological potential is underestimated. The fruits of the strawberry tree (Arbutus unedo L.) are an underappreciated fruit species in Croatia, whose potential for cultivation and consumption has not yet been sufficiently utilized. Therefore, it can be considered as an excellent raw material for the production of functional foods, which are under increasing demand by consumers. The aim of this study was to investigate the chemical and biological value of strawberry tree fruit collected in October 2021 in island Mali Lošinj, Croatia. Among the chemical properties, soluble solids content (SSC), pH, total carbohydrates, lipids, crude protein, crude cellulose, crude energy and sugar content were determined. Among bioactive compounds, total phenolic content (TPC), total flavonoids (TF), anthocyanins (ANT), hydroxycinnamic acids (HCA), flavonols (FL) and condensed tannins (CT) were analyzed. The obtained results showed that strawberry fruit samples contained high SSC (38 %), pH 4.6, total carbohydrates 30, 28 g/100 g, lipids

The strawberry tree (Arbutus unedo L.) is a Mediterranean plant known for the traditional use of its fruits and leaves due to their health benefits. Thus, it has been used for years in folk medicine to relieve various health conditions such as urological and kidney problems, dermatological, cardiovascular and gastrointestinal diseases. The fruits are traditionally used for making jams, jellies, and strong alcoholic beverages, while the leaves are mostly used for preparing tea. Since the leaves were more researched, previous results indicated that they have important biological effects, so further research should focus on the fruits. Due to its chemical composition, rich polyphenolic profile and the biological potential derived from it, the plant has great prospects for the production of functional foods and nutraceuticals. However, the plant’s potential is underutilized in terms of processing. Therefore, this review summarizes the properties and the potential of the fruits and leaves of A. unedo and their possible benefits for processing with respect to agricultural, nutritive, biological and economic values.

Kocayemiş (Arbutus unedo L.) meyvesinin ve yaprağının, su ve etil alkol ekstraktlarının içerdiği fenolik madde miktarı, fenolik madde çeşitleri, antioksidan aktiviteleri; farklı konsantrasyon ve farklı metodlarla incelendi. Kocayemişin en yüksek ekstrakt verimi, meyve-su ekstraktında (483,44 mg/g) gözlenmiştir. Toplam fenolik bileşik ve toplam flavonoid madde içeriği olarak, en yüksek madde içeriğinin kocayemiş yaprak-etil alkol ekstraktında olduğu belirlenmiştir. Demir (II) iyonlarını şelatlama aktivitesi, kocayemişin diğer ekstrakları ile karşılaştırıldığında, en fazla şelatlama aktivitesinin, kocayemiş meyve-etil alkol ekstraktında olduğu gözlenmiştir. H2 O2 giderme aktivitesi bakımından, kocayemişin ekstraktları arasında en yüksek H2 O2 giderme aktivitesini, kocayemişin meyve-su ekstraktı göstermiştir. Süperoksit anyon radikali giderme aktivitesi tayininde, ekstraktlar arasında kocayemiş meyve-etil alkol ekstraktının, daha yüksek süperoksit radikali giderme aktivitesine sahip olduğu tespit edilmiştir. DPPH• radikali giderme aktivitesi açısından incelendiğinde, ekstraktlar arasında en yüksek aktivite değeri, kocayemiş meyve- etil alkol ekstraktında gözlenmiştir. ABTS• + radikali giderme aktivitesinin tayininde, kocayemiş ekstraktları arasında, kocayemiş meyve-su ekstraktının daha yüksek aktivite göstermiştir. Toplam ferrik iyonlarını (Fe 3+ ) indirgeme kapasitesi incelendiğinde, ekstraktlar arasında en yüksek demir indirgeme kapasitesi, kocayemiş yaprak-etil alkol ekstraktında bulunmuştur. Kocayemiş meyvesinde, karotenoid miktarının kuru ağırlığın mg’ı başına düşen % toplam karotenoid miktarının %4,46 olduğu belirlenmiştir. Linoleik asit sisteminde ferrik tiyosiyanat (FTC) metodu kullanılarak lipid peroksidasyonunu inhibe etme değeri için, ekstraktlar içinde en yüksek değer ise kocayemiş yaprak-etil alkol (%45,71±0,34) ekstraktında gözlenmiştir. LC-MS/MS ile fenolik madde miktarı analizinde, kocayemiş meyvesinde su ve etil alkol ekstraktlarında, en yüksek fenolik madde konsantrasyonlarının; gallik asit, izohamnetin, quersetinde olduğu, kocayemiş yaprağının etil alkol ekstraktında ise en yüksek fenolik madde konsantrasyonunun kuersetinde olduğu bulunmuştur. Kocayemiş (Arbutus unedo L.) meyvelerinden ve yaprağından elde edilen tüm ekstraktların antioksidan aktivite gösterdiği, kocayemiş meyvelerinin ve yaprağının doğal bir antioksidan kaynağı olarak kullanılabileceği, üretiminin teşvik edilerek insanların direkt tüketebileceği, in vivo çalışmalarla desteklenerek eczacılık alanında faydalı çalışmalar yapılabileceği, sonucuna varılmıştır. The antioxidant activity of arbutus berry (Arbutus unedo L.) fruit and leaf and phenolic substance amount and types in water and ethyl alcohol extracts were examined using different concentrations and methods. The highest extract yield of erbutus berry was in fruit – water. (483,44 mg/g) The highest total phenolic compaund and total flavonoid contents were found in the ethyl alcohol extract. Comparing otherextracts of arbutus berry, the highest chelating activity of ferric (II) ions was observed in ethyl alcohol extract. In determination of H2 O2 clearance activity, the highest rate was in fruit – water extract of arbutus berry. In determining the superoxide anion radical clearance activity, the highest rate was identified in the arbutus berry fruit – ethyl alcohol extract among the other extracts. From the perspective of radical DPPH• clearance activity, the highest activity was observed in arbutus berry fruit – ethyl alcohol extract among the other extracts. In determination of ABTS• clearance activity, the highest activity was observed in arbutus berry fruit – water extract among other extarcts of the fruit. + In determination of total ferric ions (Fe 3+ ) reduction capacity, the highest capacity was found in arbutus berry leaf – ethyl alcohol extract. In determining carotenoid content in arbutus berry fruit, it is identified to contain % 4,46 per mg of dry weigh. As for the rate of lipid peroxidation inhibition by using the ferric thiocyanate method in linoleic acid system, the highest score was observed in arbutus berry fruit – ethyl alcohol extract (%45,71±0,34) among the other extracts. In the analysis of phenolic substance matter by LC-MS/MS it is determined that the water and ethyl alcohol extracts in arbutus berry, the highest phenolic substance concentrations were gallic acid, isohamnetin and quarcetin, in leaf and ethyl alcohol extract the highest phenolic substance concentration was found as quercetin. It was concluded that all the extracts obtained from arbutus berry fruit and leaf have antioxidant activity and both can be used as natural antioxidant sources. In addition to these by supporting the production people can use it directly or by strenghtening with “in vivo” works in pharmaceutics, functional activities can be achievable.

Arbutus unedo L. is a small Mediterranean plant, being fgound mostly in southern Europe, north eastern Africa, Ireland, Palestine and the Canary Islands, and have been studied for its chemical composition. info:eu-repo/semantics/publishedVersion

The strategies of two evergreen Mediterranean tree species (Arbutus unedo L. and Quercus ilex L.) to face annual temperature variability and extreme event responses has been monitored in continuum by leaf fluorescence and reflectance. The effect of chilling spells and the accumulation of cold days with chilling and freezing temperatures on winter photo-inhibition was discussed in terms of modulation capacity for fluorescence parameters to daily temperature and irradiance conditions. Throughout the winter Q. ilex resulted less photoinhibited than A. unedo, showing a more dynamic response in all fluorescence parameters and a higher non-photochemical quenching capacity. These characteristics effectively act to maintain a higher electron transport capacity in Q. ilex than in A. unedo.

