Multiple liquid-liquid extraction of dissolved compounds in immortelle hydrosol with four different solvents.

Immortelle, a revered Mediterranean medicinal plant, is celebrated for its potent essential oil renowned in the cosmetic industry for its skin-enhancing properties. Yet, immortelle hydrosol, an often-overlooked byproduct, holds promise in cosmetics due to its compatibility with polar active ingredients. This study investigates the chemical composition of immortelle hydrosol by employing liquid-liquid extraction (LLE) to transfer volatile organic components into nonpolar solvents. Four solvents - chloroform, dichloromethane, hexane, and benzene - were assessed through ten consecutive extractions from industrially produced immortelle hydrosol. Quantification was achieved using GC analysis with tetradecane as an internal standard. Chloroform emerged as the most efficient solvent, yielding 2447.0 mg/L of volatile compounds, surpassing dichloromethane, hexane, and benzene. Key compounds in immortelle hydrosol included 3-pentanone, 2-methyl-1-butanol, and γ-terpineol. Importantly, the study revealed that a portion of essential oil compounds persists in the hydrosol even after ten LLE cycles, with optimal results achievable in five extractions (~92 % in most cases).

Leaves micromorphology, chemical profile, and bioactivity of in vitro-propagated Nepeta cyrenaica (Lamiaceae).

INTRODUCTION: The endemic species Nepeta cyrenaica Quézel & Zaffran, native to northeastern Libya, is valued as an important honey-bearing plant. OBJECTIVES: This study was aimed to examine the micromorphology, phytochemistry, and bioactivity of in vitro-propagated N. cyrenaica for the first time. MATERIALS AND METHODS: The leaf indumentum was examined using light and scanning electron microscopy and further characterised for histochemistry. The chemical composition of essential oil (EO) was performed using GC-MS analysis, while dichloromethane (DCM), methanol (ME), ethanol (ET), and aqueous (AQ) extracts were analysed using qualitative and quantitative LC/MS analyses. The antioxidant activities of EO and extracts were assessed using three parallel assays, while enzyme-inhibiting effects were evaluated against four enzymes. RESULTS: The leaves bear various types of glandular trichomes, with lipophilic secretion predominating. The main EO component of EO was 1,8-cineole. A considerable number of phenolics and iridoids were tentatively identified in the ME extract. Quantitative LC/MS analysis confirmed that ferulic acid, rosmarinic acid, and epigallocatechin gallate were present in the highest amount in the extracts, in which three iridoids were also quantified. Although the ME extract contained the highest amount of polyphenolics and iridoids, the DCM extract showed the best overall biological potential. Additionally, EO exerted the strongest acetylcholinesterase and tyrosinase inhibition. CONCLUSION: This study demonstrated that the endemic N. cyrenaica can be efficiently grown under in vitro conditions, where it develops various glandular trichomes that are thought to secrete and/or accumulate bioactive compounds with valuable medicinal potential.

Antifungal Activity of Lavandula angustifolia Essential Oil against Candida albicans: Time-Kill Study on Pediatric Sputum Isolates.

The aim of our study was to determine the susceptibility of 15 Candida albicans sputum isolates on fluconazole and caspofungin, as well as the antifungal potential of Lavandula angustifolia essential oil (LAEO). The commercial LAEO was analyzed using gas chromatography-mass spectrometry. The antifungal activity was evaluated using EUCAST protocol. A killing assay was performed to evaluate kinetics of 2% LAEO within 30 min treatment. LAEO with major constituents' linalool (33.4%) and linalyl acetate (30.5%) effective inhibited grows of C. albicans in concentration range 0.5-2%. Fluconazole activity was noted in 67% of the isolates with MICs in range 0.06-1 µg/mL. Surprisingly, 40% of isolates were non-wild-type (non-WT), while MICs for WT ranged between 0.125-0.25 µg/mL. There were no significant differences in the LAEO MICs among fluconazole-resistant and fluconazole-susceptible sputum strains (p = 0.31) and neither among caspofungin non-WT and WT isolates (p = 0.79). The 2% LAEO rapidly achieved 50% growth reduction in all tested strains between 0.2 and 3.5 min. Within 30 min, the same LAEO concentration exhibited a 99.9% reduction in 27% isolates. This study demonstrated that 2% solution of LAEO showed a significant antifungal activity which is equally effective against fluconazole and caspofungin susceptible and less-susceptible strains.

