Green Approach to Enhance the Recovery of Polyphenols from Blackcurrant and Bilberry Leaves: Evaluation of Microwave-Assisted and Pressurized Liquid Extraction.

The aim of the present study was to evaluate microwave-assisted (MAE) and pressurized liquid extraction (PLE) for the recovery of polyphenols from blackcurrant and bilberry leaves and the preservation of their antioxidant activity. The extractions were carried out varying the solvent/solid (SS) ratio, temperature and time. During MAE, increasing the SS ratio increased the polyphenol concentration in the extracts from blackcurrant and bilberry leaves, while increasing the temperature had a positive effect only on bilberry polyphenols. During PLE, only a temperature increase was a determining factor for the isolation of blackcurrant leave polyphenols. Based on polyphenol recovery, optimal extraction parameters were established resulting in a yield of 62.10 and 56.06 mg/g dw in the blackcurrant and bilberry MAE extracts and 78.90 and 70.55 mg/g dw in the PLE extracts. The optimized extracts were profiled by UPLC ESI MS2, and their antioxidant capacity was evaluated through FRAP, DPPH, ABTS and ORAC assays. The characterization of the extracts by UPLC ESI MS2 confirmed flavonols as the predominant compounds in both blackcurrant and bilberry leaves, while flavan-3-ols and procyanidins were the main compounds responsible for high antioxidant capacity as confirmed by the ABTS and ORAC assays. Due to the extract composition and antioxidant capacity, PLE proved to be a technique of choice for the production of blackcurrant and bilberry leave extracts with high potential for use as value-added ingredients in the food and nutraceutical industry.

Microencapsulation of Laurus nobilis L. Leaf Extract in Alginate-Based System via Electrostatic Extrusion.

Bay leaves (L. nobilis L.) are a rich source of polyphenols that hold great potential for application in functional food products in which where the main challenges are the polyphenols' low stability and bioaccessibility, which can be overcome through different microencapsulation techniques, such as electrostatic extrusion, which hasn't been applied for the encapsulation of bay leaf polyphenols (BLP) to date. Therefore, the main goal of this research was to evaluate the potential of this technique through monitoring the polyphenolic content, antioxidant activity, release kinetics, and bioaccessibility of the encapsulated BLP. The results showed that electrostatic extrusion was suitable for the encapsulation of BLP, where 1% alginate and 1.5% CaCl2 with 0.5% chitosan resulted in the highest encapsulation efficiency (92.76%) and antioxidant activity in vitro. The use of 1.5% or 2% alginate with 5% CaCl2 + 0.5% chitosan showed the most controlled release of polyphenols, while encapsulation generally increased the bioaccessibility of BLP. The results showed that electrostatic extrusion can be considered an efficient technique for the microencapsulation of BLP.

Sage, Rosemary, and Bay Laurel Hydrodistillation By-Products as a Source of Bioactive Compounds.

Essential oils from Mediterranean wild plants are widely used, but the hydrodistillation residues produced in parallel with these essential oils are significantly understudied and underexploited. Since there are only fragmentary data in the literature, we have, for the first time, systematically analyzed the chemical composition of the by-products obtained after hydrodistillation of sage, bay laurel, and rosemary leaves, i.e., hydrolates, water residues, and solid residues. The chemical composition of the hydrolates changed compared to their respective essential oils towards the dominance of more hydrophilic, oxygenated compounds, such as camphor in sage, 1,8-cineole in bay laurel, and berbenone in rosemary. However, some compounds, mostly sesquiterpenes, which were present in considerable amounts in essential oils, were absent or only present in very small amounts in the hydrolates. Furthermore, both the water and the solid residues were rich in polyphenols, such as procyanidins in bay laurel and rosmarinic acid in rosemary and sage. In conclusion, we demonstrate the valuable chemical composition of sage, rosemary, and bay laurel hydrodistillation by-products and discuss a wide range of their possible applications.

Optimization of the Spray-Drying Encapsulation of Sea Buckthorn Berry Oil.