The strawberry tree (Arbutus unedo L.), grown throughout the Mediterranean, produces edible fruit; as it is easily bruised, the sweet, reddish fruit is used mostly to prepare jams, marmalades and alcoholic beverages. As the genus is paraphyletic, phytochemical analysis can assist in defining the fruit composition with the species Arbutus unedo L. (A. unedo). Here we report on the carbohydrate, total sugar, protein, fat, fiber, ash, and mineral content of wild fruit, harvested from 45 specimens from five locations. The dominant nutrients were carbohydrates (78.2–84.8 g/100 g), total sugars (52.1–67.2 g/100 g) and dietary fiber (11.0–20.1 g/100 g). Other important nutrients supplied by A. unedo fruit include P, K, and Fe. The fruit was observed to contain health-promoting components providing 42 and 36%, of recommended daily allowance (RDA) for fiber and zinc, respectively, as well as iron and manganese, at levels exceeding minimum RDA. The free-sugar profile revealed high glucose followed by fructose content with minor amounts of sucrose (14, 11, and 6 g/100 g, respectively). Significant differences both between regions and within individuals were observed for several traits. The richness of fruit nutrients in A. unedo confers nutritional value and as such, a promising alternative fruit source.

An extract from Arbutus unedo fruits, rich in anthocyanins, was studied as a natural colorant with bioactive properties (antioxidant, antimicrobial and cytotoxic). The aqueous stability of the extract was monitored using the anthocyanins’ content as response (determined by HPLC-DAD) in function of time, temperature and pH. Aided by mechanistic/phenomenological models, the conditions that favours the stabilization of the extract were provided, highlighting the suitability of the colorant for pastry/bakery products. As a case study, the extract was incorporated into wafers and the changes on the nutritional profile, free sugars, fatty acids and antioxidant properties were monitored during 6 days of storage. The results provide information for: i) potential application of the rich extract in anthocyanins for producing a natural colorant with bioactive properties; and ii) shelf-life predictions. The extract incorporation did not cause changes in the nutritional components of wafers but added colorant and antioxidant properties. 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/00690/2013), L. Barros contract and C. Caleja (SFRH/BD/93007/2013) grant. This work is 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 also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. To Xunta de Galicia for financial support for the post-doctoral researcher of M.A. Prieto. info:eu-repo/semantics/publishedVersion

Mediterranean trees and shrubs form intra-annual density fluctuations (IADFs) in tree rings as a sign of their plasticity in wood formation in response to intraseasonal variations of environmental conditions. Different species show a different aptitude to form IADFs, due to their diverse ability to cope with climate stressors, since the occurrence of IADFs may affect plant hydraulics. Dendroecology and quantitative wood anatomy were used to characterise IADFs in Pinus pinea and Arbutus unedo co-occurring at a Mediterranean site in Italy. The relations between climate parameters (i.e. temperature and precipitation) and intra-annual tree-ring traits (i. e. IADF frequency and conduit size) were analysed to highlight the main triggers for IADF formation and their functional role.Data showed that both species are characterised by a high plastic response to climate and formed a high frequency of L-IADFs (occurrence of earlywoodlike conduits in latewood). The two species, although forming the same type of IADFs, showed different sensitivity to environmental factors. Pinus pinea showed a high dependence of tracheid size on temperature, while Arbutus unedo was more sensitive to precipitation in spring and autumn. Arbutus unedo promptly developed more than one IADF per year in response to rainfall events following drought periods.The overall results were useful to compare the aptitude of the two species in forming IADFs and to highlight the factors priming their formation. This is useful to understand wood growth reactions to environmental drivers and to evaluate the adaptive capabilities in these two species, and thus to predict forest reactions after climate changes.

The strawberry tree fruit and leaf are a rich source of minerals, easily digestible sugars, dietary fibers, vitamins (especially vitamin C) and many bioactive compounds of significant functional value. Due to their favorable chemical composition, fruits have recently become increasingly popular in consumption. The aim of this study was to determine the physical-chemical composition, content of bioactive compounds, and also the antioxidant capacity of the fruit and leaves of wild strawberry tree populations among the Adriatic coast in Croatia, as well as to investigate the influence of location on the content of specific bioactive compounds. According to the obtained results, both fruit and leaves are pronouncedly high in vitamin C content, the average value for fruits amounted to 224.21 mg/100 g FW, while that for leaves amounted to 138.08 mg/100 g FW. Additionally, significantly high values of total polyphenolic compounds were recorded both in fruits (average value of 637.94 mg GAE/100 g FW) and especially in leaves (average value of 2157.01 mg GAE/100 g FW). Several pigments from different categories were determined in the fruit depending on fruit maturity, including: total anthocyanins, β-carotene and lycopene; while in leaves chlorophylls and carotenoids. Given the high content of different bioactive compounds high values of antioxidant capacity were determined (the average value for fruits was 2269.96 µmol TE/kg and for leaves, 2237.16 µmol TE/kg). Location strongly influenced the physical-chemical composition and also the content of specialized metabolites; populations collected from southern areas (central and south Dalmatia) of the Adriatic coast tended to have higher amounts of vitamin C, total phenols, total anthocyanins and β-carotene.

U okviru ovoga diplomskog rada provedena je fitokemijska analiza biološki aktivnih tvari listova i plodova vrste Arbutus unedo L. (obična planika, maginja), prikupljenih s otoka Lošinja (lokalitet Mali Lošinj) i Koločepa. Prisutnost polifenolnih spojeva dokazana je kemijskim reakcijama stvaranja obojenih produkata i taloga. Kvalitativna analiza fenolnih kiselina (ferulične, kavene, klorogenske i ružmarinske) te flavonoida (rutin, kvercetin, naringenin) provedena je primjenom tankoslojne kromatografije (TLC). Dokazana je prisutnost flavonoidnih tvari te fenolnih kiselina i/ili iridoida, a sa sigurnošću je detektiran izokvercitrin. Kvalitativna analiza trjeslovina provedena je općim reakcijama stvaranja boje i taloga te je potvrđena prisutnost trjeslovina koje hidroliziraju i kondenziranih trjeslovina u svim ispitanim uzorcima. Kvalitativna analiza triterpena, sterola i triterpenskih kiselina provedena je TLC analizom i potvrđena je njihova prisutnost u svim ispitanim uzorcima. Nije uočena značajna varijabilnost ispitivanih bioaktivnih tvari obične planike s obzirom na različite lokalitete sabiranja biljnog materijala. Provedena fitokemijska karakterizacija predstavlja doprinos dosadašnjem istraživanju roda Arbutus te upotpunjuje znanstvene spoznaje o fitoterapijskom potencijalu maginje. A phytochemical analysis of bioactive compounds of leaves and fruits of Arbutus unedo L. gathered from the islands of Lošinj (population Mali Lošinj) and Koločep has been carried out in this graduate thesis. The presence of polyphenols has been demonstrated by general chemical reactions of developing colored products and precipitates. The qualitative analysis of phenolic acids (ferulic, caffeic, chlorogenic and rosmarinic acid) and flavonoids (rutin, quercetin, naringenin) was perfomed using thin layer chromatography (TLC). The presence of various flavonoid substances, as well as phenolic acids and/or iridoids was noticed, but it has only been confirmed for isoquercitrin. A qualitative analysis of tannins was conducted by general reactions of color and precipitate formation, which confirmed the presence of hydrolysable and condensed tannins in all examined samples. A qualitative analysis of sterols, triterpenic acids and triterpenes was performed by TLC analysis and their presence has been confirmed in all examined samples. Considering the place of collecting, no significant variability in the content of the bioactive substances occurs. The conducted phytochemical characterization represents a contribution to the scientific research of the genus Arbutus, expanding the existing knowledge of biological activity and phytotherapeutic potential of Arbutus unedo L.

