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In the paper, the identification and quantification of α- and β-amyrin in five varieties of Humulus lupulus is described. The plant samples were extracted with ethyl acetate and analyzed using of high performance liquid chromatography (HPLC) with PAD detection. The separation was achieved on RP 18 column, at 2 mL/min flow rate and at temperature of 35°C. Acetonitrile was used as a mobile phase. The established calibration curves and the other validation parameters: linearity (correlation coefficient r > 0.9988) and precision (RSD values ranged from 0.14 to 1.81%) were found to be satisfactory for the proposed method. The content of α- and β-amyrin strongly depended on varieties of H. lupulus and amounted from 72.7 to 232.5 μg/g and from 77.9 to 176.9 μg/g of dry plant material, respectively.

Cimicifuga racemosa, is a plant with a diverse and long history of medicinal use. Caffeic acid, bioactive compound, which often occurs with other polyphenols can influence the biological activity of this plant. The aim of our work was quantitative analysis of caffeic acid in roots and rhizomes of two varieties of C. racemosa. Analysis was performed by HPLC method. The extracts were separated on C18 reversed-phase column using mixture of methanol, water and formic acid (25:75:0.5 v/v/v) as a mobile phase. The flow rate of eluent was 1.0 ml·min-1. The obtained validation parameters such as linearity, linear regression equation and precision expressed as a relative standard deviation were adequate for quantitative determination. Caffeic acid was found in all tested extracts. The highest total amount of caffeic acid was determined in C. racemosa var. racemosa (255.3 μg·g-1) while its concentration in C. racemosa var. cordifolia was significantly lower (213.0 μg·g-1).

Lamium album, commonly known as "white dead nettle," is a perennial herb widely used in folk medicine. The present paper presents the toxic, anti-proliferative, and free radical (DPPH·) scavenging activities of methanol and ethyl acetate extracts of that plant. In order to determine the biologically active compounds, the plant extracts were separated by high performance thin layer chromatography (HPTLC) on silica gel Si 60 F254 and high performance liquid chromatography (HPLC) combined with densitometry. Methanol extract was rich mainly with flavonoids and phenolic acids. Ethyl acetate extract contained mainly triterpenes. Both extracts showed no toxic effects against normal human skin fibroblasts (HSF) in the range of applied concentrations (25-225 μg/ml). Anti-proliferative activity revealed that methanol extract expressed lower inhibitory properties than ethyl acetate one. The MTT test was, however, less sensitive than Neutral Red (NR) assay. Ethyl acetate extract did not exhibit DPPH· radical scavenging activity. Methanol extract reduced the radical of about 29% at the highest applied concentration (225 μg/ml). Both extracts slightly influenced cellular cytoskeleton organization and amount, and size of agyrophilic nucleolar organizer regions (AgNOR) protein deposits. These findings suggest that extracts of Lamium album exhibit potential usefulness in preparation of new natural formulations. [ABSTRACT FROM AUTHOR]

