A Novel Approach to Serving Plant-Based Confectionery-The Employment of Spray Drying in the Production of Carboxymethyl Cellulose-Based Delivery Systems Enriched with Teucrium montanum L. Extract.

In this study, spray drying was used as a technological solution for the valorization of Teucrium montanum extract into carboxymethyl cellulose-based delivery systems (CMC), individually or in combination with collagen, guar gum, gum arabic, and kappa-carrageenan. The results showed that the process yield and morphological properties were positively influenced by the introduction of CMC binary blends. The employment of CMC resulted in a high encapsulation efficiency (77-96%) for all phenylethanoid glycosides (PGs) analyzed. Due to the low wettability of the microparticles, a relatively gradual in vitro release of the PGs was achieved. Infusion of the filling with hydrophilic T. montanum extract encapsulated in microparticles with high hydrophobic surface area proved to be a practical route for significant confectionery fortification (5-9 mg PGs per dw serving), ensuring prolonged interaction between the food matrix used and the extract under simulated gastrointestinal conditions. Based on sensory evaluation, the introduction of kudzu starch into the jelly matrix has shown a texture-modifying potential.

Physicochemical and Sensory Stability Evaluation of Gummy Candies Fortified with Mountain Germander Extract and Prebiotics.

Health-conscious consumers seek convenient ways of incorporating different functional ingredients into their diets. Gummy candies are among the most popular confectionery products but generally regarded as nutritionally empty. A gelatin-sugar matrix, providing a highly appreciated sensory experience of sweetness and chewiness, could be used to deliver various bioactive compounds, especially those carrying an unpleasant taste. This work aimed to formulate gelatin gummies based on the mountain germander extract (MGe) as a source of phenylethanoid glycosides (PhEG). Sucrose and glucose syrup contents were partially or completely substituted with combinations of xylitol, maltitol and prebiotic poly- and oligosaccharides. Chemical, textural and sensory parameters were evaluated after production and 2 months of storage. Formulations containing fructooligosaccharides and xylooligosaccharides maintained a characteristic appearance during storage at all three levels of sugar (high, low and none), whereas inulin-added and plain (i.e., without prebiotic) candies suffered from mold contamination or appearance/textural changes. The color of the candies noticeably changed and appeared darker. The PhEG were shown to be stable during the candies' production (approximately 90%) and generally maintained their contents during storage. Texture parameters, except hardness, exhibited high positive correlations and resembled the commercial product. Sensory-wise, a moderate bitterness intensity with a decreasing tendency, along with the high transparency and preservation of the characteristic shape facilitated high general acceptance. Gummy candies with prebiotics were shown to be a highly suitable matrix for the bitter MGe, delivering up to 40 mg of PhEG and 4.5 g of prebiotics in one serving size. This study provides a reference for implementing herbal extracts and emerging prebiotics (XOS) in functional confectionery.

Studying the Functional Potential of Ground Ivy (Glechoma hederacea L.) Extract Using an In Vitro Methodology.

Glechoma hederacea L., known as ground ivy, has a long history of use in folk medicine. The main bioactive compounds in ground ivy are polyphenolic compounds known for their potent antioxidant and antimicrobial activities and thus have high potential as functional ingredients against bacterial infections and the occurrence of chronic diseases associated with oxidative stress in the human body. The aim of the present study was to determine the biological activity of ground ivy extract on selected human cell lines, including hepatic (HepG2), tongue (CAL 27), gastric (AGS) and colon (Caco-2) cancer cell lines by evaluating cytotoxicity, formation of reactive oxygen species and genotoxicity. The antioxidant capacity of the extract was additionally evaluated using cellular model macromolecules of protein and DNA, bovine serum album and plasmid phiX174 RF1 DNA. The effect of ground ivy extract on representatives of human microflora, including L. plantarum, E. coli and S. aureus, was also studied. The cytotoxicity of the extract depended on the type of cells treated, and the pro-oxidant effect generally decreased with increasing exposure time. The most pronounced genoprotective effect against hydroxyl radical damage was monitored in model plasmid DNA and occurred at the highest tested concentration (0.25 mg mL-1), with 95.89% preservation of the supercoiled form of the plasmid. This concentration also had the most significant antioxidant activity on the model protein-14.01% more than the positive control prepared using Trolox. The ground ivy extract showed high antimicrobial potential against the pathogenic bacteria E. coli and S. aureus.

