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.

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.

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.

Antioxidants and Bioactive Compounds in Food: Critical Review of Issues and Prospects.

This review paper gives an insight into the effective delivery mechanisms for health-promoting substances and highlights the challenges of using antioxidants and bioactives in foods. The selection criteria for choosing bioactives and their extraction in bioavailable form with their adequate incorporation techniques and delivery mechanisms are covered. Moreover, an overview of existing methods for determination of bioactivity is given. The importance of scientifically evaluating the effects of foods or food components on consumer health before making claims about the healthiness is aligned. Finally, a scientific perspective on how to respond to the booming demand for health-promoting products is given, and we acknowledge that despite the work done, there are still many challenges that need to be overcome.

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.

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.

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.

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.

UPLC-MS2 Profiling of Blackthorn Flower Polyphenols Isolated by Ultrasound-Assisted Extraction.

Ultrasound-assisted extraction (UAE) was optimized for fast and effective isolation of blackthorn flower polyphenols. The effects of ethanol concentration (50% and 70%) in extraction solvent, ultrasound amplitude (50%, 75%, and 100%) and extraction time (3, 6, and 9 min) were evaluated. UPLC-MS2 was used to determine phenolic profile of blackthorn flowers extracts. A total of 28 different phenolic compounds were identified, belonging to the classes of hydroxycinnamic acids, flavonol glycosides and flavanols. Kaempferol and quercetin derivatives were the most abundant compounds represented by kaempferol-pentoside and rhamnoside in concentrations up to 494.94 and 436.62 mg/100 g and quercetin-pentoside in concentration up to 226.75 mg/100 g. Observed parameters of UAE did not affect the qualitative phenolic composition of blackthorn flower extracts. On the contrary, the concentration of polyphenols increased with higher ethanol concentration in solvent, higher ultrasound amplitude and prolongation of extraction time. The established conditions for optimal extraction of blackthorn flower polyphenols using ultrasound were: 70% ethanol as extraction solvent, 75% of ultrasound amplitude and 9 min extraction time, resulting in extract rich in polyphenols with a great potential for use in pharmaceutical and food industry. PRACTICAL APPLICATION: In this study, ultrasound-assisted extraction (UAE) was optimized in order to establish fast and effective method for production of crude blackthorn flower extract rich in polyphenols. UPLC-MS2 analysis showed 28 different phenolic compounds belonging to the classes of hydroxycinnamic acids, flavonol glycosides, and flavanols with quercetin and kaempferol glycosides in the highest concentrations, implicating high potential of blackthorn flowers' extract for use in food and pharmaceutical industries.