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Four new caffeoyl -glucaric and -altraric acid derivatives along with eleven known compounds were isolated from aerial parts of Galinsonga parviflora. Their structures were elucidated by high-resolution spectroscopic studies. The four new compounds were determined as being 2,3,4,5-tetracaffeoylglucaric acid (1), 2,4,5-tricaffeoylglucaric acid (2), 2,3,4- or 3,4,5-tricaffeoylaltraric acid (3) and 2,3(4,5)-dicaffeoylaltraric acid (4). A reliable criterion for the determination of the linkage position of caffeic acids moieties in glucaric acid derivatives has been proposed, on the basis of detailed analysis of the respective J-couplings, including substitution and solvent influence on the observed values. All hexaric acids derivatives appeared as inhibitors of reactive oxygen species production by stimulated neutrophils.

In this study, the flowers of Forsythia × intermedia, Forsythia suspensa and Forsythia ovata are presented as valued raw materials for isolation of rutin, an industrial and pharmacologically active flavonoid. An efficient strategy based on three high-performance chromatographic techniques was established to screen and purify the target compound from the plant matrices. In the first step, an UHPLC-PDA-ESI-MS 3 assay documented the relatively simple phytochemical profiles of the flower extracts and led to the identification of ten polyphenols classified as caffeoyl acid derivatives, flavonoids, phenylethanoids and lignans, with rutin as the major constituent. Significant levels of rutin (20.0–57.6 mg/g dw) were found in the flowers by the validated HPLC-PDA method, with the highest values observed for F. suspensa . Finally, a novel, fast, single-step HPCCC method was developed and successfully applied for the separation of rutin from the plant samples. The ethyl acetate- n -butanol-water (4:1:5, v / v / v) was selected as the optimum two-phase solvent system for both analytical and preparative normal (NP) and reversed-phase (RP) HPCCC separations. Under optimal RP-HPCCC conditions, 155 mg of the crude methanol extract was separated in a single 50 min run, yielding 14.6 mg of rutin with 97.2% HPLC purity and 95.9% recovery. The process efficiency of the HPCCC rutin isolation was demonstrated to be superior to the existing HSCCC methods. The results indicated the usefulness of Forsythia flowers and the developed RP-HPCCC methodology for practical and commercial applications.

The gut microbiota-derived metabolites of ellagitannins and green tea catechins, urolithin A (uroA) and 5-(3′,4′,5′-trihydroxyphenyl)-γ-valerolactone (M4), respectively, are among the main compounds absorbed into human system after ingestion of these polyphenols. The aim of this study was to establish the effects of M4, uroA, and their combinations on LNCaP cells, an androgen dependent prostate cancer in vitro model.. The LNCaP cells were incubated with increasing concentrations of tested metabolites. The cell proliferation was determined by measurement of DNA-bisbenzimide H 33 258 complexes fluorescence. The isobolographic analysis was used to establish the type of interaction between metabolites. The apoptosis, androgen receptor (AR) localization, and phosphorylation of Akt kinase were measured by flow cytometry. Prostate-specific antigen (PSA) secretion was determined by ELISA. M4 showed modest antiproliferative activity in LNCaP cells (IC50 = 117 µM; CI: 81 – 154). UroA decreased proliferation (IC50 = 32.7 µM; CI: 24.3 – 41.1) and induced apoptosis of LNCaP cells. The mixture of M4 with uroA had synergistic antiproliferative effect. Moreover, M4 potentiated inhibition of PSA secretion and enhanced retention of AR in cytoplasm caused by uroA. Interestingly, uroA increased levels of pSer473 Akt in LNCaP cells. These results show that colonic metabolites may contribute to chemoprevention of prostate cancer by varied polyphenol-rich diet or composite polyphenol preparations.

The current study estimates the effect of different cytokinins on shoot proliferation and biosynthesis of caffeic acid derivatives in Dracocephalum forrestii in vitro culture. The shoots were grown on Murashige and Skoog (MS) agar medium with 1 µM indole-3-acetic acid (IAA) and different content of 6-benzyloaminopurine (BAP), zeatin, kinetin (1, 2, 4, 8, 18 µM) or thidiazuron (TDZ) (0.1, 0.2, 0.5, 1, 2 µM). The highest multiplication rate (about seven shoots and/or buds per explant) was obtained after 4 weeks of culture on MS medium with 1 µM IAA and 8 or 16 µM BAP. Optimal biomass of plant material was also received on the same media. The identity of the compounds present in the hydromethanolic extracts from D. forrestii shoots grown on cytokinin-supplemented media was confirmed using UPLC–PDA–ESI–MS method. The analysis revealed the presence of nine metabolites recognized as caffeic acid derivatives. The content of the predominant phenolic acids in the extracts, i.e. rosmarinic acid (RA) and salvianolic acid B (SAB), was determined with UHPLC. The highest yield of RA was found in shoots cultivated in the medium containing 1 µM IAA and 2 µM BAP (18.7 mg/g DW). The highest level of SAB (5.3–5.9 mg/g DW) was identified in multiple shoots grown in the presence of 1 µM IAA and 0.5–1 µM TDZ or 2 µM BAP.

Salvia viridis L. is an annual herb used in Mediterranean medicine. The purpose of this study was to determine the polyphenol profile of aqueous (decoction and infusion) and hydroethanolic extracts of aerial parts of field-grown S.viridis and to evaluate their antioxidant activity. The polyphenol profiling was performed via UPLC-DAD/ESI-MS. Additionally, the total polyphenol content in extracts tested were determined by UV-Vis spectrophotometry using the Folin-Ciocalteu assay. The antioxidant effect was evaluated by the FRAP, DPPH, ABTS, O2&bull, &minus, scavenging and TBARS methods. The hydroethanolic extract gave the highest content of total phenolic compounds, followed by the infusion. The UPLC-DAD/ESI-MS analysis of extracts showed a total of 19 phenolic compounds identified as flavonoids (four compounds), phenylethanoids (eight compounds) and phenolic acids (seven compounds). Rosmarinic acid was the predominant phenolic acid, verbascoside was the predominant phenylethanoid, while apigenin glucuronide or methylluteolin glucuronide, depending on the sample, were the predominant flavonoids in the analyzed extracts. The presence of a high polyphenol level indicated a high antioxidant activity of both the infusion and the hydroalcoholic extract. These results indicate that S. viridis is a rich resource of phenolic compounds and can be used in dietary applications with the potential to reduce oxidative stress.