Strontium is an unavoidable element occurring in plants due to its abundance in the soil and similarity with calcium. To mimic natural conditions, impacts of additional inorganic (nitrate) or organic (urea and allantoin) nitrogen sources (1 mM of each N form in addition to 3.53 mM N in the basic cultivation solution) or N deficit on strontium-induced changes (100 µM Sr) in the widely used medicinal plant Hypericum perforatum L. were studied. Though various effects of Sr on primary (stimulation of amino acids but depression of most Krebs acids, ascorbic acid and thiols) and secondary metabolites (stimulation of phenols but no change of pseudo/hypericin) or mineral elements were observed (reduction of Ca amount in both shoots and roots), organic N forms often mitigated negative action of Sr or even combined stimulatory impact was observed. Organic N forms also elevated shoot accumulation of Sr while N deficit reduced it. Additional N forms, rather than Sr itself, modulated reactive oxygen species and nitric oxide formation in the root tissue. Germination experiment showed no toxicity of Sr to H. perforatum up to 1 mM Sr and even stimulated accumulation of amino acids and phenols, indicating similar ontogenetic-related responses., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Responses of common medicinal herb Matricaria chamomilla to short-term cadmium (Cd) exposure (48 h) alone or in combination with nitric oxide (NO) scavenger (PTIO) or donor (SNP) were studied. Modulators revealed expected impact on NO formation (depletion under PTIO but elevation under SNP) and confirmed anomalous impact on Cd accumulation (stimulation by both modulators). Changes of proline and free amino acids in the roots and fatty acids (mainly α-linolenic acid) in the shoots indicate rather relation to elevated Cd uptake than to altered NO formation. On the contrary, root profile of fatty acids revealed the most prominent changes in response to PTIO (elevation of monounsaturated and polyunsaturated fatty acids) which confirm that NO depletion stimulates their biosynthesis. Soluble phenols were elevated by SNP co-application in the roots while accumulation of chlorogenic acid and umbelliferone revealed clear regulation by NO (i.e. enhancement in response to SNP and depletion in response to PTIO). Data indicate that quantities of some metabolites are affected by accumulated Cd or NO formation and that the opposite NO modulators do not automatically lead to the opposite changes of plant metabolites., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Roots and leaves of Carlina acaulis L. are still used in ethnomedicine in many European countries; however, the limited occurrence of the plants and protection of this species necessitate a search for alternative ways for obtaining this plant material. In this study, in vitro cultures, hydroponic cultures, and field cultivation were applied to obtain the C. acaulis plant material. Its quality was evaluated using antioxidant activity tests and high performance liquid chromatography analysis. Our study showed that the antioxidant activity and the content of chlorogenic and 3,5-di-caffeoylquinic acid in roots of plants cultivated in hydroponics and field conditions were comparable. However, the amount of carlina oxide was significantly higher in plants from the field. The flavonoid content in leaves obtained from both cultivation systems was at the same level; however, the antioxidant activity and the content of the investigated metabolites were higher in the soil cultivation system. The callus line exhibited high differentiation in phytochemical compositions depending on the treatments and medium compositions.

Silver is one of the most toxic heavy metals for plants, inducing various toxic symptoms and metabolic changes. Here, the impact of Ag(I) on Carlina acaulis physiology and selected metabolites was studied using two Ag concentrations (1 or 10 µM) after 14 days of exposure. The higher concentration of Ag(I) evoked reduction of growth, while 1 µM Ag had a growth-promoting effect on root biomass. The translocation factor (<0.04) showed that Ag was mainly retained in the roots. The 1 µM Ag concentration increased the level of low-molecular-weight organic acids (LMWOAs), while 10 µM Ag depleted these compounds in the roots. The increased concentration of Ag(I) elevated the accumulation of phytochelatins (PCs) in the roots and reduced glutathione (GSH) in the shoots (but not in the roots). At 1 µM, Ag(I) elevated the level of phenolic and triterpene acids, while the 10 µM Ag treatment increased the carlina oxide content in the roots. The obtained results indicate an alteration of metabolic pathways of C. acaulis to cope with different levels of Ag(I) stress. Our data imply that the intracellular binding of Ag(I) and nonenzymatic antioxidants contribute to the protection against low concentrations of Ag ions., Competing Interests: The authors declare no conflict of interest.

The impact of long-term exposure to Sr 2+ (LTE, four doses, 43.5 mg Sr 2+ per pot, with a total of 174 mg Sr 2+ per pot during the entire period of cultivation) and short-term exposure to Sr 2+ (STE, one dose, 870 mg Sr 2+ per pot four days before harvest) on the content of phytoestrogens and allantoin in soybeans were compared. Sr 2+ accumulation, the effect on the concentration of macroelements, and basic physiology were also analyzed. LTE reduced the content of malonyldaidzin and malonylgenistin in the roots (58% and 50% compared to the control, respectively). STE increased the amount of all isoflavones in the stem and genistein in the leaves and decreased the content of malonyldaidzin and malonylgenistin in the leaves (55% and 48% compared to the control, respectively) and roots (69% and 62% of the control, respectively) as well as genistein and coumestrol in the roots (both 50% compared to the control). Sr 2+ presence stimulated the accumulation of allantoin in the roots (three-fold higher than in the control), but only STE had similar effects on the shoots. In contrast to LTE, Sr 2+ was transported extensively from the roots to the leaves under STE. In comparison to the control, LTE resulted in an increase in the Ca content in the stem by 36%, whereas Ca 2+ accumulation in the leaves, stems, and roots increased by 60%, 80%, and 36%, respectively, under STE. Additionally, a significant accumulation of K was found only in the roots of the LTE group. The chlorophyll content did not differ between the treatments. Overall, the production of phytoestrogens and Sr accumulation were affected by both the applied dose and the duration of exposure to Sr., Competing Interests: The authors declare no conflicts of interest.