There are not many exhaustive works emphasizing the amount of genetic diversity among the strawberry tree (Arbutus unedo L) genotypes in Morocco. This work aims to assess the biochemical composition of strawberry tree fruits, as well as to establish the variation of this composition among them. In this study, total phenols (TP), total flavonoids (TF), condensed tannins (CT) and hydrolyzable tannins (HT), total anthocyanins (TA), and free radical scavenging activity through ABTS were investigated in strawberry tree fruits. Furthermore, qualitative and quantitative analyses of individual phenolic compounds by high-performance liquid chromatography (HPLC) were carried out. Color parameters such as lightness (L *), Chroma (c *), and hue angle (h°) were also investigated. All studied variables showed highly significant differences among all samples with the exception of hydrolyzable tannins and chromatic coordinates. TP varied from 22.63 ± 1.74 to 39.06 ± 2.44 mg GAE/g DW, TF varied from 3.30 ± 0.60 to 8.62 ± 1.10 mg RE/g DW, and TA ranged between 0.12 ± 0.06 and 0.66 ± 0.15 mg cya-3-glu/100 g DW. In addition, CT and HT amounts were in the range of 10.41 ± 1.07-16.08 ± 1.50 mg TAE/g DW and 4.08 ± 2.43-6.34 ± 3.47 TAE/g DW, respectively. Moreover, the IC50 value (ABTS) ranged between 1.75 and 19.58 mg AAE/g DW. 17 phenolic compounds were detected in strawberry tree fruits. Gallocatechol and catechin were the most abundant phenolic compounds. Matrix of correlations revealed significant positive and negative correlations among variables particularly c *, a *, and b *. Principal component analysis (PCA) showed that the first three components formed than 68% of the total inertia. The following variables gallic acid, protocatechuic, gallocatechin, gallic acid derivative, chlorogenic acid, syringic acid, ellagic acid derivative II, L *, and h * were the most involved in the total variance explained. Hierarchical clustering classified samples into one main cluster, with a single branch. The results highlight a high biochemical diversity within studied strawberry genotypes, which is probably more genetically related.

Phenolic compounds are well-known bioactive compounds in plants that can have a protective role against cancers, cardiovascular diseases and many other diseases. To promote local food development, a comprehensive overview of the phenolic compounds’ composition and their impact on human health from typical Mediterranean plants such as Punica granatum L., Ziziphus jujuba Mill., Arbutus unedo L., Celtis australis L., Ficus carica L., Cynara cardunculus var. Scolymus L is provided. Moreover, the potential use of these data for authenticity determination is discussed. Some of the plants’ phenolic compounds and their impact to human health are very well determined, while for others, the data are scarce. However, in all cases, more data should be available about the content, profile and health impacts due to a high variation of phenolic compounds depending on genetic and environmental factors. Quantifying variation in phenolic compounds in plants relative to genetic and environmental factors could be a useful tool in food authentication control. More comprehensive studies should be conducted to better understand the importance of phenolic compounds on human health and their variation in certain plants.

A natural holm oak forest was selectively thinned to test thinning as a tool to reduce tree mortality, increase productivity, and reverse the recent regression of the dominant species (Quercus ilex) induced by climate change. Thinning increased aboveground productivity and reduced stem mortality in this Mediterranean forest during four years just after thinning, contributing to the maintenance of forest functioning under changing climatic conditions. Q. ilex was the only species positively affected by the thinning: stem growth increased for all stem sizes, and mortality was significantly lower in thinned plots. On the contrary, mortality rates of Phillyrea latifolia and Arbutus unedo were not significantly lower. Stem growth increased for P. latifolia only in the smallest stem-size class. Our results highlight the suitability of selective thinning for improving the forest productivity and ensuring the conservation of Mediterranean coppices. Other benefits of selective thinning, such as a decrease in the risk of fire dispersion and an increase in the water supply for human populations, are also discussed.

Navadna jagodičnica (Arbutus unedo L.) je v Sredozemlju pogosta evmediteranskavrsta, katere areal sega tudi v Istro. Do leta 1971, ko je bila odkrita na rtu Ronek, kjer je danes edino verjetno naravno nahajališče v Sloveniji in je najbolj severna populacija na vzhodni jadranski obali, je bilav Tržaškem zalivu le sajena. Glavni cilji naloge so bili ugotoviti stanje in tendenco razvoja populacije na rtu Ronek ter jo primerjati s populacijo z rta Dente pri izlivu reke Mirne na Hrvaškem, najbližjo naravno populacijo. Populacija na rtu Ronek obsega 19 relativno vitalnih osebkov z dobrimi svetlobnimi razmerami, zmernim cvetenjem in slabim obrodom. Pomlajevanja se niugotovilo. Z morfometrijsko analizo listov se ni odkrilo značilnih razlik med populacijama. Jagodičnica je vrsta vmesnih sukcesijskih stadijev. Ker ji za svetlobo konkurirajo druge drevesne vrste in vzpenjalke, je za njeno ohranitev na rtu Ronek potrebno ukrepanje. V nalogi so bile raziskale prednosti in slabosti petih strategij, ki se jih je analiziralo po SWOT metodi. Arbutus unedo L. is a common tree species in the Mediterranean whose natural distribution range extends as far north as the Istrian peninsula. Historically, A. unedo was cultivated in the Trieste Gulf, however, there wereno known natural locations of this species. Until 1971, the only probable northern-most natural location in the eastern Adriatic Sea was in cape Dente, located at the mouth of the Mirna River in Croatia. In 1971, a small population of A. unedo was discovered in Slovenia, and is now believed to be the northern-most natural population of the eastern Adriatic coast. The goal of this project was to assess the situation, state, and development trend of A. unedo in cape Ronek, and to determine whether there were significant differences between this and the cape Dente population. The cape Ronek population comprises of 19 relatively viable specimens with good light conditions, poor to moderate flowering and poor fruiting. There were no signs of any regeneration and leaf morphometric analysis revealed no differences between populations. Because A. unedo is a species of intermediate successional stages with strong competition for light from other tree species and climbers, action is necessary for its conservation on cape Ronek. The advantages and disadvantages of five strategies for conserving the population has been researched and presented in SWOT table analyses.

La présente étude a pour objectif d’évaluer l’effet du stade de maturation sur le contenu en substances bioactives, en particulier les composés antioxydants de nature phénolique extraites par des solutions hydroéthanoliques et l’estimation de l\'évolution de l’activité antioxydante du fruit de l\'Arbousier (Arbutus unedo L.).Les fruits de l\'Arbousier ont été classés en 4 stades, le premier stade est quand le fruit est de couleur verte, le stade 2 comprend les fruits jaunes, le troisième stade est lorsque le fruit devient orange, alors que le stade 4 représente le dernier stade où les fruits sont en pleine maturité c\'est à dire rouges. Quatre extraits sont obtenus, les fruits à maturité; fruits oranges et fruits rouges ont donné des quantités élevées en terme de rendement d\'extraction par le solvant hydroalcoolique.Les différents extraits ont été testés pour leur teneur en composés phénoliques totaux, flavonoïdes, tannins condensés, chlorophylle et caroténoïdes, ainsi que leur activité antioxydante (anti-radicalaire). Les résultats obtenus montrent que le stade de maturation affecte significativement la teneur en antioxydants ainsi que l’activité antioxydante. On observe une réduction des tannins au cours de la maturation; de 124,9 mgCE/g dans le fruit non mûrs à 63,8 mgCE/g dans les fruits en pleine maturité. A l\'opposé, le contenu en flavonoïdes augmente; de 11,5 mgRE/g dans le fruit vert à 38,9mgRE/g dans le fruit orange. Les anthocyanes sont présentes dans le fruit à maturité et atteignent un seuil élevé en pleine maturité (12,7 µg/g). Nos résultats sur l\'activité antiradicalaire montrent que le stade de maturation affecte sensiblement le pouvoir anti-radicalaire du fruit. Tous les fruits ont la capacité de piéger efficacement le radical libre DPPH, mais le potentiel antioxydant le plus puissant (EC50: 0,5 mg/ml) est observé quand le fruit arrive à sa pleine maturation., The objective of this study is to evaluate the effect of the ripening stage on phytochemical composition, in particular the phenolic antioxidant compounds and antioxidant activity of Arbutus unedo L. fruits.The fruits of the strawberry tree have been classified in 4 stages, the first stage is when the fruit is green, stage 2 includes yellow fruits, the third stage is when the fruit turns orange, while stage 4 represents the last stage where the fruit so ripe, i.e. red. Four extracts are obtained, the ripe orange and red fruits yielded high amounts in terms of extraction yield by a hydroethanolic solvent.Extracts were tested for their content of total phenolic, flavonoids, condensed tannins, chlorophyll, and carotenoids, as well as their antiradical activity. The results obtained show that the ripening stage significantly affects antioxidant content as well as antiradical activity. Tannins are reduced during the ripening period; 124,9 mgCE/g in unripe fruit to 63,8 mgCE/g in fully ripe fruit. In contrast, the content of flavonoids increases; 11,5 mgRE/g in green fruit to 38,9 mgRE/g in orange fruit. Anthocyanins are present in the ripe fruit and reach a high value at the full ripe stage (12,7 µg/g). Our results on antiradical activity show that the ripening stage significantly influences the antiradical power of the fruit. All the fruits have the ability to scavenge free radical DPPH, whereas the most potent antiradicalar potential (EC50: 0,5mg/ml) is observed when the fruit reaches full ripe stage.

