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.

Optimisation of Almond-Based Dairy-Free Milk Alternative Formulation Fortified with Myrtle, Bay Leaf and Fennel Extracts§.

Research background: Herbs and spices used in traditional medicine are nowadays increasingly used in combinations to create functional food formulations aimed at treating specific symptoms and disorders. Among herbs originating from the Mediterranean region, extracts of myrtle (Myrtus communis L.), bay leaf (Laurel nobilis L.) and fennel (Foeniculum vulgare Mill.) are traditionally used for gastrointestinal disorders. When considering how to incorporate these extracts into products, dairy-free milk alternatives provide an excellent base with almond-based drinks being among the most popular within this group. Experimental approach: The aim of this study is therefore to optimise the formulation of an almond drink fortified with a 25 % (on dry mass basis) aqueous herbal extract containing myrtle, bay leaf (25 % each) and fennel seed (50 %) extracts. A central composite design with 20 formulations varied the content of φ(aqueous herbal extract)=2-6 %, lecithin as emulsifier 0.15-0.45 and xylitol as sweetener 2-5 % (m/V), while antioxidant activity, total phenolic content and sensory properties were determined as dependent variables. Results and conclusions: The antioxidant activity and total phenolic content of the prepared almond drink formulations increased with the amount of added concentrated aqueous herbal extracts, as did the colour, herbal odour and flavour, bitterness and aftertaste. The addition of lecithin resulted in a darker colour and the sweetness was increased by the xylitol content. All formulations had good overall attractiveness, which increased with higher xylitol content. Novelty and scientific contribution: The current work offers new insights into the optimisation of fortified dairy-free alternatives. The addition of myrtle, bay leaf and fennel seed extracts to almond drink-based formulations resulted in a 12-fold increase in antioxidant activity. Xylitol masks the potential bitterness of the phenolic compounds so that higher amounts of extracts can be added.

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.

The Effect of a High-Protein Diet Supplemented with Blackthorn Flower Extract on Polyphenol Bioavailability and Antioxidant Status in the Organs of C57BL/6 Mice.

The health benefits of polyphenols are based on their bioavailability, which is why a significant portion of research focuses on factors that affect their bioavailability. Previous studies suggest that the intake of polyphenols along with macronutrients in food represents one of the key factors influencing the bioavailability of polyphenols and, consequently, their biological activity in the organism. Since polyphenols in the human diet are mainly consumed in food together with macronutrients, this study investigated the in vivo absorption, metabolism, and distribution of polyphenolic compounds from the water extract of blackthorn flower (Prunus spinosa L.) in combination with a protein-enriched diet in the organs (small intestine, liver, kidney) of C57BL/6 mice. The bioaccumulation of polyphenol molecules, biologically available maximum concentrations of individual groups of polyphenol molecules, and their effect on the oxidative/antioxidative status of organs were also examined. The results of this study indicate increased bioabsorption and bioavailability of flavan-3-ols (EC, EGCG) and reduced absorption kinetics of certain polyphenols from the groups of flavonols, flavones, and phenolic acids in the organs of C57BL/6 mice after intragastric administration of the water extract of blackthorn flower (Prunus spinosa L.) in combination with a diet enriched with whey proteins. Furthermore, subchronic intake of polyphenols from the water extract of blackthorn flower (Prunus spinosa L.) in combination with a diet enriched with whey proteins induces the synthesis of total glutathione (tGSH) in the liver and superoxide dismutase (SOD) in the liver and small intestine. The results of this study suggest potential applications in the development of functional foods aimed at achieving the optimal health status of the organism and the possibility of reducing the risk of oxidative stress-related disease.

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.

Sterols and pentacyclic triterpenoids from nettle root: content and composition as affected by pressurized liquid extraction.

BACKGROUND: Nettle is a medicinal plant rich in bioactive molecules. The composition of nettle leaves and stems has been extensively studied, whereas the root has been insufficiently investigated. Therefore, the present study aimed to optimize the parameters of advanced extraction technique, pressurized liquid extraction (PLE), for the lipid fraction of nettle root rich in triterpenoid derivatives and to compare the efficiency of isolation under optimal conditions with conventional Soxhlet extraction (SE). RESULTS: The PLE yields ranged from 0.39-1.63%, whereas the total content of triterpenoid derivatives ranged from 43.50-78.26 mg 100 g-1 , with nine sterols and three pentacyclic triterpenoids identified and quantified within a total range of 42.81-76.57 mg 100 g-1 and 0.69-1.68 mg 100 g-1 dried root, respectively. The most abundant sterol and pentacyclic triterpenoid were ß-sitosterol and ß-amyrin acetate, with mean values of 50.21 mg 100 g-1 and 0.56 mg 100 g-1 dried root. CONCLUSION: The optimal PLE conditions were 150 °C/5 min/four cycles and showed significantly better performance compared to SE (68 °C, 8 h), establishing an excellent technique for the isolation of the nettle root lipid fraction. Also, triterpenoid derivatives from nettle could be used as functional ingredients for the development of new foods and dietary supplements. © 2022 Society of Chemical Industry.

The Effects of Nettle Extract Consumption on Liver PPARs, SIRT1, ACOX1 and Blood Lipid Levels in Male and Female C57Bl6 Mice.