Isoquinoline alkaloids are the main group of secondary metabolites present in Chelidonium majus extracts, and they are still the object of interest of many researchers. Therefore, the development of methods for the investigation and separation of the alkaloids is still an important task. In this work, the application potential of a silica-based monolithic column for the separation of alkaloids was assessed. The influence of the organic modifier, temperature, salt concentration, and pH of the eluent on basic chromatographic parameters such as retention, resolution between neighboring peaks, chromatographic plate numbers, and peak asymmetry were investigated. Based on the obtained results, a gradient elution program was developed and used to separate and quantitatively determine the main alkaloids in a Chelidonium majus root extract., Competing Interests: The authors declare no conflict of interest.

Diosmin is a natural compound with a wide range of biological activity, e.g., it improves lymphatic drainage, supports microcirculation, and increases venous tone, and venous elasticity, hence, it is applied in the pharmacotherapy of chronic venous disorders (CVD). The aim of this study was to assess the correlation between diosmin administration (2 × 600 mg daily) in patients suffering from CVD and the levels of selected factors influencing angiogenesis, which are involved in CVD pathophysiology. Thirty-five CVD patients were examined. Levels of plasma tumor necrosis factor alpha (TNF alpha), vascular endothelial growth factor (VEGF-A and VEGF-C); angiostatin, interleukin 6 (IL-6), fibroblast growth factor 2 (FGF2); and plasminogen (PLG) were measured with an Elisa assay before and after three months of diosmin administration. The clinical symptoms of CVD were monitored using ultrasound images, echo Doppler assay, visual analogue scale (VAS), and measurement of the leg circumference. The average content of TNF alpha, VEGF-C, VEGF-A IL-6, and FGF2 decreased after the therapy with diosmin in a significant manner; with p < 0.001, p < 0.05, p < 0.05, p < 0.01, and p < 0.01, respectively, and a significant ( p < 0.05) increase in the plasma angiostatin level after the three-month treatment was found. A significant ( p < 0.05) decrease in edema and the average leg circumference of the patients was observed after the therapy. Diosmin influences the angiogenic and inflammatory mechanisms involved in the pathophysiology of edema presented in patients with a different class of CVD.