The aim of this study was to evaluate the effect of spray-drying parameters on the physicochemical properties of encapsulated sea buckthorn berry oil. Different carriers (gum arabic, ß-cyclodextrin, and their mixture (1:1, w/w)), inlet air temperatures (120, 150, and 180 °C), and carrier-to-oil ratios (2, 3, and 4, w/w) were evaluated. The obtained powders were characterized in terms of the product yield (36.79-64.60%), encapsulation efficiency (73.08-93.18%), moisture content (0.23-3.70%), hygroscopicity (1.5-7.06 g/100 g), solubility (19.55-74.70%), bulk density (0.25-0.44 g/L), total carotenoid content (mg/100 g dm), and antioxidant capacity (871.83-1454.39 µmol TE/100 g dm). All physicochemical properties were significantly affected by the carrier-to-oil ratio and inlet air temperature. Higher carrier-to-oil ratios increased the product yield, encapsulation efficiency, solubility, and bulk density and decreased the powder hygroscopicity. Elevating the drying temperatures during spray drying also increased the product yield, encapsulation efficiency, and solubility, while it decreased the powder moisture content, total carotenoid content, and antioxidant capacity. Based on the physicochemical properties, the use of ß-cyclodextrin as a carrier, a drying temperature of 120 °C, and a carrier-to-oil ratio of 4 were selected as optimal conditions for the production of sea buckthorn berry oil powder. The obtained powder is a valuable material for a wide range of applications in the food and nutraceutical industries.

Physicochemical Properties, Antioxidant Capacity, and Bioavailability of Laurus nobilis L. Leaf Polyphenolic Extracts Microencapsulated by Spray Drying.

Laurel (Laurus nobilis L.) leaves are a rich source of polyphenols with the potential for use in functional foods, where the main obstacle is their low stability and bioavailability, which can be improved by spray drying (SD). This research examined the influence of SD parameters, including inlet temperature (120, 150, and 180 °C), carrier type (ß-cyclodextrin (ß-CD); ß-CD + maltodextrin (MD) 50:50; ß-CD + gum arabic (GA) 50:50), and sample:carrier ratio (1:1, 1:2 and 1:3) on the physicochemical properties, encapsulation efficiency, polyphenolic profile, antioxidant capacity and bioaccessibility of laurel leaf polyphenols. The highest encapsulation efficiency was achieved at a sample:carrier ratio 1:2 and the temperature of 180 °C by using either of the applied carriers. However, the application of ß-CD + MD 50:50 ensured optimal solubility (55.10%), hygroscopicity (15.32%), and antioxidant capacity (ORAC 157.92 µmol Trolox equivalents per g of powder), while optimal moisture content (3.22%) was determined only by temperature, demanding conditions above 150 °C. A total of 29 polyphenols (dominantly flavonols) were identified in the obtained powders. SD encapsulation increased the bioaccessibility of laurel flavonols in comparison to the non-encapsulated extract by ~50% in the gastric and ~10% in the intestinal phase, especially for those powders produced with carrier mixtures.

Phenolic, Headspace and Sensory Profile, and Antioxidant Capacity of Fruit Juice Enriched with Salvia officinalis L. and Thymus serpyllum L. Extract: A Potential for a Novel Herbal-Based Functional Beverages.

Since certain constituents are not naturally present in pure fruit juices, incorporating herbal extracts can provide specific sensory properties to the beverages and improve their biopotential. In our previous research, it was found that sage (Salvia officinalis L.), wild thyme (Thymus serpyllum L.), and combinations of their extracts had the highest total phenolic content and a unique composition of volatile compounds, which can contribute to the aromatic and antioxidant qualities of functional products. Therefore, this research aimed to investigate the potential of sage and wild thyme extracts, as well as their mixture (wild thyme:sage at 3:1, v/v), to enrich fruit juices (apple, pineapple, and orange). Obtained beverages were evaluated for sensory properties as well as phenolic and headspace composition (UPLC-MS/MS and HS-SPME/GC-MS analysis) and antioxidant capacity (ORAC assay). The incorporation of wild thyme extract in pineapple juice provided the most harmonious flavor and the highest content of volatile compounds (on PDMS/DVB fiber). The orange juice formulations were the most enriched with phenolic and volatile compounds (on DVB/CAR/PDMS fibers). The highest antioxidant capacity was observed in the formulation with orange juice and sage extract (22,925.39 ± 358.43 µM TE). This study demonstrated that enriching fruit juices with sage and wild thyme extracts could create functional beverages with improved sensory and health-promoting properties, providing valuable insights for the food and beverage industry to meet the growing demand of health-conscious consumers for natural and functional products.

Effect of Spray Drying Encapsulation on Nettle Leaf Extract Powder Properties, Polyphenols and Their Bioavailability.

