Acmella oleracea (L.) R.K. Jansen: Alkylamides and phenolic compounds in aerial parts and roots of in vitro seedlings.

Acmella oleracea L. is an edible and medicinal plant commonly known for its local anaesthetic effect induced by the alkylamide spilanthol. It is also rich of secondary metabolites of biological interest, mainly phenolic acids and glycosylated flavonoids. This study evaluated for the first time alkylamides and phenolic compounds in aerial parts and roots of seedlings grown in vitro and produced from single seed and from regenerating lines. The extracts obtained showed similar chemical profiles and the caffeic acid derivatives were the most abundant phenolic compounds. Spilanthol was prevalent in the aerial parts, especially in samples of seedlings obtained from regenerating lines, in which reached maximum value of 1.72 mg/g dried matter (DM). The roots contained a lower content of alkylamides, while showing twice as much total phenols (11.19 mg/g DM) as the aerial parts. A hexane fractionation step allowed to recover spilanthol and its derivatives in a concentrated extract, which reached a value of 63.4 mg/g dried extract in the aerial parts from seedlings from regenerating lines. Hydroalcoholic dried extracts showed high yields (30-45 % on dried matter) and those obtained from aerial parts contained up to 5.69 mg/g of spilanthol.

Formulation of a Phenol-Rich Extract from Unripe Olives (Olea europaea L.) in Microemulsion to Improve Its Solubility and Intestinal Permeability.

The beneficial properties of phenolic compounds from Olea europaea L. are well-known. An olive extract (OE) was prepared from unripe olives (Moraiolo cultivar). The study aimed to formulate OE into a microemulsion (ME) in oral dosage form. OE was extracted from olives with EtOH:H2O (80:20) and characterized by HPLC-DAD. ME composition was stated by a solubility and pseudo-ternary diagram. The ME was chemically and physically characterized, and its stability at 4 °C was analyzed for three months. The ability of the formulation to ameliorate the solubility and the intestinal permeability of OE was evaluated by a Parallel Artificial Membrane Permeability Assay (PAMPA) assay and Caco-2 cells. The total phenolic content of the extract was 39% w/w. The main constituent was oleuropein (31.0%), together with ligstroside (3.1%) and verbascoside (2.4%). The ME was prepared using Capryol 90 as the oily phase, and Cremophor EL and Transcutol (2:1) as surfactant and co-surfactant, respectively. ME droplet size was 14.03 ± 1.36 nm, PdI 0.20 ± 0.08, ζ-potential -1.16 ± 0.48. Stability of ME was confirmed for at least three months. The formulation was loaded with 35 mg/mL of OE, increasing the solubility of the extract by about four times. The enhanced permeability of OE was evaluated by PAMPA, as demonstrated by the Pe value (1.44 ± 0.83 × 10-6 cm/s for OE hydroalcoholic solution, 3.74 ± 0.34 × 10-6 cm/s for OE-ME). Caco-2 cell transport studies confirmed the same results: Papp was 16.14 ± 0.05 × 10-6 cm/s for OE solution and 26.99 ± 0.45 × 10-6 cm/s for OE-ME. ME proved to be a suitable formulation for oral delivery.

A Study on the Biodiversity of Pigmented Andean Potatoes: Nutritional Profile and Phenolic Composition.

The characterization of six varieties of native Andean potatoes with a wide biodiversity in tuber shape, flesh, and skin color was performed, through the determination of their proximate composition, mineral content, and phenolic profile. Minerals concentration revealed significant genotypic variation. Potassium was the most abundant element in all varieties, ranging from 7272.9 to 13,059.9 µg/g and from 12,418 to 17,388.6 µg/g dried weight for the flesh and skin samples, respectively. Iron content was relevant, ranging from 20.5 to 39.9 µg/g and from 112.2 to 288.8 µg/g dried weight in flesh and skin samples, respectively. Phenolic compounds were consistently higher in the skin than in the flesh. The total content varied greatly from 19.5 to 2015.3 µg/g and from 1592.3 to 14807.3 µg/g dried weight for flesh and skin tissues, respectively. 5-caffeoylquinic acid was 74% of the total phenolic acids. Different pattern of anthocyanins was found, depending on the color of the variety; the red genotypes contained predominantly pelargonidin derivatives, while the purple samples had petunidin as a major anthocyanidin. This study increases the knowledge of the composition of the local Andean varieties (which are only scarcely studied so far), helping to enhance these genotypes and the conservation of biodiversity.