Aim of the study: Taking into account that overactivated leukocytes are an important factor in the development of many chronic diseases, we investigated the activity of phytochemically characterized (HPLC-DAD-MSn) extracts from forsythia leaves and flowers on the pro- and anti-inflammatory functions of leukocytes (effects on IL-1β, IL-8, TNF-α, and TGFβ release) and their adherence to endothelial cells. Using bio-guided fractionation, we isolated the active compounds and determined their biological activity, and we included the positive control quercetin. Methods: The effect on IL-1β, TNF-α, IL-8, and TGF-α production by leukocytes was measured by enzyme-linked immunosorbent assay (ELISA). The surface expression of adhesion molecules was analyzed with flow cytometry, and the neutrophil attachment to the endothelial cells was assessed fluorimetrically. The effects on p38MAPK, ERK1/2 and JNK phosphorylation were determined using western blots. Results: Leaf extracts had the effect of decreasing TNF-α production in neutrophils and monocyte/macrophage cells. The bio-guided fractionation led to the isolation of the following lignan aglycones: (+)-pinoresinol, (+)-epipinoresinol, (−)-matairesinol, (+)-phillygenin, and (−)-arctigenin. Only phillygenin was able to stimulate the anti-inflammatory function of macrophages by inducing TGF-β release and IL-10 receptor surface expression. Arctigenin, phillygenin, and a metabolite produced by the gut microbiota, enterolactone, decreased TNF-α and IL-1β production and neutrophil adhesion to endothelial cells, probably by attenuating the p38 and ERK kinase pathways. Conclusion: Forsythia x intermedia is a valuable source of active lignans, which may be potential candidates for treating inflammatory diseases that are associated with the excessive production of cytokines such as TNF-α and IL-1β.

In an initial screening, the dichloromethane extract from the leaves of Melodorum fruticosum showed distinct inhibitory effects on the release of interleukin‐8 (IL‐8) in human neutrophils. Therefore, the aim of the present study was the phytochemical and pharmacological investigation of this extract, to better understand which compounds might be responsible for the anti‐inflammatory effect. Phytochemical analysis led to the isolation of 12 known compounds and two new natural products, 5‐hydroxy‐6‐(2‐hydroxybenzyl)‐4′,7‐dimethoxyflavanone (13) and 2′,4′‐dihydroxy‐3′‐(2‐hydroxybenzyl)‐4,6′‐dimethoxychalcone (14). The influence of the isolated compounds on the production and release of the pro‐inflammatory factors IL‐8, tumor necrosis factor alpha (TNF‐α), reactive oxygen species (ROS), and adhesion molecules (CD62L and CD11b) in human neutrophils was evaluated. Three constituents, melodamide A, 2′,4′‐dihydroxy‐4,6′‐dimethoxychalcone, and 2′,6′‐dihydroxy‐4′‐methoxychalcone, showed significant inhibition of IL‐8 release (IC50=6.6, 8.6, and 11.6 μm, respectively) and TNF‐α production (IC50=4.5, 13.3, and 6.2 μm, respectively).

Scope Ellagitannin-rich food products and medicinal plant materials were shown to have beneficial effects toward intestinal inflammation. Due to the questionable bioavailability of ellagitannins their gut microbiota metabolites-urolithins have come to be regarded as potential factors responsible for biological activities observed in vivo. The aim of the study was to determine the influence of the three most abundant bioavailable ellagitannin gut microbiota metabolites-urolithins A, B, and C on inflammatory responses in RAW 264.7 murine macrophages, which are involved in the pathogenesis of intestine inflammation. Methods and results Urolithins A, B, and C decreased NO production via inhibition of the iNOS protein and mRNA expression. They decreased the expression of IL-1β, TNF-α, and IL-6 mRNA in LPS challenged RAW 264.7 murine macrophages. A clear inhibition of NF-κB p65 nuclear translocation and p50 DNA-binding activity was associated with the observed anti-inflammatory activities of urolithins. Among the tested compounds urolithin A had the strongest anti-inflammatory activity. Conclusion The anti-inflammatory effects of urolithins at concentrations that are physiologically relevant for gut tissues (≥40 μM), as revealed in this study, support the data from in vivo studies showing the beneficial effects of ellagitannin-rich products toward intestinal inflammation.

Ethnopharmacological relevance Purple loosestrife -Lythrum salicaria L. is a herbaceous perennial plant belonging to the Lythraceae family. It has been used for centuries in European traditional medicine. Despite Lythri herba being a pharmacopoeial plant material (Ph. Eur.), L. salicaria popularity as a medicinal plant has recently declined. The aim of the paper is to recall a traditional and historical use of L. salicaria and juxtapose it with comprehensive view on the current knowledge about its chemical composition and documented biological activities in order to bring back the interest into this valuable plant and indicate reasonable directions of future research and possible applications. Materials and methods Systematic survey of historical and ethnopharmacological literature was carried out using sources of European and American libraries. Pharmacological and phytochemical literature research was performed using Scopus, PubMed, Web of Science and Reaxys databases. Results The review of historical sources from ancient times till 20th century revealed an outstanding position of L. salicaria in traditional medicine. The main applications indicated were gastrointestinal tract ailments (mainly dysentery and diarrhea) as well as different skin and mucosa affections. The current phytochemical studies have shown that polyphenols ( C -glucosidic ellagitannins and C -glucosidic flavonoids) as well as heteropolysaccharides are dominating constituents, which probably determine the observed pharmacological effects. The extracts and some isolated compounds were shown to possess antidiarrheal, antimicrobial, anti-oxidant, anti-inflammatory and anti-diabetic activities. Conclusions The intrinsic literature overview conclusively demonstrates that L. salicaria L. used to be considered as an exceptionally effective remedy in European traditional medicine. Despite its unquestionable important position from unknown reasons its popularity has been weakened during the past few decades. Unfortunately the contemporary pharmacological research is still insufficient to support its thoroughly described traditional uses. The necessity of complex studies regarding modes of action, which would directly refer to L. salicaria main traditional applications—gastrointestinal tract ailments, is strongly underlined.

Galinsoga ciliata Raf. Blake like Galinsoga parviflora Cav., comes from the Andes region. The chemical composition, activity and use are similar for both species. Galinsoga species are used in folk medicine as anti-inflammatory agents and accelerators for wound healing. Extracts are applied topically onto the skin to treat dermatological diseases, eczemas, lichens and hard-healing-wounds, and also to treat snakebites. Orally they used to cure flu and colds. In the studies using HPTLC method, different stationary phases, including unmodified silica gel, silica gels modified with CN, NH2, DIOL and RP18 groups were tried. The best separation of the tested compounds was achieved on silica gel plates, when as mobile phases mixtures – ethyl acetate–acetic acid–formic acid–water (100:11:11:26, v/v/v/v), ethyl acetate–methanol–formic acid–water (50:3:4:6, v/v/v/v) and ethyl acetate–methyl ethyl ketone–formic acid–water (30:9:3:3, v/v/v/v) – were used. Using reference substances, in the examined extracts the presence of flavonoids: patulitrin, quercimeritrin, quercitagetrin, and phenolic acids – caffeic and chlorogenic acids was found. HPLC analyses of extracts were carried out on a reversed-phase Zorbax SB column (150 mm × 2.1 mm, 1.9 μm). The mobile phase (A) was water/acetonitrile/formic acid (95:5:0.1, v/v/v) and the mobile phase (B) was acetonitrile/formic acid (100:0.1, v/v). A linear gradient system was used: 0–30 min, 1–30% B. Application of HPLC-DAD-MS method confirmed the results obtained by HPTLC method. Moreover, in the tested extracts the presence of caffeoylglucaric acids as dominating compounds was detected.