Nettle (Urtica dioica L.) is a plant rich in a health-promoting compounds such as polyphenols, which are sensitive and unstable compounds with low bioavailability, that need to be stabilized and protected from external influences. Therefore, the aim of this study was to examine how the temperature, type of carrier and sample to carrier ratio influence the physicochemical properties and encapsulation and loading capacity of the nettle leaf extract powder and examine the effect of encapsulation on the antioxidant capacity and bioavailability of polyphenols. The process yield ranged from 64.63-87.23%, moisture content from 1.4-7.29%, solubility from 94.76-98.53% and hygroscopicity from 13.35-32.92 g 100 g-1. The highest encapsulation (98.67%) and loading (20.28%) capacities were achieved at 160 °C, ß-CD:GA (3:1) and sample:carrier ratio of 1:3. Extracts encapsulated at selected conditions showed high antioxidant capacity and distinct polyphenolic profile comprised of 40 different compounds among which cinnamic acids were the most abundant. Moreover, the encapsulation increased the bioavailability of nettle leaf polyphenols, with the highest amount released in the intestinal phase. Thus, the obtained encapsulated extract represents a valuable source of polyphenols and may therefore be an excellent material for application in value-added and health-promoting products.

Effect of UV-C Irradiation on the Shelf Life of Fresh-Cut Potato and Its Sensory Properties after Cooking.

Research background: Potato tissue is damaged during fresh-cut production, which makes fresh-cut potato susceptible to the quality loss and microbiological spoilage. At the same time, such products are desirable due to their convenience; however, they are extremely sensitive and have short shelf life. The main challenge of the fresh-cut potato industry is to find possibilities to overcome these drawbacks. UV-C treatment, known for its antibacterial activity, is a promising technique and it shows a potential to improve shelf life of fresh-cut potato products. Experimental approach: The influence of the UV-C treatment on the safety and quality, as well as sensory traits of fresh-cut potato (Solanum tuberosum L. cv. Birgit) during storage was examined. For this purpose, 0-, 3-, 5- and 10-min UV-C irradiation was applied on vacuum-packed potato slices pretreated with sodium ascorbate solution. During 23 days of storage at (6±1) °C, microbiological, physicochemical and sensory properties of raw samples were monitored, along with sensory properties of boiled and fried fresh-cut potatoes. Results and conclusions: The 5- and 10-min UV-C treatments significantly reduced microbial growth, increased total solids and lightness (L*), and positively affected odour and firmness of raw potatoes. Cooked UV-C-treated samples were described with more pronounced characteristic potato odour and taste. Overall, UV-C-treated fresh-cut potato retained its good quality and sensory traits up to 15 days at (6±1) °C. Novelty and scientific contribution: To the best of our knowledge, this is the first scientific article dealing with the effect of UV-C light on durability (safety, quality and sensory traits) of fresh-cut potato cv. Birgit and its suitability for boiling and frying. In general, UV-C treatment is a known antimicrobial technique, but its application on fresh-cut potato is poorly explored. Results confirmed that vacuum-packed fresh-cut potato treated only with UV-C and sodium ascorbate as anti-browning agent, without the addition of chemical preservatives, had twofold longer shelf-life at (6±1) °C than the fresh-cut potato not treated with UV-C. Fresh-cut potato treated with UV-C retained good overall quality and sensory properties either raw, boiled or fried. Results of this study could also be useful for producers in terms of potential UV-C application as a strategy for prolonging the shelf-life of fresh-cut potato.

Pressurized Liquid Extraction as a Novel Technique for the Isolation of Laurus nobilis L. Leaf Polyphenols.

Laurus nobilis L., known as laurel or bay leaf, is a Mediterranean plant which has been long known for exhibiting various health-beneficial effects that can largely be attributed to the polyphenolic content of the leaves. Pressurized liquid extraction (PLE) is a green extraction technique that enables the efficient isolation of polyphenols from different plant materials. Hence, the aim of this research was to determine optimal conditions for PLE (solvent, temperature, number of extraction cycles and static extraction time) of laurel leaf polyphenols and to assess the polyphenolic profile of the optimal extract by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) as well as to evaluate the antioxidant activity determined by FRAP, DPPH and ORAC assays. The optimal PLE conditions were 50% ethanol, 150 °C, one extraction cycle and 5 min static time. The polyphenolic extract obtained at optimal PLE conditions comprised 29 identified compounds, among which flavonols (rutin and quercetin-3-glucoside) were the most abundant. The results of antioxidant activity assays demonstrated that PLE is an efficient green technique for obtaining polyphenol-rich laurel leaf extracts with relatively high antioxidant activity.