Strawberry-tree (Arbutus unedo L.) honey is expensive and difficult to characterize by melissopalinology. This study aimed to authenticate strawberry-tree (A. unedo L.) honeys from southern Europe, determining arbutin, groups of polyphenols, volatile and semivolatile compounds, as well as biological activities such as trolox equivalent antioxidant capacity, antioxidant activities against both hydroxyl (AOA) and superoxide radicals (SRS), oxygen radical absorbance capacity, anti-inflammatory activity, and antimicrobial activity against 7 microorganisms (Escherichia coli, Streptococcus mutans, Staphylococcus aureus, Aspergillus flavus, Aspergillus niger, Fusarium sp., and Penicillium commune). Arbutin, analyzed using HPLC-UV, was quantified in 83% of the samples. Polyphenols' contents were high. Norisoprenoids and benzene derivatives were the major compounds determined using gas chromatography-mass spectrometry. Theobromine was detected in 67% of samples. 2,6,6-Trimetyl-4-oxo-2-cyclohexen-1-carboxaldehyde, 3,4,5 trimethylphenol and 2-hydroxycyclopent-2-en-1-one were proposed as potential strawberry-tree floral markers. Antioxidant and anti-inflammatory activities were significant. Honeys’ extracts showed higher AOA and SRS, and better antimicrobial activities than the honeys. This study highlights the potential of strawberry-tree honeys and/or their phenolic extracts for food, pharmaceutical and cosmetic uses., Spanish Junta de Castilla y León, Spain (BU041G18)

Ethnopharmacological relevance Arbutus unedo L., the strawberry tree (Ericaceae family) is of increasing interest because of its common traditional, industrial, chemical and pharmaceutical uses. The plant is a typical evergreen plant of the Mediterranean basin, as well as of other regions with hot summers and mild rainy winters. This review covers the studies relevant to Arbutus unedo L. utilization in the current pharmacological therapy. Materials and methods The available information on traditional uses, phytochemistry and biological activities of Arbutus unedo L. was collected from scientific databases through a search using the keywords ‘Arbutus unedo L.’ and/or ‘strawberry tree’ in ‘Google Scholar’, ‘Pubmed’, ‘Sciencedirect’, ‘SpringerLink’, ‘Web of Science - Clarivate Analytics’ and ‘Wiley’. Unpublished Ph.D. and M.Sc. dissertations were also consulted for chemical composition, biological activities and traditional uses of Arbutus unedo L. and for manual search of additional references. Results The fruits of the plant have been traditionally used as antiseptics, diuretics and laxatives in folk medicine, while the leaves have been used due to their diuretic, urinary antiseptic, antidiarrheal, astringent, depurative and antihypertensive properties. According to the scientific literature survey, different extracts obtained from Arbutus unedo L. have demonstrated a high pharmacological potential due to their in vitro and preclinical antibiotic, antifungal, antiparasitic, antiaggregant, antidiabetic, antihypertensive, anti-inflammatory, antitumoral, antioxidant, and spasmolytic properties. Conclusion This review suggests that A. unedo is a promising source of phytopharmaceutical products. The potential advantages of Arbutus unedo are related with the presence of polyphenolic compounds in its composition. However, further studies are needed to ascertain some profitable effects in humans. The beneficial effects associated with this shrub suggest that Arbutus unedo can be used for the development of new drugs to treat diseases such diabetes, hypertension, among others. Nonetheless, the safety of the Arbutus unedo compounds should also be examined.

BACKGROUND AND AIMS: Mediterranean trees have patterns of cambial activity with one or more pauses per year, leading to intra-annual density fluctuations (IADFs) in tree rings. We analysed xylogenesis (January 2015–January 2016) in Pinus pinea L. and Arbutus unedo L., co-occurring at a site on Mt. Vesuvius (southern Italy), to identify the cambial productivity and timing of IADF formation. METHODS: Dendrochronological methods and quantitative wood anatomy were applied and enabled IADF identification and classification. KEY RESULTS: We showed that cambium in P. pinea was productive throughout the calendar year. From January to March 2015, post-cambial (enlarging) earlywood-like tracheids were observed, which were similar to transition tracheids. The beginning of the tree ring was therefore not marked by a sharp boundary between latewood of the previous year and the new xylem produced. True earlywood tracheids were formed in April. L-IADFs were formed in autumn, with earlywood-like cells in latewood. In A. unedo, a double pause in cell production was observed, in summer and winter, leading to L-IADFs in autumn as well. Moreover, the formation of more than one IADF was observed in A. unedo. CONCLUSIONS: Despite having completely different wood formation models and different life strategies, the production of earlywood, latewood and IADF cells was strongly controlled by climatic factors in the two species. Such cambial production patterns need to be taken into account in dendroecological studies to interpret climatic signals in wood from Mediterranean trees.

The strawberry tree (Arbutus unedo L.), grown throughout the Mediterranean, produces edible fruit, as it is easily bruised, the sweet, reddish fruit is used mostly to prepare jams, marmalades and alcoholic beverages. As the genus is paraphyletic, phytochemical analysis can assist in defining the fruit composition with the species Arbutus unedo L. (A. unedo). Here we report on the carbohydrate, total sugar, protein, fat, fiber, ash, and mineral content of wild fruit, harvested from 45 specimens from five locations. The dominant nutrients were carbohydrates (78.2–84.8 g/100 g), total sugars (52.1–67.2 g/100 g) and dietary fiber (11.0–20.1 g/100 g). Other important nutrients supplied by A. unedo fruit include P, K, and Fe. The fruit was observed to contain health-promoting components providing 42 and 36%, of recommended daily allowance (RDA) for fiber and zinc, respectively, as well as iron and manganese, at levels exceeding minimum RDA. The free-sugar profile revealed high glucose followed by fructose content with minor amounts of sucrose (14, 11, and 6 g/100 g, respectively). Significant differences both between regions and within individuals were observed for several traits. The richness of fruit nutrients in A. unedo confers nutritional value and as such, a promising alternative fruit source.