What kind of coffee do you drink? An investigation on effects of eight different extraction methods.

The chemical composition of brewed coffee depends on numerous factors: the beans, post-harvest processing and, finally, the extraction method. In recent decades, numerous coffee-based beverages, obtained using different extraction techniques have entered the market. This study characterizes and compares eight extraction coffee methods from a chemical-physical point of view, starting from the same raw material. Specifically, three types of Espresso, Moka, French Press, and 3 filter coffee that for the first time are reported in the scientific literature Cold Brew, V60, and Aeropress are compared. Physical measurements included the quantification of total dissolved solids, density, pH, conductivity, and viscosity. Chemical analyses identified 15 chlorogenic acids (CGAs): six caffeoylquinic acids, one p-Coumaroylquinic acid, one Feruloylquinic Acid, four Caffeoylquinic lactones, and three Dicaffeoylquinic acids. Maximum caffeine and CGA concentrations were found in Espresso coffees, while Moka and filtered coffees were three to six times less concentrated. The classic Espresso method was most efficient for caffeine and CGA recovery, with a yield almost double that of other methods. Per-cup caffeine and CGAs were higher in Cold Brew than Espresso coffees, as a function of the volume of beverage, which ranged from 30 mL (for espresso) to 120 mL (for filtered coffees). In light of these results, it is not possible to establish how many cups of coffee can be consumed per day without exceeding the recommended doses, since according to the applied brewing method, the content of the bioactive substances varies considerably.

In depth study of phenolic profile and PTP-1B inhibitory power of cold-pressed grape seed oils of different varieties.

This paper investigates the phenolic composition of 17 monocultivar commercial cold-pressed grape seed oils. Chromatographic profiles showed the presence of more than 28 molecules, 11 of which were successfully identified by HPLC-DAD-MS-TOF and HPLC-FLD analysis. Pinoresinol, ethyl caffeate and ethyl gallate were detected for the first time in these oils. The total phenolic content ranged between 0.83 mg/kg for Viognier sample to 15.16 mg/kg for Merlot org sample. The detected ethyl esters can be suggested as markers to evaluate the intensity of fermentation in grape seeds before oil extraction, and to control the sensorial quality of the produced oils. In addition, the inhibitory power of these phenolic extracts against Protein Tyrosine Phosphatase 1B enzyme (PTP-1B), overexpressed in type-two diabetes, was investigated for the first time. Data highlighted a good correlation between total phenolic content and inhibitory power, with pinoresinol, p-coumaric acid and quercetin making the greater contributions.

Effects of ionizing radiation on bio-active plant extracts useful for preventing oxidative damages.

Humans are exposed to ionizing radiations in medical radiodiagnosis and radiotherapy that cause oxidative damages and degenerative diseases. Airplane pilots, and even more astronauts, are exposed to a variety of potentially harmful factors, including cosmic radiations. Among the phytochemicals, phenols are particularly efficient in countering the oxidative stress. In the present study, different extracts obtained from plant food, plant by-products and dietary supplements, have been compared for their antioxidant properties before and after irradiation of 140 cGy, a dose absorbed during a hypothetical stay of three years in the space. All the dry extracts, characterized in terms of vitamin C and phenolic content, remained chemically unaltered and maintained their antioxidant capability after irradiation. Our results suggest the potential use of these extracts as nutraceuticals to protect humans from oxidative damages, even when these extracts must be stored in an environment exposed to cosmic radiations as in a space station.

Characterization and comparison of cold brew and cold drip coffee extraction methods.

BACKGROUND: Each region of the world has its own methods, protocols, instruments and procedures regarding how to brew coffee. The final result in the cup is strongly affected by the extraction method, and many studies have focused on this subject. However, few studies have investigated slow, cold extraction methods, despite their popularity among baristas. Therefore, the present study aimed to characterize and compare two cold extraction methods: cold brew and cold drip. RESULTS: Physical and chemical analyses were used to describe coffee beverages in terms of pH, total solids, refractive index, density and viscosity. Caffeine and cinnamic acids were quantified using high-performance liquid chromatography (HPLC)/diode array detector and HPLC/mass spectrometry. A sensory evaluation included aroma, flavor and textural attributes. CONCLUSIONS: Significant differences were found in the chemical and physical parameters, both between and within the two methods, as a function of the extraction temperature and contact time. Similarly, the sensory evaluation found differences in flavor profiles, as measured in terms of bitterness, sweetness, sourness and global intensity. © 2018 Society of Chemical Industry.

