Search

Eranthis longistipitata Regel is an endemic plant of Central Asia. The flavonoid profile of E. longistipitata leaves was studied by mass spectrometry for the first time (natural populations of Kyrgyzstan and Uzbekistan, in 70% aqueous–ethanol extracts by liquid chromatography coupled with high-resolution mass spectrometry). Mass spectrometry revealed 18 flavonoid compounds. Flavonols featured the highest diversity, and 10 such substances were identified: 2 free aglycones (quercetin and kaempferol), 6 quercetin glycosides (peltatoside, hyperoside, reynoutrin, quercetin 3-sambubioside, rutin, and isoquercitrin), and 2 kaempferol glycosides (juglalin and trifolin). Two flavans (cianidanol and auriculoside), two hydroxyflavanones (6-methoxytaxifolin and aromadendrin), and one C-glycoside flavone—carlinoside—were identified. Dihydroxychalcones aspalathin, phloridzin, and phloretin were found too. Levels of rutin, quercetin, kaempferol, and hyperoside were confirmed by means of standards and high-performance liquid chromatography. Rutin concentration was the highest among all other identified flavonoid compounds: in the leaf samples from Kyrgyzstan, it ranged from 2.46 to 3.20 mg/g, and in those from Uzbekistan, from 1.50 to 3.01 mg/g. The diversity of flavonoid compounds in E. longistipitata leaves is probably due to external ecological and geographic factors and adaptive mechanisms.

Leaves of Crataegus sanguinea Pall. afforded for the first time ergosterol 3-O-β-D-glucopyranoside, p-coumaric acid 4-O-β-D-glucopyranoside, trifolin, quercitrin, the new compound sanguineoside (5,7,3′,5′-tetrahydroxyflavanone 7-O-β-D-glucopyranoside), and compounds known from this species, e.g. oleanolic acid, caffeic acid, hyperoside, vitexin, and vitexin 2″-O-rhamnoside, which was the dominant flavonoid.

Zeylanicobdella arugamensis (Hirudinea), a marine parasitic leech, not only resulted in the mortality of the host fish (Groupers) but also caused economic losses. The current study aimed to elucidate the antiparasitic efficacy of the aqueous extract of the Azadirachta indica leaves against Z. arugamensis and to profile the composition via LC-Q Exactive HF Orbitrap mass spectrometry. Different concentrations (25, 50 and 100 mg/mL) of A. indica extract were prepared and tested on the parasitic leeches. The total mortality of leeches was noticed with an exposure to the A. indica aqueous extract. The average times required for the aqueous extract at concentrations of 25, 50 and 100 mg/mL to kill the leeches were 42.65 ± 9.20, 11.69 ± 1.11 and 6.45 ± 0.45 min, respectively, in a dose-dependent manner. The Orbitrap mass spectrometry analysis indicated the presence of five flavonoids (myricetin 3-O-galactoside, trifolin, isorhamnetin, quercetin and kaempferol), four aromatics (4-methoxy benzaldehyde, scopoletin, indole-3-acrylic acid and 2,4-quinolinediol), three phenolics (p-coumaric acid, ferulic acid and phloretin) and two terpenoids (pulegone and caryophyllene oxide). Thus, our study indicates that A. indica aqueous extract is a good source of metabolites with the potential to act as a biocontrol agent against the marine parasitic leech in aquaculture.

Schisandra rubriflora is a dioecious, underestimated medicinal plant species known from traditional Chinese medicine. The present study was aimed at characterising the polyphenolic profile composition and the related antioxidant capacity of S. rubriflora fruit, stem and leaf and in vitro microshoot culture extracts. Separate analyses of material from female and male specimens were carried out. This study was specifically aimed at detailed characterisation of the contribution of phenolic compounds to overall antioxidant activity using ultra-high-performance liquid chromatography with a photodiode array detector coupled to electrospray ionization ion trap mass spectrometry (UHPLC-DAD-ESI-MS3) and a high-performance liquid chromatography-diode array detector (HPLC-DAD). Using UHPLC-DAD-ESI-MS3, twenty-seven phenolic compounds from among phenolic acids and flavonoids were identified. Concentrations of three phenolic acids (neochlorogenic, chlorogenic and cryptochlorogenic acids) and eight flavonoids (hyperoside, rutoside, isoquercitrin, guaijaverin, trifolin, quercetin, kaempferol, and isorhamnetin) were determined using HPLC-DAD using reference standards. The highest total phenolic content was confirmed for the stem and leaf extracts collected in spring. The contents of phenolic compounds of in vitro biomasses were comparable to that in the fruit extracts. The methanolic extracts from the studied plant materials were evaluated for their antioxidant properties using various in vitro assays, namely free radicals scavenging estimation using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), ferric-reducing antioxidant power (FRAP) and cupric-reducing antioxidant capacity (CUPRAC) as well as QUick, Easy, New, CHEap, and Reproducible CUPRAC (QUENCHER-CUPRAC) assays. A close relationship between the content of polyphenolic compounds in S. rubriflora and their antioxidant potential has been documented.