Teucrium montanum L.-Unrecognized Source of Phenylethanoid Glycosides: Green Extraction Approach and Elucidation of Phenolic Compounds via NMR and UHPLC-HR MS/MS.

Health-oriented preferences, a demand for innovative food concepts, and technological advances have greatly influenced changes in the food industry and led to remarkable development of the functional food market. Incorporating herbal extracts as a rich source of bioactive compounds (BC) could be an effective solution to meet the high demand of consumers in terms of expanding the high-quality range of functional foods. The aim of this study is the valorization of the bioactive potential of T. montanum L., an understudied Mediterranean plant species, and the in-depth elucidation of a polyphenolic profile with a UHPLC-HR MS/MS and NMR analysis. The total phenolic content (TPC) and antioxidant capacity (AC) were determined on heat-assisted (HAE), microwave-assisted (MAE) and subcritical water (SWE) extracts. In terms of antioxidant capacity, SWE extracts showed the most notable potential (ABTS: 0.402-0.547 mmol eq Trolox g-1 dw, DPPH: 0.336-0.427 mmol eq Trolox g-1 dw). 12 phenolic compounds were identified in the samples of T. montanum from six microlocations in Croatia, including nine phenylethanoid glycosides (PGs) with total yields of 30.36-68.06 mg g-1 dw and 25.88-58.88 mg g-1 dw in HAE and MAE extracts, respectively. Echinacoside, teupolioside, stachysoside A, and poliumoside were the most abundant compounds HAE and MAE extracts, making T. montanum an emerging source of PGs.

Reduction of oral pathogens and oxidative damage in the CAL 27 cell line by Rosmarinus officinalis L. and Taraxacum officinale Web. extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Dandelion (Taraxacum officinale Web.) and rosemary (Rosmarinus officinalis L.) are treasured botanicals with a long usage history in traditional herbal practices worldwide. Dandelion was used to treat kidney, spleen, and liver disease, as well as cardiovascular disease, diabetes, and bacterial infections, whereas rosemary was used to treat pain, spasms, and to improve blood circulation. AIM OF THE STUDY: The aim of this study was to determine the influence of rosemary and dandelion leaves aqueous extracts on the human tongue epithelial carcinoma cell line (CAL 27) at the level of interaction between oral microbiota and tongue epithelial cells, genomic damage, and H2O2 - induced oxidative damage protection. MATERIALS AND METHODS: The polyphenolic composition of the extracts was determined by spectrophotometric and HPLC analyses. After extract treatment, cytotoxic impact and ROS generation in CAL 27 cells were measured using the MTT assay and the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay, respectively. Microdilutions were applied to investigate the antimicrobial and adhesive properties against representatives of the oral microbiota. The single-cell gel electrophoresis (comet assay) and cytokinesis-blocked micronucleus cytome assay (CBMN cyt) were used to detect induced genomic damages. RESULTS: Both extracts increased the adhesion of the lactic acid bacteria L. plantarum but decreased the adhesion of the bacterial pathogens S. enterica serovar Typhimurium LT21 and E. coli K-12 MG1655 adhesion onto CAL 27 cells. 1 h treatment with 5x concentrated dandelion extract and 1x, 2.5x, and 5x of rosemary extract caused an increase in comet tail intensity. CBMN cyt results demonstrated a significant increase in micronucleus formation even at concentrations several times lower than the usual bioactive compound concentrations found in a cup of beverage, with higher concentrations also inducing cell apoptosis and necrosis. Rosemary extract showed a protective effect against H2O2 - induced oxidative damage by decreasing the apoptotic cell number, probably preventing mutations leading to tumor aggressiveness, invasion, and metastasis. CONCLUSIONS: Both tested extracts demonstrated their usefulness in maintaining good oral bacteria balance and their protective capability as powerful antitumor agents by causing a protective apoptotic effect in tumor cell line already at the dosage of an average daily cup.

Microencapsulation of Dandelion (Taraxacum officinale L.) Leaf Extract by Spray Drying.