The antimicrobial effects of three phenolic extracts from Rosmarinus officinalis L., Vitis vinifera L. and Polygonum cuspidatum L. on food pathogens.

In this study three phenolic extracts were examined, without volatile fraction, against common food pathogens. The samples, all suitable for food application, were from the leaves of Rosmarinus officinalis L., Vitis vinifera L., and the root of Polygonum cuspidatum L. The microorganisms tested were Escherichia coli O157:H7, Salmonella Enteritidis, Salmonella Typhi, Yersinia enterocolitica and Listeria monocytogenes, well-known as important food pathogens. The results demonstrated a microbicidal activity of all the tested compounds at different concentrations; the rosemary extract showed greater efficacy than the other compounds against the tested microorganisms. In particular, the best results were obtained with rosemary extract against E. coli O157:H7 and L. monocytogenes with values of 200 and 270 µg/mL, respectively. Our results show that rosemary extract, often present as a natural antioxidant in food, can also be proposed as a natural disinfectant in the food field.

Recovery and stability over time of phenolic fractions by an industrial filtration system of olive mill wastewaters: a three-year study.

BACKGROUND: The recovery of phenolic compounds from olive milling is recognized as strategic for producers. The aim of this work was to evaluate the quality and stability of retentates obtained from olive mill wastewaters treated with a membrane filtration system constituted by a micro-, ultra- and nanofiltration, followed by a final reverse osmosis, over three crop seasons. Efficiency was evaluated in terms of phenolic amount in the retentates and of organic load in the final discarded waters. Phenolic compounds were quantified using tyrosol as external standard. RESULTS: Our study highlighted a reproducibility of the process over years and a low organic load in permeates from reverse osmosis. Hydroxytyrosol was very stable in the liquid products at 18-28 °C over 24 months of storage. The retentates from reverse osmosis showed the highest phenolic content (78.6 mg g-1 dry matter in 2015), associated with a potassium content of 22 g kg-1 . CONCLUSIONS: The liquid concentrated retentates showed an unexpected stability over time of their bioactive phenolic compounds, particularly of hydroxytyrosol. These samples recovered from olive mill wastewaters can be good sources of natural antioxidants and potassium to guarantee the correct intake and to formulate new food ingredients or food products. © 2017 Society of Chemical Industry.

Polysaccharides from by-products of the Wonderful and Laffan pomegranate varieties: New insight into extraction and characterization.

The main crude polysaccharides (CPS), extracted from two widely cultivated pomegranate varieties, Laffan and Wonderful, were studied and characterized. We obtained the highest CPS extraction yield (approximatively 10% w/w on dried matter) by 1h of decoction (ratio 1/40w/v). The predominant polymers (75-80%) of the CPS samples showed a hydrodynamic volume close to 2000kDa by size exclusion chromatography and the exocarp and mesocarp profiles were very similar. The proton spectra (1H NMR), according to sugar composition and gelling ability, confirmed the main polysaccharide fractions were pectin with different acylation and methylation degree. The CPS from Laffan and Wonderful mesocarp showed prebiotic properties in vitro with Lactobacillus and Bifidobacterium strains. The composition of the decoction (12% ellagitannins and 10% of CPS) obtained by a green extraction process of pomegranate by-products, makes it a suitable component of functional food formulations.

Chemical constituents, radical scavenging activity and enzyme inhibitory capacity of fruits from Cotoneaster pannosus Franch.

Cotoneaster pannosus (Rosaceae) is a semievergreen shrub, producing globose dark red pomes, native to China and widely used as an ornamental plant all over the world. Despite its extensive cultivation, little information is available on the chemical composition and biological activities of its fruits. In this work, the analysis of the chemical composition of C. pannosus fruits, in terms of phenolic components, carotenoids and ascorbic acid by HPLC/DAD, HPLC/ESI-MS and MS/MS as well as in terms of macro- and micro-nutrients was performed. The fruits proved to be a good source of shikimic acid and caffeoylquinic acids, whereas ß-carotene, pelargonidin-3-O glucoside and cyanidin-3,5-rutinoside gave an important contribution to the color of the fruit. Both the polar and apolar fruit extracts showed noteworthy radical scavenger activity and inhibitory effects against monoamine oxidase A (MAO-A), tyrosinase (TYR) and α-glucosidase, making C. pannosus red pomes a promising candidate ingredient in functional foods and dietary supplements.