Ethnopharmacological Relevance: The genus Carlina (Asteraceae) comprises approx. 30 plant species growing in Europe and Asia. Carlina acaulis L. was widely used as medicine, especially in Germany and Poland. The plant root was applied e.g. to treat various skin diseases and as a diaphoretic, diuretic, and anthelmintic agent. At the end of the 19th century, the medicinal use of the plant ceased though C. acaulis roots are still used in folk medicine, especially in rural areas of the Balkans where the plant grows in the wild., Aim of the Review: The aim of this review is to compile historical and current ethnomedical uses of C. acaulis and compare these applications with the results of modern chemical and pharmacological research of this plant. This analysis may give a lead for further research on this old, almost forgotten medicinal plant., Methodology: Historical data on the medicinal application of C. acaulis over many centuries were collected. The historical uses were compared with the contemporary indications. The present day knowledge on the chemistry and biological activity of C. acaulis is also reviewed. This information is compared with the ethnopharmacological information., Results: Plants from the genus Carlina were already described by Theophrastus of Eresus, Dioscorides, Pliny the Elder, and Galen of Pergamum. Carlinae radix was still featured in Renaissance botanical books and in official pharmacopoeias, but disappeared from the scene in the 19th century. The present day information on the phytochemistry and pharmacological activity of these plants is limited. There are some data on the occurrence of common plant ingredients such as essential oil, flavonoids, phenolic acids, triterpenes, inulin, and fatty acids. However, most concern the green parts of the plant. Its medicinal use remains poorly understood, as there are only very limited results of in-vitro studies on the roots., Conclusions: The historical uses of the plant can still be found in rural areas. Yet, detailed phytochemical and pharmacological research is needed to evaluate the effectiveness and safety of the medical use of Carlina acaulis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Chelidonium majus ( Papaveraceae ) extracts exhibit antimicrobial activity due to the complex alkaloid composition. The aim of the research was to evaluate the antimicrobial potential of extracts from wild plants and in vitro cultures, as well as seven major individual alkaloids. Plant material derived from different natural habitats and in vitro cultures was used for the phytochemical analysis and antimicrobial tests. The composition of alkaloids was analyzed using chromatographic techniques (HPLC with DAD detection). The results have shown that roots contained higher number and amounts of alkaloids in comparison to aerial parts. All tested plant extracts manifested antimicrobial activity, related to different chemical structures of the alkaloids. Root extract used at 31.25-62.5 mg/L strongly reduced bacterial biomass. From the seven individually tested alkaloids, chelerythrine was the most effective against P. aeruginosa (MIC at 1.9 mg/L), while sanguinarine against S. aureus (MIC at 1.9 mg/L). Strong antifungal activity was observed against C. albicans when chelerythrine, chelidonine, and aerial parts extract were used. The experiments with plant extracts, individually tested alkaloids, and variable combinations of the latter allowed for a deeper insight into the potential mechanisms affecting the activity of this group of compounds.

The aim of the study was to demonstrate the potential of the promotion and regulation of plant physiology and growth under control and copper stress conditions, and the impact of the exogenous application of methyl jasmonate on this potential. Runner bean plants were treated with methyl jasmonate (1 or 10 µM) (J; J1 or J10) and Cu (50 µM), and inoculated with a bacterial isolate (S17) originating from Spitsbergen soil, and identified as Pseudomonas luteola using the analytical profile index (API) test. Above- and under-ground plant parts were analyzed. The growth parameters; the concentration of the photosynthetic pigments, elements, flavonoids (FLAVO), phenolics (TPC), allantoin (ALLA), and low molecular weight organic acids (LMWOAs); the activity of antioxidant enzymes and enzymes of resistance induction pathways (e.g., superoxide dismutase (SOD), catalase (CAT), ascorbate (APX) and guaiacol (GPX) peroxidase, glucanase (GLU), and phenylalanine (PAL) and tyrosine ammonia-lyase (TAL)), and the antioxidant capacity (AC) were studied. The leaves exhibited substantially higher ALLA and LMWOA concentrations as well as PAL and TAL activities, whereas the roots mostly had higher activities for a majority of the enzymes tested (i.e., SOD, CAT, APX, GPX, and GLU). The inoculation with S17 mitigated the effect of the Cu stress. Under the Cu stress and in the presence of J10, isolate S17 caused an elevation of the shoot fresh weight, K concentration, and TAL activity in the leaves, and APX and GPX (also at J1) activities in the roots. In the absence of Cu, isolate S17 increased the root length and the shoot-to-root ratio, but without statistical significance. In these conditions, S17 contributed to a 236% and 34% enhancement of P and Mn, respectively, in the roots, and a 19% rise of N in the leaves. Under the Cu stress, S17 caused a significant increase in FLAVO and TPC in the leaves. Similarly, the levels of FLAVO, TPC, and AC were enhanced after inoculation with Cu and J1. Regardless of the presence of J, inoculation at Cu excess caused a reduction of SOD and CAT activities, and an elevation of GPX. The effects of inoculation were associated with the application of Cu and J, which modified plant response mainly in a concentration-dependent manner (e.g., PAL, TAL, and LMWOA levels). The conducted studies demonstrated the potential for isolate S17 in the promotion of plant growth.

