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The work is aimed at phytochemical characterization and In Vitro evaluation of antioxidant actions, anti-inflammatory effects, and cytotoxicity of purified extracts from three rupturewort (Herniaria L.) species, i.e., Herniaria glabra (HG), H. polygama (HP), and H. incana herb (HIh). The total phenolic content established in the purified extracts (PEs) of HIh, HP, and HG was 29.6, 24.0, and 13.0%, respectively. Thirty-eight non-saponin metabolites were identified using LC-HR-QTOF-ESI-MS; however, only 9 were common for the studied Herniaria species. The most abundant phenolic compound in HG-PE was narcissin (7.4%), HP-PE shared 3 major constituents, namely cis-2-hydroxy-4-methoxycinnamic acid 2-O-β-glucoside (cis-GMCA, 5.8%), narcissin (5.4%), and rutin (5.3%). Almost half of HIh phenolic content (14.7%) belonged to oxytroflavoside A (7-O-methylkaempferol-3-O-[3-hydroxy-3-methylglutaryl-(1→6)]-[α-rhamnopyranosyl-(1→2)]-β-galactopyranoside). Antioxidant properties of the Herniaria PEs were evaluated employing an experimental model of human blood plasma, exposed to the peroxynitrite-induced oxidative stress. The assays demonstrated significant reduction of oxidative damage to protein and lipid plasma components (estimated by measurements of 3-nitrotyrosine, protein thiol groups, thiobarbituric acid-reactive substances), and moderate protection of its non-enzymatic antioxidant capacity. Anti-inflammatory properties of the Herniaria PEs were evaluated In Vitro as inhibitory effects against cyclooxygenases (COX-1 and -2) and concanavalin A-induced inflammatory response of the peripheral blood mononuclear cells (PBMCs). None of the studied plants showed inhibitory effects on COXs but all purified extracts partly reduced the release of interleukin 2 (IL-2) and tumor necrosis factor-alpha (TNF-α) from PBMCs, which suggested their prospective ability to up-regulate inflammatory response of the cells. The purified extract from H. glabra turned out to be the most efficient suppressor of PBMCs’ inflammatory response. Additionally, cytotoxicity of purified Herniaria extracts on PBMCs was ruled out based on In Vitro studies.

In the study, Trifolium medium L. and T. pratense L. were used to obtain lyophilisates which were named as TML and TPL, respectively. The former clover taxon represents a little explored species, while the latter one is often found in dietary supplements and functional foods due to the content of isoflavones that alleviate various menopausal symptoms. Detailed phytochemical profiles of both lyophilisates were examined and compared using coupled chromatographic (RP-LC) and spectroscopic PDA/ESI-QToF/MS-MS methods. A total of 54 and 55 compounds were identified in TML and TPL, respectively. Close chemotaxonomic similarities were confirmed for both clover taxa examined, especially in terms of the most abundant isoflavones and hydroxycinnamates. For the first time, neurotropic polyphenols (AChE inhibitors) were identified in clover extracts using TLC-bioautography. In this group, caffeoylmalic acid exhibited significant AChE inhibitory effect confirming the potential of TPL and TML to support physiological functions of the endocrine and nervous systems.

Dried leaf samples of Pyrus communis L. var. 'Conference' and Pyrus pyrifolia Burm. f. (Nakai) var. 'Shinseiki' were subjected to the successful extraction procedures using various solvents, followed by filtering and/or drying liquid plant preparations under reduced pressure. As a result of this, for each Pyrus leaf sample examined, four dried residues were obtained, including methanolic (EA), ethyl acetate (EC), water (EB), and the residue obtained from aqueous solution (ED). Antiradical activity of these preparations was measured using the ABTS+• assay, and antimicrobial activity was examined using various strains of bacteria and yeasts. The highest antiradical activity was observed for EC from leaves of P. communis var. 'Conference' collected in May, but the highest average antibacterial activity was noted for EC residues from P. pyrifolia var. 'Shinseiki' collected in May. Antibacterial activity positively correlated with concentration of hydroquinone in extracts. No antifungal activity was observed for any extract. In addition, qualitative and quantitative analyses of active polyphenolic components in extracts from Pyrus were performed. Hydroquinone and hydroxycinnamic acid derivatives were analyzed using a new optimized method comprising reversed-phase high-performance liquid chromatography (RP-LC) coupled with simultaneous photodiode-array and fluorescence detection.