Anti-Dermatophyte and Anti-Malassezia Activity of Extracts Rich in Polymeric Flavan-3-ols Obtained from Vitis vinifera Seeds.

Several human skin diseases are associated with fungi as dermatophytes and Malassezia. Skin mycoses are increasing and new alternatives to conventional treatments with improved efficacy and/or safety profiles are desirable. For the first time, the anti-dermatophytes and the anti-Malassezia activities of Vitis vinifera seed extracts obtained from different table and wine cultivars have been evaluated. Geometric minimal inhibitory concentration ranged from 20 to 97 µg/mL for dermatophytes and from 32 to 161 µg/mL for Malassezia furfur. Dried grape seed extracts analyzed by HPLC/DAD/ESI/MS showed different quali-quantitative compositions in terms of monomeric and polymeric flavan-3-ols. The minimal inhibitory concentrations for Trichophyton mentagrophytes and for M. furfur were inversely correlated with the amount of the polymeric fraction (r = -0.7639 and r = -0.7228, respectively). Differently, the antifungal activity against T. mentagrophytes was not correlated to the content of flavan-3-ol monomers (r = 0.2920) and only weakly correlated for M. furfur (r = -0.53604). These results suggest that extracts rich in polymeric flavan-3-ols, recovered from V. vinifera seeds, could be used for the treatment of skin fungal infections. Copyright © 2016 John Wiley & Sons, Ltd.

Anti-neuropathic effects of Rosmarinus officinalis L. terpenoid fraction: relevance of nicotinic receptors.

Traditional uses and current results highlight the neuroprotective properties of Rosmarinus officinalis L. The compelling need for novel strategies able to relieve neuropathic pain encouraged us to analyze different rosemary leaf extracts in rats following chronic constriction injury (CCI) of sciatic nerve. Ethanol, acetone, and the innovative ultrasound-hexane extractive methods were used to obtain: EE, AE, and for hexane extracts UREprel and URE. Extracts were characterized in terms of typical constituents and repeatedly administered to CCI-rats (13-days treatment, from the day of surgery). URE showed the best efficacy and potency in reducing hypersensitivity to noxious- and non-noxious stimuli and spontaneous pain. URE contained the higher quantity of the terpenoid carnosic acid (CA) and its efficacy was compared to pure CA. Histological analysis of the sciatic nerve revealed that URE prevented axon and myelin derangement, edema and inflammatory infiltrate. In the dorsal horn of the spinal cord, URE did not reduce astrocyte activation. Both the pain reliever and the neuroconservative effects of URE were significantly prevented by the nicotinic receptor (nAChR) antagonist mecamylamine. In conclusion, the hexane-ultrasound rosemary extract is able to reduce neuropathic hypersensitivity and protect nervous tissues. Effectiveness is mainly related to the terpenoid fraction by mechanisms involving nAChRs.

Dietary supplement based on stilbenes: a focus on gut microbial metabolism by the in vitro simulator M-SHIME®.

Polyphenols and intestinal microbiota can influence each other, modifying metabolism and gut wellness. Data on this mutual effect need to be improved. Several studies on the biological activities of resveratrol and derivatives have been carried out, but the effects of a continuous administration of stilbenes on gut microbiota have not yet been investigated. This study evaluated the effects of an extract from Vitis vinifera, containing a combination of t-resveratrol and ε-viniferin, on intestinal microbiota, using the advanced gastrointestinal simulator M-SHIME®. A triple M-SHIME® experiment was performed using two concentrations of the extract (i.e. 1 and 2 g L-1), simulating a continuous daily intake. The effects were evaluated in terms of microbial functionality (SCFA and NH4+) and composition (DGGE and Illumina sequencing), since the microbiological aspect has been less considered so far. The treatment induced changes in microbial functionality and composition. In fact, the levels of SCFA and NH4+ suffered a strong decrease (i.e. inhibition of the saccharolytic and proteolytic activity), while DGGE and Illumina showed important modifications of the microbiota composition, associated with an imbalance of the colonic microbiota (i.e. increase in the relative abundance of Enterobacteriaceae). HPLC-DAD-TOF-MS analyses demonstrated that the metabolism of t-resveratrol and other stilbenes was inhibited by continuous administration. Our results suggest M-SHIME® as an explorative tool to define the dosage of food supplements, in particular to simulate effective continuous administration in humans.