We initiated and optimized in vitro culture conditions of the endemic Chinese plant species—Schisandra henryi C. B. Clarke. Different types of in vitro solid cultures (microshoot and callus), cultivation periods (10, 20, and 30 days), and selected concentrations of BA, IBA, GA3 (0 to 3 mg/l) in the Murashige and Skoog (MS) medium were tested. The presence of dibenzocyclooctadiene lignans (schisandrin, gomisin G, schisantherin A and B, deoxyschisandrin and schisandrin C), dibenzylbutane lignans (hernicine B), aryltetralin lignans (wulignan A1 and A2, epiwulignan A1, enshicine, epienshicine and dimethylwulignan A1), and triterpenoids: kadsuric acid and schisanhenric acid was confirmed by UHPLC–MS/MS analysis. Using HPLC–DAD, the qualitative and quantitative profiles of dibenzocyclooctadiene lignans, phenolic acids and flavonoids in methanolic extracts from biomass were estimated. The maximum total amounts of these groups of metabolites were 873.71, 840.89 and 421.98 mg/100 g DW, respectively. The main compounds were: schisantherin B (max. 622.59 mg/100 g DW), schisantherin A (max. 143.74 mg/100 g DW), neochlorogenic acid (max. 472.82 mg/100 g DW), caftaric acid (max. 370.81 mg/100 g DW), trifolin (max. 138.56 mg/100 g DW) and quercitrin (max. 122.54 mg/100 g DW). The highest total amounts of secondary metabolites estimated in the extracts from in vitro cultures were, respectively, 13.0, 7.0, and 1.4 times higher than in the leaf extracts analyzed for comparison. This is the first report on the biosynthetic potential of cells from Schisandra henryi in vitro cultures. The initiation, broad optimization and comprehensive phytochemical studies of lignans, phenolic acids and flavonoids in the extracts from Schisandra henryi microshoot and callus in vitro cultures were elaborated for the first time.

Spiraea L. belongs to a genus of deciduous-leaved shrubs in the Rosaceae family that is abundant in Eastern Siberia. The study of six species of Spiraea growing in the Baikal region revealed that they were characterized with high content of phenolic compounds and their extracts demonstrated an inhibitory effect on α-amylase. It was established using correlation analysis that flavonoids were the leading components associated with this biological effect of the extracts. The extracts from S. salicifolia leaves demonstrated the highest activity with IC50 69.30 μg/mL. Fractionation and chromatographic separation of the extracted compounds from the ethyl acetate fraction of S. salicifolia allowed isolation of 18 compounds with 15 of those isolated for the first time from this species including trifolin, 6′′-O-caffeoyl-hyperoside, 6′′-O-caffeoylisoquercitrin, 6′′-O-caffeoyl-astragalin, 1-O-p-hydroxybenzoyl-6-O-p-coumaroyl-β-d-glucopyranoside, 3,4,5-tri-O-caffeoylquinic acid, isoramnetinin-3-O-β-d-glucopyranoside, tiliroside, isoramnetin-3-O-α-lrhamnopyranoside, 1-O-cis-cinnamoyl-6-O-(2′-methylene-4′-hydroxybutyroyl)-β-d-glucopyranose, 1-О-(4′′-hydroxy-3′′-methylfurane-2′′-one)-6-О-trans-cinnamoyl-β-d-glucopyranose, 1-О-(4′′-hydroxy-3′′-methylfurane-2′′-one)-6-О-cis-cinnamoyl-β-d-glucopyranose, 6-tuliposide A, and tulipalin A. Flavonoid caffeoyl glycosides were identified as the most active inhibitors of α-amylase with 6′′-O-caffeoyl-hyperoside demonstrating the maximum IC50 of 46.18 μg/mL that determined total anti-α-amylase effect of the S. salicifolia extract. The total content of 6′′-О-caffeoyl-hyperoside in S. salicifolia leaves was 0.60–10.53 mg/g and of flavonoids, 12.02–23.17 mg/g. The study showed that the acylated flavonoids from Spiraea were effective inhibitors of α-amylase.

A novel acylated quercetin glycoside, floralpanasenoside A (1) and five known flavonoid glycosides, panasenoside (2), quercetin 3-O-(2''-β-D-glucopyranosyl)-β-D- galactopyranoside (3), trifolin (4) kaempferol 7-O-α-L-rhamnoside (5), and afzelin (6) were isolated from the flower buds of Panax ginseng. Their structures were established by spectroscopic data and comparison with the literature values. Four of the six isolated compounds including 1 (IC50 = 62.4) exhibited α-glucosidase inhibitory activity with IC50 values lower than acarbose (385.2 μM). The molecular docking study indicated that 1 bound to the active site of α-glucosidase with numerous hydrogen bond interactions.