Non-Volatile and Volatile Bioactives of Salvia officinalis L., Thymus serpyllum L. and Laurus nobilis L. Extracts with Potential Use in the Development of Functional Beverages.

Functional beverages based on herbal extracts are highly demanded products due to the presence of bioactives with promising health benefits and interesting and characteristic sensory properties. Mediterranean medicinal and aromatic herbs contain a wide range of bioactives (non-volatile polyphenols, volatile terpenes) that are important constituents of herbal extracts and essential oils. The antioxidant capacity and potential health benefits of these bioactives could be associated with their synergistic effects. Therefore, this study aimed to characterize the non-volatile and volatile bioactives of sage (Salvia officinalis L.), wild thyme (Thymus serpyllum L.) and laurel (Laurus nobilis L.) aqueous extracts and their two- and three-component mixtures as well as their antioxidant capacity. The content of total phenols, flavonoids, hydroxycinnamic acids and flavonols was determined spectrophotometrically. Individual polyphenols were analyzed by LC-MS/MS, the volatiles were analyzed by HS-SPME/GC-MS, and the antioxidant capacity was analyzed by ORAC and DPPH assays. The results showed that aqueous extracts of all examined herbs and their mixtures contained a high content of phenolic compounds ranging from 0.97 to 2.79 g L-1 of the sample, among which the most common were flavonols. At the same time, mono- and sesquiterpenes were the main volatiles. All extracts showed high antioxidant capacity, especially L. nobilis (781.62 ± 5.19 µmol TE mL-1 of the sample in the DPPH assay; 1896.10 ± 8.77 µmol TE mL-1 of the sample in the ORAC assay) and the two-component mixture of L. nobilis and T. serpyllum (679.12 ± 5.19 µmol TE mL-1 in the DPPH assay; 1913.38 ± 8.77 µmol TE mL-1 in the ORAC assay). Mixtures of herbal extracts have been shown to possess additive or synergistic effects, consequently contributing to higher antioxidant capacity. Therefore, two-component mixtures of herbal extracts showed promising potential for the production of functional beverages.

Bioprospecting of Microalgae Isolated from the Adriatic Sea: Characterization of Biomass, Pigment, Lipid and Fatty Acid Composition, and Antioxidant and Antimicrobial Activity.

Marine microalgae and cyanobacteria are sources of diverse bioactive compounds with potential biotechnological applications in food, feed, nutraceutical, pharmaceutical, cosmetic and biofuel industries. In this study, five microalgae, Nitzschia sp. S5, Nanofrustulum shiloi D1, Picochlorum sp. D3, Tetraselmis sp. Z3 and Tetraselmis sp. C6, and the cyanobacterium Euhalothece sp. C1 were isolated from the Adriatic Sea and characterized regarding their growth kinetics, biomass composition and specific products content (fatty acids, pigments, antioxidants, neutral and polar lipids). The strain Picochlorum sp. D3, showing the highest specific growth rate (0.009 h-1), had biomass productivity of 33.98 ± 0.02 mg L-1 day-1. Proteins were the most abundant macromolecule in the biomass (32.83-57.94%, g g-1). Nanofrustulum shiloi D1 contained significant amounts of neutral lipids (68.36%), while the biomass of Picochlorum sp. D3, Tetraselmis sp. Z3, Tetraselmis sp. C6 and Euhalothece sp. C1 was rich in glycolipids and phospholipids (75%). The lipids of all studied microalgae predominantly contained unsaturated fatty acids. Carotenoids were the most abundant pigments with the highest content of lutein and neoxanthin in representatives of Chlorophyta and fucoxanthin in strains belonging to the Bacillariophyta. All microalgal extracts showed antioxidant activity and antimicrobial activity against Gram-negative E. coli and S. typhimurium and Gram-positive S. aureus.

Phenolic Profile, Antioxidant Capacity and Antimicrobial Activity of Nettle Leaves Extracts Obtained by Advanced Extraction Techniques.