Introduction The genus Ligustrum (Oleaceae) is distributed in Europe and Asia (south China and Korea), where it is used to prevent hypertension, sore throats, inflammation and diabetes. The main groups of compounds in extracts of Ligustrum vulgare are biologically active secoiridoids and phenylpropanoids. Objectives The aim of the study was primarily the development and validation of a HPTLC–photodensitometry method for separation and determination of secoiridoids (oleacein, oleuropein) and phenylpropanoids (echinacoside) in different extracts prepared from leaves of L. vulgare. A secondary issue was the quantitative screening of oleacein, oleuropein and echinacoside in extracts from leaves collected at different stages of plant growth (from May to September). Methods A HPTLC–photodensitometry method was developed and validated for quantification of oleuropein, oleacein and echinacoside in plant extracts (aqueous and ethanolic extract, decoction, infusion). Silica gel was used as the stationary phase and dichloromethane:methanol:formic acid:water (80:25:1.5:4, v/v/v/v) as the mobile phase. Results The HPTLC–photodensitometry method developed for quantification of oleacein, oleuropein and echinacoside was specific, accurate and precise. The presence of oleacein was detected in aqueous extracts, whereas oleuropein was present, in particular, in ethanolic extracts, decoctions and infusions. Echinacoside was detected in all the extracts prepared. The content of secoiridoids was variable from May to September, whereas the amount of echinacoside increased in this term. Conclusion The developed and validated HPTLC–photodensitometry method allowed performing fast screening of quantitative profiles of oleacein, oleuropein and echinacoside in preparations of privet leaves. Copyright © 2015 John Wiley & Sons, Ltd.

The aim of the study was to obtain transformed roots ofRhaponticum carthamoidesand evaluate their phytochemical profile. Hairy roots were induced from leaf explants by the transformation ofAgrobacterium rhizogenesstrains A4 and ATCC 15834. The best response (43%) was achieved by infection with A4 strain. The effects of different liquid media (WPM, B5, SH) with full and half-strength concentrations of macro- and micronutrients on biomass accumulation of the best grown hairy root line (RC3) at two different lighting conditions (light or dark) were investigated. The highest biomass (93 g L−1of the fresh weight after 35 days) was obtained in WPM medium under periodic light. UPLC-PDA-ESI-MS3and HPLC-PDA analyses of 80% aqueous methanol extracts from the obtained hairy roots revealed the presence of eleven caffeoylquinic acids and their derivatives and five flavonoid glycosides. The production of caffeoylquinic acids and their derivatives was elevated in hairy roots grown in the light. Only light-grown hairy roots demonstrated the capability for the biosynthesis of such flavonoid glycosides as quercetagetin, quercetin, luteolin, and patuletin hexosides. Chlorogenic acid, 3,5-di-O-caffeoylquinic acid and a tentatively identified tricaffeoylquinic acid derivative were detected as the major compounds present in the transformed roots.

The antioxidant activity of the crude extract (60% ethanol) from the leaves of Desmodium adscendens (Sw.) DC. (Fabaceae) was observed in DPPH, xanthine/xanthine oxidase, lipid peroxydation and neutrophils burst tests. Further activity-guided fractionation on C18 column (water, 20% methanol, 50% methanol and 100% methanol) resulted in the separation of the fraction (50% methanol) with the highest antioxidant capacity. HPLC-DAD analysis of biologically active fraction revealed the presence of two pairs of flavonoid isomers as the dominant constituents. Those compounds were isolated and purified by multi-step liquid column chromatography (Sephadex LH20). Their structures were elucidated by various spectroscopic techniques, including NMR, UV and MS. Based on 1D and 2D NMR spectra as well as ion fragmentation, flavonoids were identified as: isovitexin 2''-O-xyloside (1), vitexin 2''-O-xyloside (2), vitexin (3) and isovitexin (4). The hybrid HSQC-DEPT technique provided very fast determination of the glycosylation positions in aglycone and the type of glycosidic bond in the flavonoid isomers. This study provides novel information concerning identity of the major compounds present in the leaves of D. adscendens cultivated in Ghana, which broadens the knowledge about anti-inflammatory, antiallergic and antioxidant properties of their extracts.

The aim of our study was to investigate the presence and quantitative contents of lignans in the tissues of Taxus ×media. The presence of the lignans: pinoresinol, matairesinol and secoisolariciresinol was assessed in needles, shoots cultures and suspension culture. Pinoresinol was the only lignan found in the tissue of T. ×media. The total pinoresinol content in the needles and in the shoots was 1.24 mg/g dry weight (dw) and 0.69 mg/g dw, respectively. Most of the pinoresinol identified was appeared glycosidically bound. In needles, the amount of glycosidically bound pinoresinol (0.81 mg/g dw) was about twice as high as that of free pinoresinol (0.43 mg/g dw). The content of free and glycosidically bound pinoresinol showed the level of 0.18 mg/g dw and 0.51 mg/g dw, respectively in the in vitro shoot cultures. In the cell culture, no pinoresinol was found.

Background Urolithins are bioavailable products of gut microbiota metabolism of ellagitannins. Their biological activity includes anti-cancer effects. Purpose The aim of this study was to explore the effects of urolithins on prostate cancer cells and activity of clinically used anti-androgen, bicalutamide. Methods Prostate cancer cells were treated with urolithin A, urolithin B, urolithin C or their combinations with bicalutamide. Cell proliferation was determined by DNA fluorescence with Hoechst 33258. The combination index method was used to examine interactions. Apoptosis and androgen receptor (AR) localization were analysed by flow cytometry. Prostate specific antigen (PSA) secretion was measured by ELISA. Results Urolithins inhibited proliferation of LNCaP prostate cancer cells. The mixtures of bicalutamide with uroA and uroB had additive anti-proliferative effect. All tested urolithins induced apoptosis of LNCaP cells. However, the combinations of bicalutamide with urolithin A and urolithin B had attenuated pro-apoptotic activity. UroA and uroC decreased DHT-induced PSA secretion. In contrast, uroB impaired PSA lowering effect of bicalutamide. UroA, individually and in combination with bicalutamide, promoted cytoplasmic localization of AR. Conclusion Urolithins might contribute to chemopreventive activity of ellagitannin rich preparations. Our results support use of ellagitannin rich preparations in prostate cancer chemoprevention, but advise caution in their potential use in complementary therapy of prostate cancer. The differences in activity profiles of urolithins indicate that possible health benefits and interactions will depend on the type of produced ellagitannins metabolite.