Concentration-dependent responses of cucumber plants to cadmium (Cd, 5-15 µM) and/or allantoin (Alla, 10-1000 µM) have been investigated to detect a possible protective role of Alla under Cd excess. After 14 days of exposure, Alla often considerably reversed Cd-induced inhibition of growth and reduction of the content of photosynthetic pigments. Higher Alla doses depleted the Cd amount in shoots, which could be related to citric acid (increase in the shoots but depletion in the roots in Cd+Alla treatments) rather than to phytochelatins (Alla had a negative impact on the phytochelatin accumulation). An increase in the Alla concentration suppressed Cd-induced spatial H 2 O 2 appearance, which does not seem to be related to antioxidative enzymes (low impact of Alla on catalase, ascorbate peroxidase, and guaiacol peroxidase). On the contrary, shoot glutathione and mainly ascorbic acid accumulation strongly increased in Cd+Alla treatments, indicating their prominent role in Alla-induced amelioration of Cd-stimulated oxidative stress and growth retardation. Similarly, phenolic metabolites (total soluble phenols and flavonols) were slightly influenced by Alla and their antioxidative action was not expected. We conclude that Alla-mediated attenuation of Cd-induced toxicity relies on enhanced accumulation of glutathione and ascorbate in the shoot tissue mainly, rather than on elevated antioxidative enzyme activities., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Secondary metabolites of the roots of Eleutherococcus spp. cultivated in Poland, or the bioactivity, are not fully known. The 75% methanol extracts of five Eleutherococcus spp. ( E. senticosus , E. divaricatus , E. sessiliflorus , E. gracilistylus , and E. henryi ) were examined for the content of polyphenols and phenolic acids as well as for antiacetylcholinesterase, antihyaluronidase, anti-DPPH ∗ , and cytotoxic activities. The richest in polyphenols were the roots of E. henryi (10.4 mg/g DW), while in flavonoids the roots of E. divaricatus (6.5 mg/g DW). The richest in phenolic acids occurred the roots of E. henryi [protocatechuic acid (1865  μ g/g DE), caffeic acid (244  μ g/g DE), and p -coumaric and ferulic acids (55  μ g/g DE)]. The highest inhibition of AChE was observed for E. gracilistylus and E. sessiliflorus (32%), at the concentration of 100  μ g/0.19 mL of the reaction mixture, while that of Hyal for the roots of E. henryi (40.7%), at the concentration of 100  μ g/0.16 mL of the reaction mixture. Among five species tested, the E. henryi extract exhibited the strongest HL-60 cell line growth's inhibition (IC 50 270  μ g/mL). The extracts reduced DPPH ∗ in a time-dependent mode, at the concentration of 0.8 mg/mL. After 90 min from 14.7 to 26.2%, DPPH ∗ was reduced. A phytochemical composition and activity of the Eleutherococcus species, cultivated in Poland, are still under research; however, on the basis of the results obtained, it may be concluded that they may become a source of phytochemicals and be useful for Europe's citizens.