The theoretical study provides an overview of current data on red clover (Trifolium pratense L.), its cultivation and breeding, other practical uses and future prospects. Particular attention is paid to this taxon as a fodder plant of global importance in traditional and modern animal husbandry systems. Evidence is presented that red clover is a leading component, influencing the pasture management on all continents due to its high biomass production, protein content and overall nutritional value associated with the ability to fix atmospheric nitrogen. Some considerations cover important agrobiological and agrotechnological issues (e.g., plant interseeding) related to increasing adaptability, sustainability and yield of red clover and other crops coexisting in the same biosystems. Interesting biotechnological approaches have been pointed out in relation to the introduction to cultivation of new varieties, which are breeding hybrids of red clover and other wild species, with greater durability and yield-forming capacity. A separate part of the review is the assessment of the phytochemical profile of red clover with particular reference to biologically active specialized metabolites. Recent data from preclinical and human studies on existing and potential applications of this taxon and individual polyphenolic components in traditional and official medicine are also discussed.

Gatunki należące do rodzaju Trifolium L. (koniczyna), wykorzystywane od wieków w etnomedycynie, stanowią dziś bogate źródło fitohormonów polifenolowych. W badaniach chemotaksonomicznych udokumentowano bowiem, że czerwono kwitnące koniczyny odznaczają się szczególnie wysoką zawartością izoflawonów. Związki te posiadają aktywność estrogenopodobną, dlatego mogą działać analogicznie do selektywnych modulatorów receptorów estrogenowych (SERMs). Dane epidemiologiczne oraz liczne doniesienia z badań przedklinicznych i klinicznych dowodzą korzystnego działania tych fitoskładników zarówno w łagodzeniu wielu psychicznych i neurowegetatywnychdolegliwości, występujących w okresie menopauzy, jak i możliwości prewencji i terapii schorzeń biodegeneracyjnych, będących skutkiem deficytów estrogenowych (osteoporoza, miażdżyca). W nieodległej przyszłości, mogą również pojawić się nowe zastosowania dla izoflawonów koniczyn w zakresie zwalczania niektórych typów schorzeń nowotworowych. Ze względu na nieliczne obserwowane działania niepożądane, fitoestrogeny występujące w rodzaju Trifolium L. traktowane są jako alternatywa nie tylko dla popularnych produktów sojowych, ale i syntetycznych estrogenomimetyków, wykorzystywanych w hormonalnej terapii zastępczej (HTZ).

Roślinne adaptogeny wykazują wiele efektów farmakologicznych w organizme ludzkim, charakteryzując się jednocześnie dużym bezpieczeństwem stosowania. Liczne substancje roślinne, obdarzone właściwościami adaptogennymi, wywodzą się z Tradycyjnej Medycyny Chińskiej (TCM). Część z nich znalazła również zastosowanie w konwencjonalnej medycynie zachodniej. Celem niniejszej publikacji jest przybliżenie potencjału terapeutycznego trzech gatunków roślin z rodzaju Scutellaria L. (tarczyca), charakteryzujących się aktywnością adaptogenną. Tarczyce: bajkalska (Scutellaria baicalensis Georgi.) oraz brodata (Scutellaria barbata D.Don.) to dwa gatunki azjatyckie wykorzystywane w TCM, natomiast tarczyca bocznokwiatowa (Scutellaria lateriflora L.) jest taksonem rodzimym flory Ameryki Północnej, stosowanym pierwotnie w medycynie ludowej Indian. Głównymi składnikami tarczyc, którym przypisuje się właściwości adaptogenne są składniki polifenolowe z grupy flawonów (głównie: bajkaleina, bajkalina, wogonina). Wyciągi z tarczyc, bogate w te związki, stanowią środki lecznicze o silnym działaniu przeciwzapalnym, wykazujące szeroką aktywność antybiodegeneracyjną (neuro- hepato- i kardioprotekcyjną) oraz przeciwnowotworową. Ponadto, jako adaptogeny, flawony tarczyc wykazują normalizujący wpływ na OUN, łagodząc dolegliwości natury psychicznej, w tym reakcje lękowe, stany napięcia nerwowego czy zaburzenia snu.