Spiraea L. is a genus of deciduous shrubs of Rosaceae family widespread on the territory of Eastern Siberia. As a result of the study of six Spiraea species growing on the Baikal region, it was shown that they characterized by a high content of phenolic compounds and their extracts had an inhibitory effect on α-amylase. The use of correlation analysis made it possible to reveal the leading role of flavonoids as carriers of the biological effect of extracts. The most active was the extract of S. salicifolia leaves with an IC50 value 69,30 μg/mL After fractionation and chromatographic separation of the extractive substances, 18 compounds were isolated from the ethyl acetate fraction of S. salicifolia and fifteen of them for the first time for the species, including trifolin, 6′′-O-caffeoyl-hyperoside, 6′′-O-caffeoyl-isoquercitrin, 6′′-O-caffeoyl-astragalin, 1-O-p-hydroxybenzoyl-6-O-p-coumaroyl-β-d-glucopyranoside, 3,4,5-tri-O-caffeoylquinic acid, isoramnetinin-3-O-β-d-glucopyranoside, tiliroside, isoramnetin-3-O-α-l-rhamnopyranoside, 1-O-cis-cinnamoyl-6-O-(2′-methylene-4′-hydroxybutyroyl)-β-d-glucopyranose, 1-О-(4′′-hydroxy-3′′-methylfurane-2′′-one)-6-О-trans-cinnamoyl-β-d-glucopyranose, 1-О-(4′′-hydroxy-3′′-methylfurane-2′′-one)-6-О-cis-cinnamoyl-β-d-glucopyranose, 6-tuliposide A and tulipalin A. The most active inhibitors of α-amylase were flavonoid caffeoyl glycosides with 6′′-O-caffeoyl-hyperoside as a most effective inhibitor with IC50 value 46.18 μg/mL. Latter compound determined the total anti-α-amylase effect of S. salicifolia. The content of 6′′-O-caffeoyl-hyperoside in leaves of S. salicifolia was 0,60-10,53 mg/g and flavonoids – 12,02-23,17 mg/g. This study demonstrated the fact that the acylated flavonoids of Spiraea are effective inhibitors of α-amylase., Spiraea L. представляет собой род листопадных кустарников семейства Rosaceae, широко распространенный на территории Восточной Сибири. В результате исследования шести видов Spiraea, произрастающих в Прибайкалье, было показано, что они характеризовались высоким содержанием фенольных соединений, а экстракты обладали ингибиторным действием на α-амилазу. Применение корреляционного анализа позволило выявить лидирующую роль флавоноидов, как носителей биологического эффекта экстрактов. Наибольшей активностью обладал экстракт листьев S. salicifolia с величиной IC50 69,30 мкг/мл. После фракционирования и хроматографического разделения экстрактивных веществ из этилацетатной фракции S. salicifolia было выделено 18 соединений, в том числе 15 – впервые для вида, включая трифолин, 6′′-О-кофеил-гиперозид, 6′′-О-кофеил-изокверцитрин, 6′′-О-кофеил-астрагалин, 1-О-п-гидрокси­бензоил-6-О-п-кумароил-β-d-глюкопиранозу, 3,4,5-три-О-кофеилхинную кислоту, изорамнетин-3-О-β-D-глюкопиранозид, тилирозид, изорамнетин-3-О-α-l-рамнопиранозид, 1-О-цис-циннамоил-6-О-(2′-метилен-4′-гидрокси-бутироил)-β-d-глюкопиранозу, 1-О-(4′′-гидрокси-3′′-метилфуран-2′′-он)-6-О-транс-циннамоил-β-d-глюкопиранозу, 1-О-(4′′-гидрокси-3′′-метилфуран-2′′-он)-6-О-цис-циннамоил-β-d-глюкопиранозу, 6-тюлипозид А и тюлипалин А. Наиболее активными ингибиторами α-амилазы были флавоноидные кофеилгликозиды, причем 6′′-О-кофеил-гиперозид с максимальным показателем IC50 46,18 мкг/мл, определял общий анти-α-амилазный эффект извлечений S. salicifolia. Суммарное содержание 6′′-О-кофеил-гиперозида в листьях S. salicifolia составило 0,60–10,53 мг/г, флавоноидов – 12,02–23,17 мг/г. Проведенные исследования показали, что ацилированные флавоноиды Spiraea являются эффективными ингибиторами α-амилазы.

Hawk tea is a rich and edible resource, traditionally used as a beverage in South China. This drink has many pharmacologic effects, such as acting as an antioxidant and reducing blood sugar and lipids. The objective of this work was to explore the active compound contents, bioactivities and their monthly changes, and optimize the harvest time. In the present study, Hawk tea from each month in 2017 was collected and extracted with 70% (v/v) ethanol. The contents of the total flavonoids and total phenols were determined using the colorimetric method. We determined the contents of seven characteristic active substances&mdash, hyperin, isoquercitrin, trifolin, quercitrin, astragalin, quercetin, and kaempferol&mdash, using high-performance liquid chromatography. The crude extract was tested for its antioxidant and inhibitory properties on enzymes involved in metabolic syndrome. Specifically, 2,2-diphenyl-1-picrylhydrazyl, 2,2&prime, azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), ferric-reducing power assay, and the inhibition capacity test on &alpha, glucosidase and lipase were conducted to determine the antioxidant effect in vitro, as well as the reduction of blood sugar and lipids. Monthly variations in activities and components were determined by numeric analysis and comparison. Correlation analysis revealed that antioxidant effects are significantly correlated with the total flavonoids. The hierarchical cluster analysis of bioactivities and their contents indicates that October and November are the best harvesting months, which differs with the habitual collection of Hawk tea.

Background Trifolin (kaempferol-3-O-galactoside), which is a galactose-conjugated flavonol, exhibits antifungal and anticancer effects. However, the mechanisms underlying its anticancer activities have not yet been examined. Purpose In this study, the anticancer effects of trifolin were examined in human lung cancer cells. Methods Cytotoxicity was determined by evaluating cell viability. Apoptosis was analyzed through flow cytometry and western blotting analysis. Death receptors and inhibitors of apoptosis were evaluated through RT-PCR. Results Trifolin induced apoptosis in NCI-H460 human non-small cell lung cancer (NSCLC) cells by inhibiting the survival pathway and inducing the intrinsic and extrinsic apoptosis pathways. Trifolin decreased levels of Akt/p-Akt, whereas levels of expression of phosphatidylinositide 3-kinase (PI3K), cyclin D1, cyclin E, and cyclin A were not altered. Trifolin initiated cytochrome c release by inducing mitochondrial outer membrane permeabilization (MOMP). Trifolin increased Bcl-2-associated X protein (Bax) levels and decreased b-cell lymphoma 2 (Bcl-2) levels, while the levels of Bcl-xL were not altered. In addition, trifolin increased the levels of the death receptor involving the Fas/Fas ligand (FasL) and Fas-associated protein with the death domain (FADD), which consequently activated caspase-8, caspase-9, caspase-3, and the proteolytic cleavage of poly (ADP-ribose) polymerase (PARP). Conclusion These results suggested that trifolin induced apoptosis via death receptor-dependent and mitochondria-dependent pathways and that trifolin can be used as a therapeutic agent in human lung cancer.

Phytochemical investigation of Lespedeza cuneata led to the isolation of seventeen compounds including three steroids (β-sitosterol 1, β-sitosterol-6′-linolenoyl-3-O-β-d-glucopyranoside 3, and β-sitosterol glucoside 13), nine flavonoids (quercetin 4, kaempferol 5, isovitexin 8, hirsutrin 9, nicotiflorin 10, vitexin 11, astragalin 12, trifolin 14, and isorhamnetin 17), two phenolics (benzyl-β-d-glucopyranoside 7 and homovanillyl alcohol 16), one carotenoid (loroxanthin 2), one lignin (7R,8S–dihydrodehydrodiconiferyl alcohol 15), and one hexose (pinitol 6) on the basis of their spectroscopic data. Among these compounds, 2, 3, 7, 15 and 16 were reported for the first time from the genus Lespedeza. The taxonomic significance of these isolated compounds was also summarized.