Research background: Due to numerous health-promoting properties, dandelion has been used in traditional medicine as a herbal remedy, but also as a food product. Dandelion health benefits are ascribed to the presence of different bioactive compounds in its tissues, among which polyphenols play a significant role. However, the low stability of polyphenols is a critical parameter for their successful implementation into products. Thus, their encapsulation using appropriate carrier vehicles is highlighted as an effective technique for their stabilization and protection. The aim of this study is to microencapsulate dandelion leaf extract using spray drying and different carrier materials for the first time. Experimental approach: In spray drying, low inlet temperature of 130 °C was employed to preserve sensitive dandelion polyphenols, while guar gum, gum arabic, inulin, maltodextrin, pectin and alginate were used as carriers. The influence of different carriers and their content on physicochemical, morphological and colour properties, polyphenolic content and encapsulation efficiency of polyphenols in dandelion powders was examined. Specific polyphenols were determined using HPLC-PAD analysis. Their release profiles and antioxidant capacity in simulated gastrointestinal conditions were also evaluated. Results and conclusions: Compared to plain dandelion powder, carrier-containing dandelion powders have favourably increased solubility, enhanced flow and cohesive properties, reduced particle size and prolonged release of polyphenols under simulated gastrointestinal conditions. Powders were characterized by low moisture content (~2-8%) and high solubility (~92-97%). Chicoric acid was the most abundant compound in dandelion powders. Pectin-dandelion powder showed to be the most effective for microencapsulation of polyphenols, especially for chicoric acid entrapment (74.4%). Alginate-dandelion powder enabled the slowest gradual release of polyphenols. Novelty and scientific contribution: Spray drying at 130 °C and the applied carriers proved to be effective for microencapsulation of dandelion extract, where polyphenolic-rich dandelion powders, due to good physicochemical and encapsulation properties, could serve for the enrichment/production of different functional food products. Also, due to the lack of data on dandelion encapsulation, the obtained results could be of great interest for researchers in the encapsulation field, but also for food industry, especially in the field of instant powders.

Development, Characterization and Incorporation of Alginate-Plant Protein Covered Liposomes Containing Ground Ivy (Glechoma hederacea L.) Extract into Candies.

Ground ivy (Glechoma hederacea L.) has been known as a medicinal plant in folk medicine for generations and, as a member of the Lamiaceae family, is characterized with a high content of rosmarinic acid. The aim of the present study was to formulate delivery systems containing bioactive compounds from ground ivy in encapsulated form and incorporated into candies. Liposomes were examined as the encapsulation systems that were additionally coated with an alginate-plant protein gel to reduce leakage of the incorporated material. Bioactive characterization of the ground ivy extract showed a high content of total phenolics (1186.20 mg GAE/L) and rosmarinic acid (46.04 mg/L). The formulation of liposomes with the high encapsulation efficiency of rosmarinic acid (97.64%), with at least a double bilayer and with polydisperse particle size distribution was achieved. Alginate microparticles reinforced with rice proteins provided the highest encapsulation efficiency for rosmarinic acid (78.16%) and were therefore used for the successful coating of liposomes, as confirmed by FT-IR analysis. Coating liposomes with alginate-rice protein gel provided prolonged controlled release of rosmarinic acid during simulated gastro-intestinal digestion, and the same was noted when they were incorporated into candies.

Comprehensive Study of Traditional Plant Ground Ivy (Glechoma hederacea L.) Grown in Croatia in Terms of Nutritional and Bioactive Composition.

In the present study, ground ivy was harvested from different natural habitats in Croatia and subjected to screening analysis for nutritional and bioactive composition. To achieve maximum recovery of phenolic compounds, different extraction techniques were investigated-heat-assisted (HAE), microwave-assisted (MAE) and subcritical water (SWE) extraction. Prepared extracts were analysed by spectrophotometric methods, LC-MS/MS and HPLC-PAD methodologies. Results regarding nutritive analyses, conducted using standard AOAC methods, showed the abundance of samples in terms of insoluble dietary fibre, protein, calcium and potassium, while rutin, chlorogenic, cryptochlorogenic, caffeic and rosmarinic acid were the most dominant phenolic compounds. In addition, LC-MS/MS analysis revealed the presence of apigenin and luteolin in glycosylated form. Maximum recovery of target phenolic compounds was achieved with MAE, while SWE led to the formation of new antioxidants, which is commonly known as neoformation. Moreover, efficient prediction of phenolic composition of prepared extracts was achieved using NIR spectroscopy combined with ANN modelling.

Development and Validation of HPLC-DAD Method with Pre-Column PMP Derivatization for Monomeric Profile Analysis of Polysaccharides from Agro-Industrial Wastes.