Bu çalışmada kocayemiş meyvesi ilavesinin dondurmanın fiziksel, kimyasal ve duyusal özelliklerine etkisi araştırılmıştır. Kocayemiş meyveli dondurma üretiminde iki farklı oranda (%15, %25) kocayemiş meyve püresi ilavesi ve bir kontrol dondurması üretilmiştir. Çalışma iki tekerrürlü yapılmıştır. Örneklerde kurumadde, kül, hacim artış tayini, erime süresi, ilk damlama ve tamamen erime süreleri, pH, titrasyon asitliği tayini, yağ, protein, renk analizleri, C vitamini (askorbik asit), antioksidan aktivite tayini, şeker tayini, su aktivite tayini, duyusal analizler yapılmış, sonuçlar istatiksel olarak değerlendirilmiştir. Kocayemiş meyveli dondurmanın kurumadde, kül, yağ, pH, protein miktarlarında meyve püresi ilavesinden kaynaklanan azalma saptanırken, meyve ilavesiyle dondurmada C vitamini miktarında, troloks cinsinden antioksidan aktivite değerinde ve bununla birlikte titrasyon asitliğinde artış meydana gelmiştir. Kocayemiş meyve püresi oranlarının artışına bağlı olarak erimeye karşı olan direnci azalmış, hacim artışı değerinde artış meydana gelmiştir. Duyusal açıdan % 15 kocayemiş meyvesi içeren dondurma örnekleri kontrol grubuna daha yakın değer almıştır. In this study, the effects of strawberry tree fruit addition to physical, chemical and sensory properties of ice cream were investigated. Strawberry tree fruit ice creams were produced with two different ratios (15%, 25%) and a control group was produced. Study was conducted in two replications. Dry matter, ash, overrun, melting time, first dripping time, complete melting time, pH, titrable acidity, fat, protein, color, vitamin C (ascorbic acid), antioxidant activity, sugar, water activity, and sensory analysis were carried out and results were evaluated with statistically. When the amount of dry matter, ash, fat, pH, and total nitrogen were decreased due to addition of pulp in ice creams, amount of ascorbic acid, antioxidant activity, and titrable acidity increased. Depending on the increase in the percentage of pulp, resistance to melting was decreased; overrun was increased. Strawberry tree fruit added ice cream with 15% ratio has sensory values near to the control group.

The strawberry tree fruit and leaf are a rich source of minerals, easily digestible sugars, dietary fibers, vitamins (especially vitamin C) and many bioactive compounds of significant functional value. Due to their favorable chemical composition, fruits have recently become increasingly popular in consumption. The aim of this study was to determine the physical-chemical composition, content of bioactive compounds, and also the antioxidant capacity of the fruit and leaves of wild strawberry tree populations among the Adriatic coast in Croatia, as well as to investigate the influence of location on the content of specific bioactive compounds. According to the obtained results, both fruit and leaves are pronouncedly high in vitamin C content, the average value for fruits amounted to 224.21 mg/100 g FW, while that for leaves amounted to 138.08 mg/100 g FW. Additionally, significantly high values of total polyphenolic compounds were recorded both in fruits (average value of 637.94 mg GAE/100 g FW) and especially in leaves (average value of 2157.01 mg GAE/100 g FW). Several pigments from different categories were determined in the fruit depending on fruit maturity, including: total anthocyanins, &beta, carotene and lycopene, while in leaves chlorophylls and carotenoids. Given the high content of different bioactive compounds high values of antioxidant capacity were determined (the average value for fruits was 2269.96 &micro, mol TE/kg and for leaves, 2237.16 &micro, mol TE/kg). Location strongly influenced the physical-chemical composition and also the content of specialized metabolites, populations collected from southern areas (central and south Dalmatia) of the Adriatic coast tended to have higher amounts of vitamin C, total phenols, total anthocyanins and &beta, carotene.

There are not many exhaustive works emphasizing the amount of genetic diversity among the strawberry tree (Arbutus unedo L.) genotypes in Morocco. This work aims to assess the biochemical composition of strawberry tree fruits, as well as to establish the variation of this composition among them. In this study, total phenols (TP), total flavonoids (TF), condensed tannins (CT) and hydrolyzable tannins (HT), total anthocyanins (TA), and free radical scavenging activity through ABTS were investigated in strawberry tree fruits. Furthermore, qualitative and quantitative analyses of individual phenolic compounds by high-performance liquid chromatography (HPLC) were carried out. Color parameters such as lightness (L*), Chroma (c*), and hue angle (h&deg, ) were also investigated. All studied variables showed highly significant differences among all samples with the exception of hydrolyzable tannins and chromatic coordinates. TP varied from 22.63 &plusmn, 1.74 to 39.06 &plusmn, 2.44 mg GAE/g DW, TF varied from 3.30 &plusmn, 0.60 to 8.62 &plusmn, 1.10 mg RE/g DW, and TA ranged between 0.12 &plusmn, 0.06 and 0.66 &plusmn, 0.15 mg cya-3-glu/100 g DW. In addition, CT and HT amounts were in the range of 10.41 &plusmn, 1.07&ndash, 16.08 &plusmn, 1.50 mg TAE/g DW and 4.08 &plusmn, 2.43&ndash, 6.34 &plusmn, 3.47 TAE/g DW, respectively. Moreover, the IC50 value (ABTS) ranged between 1.75 and 19.58 mg AAE/g DW. 17 phenolic compounds were detected in strawberry tree fruits. Gallocatechol and catechin were the most abundant phenolic compounds. Matrix of correlations revealed significant positive and negative correlations among variables particularly c*, a*, and b*. Principal component analysis (PCA) showed that the first three components formed than 68% of the total inertia. The following variables gallic acid, protocatechuic, gallocatechin, gallic acid derivative, chlorogenic acid, syringic acid, ellagic acid derivative II, L*, and h* were the most involved in the total variance explained. Hierarchical clustering classified samples into one main cluster, with a single branch. The results highlight a high biochemical diversity within studied strawberry genotypes, which is probably more genetically related.

Arbutus unedo L., commonly known as strawberry-tree, is a shrub used as ornamental plant in the gloomy Mediterranean areas which fruits are popularly eaten and used to produce beverages . Beyond these common applications, these fruits could also be explored for their phenolic composition, once they are a source of anthocyanin compounds with great colouring capacity to be potentially used in food industry, in detriment of artificial colorants. In this context, the present study aimed to optimize the extraction of these compounds from A. unedo fruits through different techniques, namely heat and ultrasound assisted extraction and, for that purpose, a response surface methodology was applied using the circumscribed central composite design of three variables with five levels. For the model application, the anthocyanin concentration and the extraction yield were used as responses, being the identification of these compounds performed by HPLC-DAD-ESI/MS. The anthocyanin profile of A. unedo fruits consisted of delphinidin-3-glucoside, cyanidin-3-glucoside, and cyanidin-3-pentoside, with cyanidin-3-glucoside as the most abundant one. Heat assisted extraction proved to be the most effective technique at 5 min, 90°C, and 80% of ethanol, yielding 51.2% of fruit dry weight, with a total anthocyanin concentration of 382.4 μg/g of dried fruit, and 744.6 μg/g of extract. Moreover, these response values were slightly improved by studying the solid/liquid ratio effect at the optimal conditions indose-response format, showing steady extraction values from 5 to 40 g/L. The obtained results showed the possibility of using A. unedo fruits as a source of anthocyanin compounds for industrial applications. European Union Regional Development Fund: Programme PT2020 (CIMO Project UID/AGR/00690/2013; LA LSRE-LCM Project POCI-01-0145-FEDER-006984; Carla Pereira grant SFRH/BPD/122650/2016); Programme Portugal-NORTE2020 (Project NORTE-01-0145-FEDER-023289: DeCodE; Project NORTE-01-0247-FEDER-024479: ValorNatural®); Programmme INTERREG V-A Espanha-Portugal (POCTEP) 2014-2020 (Project 0377_Iberphenol_6_E). And Xunta de Galicia, Spain, for M.A. Prieto grant. info:eu-repo/semantics/publishedVersion