Tamjanika, a Balkan native variety of Vitis vinifera L.: Chemical characterization, antibacterial, and anti-dermatomycosis potential of seed oil.

This study was designed to explore functional food properties of edible seed oil obtained from Tamjanika seeds-autochthonous grape variety of Balkan Peninsula. In order to accomplish our goals, seed oil was isolated by Soxhlet apparatus and chemically characterized regarding fatty acids, carotenoids, tocopherols, and tocotrienols. Antimicrobial activity of the isolated oil was tested by microdilution method. For that purposes, six bacterial species were used, belonging to human infectious agents and food contaminants. Furthermore, the activity of the oil was investigated against clinical isolates of dermatomycetes. Our study has shown that oil of Vitis vinifera L. Tamjanika variety was an abundant source of polyunsaturated fatty acids (81.43%) with predominant linoleic acid. HPLC analysis revealed the presence of carotenoid lutein (0.15 mg/100 g). The seed oil was rich in tocotrienols (85.04 mg/100 g) predominating over tocopherols (8.37 mg/100 g). The oil possessed microbicidal activity against all the tested microbes. Bacteria were more sensitive to the effect of the oil (minimum inhibitory concentration [MIC] 7.7-15.4) when compared with oil effect on tested dermatomycetes (MIC 20-40). Our investigation has shown for the first time that grape oil could be active against wide spectrum of bacteria and clinically isolated dermatomycetes. The significance of this study lies in the fact that it pointed out the functional food properties of grape seed oil that was fully chemically characterized.

Population structure and adaptive variation of Helichrysum italicum (Roth) G. Don along eastern Adriatic temperature and precipitation gradient.

Immortelle (Helichrysum italicum (Roth) G. Don; Asteraceae) is a perennial plant species native to the Mediterranean region, known for many properties with wide application mainly in perfume and cosmetic industry. A total of 18 wild H. italicum populations systematically sampled along the eastern Adriatic environmental gradient were studied using AFLP markers to determine genetic diversity and structure and to identify loci potentially responsible for adaptive divergence. Results showed higher levels of intrapopulation diversity than interpopulation diversity. Genetic differentiation among populations was significant but low, indicating extensive gene flow between populations. Bayesian analysis of population structure revealed the existence of two genetic clusters. Combining the results of FST - outlier analysis (Mcheza and BayeScan) and genome-environment association analysis (Samßada, LFMM) four AFLP loci strongly associated with the bioclimatic variables Bio03 Isothermality, Bio08 Mean temperature of the wettest quarter, Bio15 Precipitation seasonality, and Bio17 Precipitation of driest quarter were found to be the main variables driving potential adaptive genetic variation in H. italicum along the eastern Adriatic environmental gradient. Redundancy analysis revealed that the partitioning of genetic variation was mainly associated with the adaptation to temperature oscillations. The results of the research may contribute to a clearer understanding of the importance of local adaptations for the genetic differentiation of Mediterranean plants and allow the planning of appropriate conservation strategies. However, considering that the identified outlier loci may be linked to genes under selection rather than being the target of natural selection, future studies must aim at their additional analysis.

Characterization, Antioxidant and Antibacterial Activity of Essential Oils and Their Encapsulation into Biodegradable Material Followed by Freeze Drying.

The study assessed the antimicrobial and antioxidant activities of commonly used and commercially available essential oils as an alternative to synthetic preservatives. The plant sources were as follows: lavender (Lavandula angustifolia), tea tree (Melaleuca alternifolia), bergamot (Citrus bergamia) and peppermint (Mentha piperita). The antioxidant activity of essential oils was tested by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2´-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods. The microdilution broth susceptibility assay revealed that lavender and bergamot essential oils were more efficient in inhibiting the bacterial growth than other tested oils, with the minimum inhibitory concentration of 5 µg/mL. This study also reports the successful implementation of an electrostatic extrusion technique for encapsulating essential oils into alginate beads, which enables the essential oils to maintain their free radical scavenging ability over time.

Sandy Everlasting (Helichrysum arenarium (L.) Moench): Botanical, Chemical and Biological Properties.