The instrumental analysis of complex mixtures of sugars often requires derivatization to enhance the method's selectivity and sensitivity. 1-Phenyl-3-methyl-5-pyrazolone (PMP) is a common sugar derivatization agent used in high-performance liquid chromatography (HPLC). Although many C18 column applications for PMP-sugar derivative analysis have been developed, their transferability is not straightforward due to variations in column chemistry and preparation technology. The aim of this study was to develop and validate an application for Zorbax Extend C18 columns for the analysis of 8 neutral and 2 acidic sugars commonly found in plant polysaccharides. The method was further compared to well-established alditol acetates and m-hydroxydiphenyl methods and employed for sugar profiling of selected agro-industrial wastes. The most influential separation factors were the mobile-phase pH and acetonitrile content, optimized at 8.0 and a 12-17% gradient, respectively. The method showed excellent linearity, repeatability and intermediate precision. High sensitivity was achieved, especially for neutral sugars, with an accuracy error range of 5-10% relative standard deviation. The sugar profiling results were highly correlated to the reference method for neutral sugars. The HPLC method was highly applicable for the evaluation of polysaccharides in selected wastes and showed advantages in terms of simplicity, accuracy in acidic sugar determination and suitability for their simultaneous analysis with neutral sugars.

Antioxidant and Sensory Assessment of Innovative Coffee Blends of Reduced Caffeine Content.

Considering the current trend in the global coffee market, which involves an increased demand for decaffeinated coffee, the aim of the present study was to formulate coffee blends with reduced caffeine content, but with pronounced antioxidant and attractive sensory properties. For this purpose, green and roasted Arabica and Robusta coffee beans of different origins were subjected to the screening analysis of their chemical and bioactive composition using standard AOAC, spectrophotometric and chromatographic methods. From roasted coffee beans, espresso, Turkish and filter coffees were prepared, and their sensory evaluation was performed using a 10-point hedonic scale. The results showed that Arabica coffee beans were richer in sucrose and oil, while Robusta beans were characterized by higher content of all determined bioactive parameters. Among all studied samples, the highest content of 3-O-caffeoylquinic acid (14.09 mg g-1 dmb), 4-O-caffeoylquinic acid (8.23 mg g-1 dmb) and 5-O-caffeoylquinic acid (4.65 mg g-1 dmb), as well as caffeine (22.38 mg g-1 dmb), was detected in roasted Robusta beans from the Minas Gerais region of Brazil, which were therefore used to formulate coffee blends with reduced caffeine content. Robusta brews were found to be more astringent and recognized as more sensorily attractive, while Arabica decaffeinated brews were evaluated as more bitter. The obtained results point out that coffee brews may represent a significant source of phenolic compounds, mainly caffeoylquinic acids, with potent antioxidant properties, even if they have reduced caffeine content.

The multiple-response modeling of heat-assisted, microwave-assisted and subcritical water extraction on selected phenolics from traditional plant species Teucrium montanum.

The growing popularity of integration of functional compounds into food and pharmaceutical products has greatly influenced the intense investigation of aromatic plants as its notable sources. One of the insufficiently explored plant species-Mountain Germander (Teucrium montanum)-may represent a prominent source of biologically active components-polyphenols. Hence, to maximally enhance their recovery, conventional (HAE) and non-thermal extraction methods-microwave-assisted extraction (MAE) and subcritical water extraction (SWE) were employed by using multiple response surface methodology (RSM). Optimization of temperature, solid-to-solvent ratio, and extraction time was evaluated by measuring total phenolic content (TPC) and antioxidant capacity (DPPH and ABTS assays), with the further estimation of the extraction efficiency of two selected phenylethanoid glycosides present in final extracts-verbascoside and echinacoside determined by HPLC analysis. The optimum HAE, MAE, and SWE extraction conditions with the highest measured dependent variables were, as follows: 100 °C, 0.5 g/50 mL and 30 min, 90 °C, 0.5 g/50 mL and 9 min, 200 °C, 0.5 g/50 mL and 15 min, respectively. Regarding examined phenylethanoid glycosides, HAE extract showed to be its richest source (16.64 mg/g dw) while SWE was estimated as at least suitable extraction technique (5.42 mg/g dw). In conclusion, this study outlined the most appropriate extraction technique and conditions for the recovery of selected phenolic compounds using RSM and gave closely insight into the antioxidative potential of valuable ethnomedicinal plant from the Balkan peninsula-Teucrium montanum.