Atualmente, existe uma preocupação da indústria alimentar na substituição de aditivos sintéticos por ingredientes naturais. Os extratos naturais obtidos a partir de plantas surgem como possíveis alternativas aos conservantes sintéticos, nomeadamente dos antioxidantes. Neste trabalho, pretendeu-se comprovar a eficácia de extratos ricos em catequina, obtido a partir de frutos de Arbutus unedo L. (medronho) e em ácido rosmarínico obtido a partir de folhas de Ocimum basilicum L. (manjericão), como conservantes naturais de pão, comparando a sua eficácia com aditivos sintéticos normalmente aplicados a produtos de panificação. Após a avaliação das propriedades antioxidantes (valores EC50 do efeito bloqueador de radicais de DPPH = 595 e 351 μg/ mL, para medronho e manjericão, respetivamente) e antimicrobianas (contra 8 bactérias e 8 fungos) e confirmação da ausência de citoxicidade (numa cultura primária de células de fígado), os extratos ricos em (+)-catequina e ácido rosmarínico foram incorporados no pão. O seu efeito foi comparado com o ácido ascórbico (E300), o sorbato de potássio (E202) e com uma amostra controlo (pão sem aditivos) imediatamente após a incorporação e após 3 e 7 dias de armazenamento, à temperatura ambiente e protegidos da luz. Os pães aditivados com os extratos naturais, especialmente de manjericão, apresentaram maior capacidade antioxidante do que os pães com ácido ascórbico e sorbato de potássio, tendo todos mantido as suas propriedades nutricionais. Em geral, os resultados obtidos demonstraram que os extratos podem ser utilizados no desenvolvimento de pão, substituindo os conservantes sintéticos, sem interferir nas características físicas e nutricionais dos produtos finais. Currently, the food industry is focused on replacing synthetic additives with natural ingredients. Natural extracts obtained from plants have emerged as possible alternatives to synthetic preservatives, namely in the class of antioxidants. In this work we intend to prove the efficacy of an extract rich in catechin, obtained from Arbutus unedo L. (strawberry tree) fruits, and of an extract rich in rosmarinic acid obtained from Ocimum basilicum L. (basil) leaves, as natural preservatives of bread and compare these results to synthetic additives which are normally used in bakery products. After evaluation of the antioxidant (EC50 values for DPPH scavenging activity = 595 and 351 μg/ mL, for strawberry tree fruits and basil leaves, respectively) and antimicrobial (against 8 bacteria and 8 fungi food contaminants) properties, and confirmation of cytotoxicity absence (in porcine liver primary cells), the extracts rich in (+)-catechin and rosmarinic acid were incorporated into the bread. Its effect was compared to ascorbic acid (E300), potassium sorbate (E202) and to a control sample (bread without additives), immediately after incorporation and in the 3rd and 7th day of storage, protected from light at room temperature. The breads incorporated with the natural extracts, especial basil, presented a higher antioxidant capacity than the breads incorporated with ascorbic acid and potassium sorbate, while the nutritional profile was not affected. Overall, the obtained results demonstrate that these extracts could be used in the development of new bread formulations replacing the synthetic preservatives, without interfering with the physical and nutritional characteristics of the final product. info:eu-repo/semantics/publishedVersion

Currently, there is a tendency for consumers to choose more natural foods [l]. Anthocyanins are secondary metabolites ofplants that have aroused industry attenüon for its coloring potential combined with confirmation ofvarious beneficiai efFects to human health [2]. However, due to their very sensitive chemical structure to several factors such as temperature, pH, oxygen and light, studies that allow its stabilization are indispensable [3]. This work aimed to study the stability of an extract obtained from Arbutus unedo L. fruits, considering 3 variables (time, temperature and pH) in aqueous solution by analyzing the content of anthocyanins. The extract was obtained by a heat-assisted extraction technique, with the optimal condiüons of 5 minutes, 90°C, 80% ethanol and 15 g/L solid/liquid ratio, obtained after an optimization process by applying a response surface methodology. The identification of three anthocyanin compounds (cyanidin- 3-0-glucoside as the major molecide) was performed by HPLC-DAD-ESI/MS, and revealed the foUowing responses, 60.9 %, with a total anthocyanin content of 500.9 ^ig/g of dry fruit and 800.9 ^ig/g of extract. Moreover, the extract was analyzed in terms of its antioxidant, antimicrobial and cytotoxic activity revealing a high bioactive potential. Finally, the anthocyanin-rich extract was stabilized by subjecting it to adverse storage conditions. Time, temperature and pH were the variables considered to evaluate the behavior of the extract and the concentration of anthocyanins from each sample wére the response factors used. In order to describe the responses, mechanical and phenomenological equations were used with a global performing fitting analysis from a multivariable perspective. The results showed that the three analyzed variables exert a direct influence on the responses obtained. It hás been found that increasing pH and T values cause an exponential increase in anthocyanin degradation. In the overall analysis of ali variables, it was concluded that the optimum conditions ofanthocyanins stability were verified at pH

Although strawberry tree (Arbutus unedo L.) leaves have long been used as a herbal remedy, insufficient information is available on their nephrotoxicity. We assessed the safety of strawberry tree water leaf extract and its key component arbutin, administered per os to Lewis rats of both genders at 200 mg/kg b.w./day for 14 and 28 days. The effects of the tested compounds on DNA integrity in renal cells was evaluated using alkaline comet assay, while kidney function was studied using serum creatinine and urea levels. Strawberry tree water leaf extract showed high biocompatibility with kidney tissue. It did not impair DNA integrity of renal cells and kidney function, either in male or female rats. However, exposure to single arbutin affected the levels of primary DNA damage in renal cells which could be related to metabolic conversion of arbutin into hydroquinone, whose nephrotoxicity has previously been proven.

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

The overwhelming growth of the world population is causing direct consequences on food demands, which has forced the food industry to apply synthetic additives in order to meet the needs of the consumer and ensuring the global quality of the food products including aspect, texture and flavor. However, due to the publication of some studies describing adverse effects associated with these synthetic additives, the natural ones have been gaining increasing interest. Arbutus unedo L. (strawberry tree) fruits and Ocimum basilicum L. (basil) leaves, are important sources of bioactive natural molecules, especially phenolic compounds, including flavan-3- ols and phenolic acids, respectively . The aim of this work was to demonstrate the antioxidant potential (free radical scavenging effect, reducing power and inhibition of lipid peroxidation) and the absence of toxicity (porcine liver primary cells) of ethanol extracts, obtained from the previously mentioned plants by using optimal conditions. Subsequently, the extracts were incorporated as natural ingredients in loaf bread samples, with two objectives: to functionalize loaf bread, introducing bioactive properties; and as natural ingredients with preservation capacity. Five groups of loaf bread samples were prepared: i) control (without natural or synthetic additives); ii) loaf bread incorporated with strawberry tree extract; iii) loaf bread incorporated with basil extract; iv) loaf bread incorporated with ascorbic acid- E300 (natural additive); and v) loaf bread incorporated with potassium sorbate- E202 (synthetic additive). Both extracts (strawberry tree fruits and basil leaves) presented antioxidant activity (EC50 values lower the 595 and 351 μg/mL, respectively), without toxicity up to the maximal tested dose, and the main compounds present were (+)-catechin and rosmarinic acid, respectively. The introduction of both natural and synthetic additives in loaf bread when compared to the control sample, did not cause significant changes, neither in pH values, nor in color or any of the evaluated nutritional parameters. The obtained results demonstrate that these extracts could be used in the development of new loaf bread replacing the synthetic preservatives, without interfering with the physical and nutritional characteristics. info:eu-repo/semantics/publishedVersion