Frangula alnus and Peganum harmala populations growing in Saudi Arabia might be rich sources of natural compounds with important biological activities. A high performance liquid chromatography diode array revealed several polyphenols in the leaf extracts for the first time, including p-coumaric acid, rosmarinic acid, chlorogenic acid, ferulic acid, quercitrin, rutoside, quercetin and trifolin in F. alnus, and hydrocaffeic acid, protocatechuic acid, rosmarinic acid, caffeic acid and cynaroside in P. harmala. F. alnus and P. harmala showed strong antioxidant effects attributed to the polyphenolic composition of leaves and reduction of reactive oxygen species (ROS) accumulation. F. alnus and P. harmala leaf extracts showed cytotoxic effects against Jurkat, MCF-7, HeLa, and HT-29 cancer cells using MTT and flow cytometry assays. These activities were attributed to the polyphenolic composition of leaves including quercitrin, trifolin and cymaroside, as well as the activation of caspase family enzymes 2, 6, 8 and 9 in treated cancer cells compared to control. The current findings of this study include a novel comprehensive investigation on the polyphenol composition and anticancer effects of leaf extracts of F. alnus and P. harmala from natural populations in Saudi Arabia.

Epidemiological studies observed that Zanthoxylum bungeanum leaf, a popular vegetable in China, could interfere progression of cardiovascular diseases, especially hyperlipidemia. In the work, the pharmacological properties of Z.bungeanum leaves extract (ZLE) containing isovitexin, vitexin, hyperoside, rutin, isoquercitrin, foeniculin, trifolin, quercitrin, astragalin, and afzelin were investigated using multi-system such as mice organ tissue, HepG2 cell and apoE-/- mice. ZLE exhibited a stronger activity of scavenging free radicals. ZLE also appeared to inhibit lipid peroxidation of mice organ tissues including the heart, liver, spleen, lung, kidney, brain and pancreas. ZLE could lower lipid accumulation in HepG2 cells induced by free fatty acids (FFAs, an inducer of lipid peroxidation and free radicals production) in a dose-dependent manner. Compared with fenofibrate, a commercial product popularly used in clinical, the treatment of ZLE in apoE-/- mice had a stronger anti-hyperlipidemia without any detectable histopathological damage. This is the first report that ZLE containing high content of flavonoids with satisfying safety could significantly inhibit the development of hyperlipidemia by preventing the oxidative damage induced by free radicals and lipid peroxidation.

The Zanthoxylum bungeanum leaf has been traditionally taken as a vegetable and seasoning in . To disclose the mechanisms of its benefit for human health, antioxidant activity and cytoprotective effect of the leaf were investigated. The ethyl acetate fraction exhibited the strongest ABTS and DPPH radicals scavenging activities as compared with chloroform and water fractions, as well as the crude extract. This fraction exhibited a protective effect against hydrogen peroxide-induced cytotoxicity in PC12 cells by MTT reduction assay. Furthermore, the cytoprotective effect was confirmed using flow cytometry analysis. Comparing with the model cells, the ethyl acetate fraction significantly increased G0/G1 and G2/M phase cells together with reducing S phase cells. Ten flavonoid glycosides were identified as isovitexin, vitexin, hyperoside, rutin, isoquercitrin, foeniculin, trifolin, quercitrin, astragalin, and afzelin from this fraction using high performance liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance spectroscopy. This is the first report for the presence of vitexin, isovitexin, astragalin, trifolin and afzelin in Z. bungeanum. Hence, the results present here suggest the potential utility of Z. bungeanum leaf as a source of natural antioxidant due to its diversity and high content of flavonoids.

Objective To separate and enrich compounds from Apocynum venetum L. leaves. Methods The core-shell nanoparticles consisting of poly (methyl methacrylate) (PMMA) cores surrounded by various chitosan shells, induced by a tert-butylhydroperoxide (TBHP) solution. And core-shell nanoparticles was applied to immobilize Alpha glycosidase enzymes. Some compound from the extrat of Apocynum venetum L. leaves were absorbed by the immobilized Alpha glycosidase enzymes. and the components absorbed were detected by HPLC. Results Hyperoside and trifolin can be adsorbed by the nanoparticles immobilized Alpha glycosidase enzymes. Conclusion The method applied to separate and preconcentrate Apocynum leaves’ active ingredients is feasible.

In the present study we demonstrated that the flavonoid derivative trifolin acetate (TA), obtained by acetylation of naturally occurring trifolin, induces apoptosis. Associated downstream signaling events were also investigated. TA-induced cell death was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the presence of the selective caspase inhibitors z-LEHD-fmk (caspase-9), z-DEVD-fmk (caspase-3) and z-VEID-fmk (caspase-6). The apoptotic effect of TA was associated with (i) the release of cytochrome c from mitochondria which was not accompanied by dissipation of the mitochondrial membrane potential (ΔΨm), (ii) the activation of the mitogen-activated protein kinases (MAPKs) pathway and (iii) abrogated by the over-expression of Bcl-2 or Bcl-xL. TA-induced cell death was attenuated by inhibition of extracellular signal-regulated kinases (ERK) 1/2 with U0126 and inhibition of p38MAPK with SB203580. In contrast, inhibition of c-Jun NH2-terminal kinase (JNK) by SP600125 significantly enhanced apoptosis. Although reactive oxygen species (ROS) increased in response to TA, this did not seem to play a pivotal role in the apoptotic process since different anti-oxidants were unable to provide cell protection. The present study demonstrates that TA-induced cell death is mediated by an intrinsic-dependent apoptotic event involving mitochondria and MAPK, and through a mechanism independent of ROS generation.