Nettle is a widely known plant whose high biological activity and beneficial medicinal effects are attributed to various bioactive compounds, among which polyphenols play an important role. In order to isolate polyphenols and preserve their properties, advanced extraction techniques have been applied to overcome the drawbacks of conventional ones. Therefore, microwave-assisted extraction (MAE) has been optimized for the isolation of nettle leaves polyphenols and it was compared to pressurized liquid extraction (PLE) and conventional heat-reflux extraction (CE). The obtained extracts were analyzed for their individual phenolic profile by UPLC MS2 and for their antioxidant capacity by ORAC assay. MAE proved to be the more specific technique for the isolation of individual phenolic compounds, while PLE produced extracts with higher amount of total phenols and higher antioxidant capacity. Both techniques were more effective compared to CE. PLE nettle extract showed antimicrobial activity against bacteria, especially against Gram-negative Pseudomonas fragi ATCC 4973 and Campylobacter jejuni NCTC 11168 strains. This suggests that PLE is suitable for obtaining a nettle extract with antioxidant and antimicrobial potential, which as such has great potential for use as a value-added ingredient in the food and pharmaceutical industry.

Composition and Antioxidant Properties of Pigments of Mediterranean Herbs and Spices as Affected by Different Extraction Methods.

This study examined the composition and properties of chlorophyll and carotenoid extracted from the leaves of several Mediterranean evergreen shrubs and subshrubs (Myrtus communis L., Pistacia lentiscus L., Thymus vulgaris L., Salvia officinalis L. and Laurus nobilis L.) commonly used as herbs and spices. In order to fully assess their composition over a wide polarity range, pigments were extracted by successive solvent extraction with hexane, 80% acetone and 96% ethanol. Agitation-assisted extraction (AAE), ultrasound-assisted extraction (UAE) and pressurized liquid extraction (PLE) were employed and compared regarding their effect on the pigments' yield and composition. Individual chlorophylls and carotenoids were analyzed by HPLC-DAD, while the content of total pigments and the extracts' antioxidant capacity were determined spectrophotometrically. Throughout the experiments, pheophytin a, b and b' were dominant chlorophyll molecules, while lutein and ß-carotene were dominant carotenoids. Overall, the extracted pigments were determined as being in the range of 73.84-127.60 mg 100 g-1 and were the lowest in T. vulgaris, with no significant differences between other species. M. communis and P. lentiscus had the highest antioxidant capacities, showing a moderate positive correlation with carotenoid and chlorophyll levels. Significant differences were found in the levels of individual pigments with most of them showing a medium level of polarity due to the dissolution in acetone as a medium polar solvent. AAE and PLE demonstrated similar efficacy in the extraction of both carotenoids and chlorophylls; however, preference can be given to PLE, being a novel method with numerous advantages, e.g., shorter extraction time and lower solvent consumption. The examined plant species certainly expressed great diversity and showed the potential for application in the production of various functional products.

Effect of UV-C Irradiation, Storage and Subsequent Cooking on Chemical Constituents of Fresh-Cut Potatoes.

UV-C irradiation successfully reduces the growth of microorganisms, but it can also affect the content of phenolics and sugars of fresh-cut potatoes (FCP). This could consequently alter antioxidant capacity of FCP or its potential for acrylamide formation. Therefore, this paper investigates the influence of UV-C irradiation on the content of phenolics [chlorogenic acid (CA)] and individual sugars during storage of FCP as well as after cooking. Acrylamide was also monitored in FCP after frying. Potato slices pre-treated with sodium ascorbate solution and vacuum-packaged were UV-C irradiated for 0, 3, 5, and 10 min in order to obtain irradiation doses of 0, 1.62, 2.70, and 5.40 kJ m-2, respectively, stored for 23 days (+6 °C), and subsequently boiled and fried. As the applied dose and storage duration increased, the CA content in raw FCP decreased (it retained for 75.53-88.34%), while the content of sugars as well as acrylamide in fried FCP increased. Although the increase was the most noticeable at the applied dose of 2.70 kJ m-2, the acrylamide content was always below proposed limit. Boiling and frying reduced the content of CA and sugars. In spite of certain alterations, applied doses of irradiation can ensure acceptable product in regard to phenolics and sugars, and acrylamide content particularly.

Microwave Assisted Extraction and Pressurized Liquid Extraction of Sulfated Polysaccharides from Fucus virsoides and Cystoseira barbata.