Mediterranean forests are fragile ecosystems vulnerable to recent global warming and reduction of precipitation, and a long-term negative effect is expected on vegetation with increasing drought and in areas burnt by fires. We investigated the spatial distribution of genetic variation of Arbutus unedo in the western Iberia Peninsula, using plastid markers with conservation and provenance regions design purposes. This species is currently undergoing an intense domestication process in the region, and, like other species, is increasingly under the threat from climate change, habitat fragmentation and wildfires. We sampled 451 trees from 15 natural populations from different ecological conditions spanning the whole species' distribution range in the region. We applied Bayesian analysis and identified four clusters (north, centre, south, and a single-population cluster). Hierarchical AMOVA showed higher differentiation among clusters than among populations within clusters. The relatively low within-clusters differentiation can be explained by a common postglacial history of nearby populations. The genetic structure found, supported by the few available palaeobotanical records, cannot exclude the hypothesis of two independent A. unedo refugia in western Iberia Peninsula during the Last Glacial Maximum. Based on the results we recommend a conservation strategy by selecting populations for conservation based on their allelic richness and diversity and careful seed transfer consistent with current species' genetic structure. info:eu-repo/semantics/publishedVersion

Mestrado em cooperação com a Universidade de Salamanca El primer paso crítico de este estudio fue obtener las condiciones óptimas que maximizan la extracción de antocianinas a partir de frutos de Arbutus unedo L., mediante la comparación de dos técnicas extractivas asistidas por diferentes energías físicas, el calor y los ultrasonidos. Para ello, se aplicó una metodología de superficie de respuesta, utilizando el diseño compuesto central circunscrito de tres variables con cinco niveles. Se identificaron tres compuestos de antocianina por HPLC-DAD-ESI/MS, siendo cianidina-3-O-glucósido la molécula principal. La extracción asistida por calor resultó ser la técnica más efectiva, que produjo un rendimiento del 60,9 %, con un contenido total de antocianinas de 500,9 μg/g de fruto seco y 800,6 μg/g de extracto siguiendo unas condiciones de 5 min, 90 ºC y 80 % de etanol acidificado. Además, estos valores se mejoraron ligeramente al estudiar el efecto de la relación sólido/líquido en un formato de dosis-respuesta (de 5 a 150 g/L), en las condiciones óptimas de extracción, manifestando valores de extracción prácticamente constantes entre los 5 y los 40 g/L. Una vez conseguida la optimización de la extracción de antocianinas (principalmente cianidina-3-O-glucósido) a partir de frutos de Arbutus unedo L., se procedió a estudiar su potencial como aditivo natural colorante, con la finalidad de ser utilizado en la industria alimentaria como substituto de colorantes sintéticos, cuya demanda va en decadencia progresiva. En primer lugar, se analizaron las propiedades bioactivas del extracto, analizando sus capacidades antioxidantes, antimicrobianas y citotóxicas. Los resultados mostraron que el extracto optimizado presentaba importantes propiedades antioxidantes, así como actividades citotóxicas (en líneas celulares tumorales) y antimicrobianas significativas. Complementariamente, se evaluó su posible toxicidad, corroborándose efectos nulos. Una vez analizadas minuciosamente las características del extracto, se procedió a su estabilización. Para ello se sometió el extracto rico en antocianinas a las condiciones más adversas de conservación (solución acuosa) y se estudió su comportamiento teniendo en cuenta las principales variables que afectan a la misma (tiempo, temperatura y pH). Los valores utilizados como respuestas fueron las concentraciones de antocianinas de cada muestra, determinadas por HPLC-DAD. Se usaron ecuaciones matemáticas fenomenológicas para describir las respuestas, y se proporcionaron patrones para la estabilización del extracto rico en antocianinas en solución acuosa. Finalmente, el extracto se incorporó en gofres como caso práctico de estudio en productos de pastelería, con el objeto de evaluar su posible aplicación en matrices alimentarias. La estabilidad completa del perfil nutricional, los azúcares libres, los ácidos grasos y la actividad antioxidante se controlaron justo después de que los gofres se hornearan y tras 3 y 6 días de almacenamiento. Por lo tanto, los resultados obtenidos proporcionan información para: i) la posible aplicación industrial del extracto de frutos de A. unedo como fuente alternativa de antocianinas para producir un aditivo natural colorante con propiedades bioactivas; ii) predicciones de vida útil en condiciones específicas de temperatura y pH; y iii) la incorporación del extracto de A. unedo rico en antocianinas no solo no causó cambios significativos en el valor nutricional de gofres en comparación con las muestras control, sino que dicha incorporación añadió propiedades colorantes y bioactivas al producto original, lo que respalda su posible aplicación en la preparación de productos de pastelería y panadería. The first critical step of this study was to obtain the optimal conditions that maximize the anthocyanins extraction from fruits of Arbutus unedo L., by comparing two extractive techniques assisted by different physical energies, heat and ultrasounds. For this, a response surface methodology was applied, using the three-variable circumscribed central composite design with five levels. Three anthocyanin compounds were identified by HPLC-DAD-ESI/MS, being cyanidin-3-O-glucoside the major molecule. Heat assisted extraction proved to be the most effective technique, yielding a 60.9 %, with a total anthocyanin content of 500.9 μg/g of dry fruit and 800.9 μg/g of extract following conditions of 5 min, 90 °C and 80 % acidified ethanol. In addition, these values were slightly improved by studying the effect of the solid/liquid ratio under optimum conditions, in the dose-response format, showing constant extraction values between 5 and 40 g/L. Once the extraction optimization of anthocyanins (mainly cyanidin-3-O-glucoside) from fruits of Arbutus unedo L. was achieved, its potential as a natural coloring additive was studied to replace, in the food industry, artificial additives, whose demand goes in progressive decay. First, the bioactive capacities of the extract were analyzed by testing its antioxidant, antimicrobial and cytotoxicity properties, revealing high antioxidant properties, as well as significant cytotoxic (in tumor cell lines) and antimicrobial activities. Complementarily, the toxicity of the extract was evaluated, validating its non-toxic effects. Afterwards, a stability study of the extract was carried out, considering the main variables that could affect the extract (time, temperature and pH) under the most unfavorable conservation conditions (aqueous solution). Responses were measured by anthocyanin contents determined by HPLC-DAD. Mechanical and phenomenological equations were used to describe the responses, and patterns were provided for the stabilization of the rich extract in anthocyanins in aqueous solution. Finally, the extract was incorporated in waffles in order to study its possible application in food matrices, in this case, in a pastry product. A complete stability of the nutritional profile, free sugars, fatty acids and antioxidant activity were monitored right after the waffles were baked and subsequently with 3 and 6 days of storage. Therefore, the results obtained provide information for: i) the possible industrial application of A. unedo fruit extract as an alternative source of anthocyanins to produce a natural coloring additive with bioactive properties; ii) predictions of shelf life under specific conditions of temperature and pH; and iii) the incorporation of A. unedo extract rich in anthocyanins, did not only cause significant changes in the nutritional value of waffles in comparison with the control samples, but also added color and bioactive properties to the original product, supporting its potential application in the preparation of pastry and bakery products.

The genetic diversity within and between four natural Arbutus unedo L. populations in Portugal was investigated using morphological characters, inter-simple sequence repeats (ISSR) and random amplified polymorphic (RAPD) markers, for a proper conservation of this resource. The results showed a high morphological variation between and within-populations. The dry-weight, length, width and peduncle length of the leaves were the most discriminating characters, being responsible for 52.6% of the total variation. The molecular analysis, based on both RAPD and ISSR, indicated moderate genetic diversity within populations, with values of Nei’s gene diversity ranging from 0.16 to 0.39 and the Shannon’s diversity from 0.24 to 0.57. The highest genetic diversity occurred within Braganc¸ a population. At the species level, the genetic variation was mainly within populations (75%), suggesting a low-moderate genetic differentiation among populations. The high level of gene flow estimated among populations (1.499) showed to be the main reason for their low differentiation. The cluster analysis showed that only one population formed a distinctive cluster. The remaining formed a second cluster due to their geographical approach, which has been showed to affect significantly the genetic distances among populations. No correlation between RAPD and ISSR markers, and between the molecular and morphological data was found. From the conservation point of view it is proposed to combine both in- and ex situ approaches. The maintenance of sufficient gene flow and the increase of the effective population sizes are the main in situ actions proposed. The ex situ conservation, should involve the collection of a high number of individuals within populations.