Four flavonoids; kaempferol-3-O-galactoside (trifolin) [1], quercetin-3-O-galactoside (hyperoside) [2], quercetin-3-O- glucoside (isoquercitrin) [3] and kaempferol-3-O-rutinoside [4] together with chlorogenic acid [5] and chlorogenic acid methyl ester [6] have been isolated and purified for the first time from the leaves of Tylophora indica by HPLC. The structures were elucidated by the application of MS, IR, UV, ID- and 2D-NMR spectroscopic analyses and by comparison with literature data. All compounds 1-6 were found to exhibit antioxidant effects, based on the scavenging activity of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical.

The crystal structure of monohydrated trifolin (kaempferol 3-O-β-D-galactopyranoside) (an important biologically active compound, which was isolated from the aerial part of Consolida oliveriana) has been determined from conventional laboratory X-ray powder diffraction data. Variable counting time technique was used during measurement and crystal structure was solved by means of Monte Carlo algorithm. The final structure was achieved by Rietveld refinement using both constraints and restraints on interatomic bond lengths and angles.

This study investigates the in vitro activity of a number of flavonoids (kaempferol, quercetin, trifolin, and ace- tyl hyperoside) and their acetylated products: kaempferol acetate, quercetin acetate, trifolin acetate, and acetyl hyperoside acetate) isolated from the aerial parts of plant Consolida oliveriana against epimastigote, amastigote and metacyclic forms of T. cruzi, their cytotoxic against a host Vero-cell line and analyse the possible mechanism by which these molecules act. Acetylated compounds were potent T. cruzi epimastigote growth inhibitors with activity levels similar to those of benzni- dazole, used as the reference drug. These compounds, at the dosage IC25, decreased the ability of metacyclic forms to in- vade mammalian cells, their intracellular replications and transformation in trypomastigotes, with no toxicity to the host cells. The cells treated presented severe damage in their ultrastructure: intense vacuolization, and appearance of lysosomes as well as other residual bodies. The mitochondrial section appeared larger in size, with a swollen matrix. In addition, these compounds changed the excretion of end metabolites, primarily affecting acetate and succinate excretion, possibly by directly influencing certain enzymes or their synthesis. The potent tripanocidal activities of the flavons described here represent an exciting advance in the search for new antiprotozoal agents.

The aerial parts of Lespedeza cuneata G. Don, perennial legume native to Eastern Asia, have been used therapeutically in traditional Asian medicine to protect the function of liver, kidneys and lungs. However, little is known about the pharmaceutical effect of extracts from this plant. In the present study, the aerial parts of L. cuneata were used to prepare an ethanol extract, which was then tested for hepatoprotective effects against injury by tert-butyl hyperoxide (t-BHP). At a dose of 20 μg/mL, the ethanol extract significantly protected HepG2 cells against the cytotoxicity of t-BHP. Further fractionation of the extract with ethyl acetate allowed the isolation of five flavonoid compounds that were structurally identified by ¹H and ¹³C NMR spectroscopy as isovitexin, hirsutrin, trifolin, avicularin and quercetin. Hirsutrin, avicularin and quercetin (10 μM) showed clear hepatoprotective activity against injury by t-BHP in HepG2 cells, whereas isovitexin and trifolin showed no protective effects. The observed hepatoprotective effect of the investigated compounds showed a high correlation with radical scavenging activity, which followed the structure-activity relationships of the flavonoid aglycones.

2( S)-3’,4'-dimethoxy-5,7-dihydroxyflavanone 7- O-neohesperidoside (1), hesperidin (2), astragalin (3), trifolin (4), nicotiflorin (5), narirutin (6), rutin (7), kaempferol 7- O-rutinoside (8), tangeretin (9) and nobiletin (10) were isolated from the MeOH extract of flowers of Citrus ‘Hebesu’. Structures of these compounds were elucidated on the basis of physical and spectral data. Absolute configuration and complete NMR spectral data for compound 1 are reported for the first time in this paper.

Maytenus ilicifolia Mart. ex Reissek é tradicionalmente usada no Brasil para o tratamento de úlcera gástrica. O presente trabalho relata a investigação fitoquímica de um extrato etanólico de folhas de M. ilicifolia (EEMIL) visando o isolamento de constituintes que foram usados como marcadores químicos para monitorar o fracionamento de um extrato aquoso liofilizado de folhas de M. ilicifolia (LAEMIL). De EEMIL, quatro flavonóides foram isolados, compreendendo o triglicosídeo flavônico mauritianina (1), trifolina (2), hyperina (4), e epi-catequina (5). O fracionamento de LAEMIL levou a 5 frações, fornecendo um derivado tetraglicosilado de canferol (3), além do galactitol (6). LAEMIL e suas frações foram avaliadas quanto aos efeitos sobre o volume e pH da secreção gástrica em ratos. Análise por CLAE (Cromatografia Líquida de Alta Eficiência) revelou que somente frações contendo o tri- e tetra-glicosídeos flavônicos 1 e 3 causaram aumento significativo de volume gástrico e pH, indicando que esses glicosídeos têm importante papel sob o efeito gastroprotetor de folhas de M. ilicifolia. Maytenus ilicifolia Mart. ex Reissek is traditionally used in Brazil for treatment of gastric ulcers. Here we report the phytochemical investigation of an ethanol extract of M. ilicifolia leaves (EEMIL) aiming at the isolation of constituents which were used as chemical markers to monitor an activity-guided fractionation of a lyophilized aqueous extract of M. ilicifolia leaves (LAEMIL). From EEMIL, four flavonoids were isolated, namely the tri-flavonoid glycosides mauritianin (1), trifolin, (2) hyperin (4), and epi-catechin (5). Fractionation of LAEMIL led to 5 fractions which afforded the tetra-glycoside kaempferol derivative (3), and galactitol (6). LAEMIL and its fractions were evaluated in rats for their effects on gastric secretion volume and pH. HPLC (High Performance Liquid Chromatography) analysis revealed that only fractions containing the tri- and tetra-flavonoid glycosides 1 and 3 caused significant increase of gastric volume and pH, thus indicating that these glycosides play an important role on the gastroprotective effect of M.ilicifolia leaves.