Background Chronic hyperalgesia and allodynia associated with progressive damage of peripheral neurons are the most prevalent complications of diabetes mellitus. Plants belonging to the family of Oleaceae were traditionally used in folk medicine for the management of diabetes. Hypothesis/Purpose The aim of this study was to investigate whether an aqueous extract from the leaves of Ligustrum vulgare (common privet) could be useful to target neuropathic pain in a rat streptozotocin (STZ) model of diabetes. Methods The chemical composition of the aqueous extract from privet leaf was characterized with the UHPLC-DAD-MS method and the analytical quantification of its constituents was performed with HPLC-DAD. Mechanical hyperalgesia and allodynia were evaluated with the Randall–Selitto and von Frey tests. Results Our investigation revealed the presence of secoiridoids: oleacein (23.48 ± 0.87 mg/g), oleocanthal (8.44 ± 0.08 mg/g), oleuropein (1.50 ± 0.01 mg/g), as well as phenylpropanoids: echinacoside (6.46 ± 0.07 mg/g), verbascoside (4.03 ± 0.04 mg/g) and p-coumaroyl glucarates in the dried aqueous extract of privet leaves. Behavioral data indicated that chronic intraperitoneal administration of the extract (50–200 mg/kg) for 21 days resulted in a decrease in diabetes-induced hyperalgesia and allodynia. Blood glucose levels remained unaltered, while body weight and water intake decreased significantly. Conclusion The aqueous privet leaf extract could serve useful in facilitating treatment of painful diabetic neuropathy. Additionally, the study showed that the antihyperalgesic activity of Ligustrum vulgare leaf extract is not likely related to its antihyperglycemic properties.

Thirteen new compounds including caffeoyl-glucaric and p-coumaroyl-altraric acid derivatives, one monoterpenoid glucoside, four secoiridoid glycosides, and three hydroxycinnamoyl phenylpropanoid glycosides esterified with an oleoside 11-methyl ester along with fifteen known compounds were isolated from flowers of Syringa vulgaris L. (Oleaceae). Their structures were elucidated by high-resolution spectroscopic methods. The tested compounds were able to decrease the production of reactive oxygen species. Moreover, oleoechinacoside (13), demethylhydroxyoleonuezhenide (14), demethyloleonuezhenide (15), syringaoleoacteoside (25) and oleoacteoside (26) at the concentration of 50μM, moderately suppressed the LPS-stimulated release of pro-inflammatory chemokine IL-8 and TNF-α from human neutrophils. Moreover, oleonuezhenide (12), oleoside 11-methyl ester (16) and oleoacteoside (26) at the concentration of 50μM were able to induce the surface expression of interleukin 10 receptor, which is suppressed by the incubation of monocyte/macrophage cells with LPS.

In traditional medicine, flowers and aerial parts of Lamium album are assigned by their anti-inflammatory, antiseptic, and mucolytic activities, and are used in chronic bronchitis and pharyngitis as well as skin, vaginal, and cervical inflammation.The aim of the present study was to compare effects of ethanolic extracts prepared from flowers and aerial parts of L. album on selected functions of human neutrophils, which are involved in an inflammatory response. In order to identify the compounds engaged in the anti-inflammatory activity of extracts, the bioassay-guided isolation of compounds was performed based on the inhibition of cytokine secretion by stimulated neutrophils.The extracts were phytochemically characterized with the HPLC-DAD-MSn method. The inhibition of reactive oxygen species production by formyl-met-leu-phenylalanine- or phorbol 12-myristate 13-acetate-stimulated neutrophils was determined using luminol- or lucigenin-dependent chemiluminescence. The effect on myeloperoxidase secretion by neutrophils was established spectrophotometrically. The levels of cytokine (interleukin 8, TNF-α) production after extract treatment was measured by ELISA.The most abundant constituents of extracts were phenylpropanoids, iridoids, flavonoids, and phenolic acids. Both extracts at concentrations of 25 and 100 µg/mL significantly inhibited reactive oxygen species production, and myeloperoxidase and interleukin 8 secretion. The phenylethanoid glycosides, such as lamiusides A, B, and C as well as 6″-O-β-D-glucopyranosylmartynoside, were isolated and identified. The cells treated with 6″-O-β-D-glucopyranosylmartynoside and lamiuside B produced 29.47 ± 7.11 % and 64.67 ± 5.25 % of interleukin 8, respectively, compared to non-treated control cells.Our results support the traditional use of L. album and indicate it as a potential source of natural anti-inflammatory constituents, such as phenylpropanoids.

In recent years, many xenobiotics derived from natural products have been shown to undergo extensive metabolism by gut microbiota. Ellagitannins, which are high molecular polyphenols, are metabolized to dibenzo[

Ethnopharmacological relevance Ellagitannin-rich plant materials are widely used in traditional medicine as effective, internally used anti-inflammatory agents. Due to the not well-established bioavailability of ellagitannins, the mechanisms of observed therapeutic effects following oral administration still remain unclear. The aim of the study was to evaluate if selected ellagitannin-rich plant materials could be the source of bioavailable gut microbiota metabolites, i.e. urolithins, together with determination of the anti-inflammatory activity of the metabolites produced on the THP-1 cell line derived macrophages model. Materials and Methods The formation of urolithins was determined by ex vivo incubation of human fecal samples with aqueous extracts from selected plant materials. The anti-inflammatory activity study of metabolites was determined on PMA differentiated, IFN-γ and LPS stimulated, human THP-1 cell line-derived macrophages. Results The formation of urolithin A, B and C by human gut microbiota was established for aqueous extracts from Filipendula ulmaria (L.) Maxim. herb (Ph. Eur.), Geranium pratense L. herb, Geranium robertianum L. herb, Geum urbanum L. root and rhizome, Lythrum salicaria L. herb (Ph. Eur.), Potentilla anserina L. herb, Potentilla erecta (L.) Raeusch rhizome (Ph. Eur.), Quercus robur L. bark (Ph. Eur.), Rubus idaeus L. leaf, Rubus fruticosus L. and pure ellagitannin vescalagin. Significant inhibition of TNF-α production was determined for all urolithins, while for the most potent urolithin A inhibition was observed at nanomolar concentrations (at 0.625 μM 29.2±6.4% of inhibition). Urolithin C was the only compound inhibiting IL-6 production (at 0.625 μM 13.9±2.2% of inhibition). Conclusions The data obtained clearly indicate that in the case of peroral use of the examined ellagitannin-rich plant materials the bioactivity of gut microbiota metabolites, i.e. urolithins, has to be taken under consideration.

Geum urbanum L. commonly known as wood avens or St. Benedict’s herb is an herbaceous perennial plant with characteristic pinnate leaves with a very large and lobed terminal leaflet. G. urbanum belongs to the family of Rosaceae and is native to humid woods of Europe and Asia (Wichtl, 2004). Infusions from the roots and rhizomes of wood avens have been traditionally used in folk medicine in the treatment of acute diarrhea and as astringent agent for inflammations of the mucosa and gums and for treatment of hemorrhoids (Wichtl, 2004; Popovic et al., 2012). This study investigates the roots of G. urbanum collected from flowering plants growing in their natural state in Umer ( Swietokrzyskie district, Poland) in September 2011. Plant material was authenticated by Izabella Kirpluk (Botanical Garden, University of Warsaw, Poland). Voucher specimen (no. GU092011) has been deposited in the Plant Collection, Department of Pharmacognosy and Molecular Basis of Phytotherapy, Medical University of Warsaw, Poland.