The aim of this study was to evaluate how nettle (Urtica dioica L.) water extract consumption would interact with regulators of peroxysomal lipid oxidation, histone deacetylase, and markers of oxidative stress in the liver and blood lipid levels in male and female C57Bl6 mice. Metabolically unchallenged (healthy) mice (n = 5 per sex) were treated with a nettle extract in a dose of 40 mg of total polyphenols in the extract per kg mice body weight. The nettle extract was applied daily along with normal diet for 15 days. The serum triglycerides, cholesterol, HDL, LDL, and liver PPAR-α, PPAR-γ, PGC-1-α, ACOX1, SIRT1, MDA, SOD, CAT, and GSH were compared between exposed and unexposed (control) animals. In males, the PPAR-α, PGC1-α, and ACOX1 levels together with systemic HDL cholesterol were significantly (p ≤ 0.05) increased while the LDL cholesterol decreased (p ≤ 0.05). In females, no changes in PPAR-α and PGC1-α or serum lipids were noted, but the ACOX1 content in the liver was significantly (p ≤ 0.05) increased. The SIRT1 activity increased (p ≤ 0.05) only in females. In both sexes, the PPAR-γ levels were not significantly (p ≤ 0.05) affected in either sex. The results indicate that nettle plant extract has the potential to modulate selected transcriptional factors and histone deacetylase in vivo, with certain sex differences, which should be studied further in similar models.

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.

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.

Antioxidant and Anti-Atherogenic Activities of Essential Oils from Myrtus communis L. and Laurus nobilis L. in Rat.

Essential oils (EOs) from aromatic and medicinal plants, such as myrtle (Myrtus communis L.) and Laurel (Laurus nobilis L.), are gaining popularity as a potential ingredient in functional foods and nutraceuticals. This study aims to investigate whether the essential oils (EOs) could be effective in weight control, antioxidative and antilipidemic status of rats by affecting microbiota and its enzymes activity and whether changes in intestinal enzyme activity affect the health of rats. The intragastric application of laurel and myrtle EOs to rats for two weeks affects weight loss, reduces glycolytic activity, lipid parameters (cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-C) and very low-density lipoprotein cholesterol (VLDL-C)) and atherogenic indicators, leading to cardiovascular protection. Laurel EO can be an excellent candidate for the treatment of drug-induced obesity and related diseases, since it affects lipid metabolism in the liver and inhibits the enzymes responsible for the metabolism of carbohydrates into glucose in the digestive tract, leading to weight loss. In contrast, myrtle EO shows a better antioxidant capacity in most tissues, except kidneys, where it causes a pro-oxidative effect, compared to laurel EO. Myrtle EO increases the permeability and instability of the erythrocyte membrane, resulting in a loss of selectivity for the entry of toxic substances into the cell. On the other hand, myrtle EO leads to intestinal inflammation by reducing the number of probiotic bacteria and increasing Enterobacter.

Isolation of Laurus nobilis Leaf Polyphenols: A Review on Current Techniques and Future Perspectives.

In recent years, the market demand for products enhanced with ingredients derived from natural products, such as polyphenols, is rapidly increasing. Laurus nobilis L., known as bay, sweet bay, bay laurel, Roman laurel or daphne is an evergreen Mediterranean shrub whose leaves have traditionally been used in cuisines and folk medicine due to their beneficial health effects, which can nowadays be scientifically explained by various biological activities of the leaf extracts. Many of these activities can be attributed to phenolic compounds present in L. nobilis leaves which include flavonoids, phenolic acids, tannins (proanthocyanidins) and lignans. In order to enable efficient industrial utilization of these valuable compounds, it is crucial to establish optimal extraction procedures resulting in the highest yields and quality of the extracts. This paper offers the first systematic review of current literature on the influence of conventional and advanced extraction techniques, including microwave-assisted, ultrasound-assisted, enzyme-assisted, supercritical-CO2 and mechanochemical-assisted extraction on the phenolic content of L. nobilis leaf extracts, allowing more efficient planning of further research and simplifying the steps towards industrial utilization of this plant.

Blackthorn Flower Extract Impact on Glycaemic Homeostasis in Normoglycaemic and Alloxan-Induced Hyperglycaemic C57BL/6 Mice.

RESEARCH BACKGROUND: The use of plants and their extracts in treatments of chronic diseases is widely known in traditional medicine. The aim of this study is to determine the effects of 10-day consumption of blackthorn (Prunus spinosa L.) flower extract on blood glucose, glycaemic load, serum α-amlyase activity and insulin concentration in normoglycaemic and hyperglycaemic (alloxan-induced) mice model. EXPERIMENTAL APPROACH: Normoglycaemic and hyperglycaemic (treated with alloxan, 150 mg per kg body mass) C57BL/6 mice were administered daily, during 10 days, blackthorn flower extract by gavage. The sugar mass concentration within the extract was determined by HPLC analysis. In mice, blood and serum blood glucose concentrations, and oral glucose tolerance test were determined by blood glucometer. Serum insulin concentration was determined by ELISA assay and α-amylase activity by colourimetric assay. RESULTS AND CONCLUSIONS: The blackthorn flower extract increased glucose concentrations in normoglycaemic mice by 30% after the 1st and 5th day and by 17% after the 10th day of consumption. It is a consequence of released sugars because sugar analysis revealed 59.8 mg/L monosaccharides, mainly fructose (55.7 mg/L) and glucose (24.3 mg/L) in the extract. On the contrary, the extract consumption reduced serum blood glucose in hyperglycaemic mice by 29% after 10 days of treatment. Oral glucose tolerance test also confirmed that in the hyperglycaemic group treated with blackthorn flower extract glucose homeostasis was improved and showed decrease in blood glucose. Serum insulin concentration increased by 49% and serum α-amylase activity by 46% after 10 days of treatment with blackthorn flower extract in hyperglycaemic group. Thus, it can be concluded that blackthorn flower extract improved glucose tolerance, enhanced insulin secretion and lowered serum α-amylase activity. NOVELTY AND SCIENTIFIC CONTRIBUTION: The obtained results show for the first time the potential of blackthorn (Prunus spinosa L.) flower extract in hyperglycaemia management.

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.

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.