Sulfated polysaccharide fucoidan isolated from brown algae shows a wide range of biological activities that are significantly dependent on its chemical composition, which is closely related to the applied technique and extraction parameters. Therefore, the objective of this study was to evaluate the influence of microwave assisted extraction (MAE) and pressurized liquid extraction (PLE) parameters (solvent, temperature, time, and number of cycles) on the Fucus virsoides and Cystoseira barbata polysaccharide yield (%PS) and chemical composition (total sugar, fucose, and sulfate group). The optimal MAE parameters that resulted in the highest polysaccharide extraction from F. virsoides and C. barbata were 0.1 M H2SO4 for 10 min at 80 °C, while the optimal PLE parameters were 0.1 M H2SO4, for two cycles of 15 min at 140 °C. Furthermore, the %PS, chemical structure, molecular properties, and antioxidant activity of the F. virsoides and C. barbata polysaccharide extracts obtained with MAE, PLE, and conventional extraction (CE) performed under previously determinate optimal conditions were compared. PLE resulted in a significantly higher %PS from F. virsoides, while for C. barbata, a similar yield was achieved with CE and PLE, as well as CE and MAE, for both algae. Furthermore, the polysaccharides obtained using PLE had the highest polydispersity index, fucose, and sulfate group content, and the lowest uronic acid content; however their antioxidant activity was lower.

Bioactive Compounds in Wild Nettle (Urtica dioica L.) Leaves and Stalks: Polyphenols and Pigments upon Seasonal and Habitat Variations.

This study evaluated the presence of bioactives in wild nettle leaves and stalks during the phenological stage and in the context of natural habitat diversity. Thus, wild nettle samples collected before flowering, during flowering and after flowering from 14 habitats situated in three different regions (continental, mountain and seaside) were analyzed for low molecular weight polyphenols, carotenoids and chlorophylls using UPLC-MS/MS and HPLC analysis, while the ORAC method was performed for the antioxidant capacity measurement. Statistical analysis showed that, when compared to the stalks, nettle leaves contained significantly higher amounts of analyzed compounds which accumulated in the highest yields before flowering (polyphenols) and at the flowering stage (pigments). Moreover, nettle habitat variations greatly influenced the amounts of analyzed bioactives, where samples from the continental area contained higher levels of polyphenols, while seaside region samples were more abundant with pigments. The levels of ORAC followed the same pattern, being higher in leaves samples collected before and during flowering from the continental habitats. Hence, in order to provide the product's maximum value for consumers' benefit, a multidisciplinary approach is important for the selection of a plant part as well as its phenological stage with the highest accumulation of bioactive compounds.

The Mastic Tree (Pistacia lentiscus L.) Leaves as Source of BACs: Effect of Growing Location, Phenological Stage and Extraction Solvent on Phenolic Content.

RESEARCH BACKGROUND: Mastic tree (Pistacia lentiscus L.) of the Anacardiaceae family is an evergreen shrub from Mediterranean countries where it is used in traditional medicine. Analysis of P. lentiscus leaf, stem, fruit and root extracts showed high concentrations of principal groups of secondary metabolites (flavonoids, phenolic acids and tannins), suggesting the plant possesses great biological potential. Therefore, the aim of this research is to evaluate the impact of environmental parameters and the extraction solvent type on the concentration of phenols in mastic tree leaf extracts grown at four different locations along the Adriatic coast (Barbariga, Lun, Hvar and Vela Luka) during three phenological stages (early flowering, early fruiting and late fruiting). EXPERIMENTAL APPROACH: Since mastic tree plant has phenolic compounds with different structures and chemical properties, ethanolic and methanolic leaf extracts were analysed using high-performance liquid chromatography (HPLC) coupled with UV/Vis PDA detector. Phenolic compounds were identified by comparing the retention times and spectral data with those of standards at 280 and 340 nm. RESULTS AND CONCLUSIONS: In all samples, phenolic acids and flavonol glycosides were quantified, while catechin was quantified only in methanolic extracts. The 5-O-galloylquinic acid was determined as a predominant phenolic compound in all samples followed by monogalloyl glucose, 3,5-di-O-galloylquinic acid, 3,4,5-tri-O-galloylquinic acid and gallic acid, respectively. Myricetin-3-O-rhamnoside was found to be the predominant flavonol glycoside followed by myricetin-3-O-glucoside, myricetin-3-O-glucuronide, quercetin-3-O-rhamnoside and derivative of flavonol glycoside. The mass concentration of these compounds significantly varied during different phenological stages, at different growing locations and used extraction solvents. The highest phenolic mass concentration was determined in the samples harvested at Hvar growing location and extracted in 80% methanol. The highest total phenolic acid mass concentration was obtained in the samples harvested during the flowering phenological stage and the highest total flavonoid mass concentration in the samples harvested during the early fruiting stage. NOVELTY AND SCIENTIFIC CONTRIBUTION: The obtained data provide a better understanding of the P. lentiscus species phenolic concentration, which can lead to further investigations regarding the valorisation of mastic tree leaves as pharmaceutical products or as food products with added value.