Dried leaf samples of

The dietary supplements with claimed antioxidant activity constitute a substantial part of the dietary supplement market. In this study, we performed the LC-QTOF-MS analysis and investigated the activity profiles of popular antioxidant dietary supplements from different chemical groups in terms of quality control. The commonly used antioxidant tests and statistical analysis revealed that substantial part of the results was comparable if 1 g sample was considered, but while comparing single and daily doses, significant differences in antioxidant values were noticed in all assays. The best antioxidant activity was obtained in ORAC assay (from 142 to 13814 μM of Trolox equivalents per 1 g of sample), and the strongest correlation occurred between TPC and ORAC. The LC-QTOF-MS analysis revealed that catechins were present in samples having the best antioxidant activity and that dietary supplements showing the weakest activity contained very small amount of any chemical constituents.

In this study, methanol, ethyl acetate, water extracts, and precipitate were obtained from leaves ofMalus domesticacultivars: Golden delicious, Jonagold, Elstar, Ligol, and Mutsu. Antiradical activity of these extracts was measured using theABTS+∙radical, and antimicrobial activity was measured with the disk-diffusion method. Phenolic compounds were measured with the colorimetric method and identified with high performance liquid chromatography (HPLC). The highest antiradical activity was observed for the Jonagold variety, and in particular strong activity was noted for ethyl acetate extracts. Antimicrobial activity was observed against strains ofStaphylococcus aureus,Enterococcus faecalis, and the fungusCandida glabrata. Particularly susceptible to the extracts activity appeared to beStaphylococcus aureus, but the growth ofCandida glabratawas inhibited in the presence of ethyl acetate extracts. With the HPLC method we identified a high amount of phloridzin (above 500 mg per g of ethyl acetate extracts), lower amounts of hyperoside, isoquercitrin, and quercitrin, and traces ofp-hydroxybenzoic and chlorogenic acids. The contribution of phloridzin to antiradical activity of methanol and ethyl acetate extracts was very high (above 90%). In water extract the contribution of phloridzin was between 38.9 and 55.2%, chlorogenic acid 22.7 and 36.1%, and hyperoside 12.2 and 13.3%.

As a new sample preparation technique, pressurized liquid extraction (PLE), in combination with reversed-phase liquid chromatography (RP-LC) and photodiode-array (PDA) detection were used for the isolation and determination of phytoestrogenic isoflavones in hydrolyzed extracts obtained from aerial parts of five Trifolium L. (clover) species. To optimize the effectiveness of PLE procedure, variable extraction parameters: methanol and acetone (or their 75% aqueous solutions), as extraction solvents, temperatures (75, 100 and 125 degrees C) and the changeable number of static extraction cycles were tested. Additionally, two other micropreparative techniques: ultrasound-assisted extraction (UAE), and conventional solvent extraction (CSE), under optimized conditions, were also used and compared. Optimum extraction efficiency, expressed in the highest yield of biochanin A, formononetin, daidzein and genistein from plant material, with PLE, using methanol-water (75:25, v/v) as an extraction solvent, an oven temperature of 125 degrees C and three 5-min static extraction cycles, was obtained.

Detailed chemotaxonomic studies were undertaken to establish the qualitative profile and real amounts of the pharmacologically active isoflavone aglycones genistein, daidzein, formononetin, and biochanin A in aerial parts of thirteen Trifolium L. (clover) species, native to Poland. A newly elaborated micropreparative technique--SPE--on BakerBond octadecyl, cyclohexyl, and phenyl cartridges was used in combination with ultrasound-assisted extraction for isolation of isoflavone aglycones from hydrolyzed samples. The effectiveness of all three SPE sorbents in the purification of plant extracts was compared and very high recoveries (96%) were documented for four isoflavones. Classical photodiode-array and very sensitive fluorescence detection, coupled with reversed-phase high-performance liquid chromatography (RP-HPLC), were employed to obtain the most reliable qualitative and quantitative results. Chemotaxonomic differences combined with flower color variability were demonstrated within thirteen clover species. Concentration levels of particular isoflavones in ten Trifolium species possessing flowers with white, pink, or purple-red corolla ranged from approximately 3 to approximately 3300 microg/g dry weight, while in three yellow flowering clovers (T. aureum, T. dubium, and T. campestre) isoflavone compounds have not been detected at all. RSD values, determined for intra- and inter-day precision of the quantitative results, were not higher than 6.2% and 7.1%, respectively.