Formulation and characterization of liposomal encapsulated systems of bioactive ingredients from traditional plant mountain germander (Teucrium montanum L.) for the incorporation into coffee drinks.

Conventional and innovative (microwave-assisted and subcritical water extraction) techniques were applied to investigate the bioactive content of traditional plant - Teucrium montanum. Verbascoside and echinacoside, identified and quantified using LC-MS/MS and HPLC-PAD, were found to be the predominant phenolics in all extracts. Infusion (30 °C, 30 min) was characterized with the highest total phenolic content and antioxidant capacity and was further used for encapsulation into liposomes. Formulation of liposomes with a high encapsulation efficiency of echinacoside (68.27%) and verbascoside (80.60%), satisfactory physical properties, including size (326.2 nm) and polydispersity index (0.34), was achieved, although determined zeta potential (-23.03 mV) indicated their instability. Formulated liposomes were successfully coated with pectin and alginate that was also proved by FTIR analysis. Liposomes coated with pectin showed the most desirable in vitro digestion release of verbascoside and echinacoside, while alginate as liposome surface layer proved to be more appropriate for their retention during storage time.

Structuring new alginate network aimed for delivery of dandelion (Taraxacum officinale L.) polyphenols using ionic gelation and new filler materials.

Alginate hydrogels are often used for immobilization of plant-derived bioactive compounds by fast and simple ionic gelation technique. However, the structure of alginate gel network is very porous and mostly result with high-diffusion rates of encapsulated compound, what limits its application as delivery vehicle. In order to prevent losses of bioactives and prepare efficient encapsulation systems, the aim of this study was to evaluate a potential of new natural fillers, cocoa powder (CP) and carob (C) for structuring alginate network aimed for encapsulation of aqueous dandelion (Taraxacum officinale L.) leaf extract using ionic gelation. Whey protein isolates served as a standard filler. The influence of different concentrations of gelling medium (2% and 3% calcium chloride) on encapsulation properties of alginate systems was also evaluated. Calcium concentration affected morphological properties (more acceptable when using 3% CaCl2), while textural properties and encapsulation efficiency of polyphenols and retained antioxidant capacity were more influenced by selected delivery materials. Alginate-whey protein isolates beads were scored with the highest loading capacity of polyphenols (>93%), while newly formulated binary mixtures (alginate-cocoa powder and alginate-carob) also enabled highly efficient entrapment of polyphenols (>88%). The slowest release of polyphenols in simulated gastrointestinal fluids were obtained when alginate was combined with CP and C, where system alginate-cocoa powder prepared with lower concentration of calcium chloride (2% CaCl2) enabled the most extended release of total polyphenols and hydroxycinnamic acids. Obtained results strongly justified implementation of new plant-derived functional fillers (cocoa powder and carob) for encapsulation purposes and opened new directions for designing of binary carrier's.

The Potential of Combined Emulsification and Spray Drying Techniques for Encapsulation of Polyphenols from Rosemary (Rosmarinus officinalis L.) Leaves.

The present study evaluates the potential of encapsulation of polyphenolic antioxidants from rosemary (Rosmarinus officinalis L.) leaves by combining emulsification and spray drying techniques. To stabilize the emulsions and prepare samples suitable for use in dry products, double emulsions encapsulating rosemary polyphenolic extract and containing polyglycerol polyricinoleate (4%), whey protein isolates (2 and 4%) as emulsifiers, and maltodextrins (MDE 10 and 21) as enhancing coatings were subjected to spray drying. The obtained results show insignificant (p>0.05) effect of used maltodextrin type and protein content on mean particle size of double emulsions containing rosemary polyphenols. Morphology analyses showed that double emulsions were successfully prepared, spherical microcapsules were obtained after spray drying of double emulsions and double emulsion form was still preserved after rehydration of spray-dried microcapsules. Regardless of used maltodextrins, significantly (p>0.05) higher encapsulation efficiencies (EE) of total polyphenols (39.57 and 42.83%) in rehydrated samples were achieved when higher protein content (4% whey protein isolate) was used, indicating the major impact of protein content on EE of rosemary polyphenols. Also, using HPLC analysis, rosmarinic and caffeic acids, apigenin and luteolin derivatives were detected among specific polyphenols, where rosmarinic acid had notable encapsulation efficiency ranging from 62.15 to 67.43%. In this way, the obtained microcapsules encapsulating rosemary polyphenols could be easily blended with various dry mixtures, and serve for delivery in different functional products.