Sandy everlasting [Helichrysum arenarium (L.) Moench] is herbaceous perennial plant belonging to Asteraceae family and it is native to Europe, Central Asia, and China. It belongs to the section HELICHRYSUM (Asteraceae family, genus Helichrysum) along with H. plicatum DC. Prodr., which very similar phenolic profile and H. italicum (Roth), which is widely used for essential oil extraction. Its flowers have a long tradition in European ethnomedicine as a cholagogue, choleretic, hepatoprotective, and detoxifying herbal drug. The flowers are rich in phenolic compounds including flavonoids, chalcones, phenolic acids, coumarins, and pyrones. Apart from polyphenols, other compounds such as sterols, lignans, and glycosides of aromatic compounds have been also isolated from H. arenarium. The majority of authors confirm that the most important group of compounds responsible for biological activities is flavonoids. Moreover, significant activities of naringenin, one of the main flavonoids of H. arenarium, were reported. On the other hand, there are no clinical data about testing the extracts or preparations based on H. arenarium. Although H. arenarium is well known in phytotherapy for its potential in the treatment of gallbladder disease and are classified as endangered species in a number of European countries, very few data about its cultivation are available in the literature.

Sorbus aucuparia and Sorbus aria as a Source of Antioxidant Phenolics, Tocopherols, and Pigments.

Due to its nutritive and medicinal properties, berries of some Sorbus species are used for the preparation of jams and jelly as well as in traditional medicine. On the other hand, their chemical composition is not much studied especially of those grown in Balkan Peninsula. We have analyzed individual phenolics, tocopherols, carotenoids and chlorophylls using HPLC in berries from Sorbus aucuparia and Sorbus aria collected in different localities in Serbia and Montenegro together with the amounts of total phenolics and proanthocyanidins as well as their radical scavenging activity against DPPH radical. Berries of S. aucuparia were richer source of polyphenolics in comparision with S. aria and, regardless the species and locality, caffeoylquinic acids such as neochlorogenic and chlorogenic acid were the most abundant compounds. Among analyzed tocopherols the most abundant in all samples was α-tocopherol (0.48 - 19.85 µg/g dw) as it was ß-carotene among carotenoids (mean concentration of 0.98 µg/g dw in S. aucuparia and 0.40 µg/g dw in S. aria, respectively). Correlation between total phenolics and DPPH radical scavenging activity was noticed. Our study represents comprehensive report on chemical composition of S. aucuparia and S. aria which could contribute to a better understanding of their quality.

Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique.

Traditional maceration method was used for the extraction of polyphenols from chokeberry (Aronia melanocarpa) dried fruit, and the effects of several extraction parameters on the total phenolics and anthocyanins contents were studied. Various solvents, particle size, solid-solvent ratio and extraction time have been investigated as independent variables in two level factorial design. Among examined variables, time was not statistically important factor for the extraction of polyphenols. The optimal extraction conditions were maceration of 0.75mm size berries by 50% ethanol, with solid-solvent ratio of 1:20, and predicted values were 27.7mgGAE/g for total phenolics and 0.27% for total anthocyanins. Under selected conditions, the experimental total phenolics were 27.8mgGAE/g, and total anthocyanins were 0.27%, which is in agreement with the predicted values. In addition, a complementary quantitative analysis of individual phenolic compounds was performed using HPLC method. The study indicated that maceration was effective and simple technique for the extraction of bioactive compounds from chokeberry fruit.

Dietary supplementation with polyphenol-rich chokeberry juice improves skin morphology in cellulite.

The aim of our study was to investigate possible beneficial effects of organic chokeberry juice (OCJ) consumption in the treatment of cellulite. Twenty-nine women aged 25-48 with a cellulite grade 2 according to the Nurnberger-Muller scale were included. Anthropometric and biochemical parameters were measured. Skin structure was analyzed by ultrasonography. All subjects consumed 100 mL of OCJ per day, during 90 days. Measurements of investigated parameters were performed at 0, 45, and 90 days of the study. A marked reduction in the subcutaneous tissue thickness was observed in all subjects, with the average reduction of 1.9 mm. The length of subcutaneous tissue fascicles (ScTFL) was reduced in 97% (28 out of 29) of subjects, with the average value of 1.18 mm. After 45 days of chokeberry juice consumption, reduction of edema was observed in 55.2% of the subjects with edema at the baseline, while at the endpoint of the study, edemas were not observed in any of the subjects involved in the study. OCJ could have beneficial effects on the cellulite condition, including the length of ScTFL, subcutaneous tissue, and dermis thickness as well as on edema reduction.