The aim of our study was to determine antioxidant activity of aqueous and hydroethanolic extracts obtained from both freeze-drying leaves and flowers, as well as juice squeezed from the fresh herb of Tropaeolum majus L. In cell-free systems the scavenging of synthetic radical (DPPH) and two radicals (O 2 − , H 2 O 2 ) was evaluated. In in vitro experiments, the effect on reactive oxygen species (ROS) production by f-MLP-stimulated neutrophils, as well as the effect on ROS generation by human skin fibroblasts after UV irradiation were determined. In studies on fibroblasts, the protective effect of the extracts/juice against cell membrane damage caused by UV irradiation was also tested. The tested extracts/juice were chemically characterized by the determination of the content of flavonoids, total phenols and ascorbic acid. Scavenging activity of the tested extracts/juice against synthetic radical-DPPH was low. Against H 2 O 2 and O 2 − the extracts showed stronger antioxidant activity, while the juice was significantly active only against O 2 − . In the human neutrophils model the hydroethanolic extract most strongly inhibited production of ROS. The weakest activity was shown by the juice. Studies on human skin fibroblasts showed no cytotoxic activity of the tested extracts/juice. We also observed inhibition of ROS production by tested extracts/juice induced by UVA as well as by UVB irradiation. The strongest inhibition of ROS production after UVA irradiation was shown by the juice, while after UVB irradiation the most potent was the hydroethanolic extract. Our results partly explain the use of extracts from nasturtium in skin diseases, such as sunburns.

Ellagitannin-rich plant materials are used as popular remedies in the treatment of various inflammatory diseases. Urolithins are gut microbiota metabolites of ellagitannins and are considered responsible for in vivo health effects. Various natural products have been studied that are known sources of urolithins. However, few studies have focused on the metabolism of ellagitannin molecules. The aim of the study was to examine the metabolic fate of select ellagitannins using ex vivo cultures of human gut microbiota. Fifteen monomeric and dimeric ellagitannins, 1-O-galloyl-4,6-(S)-HHDP-β-d-glucose (2), pedunculagin (3), potentillin (4), casuarictin (5), coriariin B (6), vescalagin (7), castalagin (8), stachyurin (9), casuarinin (10), stenophyllinin A (11), stenophyllanin A (12), salicarinin A (13), gemin A (14), agrimoniin (15), and oenothein B (16), and ellagic acid (1) were studied. The formation of the metabolites in ex vivo human microbiota cultures was monitored using UHPLC-DAD-MS/MS. Ellagitannins possessing hexahydroxydiphenoyl moieties were metabolized to 6H-dibenzo[b,d]pyran-6-one derivatives, i.e., urolithins. The observed differences in amounts of produced urolithins indicated that the individual microbiota composition and type of ingested ellagitannins could determine the rate of urolithin production. When the oral ingestion of natural products containing ellagitannins with hexahydroxydiphenoyl groups is considered, the formation of urolithins and their bioactivity should be addressed.

Carpesium divaricatum Sieb. & Zucc. has a long history of use as both a medicinal and a food plant. However, except for terpenoids, its chemical constituents have remained poorly investigated. The composition of hydroalcoholic extract from aerial parts of C. divaricatum was analyzed by HPLC-DAD-MSn, revealing the presence of numerous caffeic acid derivatives that were formerly unknown constituents of the plant. In all, 17 compounds, including commonly found chlorogenic acids and rarely occurring butyryl and methylbutyryl tricaffeoylhexaric acids, were tentatively identified. Fractionation of lipophilic extract from cultivated shoots led to the isolation of 12-oxo-phytodienoic acid (12-OPDA), which is a newly identified constituent of the plant. The compound, at concentrations of 0.5, 1.0, and 2.5 μM, significantly reduced IL-8, IL-1β, TNFα, and CCL2 excretion by lipopolysaccharide (LPS)-stimulated human neutrophils. Reactive oxygen species (ROS) production induced by f-MLP was also significantly diminished in the neutrophils pretreated by 12-OPDA. The newly identified constituents of the plant seem to be partly responsible for its pharmacological activity and elevate the value of C. divaricatum as a potential functional food.

Introduction Lythri salicariaeherba is a pharmacopoeial plant material used by patients in the form of infusions in the treatment of acute diarrhoea. According to its pharmacopoeial monograph it is standardised for total tannin content, which should be not less than 5.0% using pyrogallol as a standard. Previous studies have shown that aqueous extracts from Lythri herba contain mainly ellagitannins among which vescalagin, castalagin and salicarinins A and B are dominating constituents. Objective To develop and validate an efficient UHPLC coupled with a charged aerosol detector (CAD) method for quantification of four major ellagitannins in Lythri salicariaeherba and in one commercial preparation. Methods Extraction conditions of ellagitannins from plant material were optimised. The relative response factors for vescalagin, castalagin and salicarinins A and B using gallic acid as an external standard were determined for the CAD detector. Then, a UHPLC method for quantification of ellagitannins was developed and validated. Results Four major ellagitannins were quantified in four samples of Lythri herba and in one commercial preparation. The sum of ellagitannins for each sample was determined, which varied from 30.66 to 48.80 mg/g of raw material and 16.57 mg per capsule for the preparation investigated. Conclusion The first validated UHPLC/CAD UHPLC-CAD method for quantification of four major ellagitannins was developed. The universality of the CAD response was evaluated and it is shown that although all compounds analysed have similar structures their CAD response differs significantly. Copyright © 2013 John Wiley & Sons, Ltd.

In this study, we synthesized several imperatorin analogs using imperatorin and xanthotoxin as substrates. The anti-cholinesterase activities of all compounds were evaluated in in vitro experiments according to the modified Ellman's method. For each synthesized compound, IC50 values for both enzymes were established. Galantamine hydrobromide was used as a positive control in the enzymatic experiments. All active compounds showed selectivity toward butyrylcholinesterase (BuChE) rather than acetylcholinesterase. The most active ones were 8-(3-methylbutoxy)-psoralen and 8-hexoxypsoralen with IC50 values for BuChE of around 16.5 and 16.4 µM, respectively. The results of our study may be considered as the beginning of a search for potential anti-Alzheimer's disease drugs based on the structure of natural furocoumarins.