In the present study, deprotonated and protonated polyaniline based sorbent obtained by in situ polymerization of aniline on silica (Si-PANI) was applied for SPE pretreatment of Chelidonium majus extracts before determination of benzophenanthridine, protoberberine and protopine alkaloids. The experimentally optimized conditions, such as protonated polyaniline impregnated with methanol as a sorbent, 2 mL of water and methanol mixture (1/1, v/v) as a washing solution, and 5 mL of 0.1 M methanolic solution of ammonia as an elution solvent, yielded the highest values of analyte recovery (above 96.5%) with simultaneous removal of undesirable plant matrix. Virtually modeled structure of polyaniline revealed possible occurrence of the mixed mode binding mechanism, based both on π-π and ion interactions on protonated form of polyaniline. Moreover, lack of significant changes of the polyaniline film quality assessed by Raman spectroscopy after series of ten experiments proved the reusability of the sorbent., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this work, a fast and low-cost voltammetric methodology for determination of carlina oxide in plant extracts was developed. The best results were obtained using a boron-doped diamond electrode (BDDE). The voltammetric measurements of carlina oxide were performed in a 0.1 mol/L solution of sulphuric acid. After 30 s of stirring the solution, differential pulse voltammograms (DPVs) were recorded from 0.5 to 1.8 V. The amplitude was 75 mV and the scan rate was 175 mV/s. Measurements were recorded in non-deaerated solutions. The background current was subtracted from each registered voltammogram; then they were cut from 0.5 to 1.5 V. The detection and quantification limits were 0.28 and 0.93 μg/L, respectively, and repeatability expressed as the relative standard deviation of 0.1 mg/L of carlina oxide was 1.9% (n = 5). The results were compared with those obtained using gas chromatography with a flame ionization detector and high performance liquid chromatography with a photodiode array detector., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Abiotic stress, including metal excess, can modify plant metabolism. Here we investigated the influence of long-term strontium exposure (12 weeks, 0.5⁻4.0 mM Sr) on the content of phytoestrogens and allantoin as well as the mineral composition in soybean. Seven phytoestrogens were identified in the soybean: daidzin, glycitin, genistin, malonyldaidzin, malonylgenistin, daidzein, and coumestrol. The results showed that both malonyldaidzin and malonylgenistin were dominant phytoestrogens; however, the roots contained a relatively high amount of daidzein. It was found that strontium reduced the phytoestrogen content and decreased the antioxidant capacity. Strontium evoked depletion of the sum of all phytoestrogens by 40⁻70% in the leaves, 25⁻50% in the stems and in the seeds, depending on the strontium concentration. In the roots, 0.5 and 4.0 mM of strontium decreased the total phytoestrogen content by 25 and 55%, respectively, while 2.0 mM of strontium did not exert an effect on their accumulation. On the other hand, strontium ions induced allantoin accumulation mainly in the roots. Strontium was preferentially accumulated in the leaves, with a slight impact on macro- and micro-nutrients. Our research showed strontium-secondary metabolites interaction in the soybean, which can be useful for obtaining a natural pharmaceutical product containing both strontium and phytoestrogens for remediation of postmenopausal osteoporosis.

Manganese (Mn) uptake and toxicity in chamomile (Matricaria chamomilla) and changes of phenolic metabolites in plants grown in the soil (1000 μM Mn 2+ ) or hydroponic culture (100 or 1000 μM Mn 2+ ) were studied. Under soil cultivation, Mn excess reduced growth and induced symptoms of oxidative stress (including total ROS, hydroxyl radical and lipid peroxidation as detected by fluorescence microscopy), concomitantly with depletion of non-protein thiols and ascorbic acid. Total soluble phenols and individual phenolic acids were rather depleted (p-coumaric, chlorogenic, and protocatechuic acids) or unaltered (vanillic and caffeic acids). Shoot Mn content reached 2806 μg/g DW with BAF 51.0 in the soil culture. In hydroponics, tetraploid plants contained less Mn in both shoots and roots than diploid ones with bioaccumulation factor and translocation factor (diploid/tetraploid) 57.1/37.9 and 0.39/0.32 in 1000 μM Mn treatment. Plants cultured in hydroponics revealed stimulation of some phenolic acids, mainly chlorogenic acid in the shoots and p-hydroxybenzoic and vanillic acids in the roots (more extensively in tetraploid ones which contained less Mn). Data indicate that excessive Mn accumulation has negative impact not only on the growth but also on phenolic metabolites in young plants mainly. Detailed comparison of the observed metabolic changes with limited literature focused on Mn physiology is provided as well., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