Chemical constituents of the leaves of Ehretia thyrsiflora were continuously investigated. Twelve compounds including six flavonoids and six phenolic acids, isoquercetrin, hyperoside, trifolin, astragalin, kaempferol 3-O-arabinosylgalactoside, quercetin 3-O-arabinosylgalactoside, rosmarinic acid, cinnamic acid, icariside E5, ferulic acid, α-hydroxydihydrocaffeic acid, lithospermic acid B were first isolated from this species. The considerable phenolic compounds existed in this species have important systematic significance in the argument of the family the Boraginaceae.

A new acylated avicularin, namely avicularin 2''-(4'''-O-n-pentanoyl)-gallate (1) along with 12 metabolites have been isolated from the ethyl acetate extract of the aerial parts of Jussiaea repens L. (Onagraceae). Their structures were established as trifolin 2''-O-gallate (2), quercetrin (3), guaijaverin (4), reynoutrin (5), juglanin (6), avicularin (7), hyperin (8), trifolin (9), hyperin 2''-O-gallate (10), rutin (11), kaempferol (12) and quercetin (13) on the basis of their chromatographic properties, chemical and spectroscopic evidences. The investigated ethyl acetate extract was found to be non-toxic (LD(50) up to the maximum soluble dose 4 g kg(-1) body wt.) and had significant antioxidant, hepatoprotective, anti-inflammatory and antidiabetic activities. Also some of the isolated flavonoids showed cytotoxic activity against Ehrlich ascitis carcinoma cells.

A new flavonol tetraglycoside, quercetin-3-O-[α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→6)]-β-D-galactopyranosyl]-7-O-β-D-glucopyranoside (1), and two new flavonol alkaloids, N-(8-methylquercetin-3-O-[α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→6)]-β-D-galactopyranosyl])-3-hydroxypiperidin-2-one (2) and N-(8-methylkaempferol-3-O-[α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→6)]-β-D-galactopyranosyl])-3-hydroxypiperidin-2-one (3), were isolated from the aerial parts of Astragalus monspessulanus ssp. monspessulanus. Two rare flavonoids with an unusual 3-hydroxy-3-methylglutaric acid moiety, quercetin-3-O-α-L-rhamnopyranosyl-(1→2)-[6-O-(3-hydroxy-3-methylglutaryl)-β-D-galactopyranoside (4) and kaempferol-3-O-α-L-rhamnopyranosyl-(1→2)-[6-O-(3-hydroxy-3-methylglutaryl)-β-D-galactopyranoside (5), were isolated from the aerial parts of A. monspessulanus ssp. illyricus. In addition, the eight known flavonoids alangiflavoside (6), alcesefoliside (7), mauritianin (8), quercetin-3-β-robinobioside (9), cosmosine (10), apigenin-4'-O-glucoside (11), trifolin (12), and rutin (13) were isolated from aerial parts of A. monspessulanus ssp. monspessulanus. Their structures were elucidated via NMR and HRESIMS data. In a model that tested t-BuOOH-induced oxidative stress on isolated rat hepatocytes, flavonoids 1-13 had statistically significant cytoprotective activity similar to that of silymarin, tested at 60 μg/mL. The most prominent effects were observed for flavonoids 1, 4, 7, and 12.

Sixteen subsequent fractions were prepared from the ethyl acetate fraction of Zanthoxylum bungeanum leaves after bio-guided chromatographic separation. The HPLC profiles and antioxidant activity of the various fractions indicated that the content of eight phenolic compounds (chlorogenic acid, epicatechin, rutin, hyperoside, trifolin, quercitrin, afzelin and quercetin) and antioxidant activity vary significantly, and high concentrations of a combination of eight phenolic compounds would result in an increase of the antioxidant activity. These results suggested that the eight compounds could be used as chemical markers for quality assessment of Z. bungeanum leaves. Correlation between chromatographic fingerprint and antioxidant activity of the fractions showed that quercitrin and hyperoside play crucial roles in the antioxidant activity, and they can be seen as the milestone for quality control. The findings also suggested that five obtained fractions (E-3-3, E-2-4, E-7, E-5 and E-4) could become useful supplements for functional food ingredients and health-related products.

Leaf spots and root rots are major fungal diseases in Camptotheca acuminata that limit cultivation of the plant for camptothecin (CPT), a promising anticancer and antiviral alkaloid. Bioassays showed that pure CPT and flavonoids (trifolin and hyperoside) isolated from Camptotheca effectively control fungal pathogens in vitro, including Alternaria alternata, Epicoccum nigrum, Pestalotia guepinii, Drechslera sp., and Fusarium avenaceum, although antifungal activity of these compounds in the plant is limited. CPT inhibited mycelial growth by approximately 50% (EC50) at 10−30 μg/mL and fully inhibited growth at 75−125 μg/mL. The flavonoids were less effective than CPT at 50 μg/mL, particularly within 20 days after treatment, but more effective at 100 or 150 μg/mL. CPT, trifolin, and hyperoside may serve as leads for the development of fungicides. Keywords: Camptotheca; camptothecin; trifolin; hyperoside; antifungal activity; chemical defense

Three kaempferol glycosides were isolated from the flower buds of Panax quinquefolium L. together with kaempferol (1). By means of chemical and spectroscopic methods (NMR, ESI-MS, 2D NMR), their structures were established as trifolin (2), panasenoside (3), and kaempferol-3-O-β-D-glucosyl(1→2)-β-D-galactoside-7-O-α-L-rhamnoside (4).Compound 4 is a new compound.