The aim of this study was comparison of antioxidant, anti-inflammatory, and antimicrobial activity, as well as chemical composition of extracts of Tropaeolum majus L. herb. Aqueous and hydroethanolic extracts derived from dried and freeze-dried nasturtium herb, prepared both at room temperature and at 90 °C, were studied. In the studies there were no significant differences between antioxidant activity of the extracts. All extracts showed scavenging activity against all the examined reactive species in a concentration-dependent manner. The strongest scavenging activity they showed against reactive nitrogen species, NO (SC50 4.54 ± 0.26–10.90 ± 1.39 μg/mL) and ONOO − (SC50 2.49 ± 1.50–6.37 ± 1.86 μg/mL). Among reactive oxygen species, they showed strong scavenging activity against H 2 O 2 (SC50 14.90 ± 3.91–38.63 ± 9.28 μg/mL). Scavenging activity against O 2 − was weaker, while against HClO the extracts showed very weak activity, practically at the level of statistical error. Against synthetic radical – DPPH scavenging activity of the tested extracts was negligible. The extracts demonstrated stronger antioxidant activity in ex vivo experiment on human neutrophils. The extracts showed no inhibitory activity on hyaluronidase, but at a concentration of 50 μg/mL they inhibited the activity of COX1 by approximately 60%. Lack of antimicrobial activity of the extracts seems to be associated with a low content of benzyl isothiocyanate. The aqueous extracts were characterized by the presence of esters of quinic acid with cinnamic acids (chlorogenic acids, p-coumaroylquinic acids) and the presence of flavonoids. Meanwhile the hydroethanolic extracts were mainly rich in the above mentioned acid esters.

Leaves of Ligustrum vulgare (common privet) have been used for treatment of oropharyngeal inflammations or as antirheumatic, diuretic, and hypotensive agents in folk medicine in southern Europe. Taking into account that neutrophils are involved in the inflammation, the aim of the study was to determine the effect of an aqueous extract prepared from leaves of Ligustrum vulgare on neutrophil functions. The extract was characterized by the HPLC-DAD-MSn method. The inhibition of reactive oxygen species production by formyl-met-leu-phenylalanine- or phorbol 12-myristate 13-acetate-stimulated neutrophils was determined using luminol- or lucigenin-dependent chemiluminescence. The effect on myeloperoxidase, metalloproteinase 9, and interleukin 8 production by neutrophils was measured by an enzyme-linked immunosorbent assay. Neutrophil elastase release was established spectrophotometrically. The expression of adhesion molecules on neutrophils was analyzed with flow cytometry. The main compounds detected were flavonoids, phenylpropanoids, hydroxycinnamates, and secoiridoids. The inhibition of oxidative burst by the extract was comparable in both stimuli models (formyl-met-leu-phenylalanine: IC50 = 18.2 ± 4.0 µg/mL; phorbol 12-myristate 13-acetate: IC50 = 19.8 ± 3.0 µg/mL). The extract in the concentration range of 5–50 µg/mL inhibited neutrophil elastase release by 23.9–34.1 % and myeloperoxidase release by 24.2–37.4 %. The inhibitory effect on metalloproteinase 9 and interleukin 8 production was around 20 %. The extract in the highest concentration modulated the expression of L-selectin and β 2 integrin. Our results partly support the traditional use of common privet leaves as an anti-inflammatory agent.

Extracts from Epilobium sp. herbs have been traditionally used in the treatment of prostate-associated ailments. Our studies demonstrated that the extracts from Epilobium angustifolium, Epilobium parviflorum and Epilobium hirsutum herbs are potent prostate cancer cells (LNCaP) proliferation inhibitors with IC50 values around 35 mg/ml. The tested extracts reduced prostate specific antigen (PSA) secretion (from 325.625.3 ng/ml to ~90 ng/ml) and inhibited arginase activity (from 65.21.1 mUnits of urea/mg of protein to ~40 mUnits of urea/mg protein). Selected constituents of extracts (oenothein B, quercetin-3-O-glucuronide, myricetin-3-O-rhamnoside) were proventobeactiveinrelationtoLNCaPcells.However,oenotheinBwasthestrongestinhibitorofcellsproliferation (IC50=7.80.8 mM), PSA secretion (IC50=21.93.2 mM) and arginase activity (IC50=19.22.0 mM). Additionally, ellagitannins from E. hirustum extract were proven to be transformed by human gut microbiota into urolithins. Urolithin C showed the strongest activity in the inhibition of cell proliferation (IC50=35.23.7 mM), PSA secretion (reduced PSA secretion to the level of 100.731.0 ng/ml) and arginase activity (reduced to the level of 27.93.3 mUnits of urea/mg of protein). Results of the work offer an explanation of the activity of Epilobium extractsand supporttheuseof Epilobium preparationsin thetreatment of prostate diseases. Copyright © 2013 John Wiley & Sons, Ltd.

Circaea lutetiana L. is a perennial herbaceous plant belonging to the family of Oenotheraceae (Onagraceae). It occurs as a common herb in forests of North America, Europe and Asia. Native Americans have used infusions of Circaea as a remedy for wounds and as a wash on injured parts of the body (Moerman, 1998). This study investigates the aerial parts of C. lutetiana L., which were collected from natural state in Bremen (Germany) in the period of blooming in August 2011. Plant material was authenticated by Dr. Anna Kiss and Dr. Wojciech Szypu1a (Department of Pharmaceutical Botany, Medical University of Warsaw) based on morphological characters. Voucher specimen (no. CL082011) has been deposited in the Plant Collection, Department of Pharmacognosy and Molecular Basis of Phytotherapy, Medical University of Warsaw, Poland.

In the present study we investigated the chemical composition of extracts prepared from aerial parts of Oenothera paradoxa Hudziok and Oenothera biennis L. and their antioxidative and anti-inflammatory activities. Ultra high pressure liquid chromatography (UHPLC)-DAD-MS/MS studies showed that both extracts contain a wide variety of polyphenols (39 identified constituents) among which macrocyclic ellagitannin turned out to be the main constituent. During the in vitro studies, using noncellular models, both extracts scavenged reactive oxygen species (ROS) in a concentration-dependent manner, and the lowest SC(50) values were obtained for O(2)(-) and H(2)O(2). Both extracts inhibited ROS production by stimulated human neutrophils. The stronger activity in the case of formyl-met-leu-phenylalanine stimulation suggests that both extracts may act through the receptor-dependent pathway. O. paradoxa extract and O. biennis extract exhibited anti-inflammatory activity by the inhibition of hyaluronidase and lipoxygenase in a concentration-dependent manner. The stronger activity of O.biennis extract toward lipoxygenase may be explained by its higher oenothein B content.

Polygonum aviculare L. (Common Knotrgrass) ( Polygonaceae Juss.) is an annual from which pharmacopoeial (Ph. Eur.) plant material Polygoni avicularis herba is obtained. Although its main active constituents are flavonoids and its standardization is based on their total content, no profound qualitative analysis has been performed, yet. The use of hyphenated analytical methods: UHPLC–DAD coupled with ion trap or time of flight mass detectors together with acidic hydrolysis products analysis allowed for a comprehensive determination of flavonoid composition. Among dominating compounds, myricetin, kaempferol, isorhamnetin and kaempferide glucuronides, which were not previously reported in P . avicularis herba , were discovered. The developed method can be used as a suitable tool for a more insightful, metabolome-based standardization of flavonoid rich pharmacopoeial plant material – P . avicularis herba .