Preparative separation and isolation of coumarins from petroleum ether and methanol extracts of the fruits and roots of Peucedanum verticillare L. Koch ex DC ( Umbelliferae = Apiaceae ) have been achieved by column chromatography and normal- and reversed-phase TLC with gradient elution. Isolated coumarins were identified by comparison with standards in analytical TLC and in RP HPLC with gradient elution, by melting point (m.p.) measurement, by UV spectroscopy, by MS, and by 1 H and 13 C NMR spectroscopy. Esculin, umbeliferone, bergapten, xanthotoxin, isoimperatorin, imperatorin, psoralen, and cis -kellactone were isolated for the first time from P. verticillare L. Koch ex DC. Two angular-type pyranocoumarins isolated from the roots and fruits of Peucedanum verticillare L. Koch ex DC. were identified as pteryxin and epoxypteryxin by MS and 1 H and 13 C NMR.

Centaurea L. is a genus of the family Asteraceae that comprises over 600 taxa. Representatives of the Centaurea genus were used as natural medications for many diseases. Methanolic-aqueous extracts from aerial parts of two Centaurea species: C. borysthenica Gruner and C. daghestanica (Lipsky) Wagenitz were studied for their polyphenolic composition and potential protective effect on cardiomyocytes treated with doxorubicin. Effectiveness of doxorubicin in cancer therapy is limited by a dose-dependent cardiotoxicity. Oxidative stress is a widely recognized mechanism of this phenomenon. One of the most important strategies has been an application of drug together with antioxidant agents. A cardioprotective effect of selected extracts of Centaurea species was suspected in this study. Cell viability, oxidative stress, and mitochondrial membrane potential analyses showed protective activity of the methanolic extract of C. borysthenica and C. daghestanica on rat cardiomyocytes treated with doxorubicin. Although C. borysthenica is more effective as a cardiomyocyte protective agent, in higher concentrations it weakened the drug activity. C. daghestanica extract did not change the doxorubicin efficacy in the evaluated experiment. Interestingly, both tested extracts were cytotoxic for myeloma cells. The detected antioxidant activity of the studied extracts can be used in the prevention of doxorubicin-induced cardiotoxicity.

This paper describes the retention behavior on silica and Florisil layers of ten coumarin compounds identified and isolated from the fruits of Libanotis dolichostyla Schischkin (Umbelliferae). The variation of retention as a function of the solvent composition has been studied. The linear relationships between retention factors (RF and RM) of coumarins and log c of various polar organic modifiers in the non-polar mobile phase were established. The elaborated model of retention was used for further optimization of separation conditions for preparative isolation of some coumarins contained in crude fractions derived from a petroleum extract obtained from the fruits of L. dolichostyla.

We have analysed the effect of some culture conditions and media components on callus growth rate and production of taxanes in callus of Taxus × media var. Hatfieldii. For callus induction and maintenance a Gamborg B5 medium and a White - Rangaswamy medium (WR) with different modifications were used. On an improved WR medium (containing 10 μM picloram) the callus growth factor increased up to 5.8 fold (fresh weight). Picloram only enhanced the growth of callus, but not taxane production. On WR medium with (100 μM) methyl jasmonate the paclitaxel content increased from 2.37 μg g-1 to 90 μg g-1 and cephalomannine from 5.14 μg g-1 to 29.14 μg g-1 (dry weight), whereas growth of the cultures ceased. The presence of paclitaxel and cephalomannine was established by high performance liquid chromatography.

Solid-phase extraction and preparative thin-layer chromatography were applied as sample preparation techniques for the purification of crude extracts from twigs and needles of various Taxus species as well as for the isolation of taxol and cephalomannine for further reversed phase high performance liquid chromatography analysis. Significant differences in the contents of taxanes examined were found. The preparative chromatographic methods used were compared and evaluated as routine and reproducible procedures for the rapid isolation and determination of taxol and cephalomannine in plant extracts.

A new method is described combining solid-phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC) for the isolation, purification as well as qualitative and quantitative determination of free phenolic acids in six Echinacea species. Plant extracts were purified and phenolic acids isolated on octadecyl and quaternary amine Bakerbond SPE columns; final eluates were analysed by RP-HPLC. Significant differences in the composition and amount of phenolic acids within Echinacea genus have been shown. The method can be used for quick screening analysis of the content of phenolic acids in plant material.