Seven compounds have been isolated from methanol extracts of whole Saxifraga stellaris (Saxifragaceae) plants. They were identified as the new compound, 6- O -galloyl fructose, together with 3- O -[2- O -(β- d -xylopyranosyl)-β- d -galactopyranosyl]-kaempferol, 3- O -[2- O -(β- d -xylopyranosyl)-β- d -galactopyranosyl]-quercetin, trifolin, hyperin, resveratrol-3- O -glucoside, triandrin, by chemical and spectroscopic methods. Their free radical scavenging properties are also described.

The present work objective are to investigate indigenous plants used in wound healing in India, we hereby reported our findings related to wound healing activities of plants  Ichnocarpus frutescens (L.) W. T. Aiton in vivo model studies. The plant hydro-alcoholic (Ethanol 70:Water 30) extracts of the leaves, stems and roots of Ichnocarpus frutescens was identified and tested wound healing activity. The important secondary metabolites like Ursolic acid, Kaempferol, Trifolin, Mannitol, Flavon, Phenolic acid, Triterpene glycoside(1,4-I²-D-glucopyranosyl-(1,3) I±-amyrin, I² –Sitosterol, alkaloid and Flavonoid identified in extracts. Ichnocarpus frutescens roots hydro-alcoholic extract showed wound healing activity significantly in the excision wound model in rats on topical application. The animals were divided into five groups with six rats in each group. Topically applied Ichnocarpus frutescens leaves, stems and roots extracts ointment. Taking povidone iodine ointment as standard. The results showed that hydro-alcoholic extract of Ichnocarpus frutescens stems on topical application was reduced the  scar area from 2.5±1.5 to 0.0±0.0 cm2, hydro-alcoholic extract of Ichnocarpus frutescens leaves on topical application was reduced the scar area from 2.5±1.5 to 0.2±0.0 cm2 , hydro-alcoholic extract of Ichnocarpus frutescens roots on topical application was reduced the scar area from 2.5±1.5 to 0.7±0.4 cm2, control on topical application was reduced the scar area from 2.5±1.5 to 1.2±1.0 cm2 and standard povidon iodine ointment on topical application was reduced the scar area from 2.5±1.5 to 0.5±0.02cm2 respectively. Significant increase in tensile strength were also observed and compared to the control and povidon iodine. The above result revealed that the hydro-alcoholic stems extract has remarkable wound healing potency and appear to justify the traditional use of Ichnocarpus frutescens in wound healing in India and offer a scientific support to the treatment of traditional healers.

The flowers and leaves of Trifolium repens L. (Fabaceae) were subjected to phytochemical investigation in order to identify their major chemical constituents and to evaluate in vitro antioxidant activity of the isolated compounds against DPPH˙. A total of 12 flavonoids, pterocarpan and methyl caffeate were isolated, then characterised by UV, MS, NMR spectroscopy and identified as quercetin and kaempferol 3-O-(6″-α-rhamnopyranosyl-2″-β-xylopyranosyl)-β-galactopyranosides (1, 2), kaempferol 3-O-(2″,6″-α-dirhamnopyranosyl)-β-galactopyranoside, mauritianin (3), quercetin and kaempferol 3-O-(2″-β-xylopyranosyl)-β-galactopyranosides (4, 5), kaempferol and quercetin 3-O-β-(6″-O-acetyl)-galactopyranosides (6, 7), trifolin (8), hyperoside (9), myricetin 3-O-β-galactopyranoside (10), quercetin (11), ononin (12), medicarpin 3-O-β-glucopyranoside (13) and methyl caffeate (14). Mauritianin, ononin, pterocarpan and methyl caffeate have been reported in this plant for the first time. The compounds 4, 7, 9, 10, and 11 were tested for their antioxidant effect against DPPH˙. All studied compounds were found to have potent activity, but the most effective in the test were compounds 9, 10 and 11 (EC(50) values in the range 7.51-9.52 µM).

Chemical investigation of the leaves of Elaeocarpus serratus yielded myricitrin (1), mearnsetin 3-O-β-D-glucopyranoside (2), mearnsitrin (3), tamarixetin 3-O-α-L-rhamnopyranoside (4) and the fruits of Filicium decipiens yielded three flavonol glycosides, kaempferol 3-O-rutinoside (5), kaempferol 3-O-robinobioside (6) and trifolin (7). Compound 1 showed strong antioxidant activity against DPPH.

The main polyphenols were isolated from the leaves of six selected persimmon cultivars. Seven compounds were obtained by reverse-phase HPLC, and their structures were elucidated by multiple NMR measurements. These compounds are hyperoside, isoquercitrin, trifolin, astragalin, chrysontemin, quercetin-3-O-(2''-O-galloyl-β-D-glucopyranoside) (QOG), and kaempferol-3-O-(2''-O-galloyl-β-D-glucopyranoside) (KOG). Their inhibitory activity was tested against tyrosinase for the oxidation of L-DOPA, and only chrysontemin showed inhibitory activity. To investigate the differences of their inhibitory effects, the tyrosinase inhibitory activities of their aglycons, cyanidin, quercetin, and kaempferol, were also tested. As a result, it was confirmed that the most influential moiety for tyrosinase inhibition was the 3',4'-dihydroxy groups of the catechol moiety. Moreover, the tyrosinase inhibitory activity of chrysontemin, which was identified in persimmon leaves for the first time, is supported by a simulated model of chrysontemin docking into mushroom tyrosinase.

A set of flavonoids from Consolida oliveriana, kaempferol (1), quercetin (2), trifolin (3), and acetyl hyperoside (5) and their O-acetyl derivatives (1a, 2a, 3a), and octa-O-acetylhyperoside (4) showed leishmanicidal activity against promastigote as well as amastigote forms of Leishmania spp. The cellular proliferation, metabolic, and ultrastructural studies showed that the acetylated compounds 2a, 3a, and 4 were highly active against Leishmania (V.) peruviana, while 2a as well as 4 were effective against Leishmania (V.) braziliensis. These compounds were not cytotoxic and are effective at similar concentrations up to or lower than the reference drugs (pentostam and glucantim).