Variation in the fatty-acid content in seeds of various black, red, and white currant varieties.

Currant seeds, a by-product of juice production, are recognized as a valuable source of oil rich in polyunsaturated fatty acids. We have evaluated 28 currant varieties for their oil content and fatty-acid composition. The oil content in the seeds ranged from 18.2-27.7%, and no statistical difference between varieties of different fruit color were recorded. Furthermore, the estimated oil yields in the field production ranged from 26.4-212.4 kg/ha. The GC and GC/MS chemical profiles of the seed oils extracted from all examined varieties were common for currants. Linoleic acid (LA) was the major component, with contents ranging from 32.7-46.9% of total fatty acids, followed by α-linolenic acid (ALA; 2.9-32.0 %), oleic acid (OA; 9.8-19.9%), γ-linolenic acid (GLA; 3.3-18.5%), palmitic acid (PA; 4.4-8.1%), stearidonic acid (SDA; 2.2-4.7%), and stearic acid (SA; 1.2-2.4%). Quantitative differences in the fatty-acid profiles between varieties of different fruit color were observed. Blackcurrant varieties showed significantly higher contents of LA, GLA, and PA than red and white currant varieties, whereas significantly higher amounts of ALA and OL were detected in the red and white varieties. Cluster analysis based on the chemical oil profiles joined the blackcurrants in one group, while most of the red and white cultivars joined in a second group at the same linkage distance.

Biochemical properties of the fresh and frozen black currants and juices.

Thirteen black currant varieties (Ribes nigrum L.) cultured in Serbia were characterized for their pomological properties and chemical composition (total phenolics, total anthocyanins, anthocyanin aglycones, sugars, and vitamin C). The average amount of vitamin C varied from 122.4 to 193.2 mg/100 g fresh weight (FW), while concentration of invert sugars ranged from 6.3% to 11.1%. The highest amounts of total phenolics and anthocyanins were detected in variety Ometa (278.9 mg of gallic acid equivalents per 100 g of FW [mg GAE/100 g FW] and 135.4 mg/100 g, respectively). Quantitative analyses of anthocyanin aglycones in berries were performed using high-performance liquid chromatography, and delphinidin was found to be dominant compound in 11 varieties. Total phenolics and anthocyanins contents decreased during the processing of berry fruits to juices, and the reduction of anthocyanins was more pronounced, 12%-80%. The radical scavenging activity of black currant juices was investigated by 1,1-diphenyl-2-picrylhydrazyl, and the IC(50) value ranged from 1.9 to 4.0 mg/mL. Our results also showed that freezing as a way of preservation and storage could save important phytochemicals and health benefits of berries and berry juices. The amount of total phenolics in berries increased during 1 year of storage by 46.09%-171.76% and in juices by even 107.58%, while the amount of total anthocyanins in berries and juices decrease by 5.63%-52.76% and 13.04%-36.82%, respectively.

High diversity of indigenous populations of dalmatian sage (Salvia officinalis L.) in essential-oil composition.

Essential oils of 25 indigenous populations of Dalmatian sage (Salvia officinalis L.) that represent nearly half of native distribution area of the species were analyzed. Plantlets collected from wild populations were grown in the same field under the same environmental conditions and then sampled for essential-oil analysis. The yield of essential oil ranged from 1.93 to 3.70% with average of 2.83%. Among the 62 compounds detected, eight (cis-thujone, camphor, trans-thujone, 1,8-cineole, ß-pinene, camphene, borneol, and bornyl acetate) formed 78.13-87.33% of essential oils of individual populations. Strong positive correlations were observed between camphor and ß-pinene, ß-pinene and borneol, as well as between borneol and bornyl acetate. The strongest negative correlation was detected between camphor and trans-thujone. Principal component analysis (PCA) on the basis of eight main compounds showed that first main component separated populations with high thujone content, from those rich in camphor, while the second component separated populations rich in cis-thujone from those rich in trans-thujone. Cluster analysis (CA) led to the identification of three chemotypes of S. officinalis populations: cis-thujone; trans-tujone, and camphor/ß-pinene/borneol/bornyl acetate. We propose that differences in essential oils of 25 populations are mostly genetically controlled, since potential environmental factors were controlled in this study.