Some plant species belonging toTrifoliumL. genus are a source of isoflavones considered to exert phytoestrogenic activities. The aim of the present study was to examine the effects of standardized extract obtained from aerial parts ofTrifolium mediumL., in comparison with the extract ofTrifolium pratenseL., on the development of estrogen deficiency-induced osteoporosis in rats. BothTrifoliumextracts, at doses corresponding to 10 and 20 mg/kg of isoflavone aglycones daily, or estradiol (0.2 mg/kg daily), were administered orally to ovariectomized (OVX) rats for 4 weeks. Serum bone turnover markers, bone mass, mineralization, and mechanical properties were studied. In OVX control rats, mechanical properties of the tibial metaphysis and femoral neck were strongly worsened in comparison with sham-operated control rats, and those of femoral diaphysis were unaffected. Estradiol counteracted the worsening of the tibial strength and increases in bone turnover markers. Both extracts significantly increased the strength of the femoral diaphysis and calcium and phosphorus content in the bone mineral, but onlyT. pratenseextract increased the strength of the tibial metaphysis. In conclusion, effects of bothTrifoliumextracts differed from those of estradiol. It is possible that other than isoflavone extract constituents contributed to their skeletal effects.

HPLC coupled with photodiode array (PDA) and fluorescence (FL) detectors has been used for the identification and determination of phytoestrogenic (isoflavones and coumestrol) and nonphytoestrogenic (flavones) compounds in hydrolyzed and nonhydrolyzed extracts obtained from aerial parts of Trifolium incarnatum L. and related clover species. The effective isolation technique of pressurized liquid extraction (PLE) was used. Various types of extraction solvents, i.e., ethanol, n-propanol, n-butanol, and water solutions (75%, v/v) of methanol or ethanol at changeable or constant temperatures were tested, taking into account the chemical character of isolated compounds. Higher PLE efficiency in relation to isoflavone aglycones was found for ethanol. Predominant isoflavone glycosides determined in clover samples were sissotrin and ononin, reaching levels above 1.4% dry weight (wt) in T. medium. The presence of flavone compounds (apigenin and luteolin) in aerial parts of T. incarnatum, occurring in amounts exceeding 0.4% dry wt, documented chemotaxonomic distinction of this species from other clovers examined. Additionally, within the group of phytoestrogenic isoflavones, only biochanin A and formononetin derivatives were identified in above-ground parts of T. incarnatum . The application of simultaneous PDA and FL detection enabled unambiguous confirmation of the lack of a strong phytoestrogen, coumestrol, in all hydrolyzed and nonhydrolyzed clover extracts.

In this study, the effectiveness of both classical and novel polymeric sorbents used in solid-phase extraction (SPE) for isolation of pharmacologically active flavones (baicalin, luteolin, apigenin, wogonin and chrysin) from aerial parts of 13 species of Scutellaria L. (Skullcap) genus was assessed. The highest recoveries of hydrophilic (baicalin, luteolin) flavones for Oasis HLB were obtained, whereas for medium hydrophobic (apigenin) and hydrophobic (wogonin, chrysin) flavones better quantitative results for BakerBond phenyl cartridges were stated. Eluates were analysed using reversed-phase high-performance liquid chromatography with diode array detection (RP-HPLC-DAD). For the five target compounds the determined concentrations ranged from approximately 4 to approximately 15,500 microg/g dry wt. Very good linearities (r(2)0.9995) of calibration curves were achieved for each flavone. The accuracy was below 5% for most compounds examined. This is the first method reported that enabled simultaneous qualification and quantitation of five flavones (being chemotaxonomic markers) in 13 species of Scutellaria L. genus.

Ten species belonging to the family Lamiaceae and representing the most popular medicinal plants used in Polish phytotherapy were examined for the content of free phenolic acids (PhAs). Two depsides, rosmarinic and chlorogenic acids, as well as eight simple PhAs, protocatechuic, gentisic, p-hydroxybenzoic, caffeic, vanillic, syringic, p-coumaric and ferulic acids, in different qualitative and quantitative proportions depending on the plant examined were determined by the rapid, selective and accurate method combining solid-phase extraction and high-performance liquid chromatography.

A new, simple and rapid solid-phase extraction method for the determination of furanochromones and pyranocoumarins in Ammi visnaga L. fruits and pharmaceuticals by reversed-phase high-performance liquid chromatography (RP-HPLC) was developed. The isolation of compounds examined was carried out on octadecyl BakerBond SPE columns using various concentrations of methanol, acetonitrile and tetrahydrofuran in water. High and reproducible recoveries were obtained. To compare the results of quantitative analysis a preparative TLC procedure was also elaborated and carried out.