Mestrado em Bioquímica - Métodos Biomoléculares Submitted by Alexandra Bastos (alexandrabastos@ua.pt) on 2016-04-18T11:14:35Z No. of bitstreams: 1 tese.pdf: 2407645 bytes, checksum: cb61870e8fba45523a3227675cdd37b8 (MD5) Made available in DSpace on 2016-04-18T11:14:35Z (GMT). No. of bitstreams: 1 tese.pdf: 2407645 bytes, checksum: cb61870e8fba45523a3227675cdd37b8 (MD5) Previous issue date: 2015-12-23

Intra-annual density fluctuations (IADFs) are anomalies of tree rings where wood density is abruptly altered after sudden changes in environmental conditions. Their characterisation can provide information about the relationship between environmental factors and eco-physiological processes during tree growth. This paper reports about the variability of anatomical traits and stable carbon isotopic composition along tree rings as resulting from the application of two different methodological approaches: (a) the separation of each ring into different regions (earlywood, latewood and IADF) and the comparison of anatomical and isotopic parameters measured in those specific sectors and (b) the analysis of such features in continuum along ring width. Moreover, different parameters of vessels (i. e. ecd-equivalent circle diameter, elongation, sphericity and convexity of vessel lumen) were considered to identify those more appropriate for the representation of intra-annual anatomical variations. The analysis was conducted on Arbutus unedo L. growing on the Elba Island (Italy); tree rings of this species form IADFs with features clearly responsive to the environmental conditions experienced during plant growth. Results showed that the first approach, although more suitable to obtain data for subsequent statistical comparisons and for the calculation of correlations with environmental parameters, suffers from elements of subjectivity due to the size and position of the selected tree-ring regions. The in continuum method allows a clearer identification of the variation of tree-ring properties along ring width. Regarding anatomical parameters, shape indexes were not suitable indicators of intra-annual variability. The overall analysis suggested that using both methodologies in synergy helps to gain complete information and avoid misleading interpretations of IADFs in tree rings. © 2011 Springer-Verlag.

Genetic analysis of plants relies on high yields of pure DNA. For the strawberry tree (Arbutus unedo) this represents a great challenge since leaves can accumulate large amounts of polysaccharides, polyphenols and secondary metabolites, which co-purify with DNA. For this specie, standard protocols do not produce efficient yields of high-quality amplifiable DNA. Here, we present for the first time an improved leaf-tissue protocol, based on the standard cetyl trimethyl ammonium bromide protocol, which yields large amounts of high-quality amplifiable DNA. Key steps in the optimized protocol are the addition of antioxidant compounds-namely polyvinyl pyrrolidone (PVP), 1,4-dithiothreitol (DTT) and 2-mercaptoethanol, in the extraction buffer; the increasing of CTAB (3%, w/v) and sodium chloride (2M) concentration; and an extraction with organic solvents (phenol and chloroform) with the incubation of samples on ice. Increasing the temperature for cell lyses to 70 °C also improved both DNA quality and yield. The yield of DNA extracted was 200.0 ± 78.0 μg/μL and the purity, evaluated by the ratio A(260)/A(280), was 1.80 ± 0.021, indicative of minimal levels of contaminating metabolites. The quality of the DNA isolated was confirmed by random amplification polymorphism DNA and by inter-simple sequence repeat amplification, proving that the DNA can be amplified via PCR.

Phytopharmaceuticals play an essential role in medicine, since the need to investigate highly effective and safe drugs for the treatment of diabetes mellitus disease remains a significant challenge for modern medicine. Arbutus unedo L. root has various therapeutic properties, and has been used widely in the traditional medicine as an antidiabetic agent. The current study aimed to isolate the pharmacologically active compound from A. unedo roots using accelerated solvent extraction technology, to determine its chemical structure using different instrumental analytical methods, and also to evaluate the α-glucosidase inhibitory activity. The roots of A. unedo were exhaustively extracted by high-pressure static extraction using the Zippertex® technology (Dionex-ASE, Paris, France), and the extract was mixed with XAD-16 resin to reach quantifiable amounts of active compounds which were identified by high-pressure liquid chromatography (HPLC), 1H NMR (300 MHz), and 13C NMR. The antidiabetic activity of the isolated compound was evaluated using the α-glucosidase inhibitory assay. The active compound was isolated, and its structure was identified as catechin using instrumental analysis.The results revealed that the isolated compound has potential α-glucosidase inhibitory activity with an IC50 value of 87.55 ± 2.23 μg/mL greater than acarbose. This was used as a positive control, which has an IC50 value of 199.53 ± 1.12 μg/mL. According to the results achieved, the roots of A. unedo were considered the best source of catechin and the Zippertex® technology method of extraction is the best method for isolation of this therapeutic active compound. In addition, the α-glucosidase inhibitory activity results confirmed the traditional use of A. unedo roots as an antidiabetic agent. Future clinical trials and investigations of antidiabetic and other pharmacological effects such as anticancer are required.

Some of the major constituents of strawberry tree are flavonoids and phenolic acids that are responsible for some of the pharmacological activities i.e. adstringent, antihypertensive, antidiarrheal and antibacterial. Objective of this work was to determine flavonoid aglycones and phenolic acids in strawberry tree leaves collected during 12 months of 2013 using RP-HPLC-DAD and assess antioxidant activity using ABTS stable free radical. Antioxidant activity of strawberry tree leaves varied between 11% and 28%. The content of individual polyphenols was: chlorogenic acid 0.8 to 6.5 mg/g, caffeic acid 0.6–1.0 mg/g, p-coumaric acid 0.2–6.6 mg/g and quercetin 0.5–10.7 mg/g. The highest content of polyphenols was during winter months. Compared to the thin-layer chromatography analysis we have determined presence of additional phenolic acids (caffeic and p-coumaric) and quercetin – aglycone of quercitrin, isoquercitrin and hyperoside.

Istraživanje je provedeno na listovima planike - Arbutus unedo L., skupljenim kroz 12 mjeseci tijekom 2013. godine u mjestu Božava na Dugom otoku. Tekućinskom kromatografijom visoke djelotvornosti (HPLC) provedena je kvalitativna i kvantitativna analiza ekstrakata listova. Kvalitativnom analizom utvrđena je prisutnost flavonoidnog aglikona kvercetina te fenolnih kiselina: klorogenske kiseline, kavene kiseline i p-kumarne kiseline tijekom cijele godine. Kvantitativna analiza je pokazala da količina kvercetina i fenolnih kiselina varira iz mjeseca u mjesec. Najveće količine kvercetina (10, 74 mg/g), klorogenske kiseline (6, 53 mg/g) i p-kumarne kiseline (6, 59 mg/g) nađene su u listovima skupljenim u siječnju, a kavene kiseline u listovima skupljenim u kolovozu (0, 96 mg/g). Najmanje količine kvercetina utvrđene su u listovima skupljenim u listopadu (0, 48 mg/g), klorogenske kiseline u prosincu (0, 83 mg/g), p-kumarne kiseline u svibnju (0, 23 mg/g), a kavene kiseline u listopadu (0, 64 mg/g).

Mestrado em Bioquímica - Bioquímica Alimentar Submitted by Alexandra Bastos (alexandrabastos@ua.pt) on 2015-04-24T13:49:25Z No. of bitstreams: 1 tese.pdf: 4915320 bytes, checksum: 659687408f1452b8631db2ed951ca072 (MD5) Made available in DSpace on 2015-04-24T13:49:25Z (GMT). No. of bitstreams: 1 tese.pdf: 4915320 bytes, checksum: 659687408f1452b8631db2ed951ca072 (MD5) Previous issue date: 2014-12-11