Evaluation of Polyphenolic Profile and Antioxidant Activity of Pistacia lentiscus L. Leaves and Fruit Extract Obtained by Optimized Microwave-Assisted Extraction.

Pistacia lentiscus L. is a Mediterranean shrub known for its health promoting effects attributed to a large extent to polyphenols accumulated in all parts of the plant. Microwave-assisted extraction is a green extraction technique enabling fast and effective isolation of plant polyphenols. Therefore, the aim of this research was to optimize the microwave-assisted extraction of polyphenols from Pistacia lentiscus L. leaves and fruit in terms of temperature, extraction time and microwave power and to evaluate their polyphenolic profile by UPLC/ESI-MS2 and antioxidant capacity by ORAC assay. Optimal extraction conditions for leaf polyphenols were 69 °C, 512 W and 12 min, while for fruit were slightly more intensive-75 °C, 602 W and 15 min. Obtained total phenolic content in leaves and fruit was similar to that obtained after 30 min of the heat-reflux method. The polyphenolic profile of extracts included 34 compounds, with myricetin glycosides being the most abundant compounds among flavonoids in Pistacia lentiscus L. leaves and fruit and gallic acid and its derivates among the phenolic acids. ORAC assay showed higher antioxidant capacity for Pistacia lentiscus L. leaves extract than for fruit, which is in correlation with their respective phenolic content.

UPLC-MS/MS Phytochemical Analysis of Two Croatian Cistus Species and Their Biological Activity.

Aqueous extracts of two Cistus species wild growing in Croatia-Cistus creticus (CC) and Cistus salviifolius (CS)-have been assessed with UPLC-MS/MS, showing 43 different phytochemicals, with flavonol glycosides: myricetin-3-hexoside and myricetin-rhamnoside, predominate ones in CC and myricetin-3-hexoside in CS. Antioxidant potential tested with the FRAP method showed no difference between CS and CC aqueous extracts, while higher phenolic content of CC comparing to CS, determined with a Folin-Cicolateu reagent correlated to its higher antioxidant capacity observed by the DPPH method. Both extracts were assessed for antimicrobial activity, using disc-diffusion and broth microdilution assays, targeting the opportunistic pathogens, associated with food poisoning, urinary, respiratory tract, blood stream and wound infections in humans. Antimicrobial assays revealed that fungi were in general more sensitive to both Cistus aqueous extracts, comparing to the bacteria where two extracts showed very similar activity. The most potent activity was observed against A. baumannii for both extracts. The extracts were tested on human lung cancer (A549) cell line using the MTT assay, showing very similar antiproliferative activity. After 72 h treatment with CC and CS aqueous extracts in concentration of 0.5 g/L, the viability of the cells were 37% and 50% respectively, compared to non-treated cells.

Advanced Technologies for the Extraction of Marine Brown Algal Polysaccharides.

Over the years, brown algae bioactive polysaccharides laminarin, alginate and fucoidan have been isolated and used in functional foods, cosmeceutical and pharmaceutical industries. The extraction process of these polysaccharides includes several complex and time-consuming steps and the correct adjustment of extraction parameters (e.g., time, temperature, power, pressure, solvent and sample to solvent ratio) greatly influences the yield, physical, chemical and biochemical properties as well as their biological activities. This review includes the most recent conventional procedures for brown algae polysaccharides extraction along with advanced extraction techniques (microwave-assisted extraction, ultrasound assisted extraction, pressurized liquid extraction and enzymes assisted extraction) which can effectively improve extraction process. The influence of these extraction techniques and their individual parameters on yield, chemical structure and biological activities from the most current literature is discussed, along with their potential for commercial applications as bioactive compounds and drug delivery systems.