The isolation and identification of eleven crystalline components from the aerial part of Cynanchum taiwanianum Yamazaki (Asclepiadaceae) are described. Their structures were determined on the basis of spectral evidence and chemical transformation. Besides caffeic acid, β-amyrin, and methyl phaeophorbide a, the isolated flavonoid components are classified into two groups, i.e. kaempferol derivatives (kaempferol, astragalin, afzelin, trifolin) and quercetin derivatives (quercetin, isoquercitrin, quercitrin, hyperin).

In the present study we demonstrated that the flavonoid derivative trifolin acetate (TA), obtained by acetylation of naturally occurring trifolin, induces apoptosis. Associated downstream signaling events were also investigated. TA-induced cell death was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the presence of the selective caspase inhibitors z-LEHD-fmk (caspase-9), z-DEVD-fmk (caspase-3) and z-VEID-fmk (caspase-6). The apoptotic effect of TA was associated with (i) the release of cytochrome c from mitochondria which was not accompanied by dissipation of the mitochondrial membrane potential (DeltaPsi(m)), (ii) the activation of the mitogen-activated protein kinases (MAPKs) pathway and (iii) abrogated by the over-expression of Bcl-2 or Bcl-x(L). TA-induced cell death was attenuated by inhibition of extracellular signal-regulated kinases (ERK) 1/2 with U0126 and inhibition of p38(MAPK) with SB203580. In contrast, inhibition of c-Jun NH(2)-terminal kinase (JNK) by SP600125 significantly enhanced apoptosis. Although reactive oxygen species (ROS) increased in response to TA, this did not seem to play a pivotal role in the apoptotic process since different anti-oxidants were unable to provide cell protection. The present study demonstrates that TA-induced cell death is mediated by an intrinsic-dependent apoptotic event involving mitochondria and MAPK, and through a mechanism independent of ROS generation.

From the underground parts of Eskemukerjea megacarpum HARA, two new stilbenes (14, 15) were isolated, together with a known coumarin, 5,7-dihydroxycoumarin (1), a tyramine derivative, trans-feruloyltyramine (2), two pyrogallol derivatives, gallic acid (3) and beta-glucogallin (4), four flavonoids, trifolin (5), hyperin (6), myricetin 3-O-beta-D-galactopyranoside (7), and myricitrin (8), five stilbenes, resveratorol (9), astringenin (10), piceid (11) astringin (12), and resveratorol 3-O-beta-D-(6-O-galloyl)glucopyranoside (13), a flavan-3-ol, (-)-epigallocatechin 3-O-gallate (16), two proanthocyanidins, catechin-(4alpha-->8)-epigallocatechin 3-O-gallate (17) and epicatechin 3-O-gallate-(4beta-->8)-epigallocatechin 3-O-gallate (18), and an anthocyanin, idaein (19). Compounds 14 and 15 were identified as (E)-3,5,3',4'-tetrahydroxystilbene 3-O-beta-D-(6-O-galloyl)glucopyranoside and (E)-3,5,4'-trihydroxystilbene 3-O-beta-D-(6-O-galloyl)glucopyranoside, respectively, based on spectral and chemical data.

From the whole plant of Lysimachia nummularia two new flavonol glycosides were isolated together with trifolin, isoquercitrin, myricitrin, mearnsitrin, syringetin 3-galactoside, kaempferol 3- O -rhamnosyl (1 → 2) galactoside, quercetin 3- O -neohesperidioside, rutin, kaempferol 3- O -(2,6-dirhamnosylgalactoside) and quercetin 3- O -(2,6-dirhamnosylgalactoside). The structures of the new compounds were established as myricetin 3- O -arabinofuranoside and syringetin 3- O -xylopyranoside.

A new tannic substance and five flavonoids were isolated from chestnut galls. The chemical structure of the tannic substance was established as formula I on the basis of spectral analyses. The flavonoids were confirmed to be isoquercitrin, hyperin, astragalin, trifolin and the presence of nicotiflorin was indicated.

A protein agglutinin, trifoliin, was purified from white clover seeds and seedling roots. Trifoliin specifically agglutinates the symbiont of clover, Rhizobium trifolii , at concentrations as low as 0.2 μg protein/ml, and binds to the surface of encapsulated R. trifolii 0403. This clover protein has a subunit with M r ≈ 50 000, an isoelectric point of 7.3, and contains carbohydrate. Antibody to purified trifoliin binds to the root hair region of 24-h-old clover seedlings, but does not bind to alfalfa, birdsfoot trefoil or joint vetch. The highest concentration of trifoliin on a clover root is present at sites where material in the capsule of R. trifolii binds. 2-Deoxy- d -glucose elutes trifoliin from intact clover-seedling roots, suggesting that this protein is anchored to root cell walls through its carbohydrate binding sites. We propose that trifoliin on the root hair surface plays an important role in the recognition of R. trifolii by clover.

Structure of substances XIV and XVIII, obtained previously among the constituents of the calyx of persimmons (Diopyros kaki, Ebenaceae), was examined, and XIV was determined as astragalin (kaempferol 3-β-D-glucopyranoside) and XVIII as n-butyl-β-D-fructopyranoside, although the latter was considered to be an artifact. Carbon-13 NMR spectra of kaempferol, quercetin, trifolin, astragalin, and hyperin were measured and their structure was proved from the assignment of NMR signals.

A new glycoindole alkaloid, rhynchophine, was isolated from the leaves of Uncaria rhynchophylla MIQ. and its structure was elucidated as 6'-feruloyl vincoside lactam (IV). A partial synthesis was achieved by the use of vincoside lactam as the starting material. Five known compounds were also newly isolated from the same plant, namely, vallesiachotamine, vincoside lactam, strictosamide, hyperin and trifolin.

SummaryFrom the leaves of Rubus ulmifolius Schott, quercetin, kaempferol, trifolin (kaempferol-3-0-galactoside), hyperin (quercetin-3-0-galactoside), chlorogenic acid, caffeic acid, a kaempferol-3-...