Introduction Lythri herba, a pharmacopoeial plant material (European Pharmacopoea), is obtained from flowering parts of purple loosestrife (Lythrum salicaria L.). Although extracts from this plant material have been proven to possess some interesting biological activities and its pharmacopoeial standardisation is based on total tannin content determination, the phytochemical characterisation of this main group of compounds has not yet been fully conducted. Objective To isolate ellagitannins from Lythri herba, determine their structures and develop chromatographic methods for their qualitative analysis. Results Five C-glucosidic ellagitannins – monomeric- vescalagin and castalagin together with new dimeric structures – salicarinins A–C, composed of vescalagin and stachyurin, vescalagin and casuarinin, castalagin and casuarinin units connected via formation of valoneoyl group, were isolated using column chromatography and preparative HPLC. Structures were determined according to 1H and 13C-NMR (one- and two-dimensional), electrospray ionisation–time of flight (ESI–TOF), electrospray ionisation-ion trap (ESI-MSn) and circular dichroism (CD) spectra, together with acidic hydrolysis products analysis. HPTLC on RP-18 modified plates and HPLC–DAD–MSn on RP-18 column methods were developed for separation of the five main ellagitannins. Copyright © 2012 John Wiley & Sons, Ltd.

Background In patients with hypertension the haemorrhage into carotid atherosclerotic plaque increases risk of plaque destabilization and rupture. Our previous study showed that oleacein, a secoiridoid present in extra virgin olive oil, enhanced uptake of haemoglobin-haptoglobin complex and change macrophage phenotype from pro-inflammatory M1 to anti-inflammatory M2. Purpose The aim this study was to investigate a potential role of oleacein in attenuation of carotid plaque destabilisation ex vivo . Methods Samples of atherosclerotic plaque were harvested from 20 patients with hypertension /11 women and 9 men/, who underwent carotid endarterectomy after transient ischemic attacks. Matching pieces of each plaque were incubated with increased concentration of pure oleacein /range 0–20 µM/ for 24 h. HMGB1, MMP-9, MMP-9/NGAL, TF and IL-10, as well as HO-1 secretion from plaque was measured by enzyme–linked immunosorbent assay /ELISA/. Statistical significance was set at P 0.05 and P 0.001. Results Oleacein at the concentrations of 10 and 20 µM significantly ( P 0.001) decreased secretion of HMGB1 (up 90%), MMP-9 (up to 80%), MMP-9/NGAL complex (up to 80%) and TF (more than 90%) from the treated plaque, as compared to control. At the same time IL-10 and HO-1 release increased by more than 80% ( P 0.001). Conclusion Our results indicate that oleacein possess ability to attenuate the destabilization of carotid plaque and could be potentially useful in the reduction of ischemic stroke risk.

The popularity of food products and medicinal plant materials containing hydrolysable tannins (HT) is nowadays rapidly increasing. Among various health effects attributable to the products of plant origin rich in gallotannins and/or ellagitannins the most often underlined is the beneficial influence on diseases possessing inflammatory background. Results of clinical, interventional and animal in vivo studies clearly indicate the antiinflammatory potential of HT-containing products, as well as pure ellagitannins and gallotannins. In recent years a great emphasis has been put on the consideration of metabolism and bioavailability of natural products during examination of their biological effects. Conducted in vivo and in vitro studies of polyphenols metabolism put a new light on this issue and indicate the gut microbiota to play a crucial role in the health effects following their oral administration. The aim of the review is to summarize the knowledge about HT-containing products’ phytochemistry and their anti-inflammatory effects together with discussion of the data about observed biological activities with regards to the current concepts on the HTs’ bioavailability and metabolism. Orally administered HT-containing products due to the limited bioavailability of ellagitannins and gallotannins can influence immune response at the level of gastrointestinal tract as well as express modulating effects on the gut microbiota composition. However, due to the chemical changes being a result of their transit through gastrointestinal tract, comprising of hydrolysis and gut microbiota metabolism, the activity of produced metabolites has to be taken into consideration. Studies regarding biological effects of the HTs’ metabolites, in particular urolithins, indicate their strong and structure-dependent anti-inflammatory activities, being observed at the concentrations, which fit the range of their established bioavailability. The impact of HTs on inflammatory processes has been well established on various in vivo and in vitro models, while influence of microbiota metabolites on silencing the immune response gives a new perspective on understanding anti-inflammatory effects attributed to HT containing products, especially their postulated effectiveness in inflammatory bowel diseases (IBD) and cardiovascular diseases.

Hydroalcoholic extract from Inula helenium callus tissue showed remarkable reducing capacity. An HPLC/DAD analysis revealed the presence of numerous hydroxycinnamic acid derivatives, including chlorogenic acid (5-O-CQA, 0.1 % dry weight) and 3,5-dicaffeoylquinic acid (3,5-DCQA, 0.3 % dry weight), which were among major constituents of the examined extract. Application of a hyphenated chromatographic method—UHPLC/DAD/MSn—allowed identification of sixteen compounds, derivatives of caffeic acid. Apart from the compounds commonly found in Inula sp., the plant material contained eight conjugates of caffeic acid with aldaric acid. The aldarates constituted over 50 % of the hydroxycinnamate fraction in the examined tissue. This is the first report on the occurrence of the caffeoylaldaric acids in Inula sp. and in a plant tissue culture.

Taking into account the role of reactive oxygen species in the development of inflammation, and the application of the plants of genus Galinsoga RuizPav. in folk medicines for inflammatory states, we investigated and compared the antioxidant activities of particular Galinsoga extracts and fractions. The compositions of the most active fractions were studied using thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) methods. The extracts and fractions from Galinsoga parviflora Cav. and Galinsoga quadriradiata Ruiz et Pav. possess dose-dependent free radical-scavenging ability against DPPH• and superoxide radicals, as well as inhibitory effects on linoleic acid peroxidation in a manner comparable to gallic acid. In the most active fractions, flavonoids, patulitrin, quercimeritrin, quercitagetrin and caffeoyl derivatives were detected. Our research demonstrates that the investigated herbs are an interesting source of preparations with significant antioxidant effects. Our results justify the use of both raw materials in inflammatory diseases, among others, due to their ability to prevent free radical-induced deleterious effects.

Penta-O-galloyl-β-D-glucose (PGG) occurrs in high concentrations in medicinal herbs such as Rhus chinensis, Paeonia suffruticosa, Acer truncatum and Terminalia chebula, which demonstrate anti-inflammatory activity. We investigated the effect of PGG on stimulated and non-stimulated neutrophils in processes which included reactive oxygen species generation (ROS), metalloproteinase-9 and interleukin-8 secretion (IL-8), β2 integrin (CD11b) and L-selectin (CD62L) expression and apoptosis. In concentrations of 5 μM–20 μM, PGG demonstrated statistically significant inhibition of ROS generation, IL-8 secretion and β2 integrin expression in stimulated neutrophils. The inhibition of L-selectin expression by PGG resulted in prevention in neutrophils’ endothelial attachment. The result obtained may explain the anti-inflammatory activity of this compound and underline the contribution of PGG in the activity of PGG rich plant extracts. Copyright © 2012 John Wiley & Sons, Ltd.