Acute effect of Capparis spinosa root extracts on rat articular pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Capparis spinosa L. originates from dry regions of Asia and Mediterranean basin. In traditional medicine of these areas, infusions from caper root are considered to be beneficial for the treatment of rheumatism, gout and against abdominal pains. AIM OF THE STUDY: To evaluate the pain relieving properties of a Syrian cultivar of Capparis spinosa roots in rat models of osteoarthritis and rheumatoid arthritis. MATERIALS AND METHODS: Decoction (DEC) and hydroalcoholic extract (EtH2O) were obtained from powdered roots; the latter was further separated in CH2Cl2 and aqueous (H2O-Res) fractions. The extracts were characterized in terms of spermidine alkaloids by HPLC/DAD/MS and stachydrine by NMR. Different amount of free and glycosilated forms of capparispine and analogues (from 0.5% w/w for DEC up to 7.6% w/w for CH2Cl2 fraction) were detected. Rat models of rheumatoid arthritis and osteoarthritis were induced by the intra-articular administration of Complete Freund's Adjuvant (CFA) or monosodium iodoacetate (MIA), respectively. RESULTS: Fourteenth days after CFA or MIA injection, the different preparations of Capparis spinosa (3, 30, 100 and 300mgkg-1) were acutely administered p.o.. Powdered roots (300mgkg-1), DEC (100mgkg-1), and EtH2O (300mgkg-1) significantly reduced hypersensitivity to mechanical noxious stimuli as well as spontaneous pain evaluated as hind limb bearing alterations in both models. The CH2Cl2 and the H2O-Res (30mgkg-1) were the most potent in reverting pain threshold alterations despite the different content of free alkaloids. CONCLUSIONS: Capparis spinosa extracts relieved pain related to rheumatoid arthritis and osteoarthritis after single administration. A synergistic effect due to a specific "phytochemical mixture" is suggested.

Blue honeysuckle fruit (Lonicera caerulea L.) from eastern Russia: phenolic composition, nutritional value and biological activities of its polar extracts.

In the present work we conducted a comprehensive chemical analysis of blue honeysuckle (Lonicera caerulea) spontaneously growing in eastern Russia. HPLC-DAD-ESI/MS analysis showed cyanidin-3-glucoside as the major constituent among phenolics, while nutritional analysis revealed fibre, protein, calcium and magnesium as the most important macro- and micronutrients, respectively. Fatty acid composition was dominated by polyunsaturated fatty acids, linoleic acid being the most abundant. Furthermore, we evaluated several in vitro biological activities such as antioxidant, antimicrobial, antiproliferative, wound healing and immunomodulatory effects of blue honeysuckle aqueous and ethanolic extracts that are often incorporated in food and nutraceutical preparations. While the fruit extracts were revealed to be potent radical scavengers with significant inhibition of ABTS radical, thus confirming the literature data, their inhibitory effects against microbial pathogens and tumor cell lines were negligible. The fruit aqueous extract did not show toxicity to human fibroblasts, but 24 h treatment with 150-200 µg per mL of extract slightly enhanced the cell migration when tested by scratched wound assay. Worth mentioning was the inhibitory effect displayed by the blue honeysuckle fruit aqueous extract on human lymphocytes.

An insight on the alkaloid content of Capparis spinosa L. root by HPLC-DAD-MS, MS/MS and (1)H qNMR.