In the present study, we have investigated the influence of low concentrations of polyphenols (5 μM) on histone acetyltransferase (HAT) and histone deacetylase (HDAC) activity, a monocyte cell system related to the inflammatory process. We selected gallic acid, ellagic acid, oenothein B, valoneic acid dilactone and penta- O -galloyl-β- d -glucose, which are known to affect inflammatory responses, such as IL-6 secretion and/or NF-κB down-regulation and polyphenol metabolites (urolithins A, B and C). TNF-α stimulation resulted in a reduction of THP-1 cell viability by 28.9%, reduced HDAC activity from 25.2 to 13.5 pmol/μg protein and increased HAT activity from 24.0 to 45.5 pmol/μg protein. The coincubation with ellagic acid and oenothein B restored the viability and reversed the effect of TNF-α on HAT and HDAC activities. Urolithins B and C, gallic acid and penta- O -galloyl-β- d -glucose only showed significant reductions of HAT activity, by 40–50%. Our results prove that polyphenols act as epigenetic modulators.

Phenolic compounds from virgin olive oil are known to play an antioxidant role in preventing biomolecule damage. One of the most abundant components of olive oil is an oleuropein derivative named oleacein (dialdehydic form of decarboxymethyl elenolic acid linked to hydroxytyrosol; 3,4-DHPEA-EDA). The aim of the study was to establish the antioxidant activity of oleacein for an array of reactive oxygen (superoxide anion, O 2 - ; hydrogen peroxide, H2O2; hypochlorous acid, HOCl) and nitrogen species (nitric oxide, NO; peroxynitrite, ONOO−) formation, using in vitro non-cellular systems, as well as cellular screening systems (human neutrophil oxidative burst, monocytes’ nitric oxide production). We used a well-known antioxidant, oleuropein, as a reference compound. Oleacein proved to be stronger in the reduction of formyl-met-leu-phenylalanine (f-MLP) and phorbol–myristate–acetate (PMA)-induced oxidative bursts in neutrophils (IC50 = 1.8 μM, IC50 = 1.5 μM, respectively) and myeloperoxidase release (IC50 = 8.8 μM) than was oleuropein (IC50 = 2.4 μM, IC50 = 16.0 μM, IC50 = 30.7 μM, respectively). Both compounds were stronger scavengers of O 2 - than H2O2. Oleuropein did not scavenge HOCl as opposed to oleacein (IC50 = 1.5 μM). Both compounds, in a concentration range 1–10 μM, significantly decreased nitrogen species formation in cell-free systems and by lipopolysaccharide (LPS)-stimulated monocytes.

Introduction Oenothein B (OeB) is a dimeric macrocyclic ellagitannin occurring mainly in the genus Oenothera. In the literature no validated HPLC method for quantification of this compound has been reported. Objective To develop and validate an efficient HPLC method for quantification of OeB in plant materials. Methodology Extraction conditions of OeB from plant material were optimised. Then the first validated HPLC method for quantification was developed. Results We have shown that ultrasonic-assisted extraction at 40 °C in 15 min using water are the most optimal conditions for OeB extraction. We have quantified OeB in several plant materials belonging to Oenotheraceae family. The lowest amount of OeB was present in Circaea lutetiana herb (27.64 ± 0.26 mg/g) and the highest was quantified in Epilobium hirsutum aerial parts (72.91 ± 1.38 mg/g). Conclusion The first HPLC method for quantification of OeB in plant materials was developed and validated. We also for the first time optimised the extraction conditions for OeB. Copyright © 2012 John Wiley & Sons, Ltd.

Biological investigations have revealed high scavenging capacity of Oenothera paradoxa defatted seed extract on reactive nitrogen species such as NO and ONOO − . The characteristics of the polyphenols present in the extracts were checked using high performance liquid chromatography coupled with electrospray negative ionization ion trap mass spectrometry. Extracts contained five groups of compound: phenolic acids (gallic acid, ethyl gallate, ellagic acid and ferulic acid pentoside), flavanols (catechin, catechin gallate) and oligomeric procyanidins, flavonols (quercetin-3- O -glucoside, quercetin-3- O -glucuronide, quercetin-3- O -pentoside and quercetin), and gallotannins (tetragalloyl glucose, pentagalloyl glucose and hexagalloyl glucose). Penta- O -galloyl-β- d -glucose were present in the extracts in concentrations from 9.44 to 16.75 mg/g, which demonstrated a significant NO and ONOO − scavenging activity with IC 50 0.20 and 0.06 μM, respectively, may be considered as an O. paradoxa extract quality marker.

Ethnopharmacological relevence The aim of the study was to examine extracts from twelve tannin-rich plant materials used in traditional Polish medicine for external treatment of skin and mucosa diseases considering their ability to inhibit hyaluronidase activity and elastase release from stimulated neutrophils in vitro. Materials and methods In vitro anti-hyaluronidase and anti-elastase assays together with phytochemical qualitative and quantitative screening were performed. Results The strongest inhibition of hyaluronidase was observed for extract from Lythri herba, with IC50 value 8.1 ± 0.8 μg/mL. The most active extract towards elastase release was from Hippocastani cortex which at concentration 10 μg/mL showed 62.0 ± 6.9% inhibition. Conclusion Anti-hyaluronidase and anti-elastase activity of chosen tannin-rich plant materials can support their traditional use in folk medicine. Strong inhibition of both enzymes by extract from Lythri herba makes this pharmacopeial plant material an interesting topic for further biological and phytochemical examination.

The qualitative analysis of the aqueous extract showed the presence of polyphenolics belonging to different chemical groups: flavan-3-ol derivatives (catechin and procyanidin B 3 ), gallic acid derivatives (gallic acid, ethyl gallate and pentagalloylglucose) and depsides (ellagic acid). The content of total phenolic and procyanidin in extracts depended on the duration of seeds storage and varied from 469 to 388 mg/g and from 436 to 372 mg/g, respectively. The HPLC quantitative analysis of the three main compounds showed that their content varied from 3.70 to 6.18 mg/g for gallic acid, from 23.43 to 18.91 mg/g for (+)-catechin and from 12.50 to 8.02 mg/g for pentagalloylglucose. During seeds storage, the quantity of pentagalloylglucose in extracts decreased by about 30%, while gallic acid content increase was about 30%. The quantity of (+)-catechin has changed less significantly. The scavenging potential of the extracts determined with the DPPH method correlated with the higher concentration of pentagalloylglucose.

Summary This article is the first report describing a new validated method to determine the content of HupA in Huperzia selago (Huperziaceae) from wild population and obtained in in vitro culture using the chaotropic mobile phase. An aqueous-organic (acetonitrile) mobile phase with an added chaotropic salt (NaPF6) was used. The system of mobile phases ensured very high selectivity and efficiency at up to N = 6683 ± 963 theoretical plates calculated for isocratic mode. A Hypercosil GOLD column, C18 250 × 4.6 mm, and a Hypercosil GOLD precolumn, 5UM 10 × 4 mm, were employed for detection at four wavelengths, 230 nm being analytical. The regression coefficient (R2) of the calibration was 0.9993 over the range 25–1252 μg mL−1. The recovery rates were 98.36–105.1% with RSD