The Capparis spinosa L. has a wide distribution in the Old World from South Europe, North and East Africa, Madagascar, Southwest and Central Asia to Australia and Oceania. The consolidated traditional use of C. spinosa root as remedy against different pains in human is well known since the antiquity. Various secondary metabolites have been found in caper plant, nevertheless, few studies have been focused to the analysis of root constituents. To date, several free and glycosilated spermidine alkaloids and a more polar alkaloid, the stachydrine, have been isolated from the root of C. spinosa. Aim of this work was to improve the knowledge on the alkaloid content of the root of a Syrian sample of C. spinosa by HPLC-DAD-MS(n) and to propose methods to quantify these molecules in different raw extracts. A decoction, an hydroalcoholic extraction and a fractionation process to selectively recover the spermidine alkaloids were applied. To our knowledge, this is the first HPLC-DAD-MS(n) profile that pointed out the co-presence of stachydrine, several isobaric forms of capparispine and/or capparisine in free and glycosylated forms and some isobars of isocodonocarpine or codonocarpine as monoglycosides in extracts of C. spinosa root. The determination by HPLC/DAD for the spermidine alkaloids expressed as p-OH-coumaric acid gave values up to 3.5mg/g dried root and the stachydrine evaluated by (1)H NMR was close to 12.5mg/g dried root. Overall, the total alkaloids were almost doubled in hydroalcoholic extract with respect to the decoction, and the stachydrine in the cortex was almost double than in the whole root.

Phenolic composition of "bud extracts" of Ribes nigrum L., Rosa canina L. and Tilia tomentosa M.

Liquid preparations such as tinctures and "bud extracts" are typical products widely used in European countries but which have been poorly studied to date. Our research was focused on obtaining a phytochemical characterization of secondary metabolites of black currant, dog rose and silver linden bud extracts and evaluating the changes of selected chemical markers over time. 16 different preparations have been analyzed by the use of HPLC-DAD-MS. Several flavonols were identified in black currant samples including quercetin derivatives (6.7-30.4mg/L). Also hydroxycinnamic acids, mainly p-coumaric acid derivatives (4.1-38.9mg/L) were identified. Flavonols such as glycosides of quercetin and kaempferol, hydroxycinnamic acids, and ellagitannins were detected in dog rose samples, with gallotannins being the main components (up to 1.7g/L). The Tilia tomentosa extracts contained flavonols (quercetin, kaempferol, apigenin derivatives) as principal components with the exception of a single commercial extract with hydroxycinnamic acids as the most abundant metabolites. In this study, without applying accelerated ageing protocols, the stability over time of these liquid preparations was evaluated for up to 10 months and demonstrated negligible variations.

An innovative approach to the recovery of phenolic compounds and volatile terpenes from the same fresh foliar sample of Rosmarinus officinalis L.

Rosmarinus officinalis L. is a plant of relevant commercial interest because of its volatile fraction and also its phenolic constituents which are both well known for their numerous properties. Nevertheless, an extractive method suitable to recovering both the aromatic and phenolic fractions from the same fresh foliar tissue has not yet been reported. In this work we have optimized a two-step procedure able to recover first the phenolic compounds and successively the volatile terpenes from the same foliar sample. The recovery of the whole phenolic fraction, partially degraded using a traditional extractive method, was guaranteed and we observed a significant increment in the amount of volatile terpenes compared to a traditional extraction procedure. We also highlight crucial information on the enzymatic activity of the endogenous oxidases that rapidly transform the phenolic substrates, mainly the rosmarinic acid. Our results suggest that this extractive procedure could also be used for other aromatic plants, thus providing a useful tool for more complete analyses of the main phytochemicals available in fresh foliar samples and creating the possibility of incrementing yields of volatile compounds.

Evaluation of anti-Candida activity of Vitis vinifera L. seed extracts obtained from wine and table cultivars.

For the first time, grape seed extracts (GSEs), obtained from wine and table cultivars of Vitis vinifera L., cultured in experimental fields of Lazio and Puglia regions of Italy and grown in different agronomic conditions, have been tested on 43 Candida species strains. We demonstrated a significant correlation between the content of the flavan-3-ols in GSEs extracts, with a polymerization degree ≥ 4, and anti-Candida activity. Moreover, we demonstrated that GSEs, obtained from plants cultured with reduced irrigation, showed a content of polymeric flavan-3-ols >250 mg/g with geometric mean MIC values between 5.7 and 20.2 mg/L against Candida albicans reference strains. GSE, showing 573 mg/g of polymeric flavan-3-ols, has been tested in an experimental murine model of vaginal candidiasis by using noninvasive in vivo imaging technique. The results pointed out a significant inhibition of Candida albicans load 5 days after challenge. These findings indicate that GSEs with high content of polymeric flavan-3-ols can be used in mucosal infection as vaginal candidiasis.