The conditions of micellar electrokinetic capillary chromatography for separation and simultaneous measurement of coumarins (coumarin, scoparone, isoscopoletin, esculin, esculetin, umbelliferone) including furanocoumarins (xanthotoxin, byakangelicin, isopimpinellin, bergapten, phellopterin, xanthotoxol) have been elaborated. The influence of different parameters, such as the pH of the buffer, sodium cholate (SC) or methanol concentration in the buffer, on the migration time, peak resolution, peak asymmetry, and number of theoretical plates was investigated. The optimum separation of the compounds was achieved using 50-µm i.d. capillaries with a total length of 64.5 cm (56 cm effective length) and a buffer system at pH 9.00 consisting of 50 mM sodium tetraborate, 45 mM SC, and 20% of methanol (v/v). The developed method ensured good repeatability of corrected peak areas and migration times (the relative standard deviations were in the range of 2.8-6.1% and 0.8-4.0%, respectively). The average limit of detection for all studied compounds was below 1.3 µg mL. Moreover, good linearity of the relationship between the peak corrected area and the concentration of the compounds was observed (correlation coefficient >0.99). The method was successfully applied in the quantitative analysis of two different types of samples, i.e. Heracleum sphondylium herb and Aesculus hippocastanum cortex., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Oxidative stress plays an important role in the pathophysiology of many human disorders, while antioxidants prevent the development of various adverse symptoms. Diosmin is a natural flavonoid applied in vascular system disorders, especially in chronic venous insufficiency (CVI), and it plays a significant part in the alleviation of CVI symptoms. Due to antioxidant activity, it also has the ability to scavenge the oxygen free radicals and hence decreases the level of oxidative stress biomarkers, such as prostaglandins and their precursors-isoprostanes. In the study, the influence of diosmin treatment on the level of isoprostanes in plasma samples of patients suffering from CVI was examined. The qualitative analysis was performed using high-performance liquid chromatography with spectrometry detection (LC-MS). The statistically significant decrease of isoprostane content after 3 months of treatment was observed within the studied group; however, the most significant changes were observed in patients who smoke.

Accumulation of selected secondary metabolites in two Hypericum species (H. perforatum and H. annulatum) was compared in their vegetative parts (stems and leaves) and in terms of the extraction solvent (80% aq. methanol or 60% aq. ethanol). The presence of chlorogenic acid and quercitrin was not detected in stem of both species. Almost all metabolites were more accumulated in the leaves than in the stems (rutin, hyperoside, quercetin and hypericin) but epicatechin showed the opposite in both species and hyperforin in H. annulatum. Extraction solvents showed rather species-specific differences with EtOH being more suitable for the extraction of hypericin, quercetin, quercitrin, and hyperoside (on average, for both the leaves and stems, extraction increased by approximately 130, 30, 25, and 15%, respectively) while MeOH for the extraction of epicatechin, rutin, and hyperforin (increased extraction by approximately 50, 40, and 35%, respectively). On the other hand, content of total soluble phenols did not differ in relation to solvent in any organ or species. Various ages of H. annulatum plants did not show dramatic impact on the amount of metabolites. Subsequently, the usefulness of capillary electrophoresis (CE) as an alternative to HPLC for the quantification of metabolites in H. perforatum was tested and results showed non-significant differences between CE and HPLC with the methods we developed (the difference did not exceed 10%)., (Copyright © 2018 Elsevier B.V. All rights reserved.)

As antique as Dioscorides era are the first records on using Chelidonium as a remedy to several sicknesses. Inspired by the " signatura rerum " principle and an apparent ancient folk tradition, various indications were given, such as anti-jaundice and cholagogue, pain-relieving, and quite often mentioned-ophthalmological problems. Central and Eastern European folk medicine has always been using this herb extensively. In this region, the plant is known under many unique vernacular names, especially in Slavonic languages, associated or not with old Greek relation to "chelidon"-the swallow. Typically for Papaveroidae subfamily, yellow-colored latex is produced in abundance and leaks intensely upon injury. Major pharmacologically relevant components, most of which were first isolated over a century ago, are isoquinoline alkaloids-berberine, chelerythrine, chelidonine, coptisine, sanguinarine. Modern pharmacology took interest in this herb but it has not ended up in gaining an officially approved and evidence-based herbal medicine status. On the contrary, the number of relevant studies and publications tended to drop. Recently, some controversial reports and sometimes insufficiently proven studies appeared, suggesting anticancer properties. Anticancer potential was in line with anecdotical knowledge spread in East European countries, however, in the absence of directly-acting cytostatic compounds, some other mechanisms might be involved. Other properties that could boost the interest in this herb are antimicrobial and antiviral activities. Being a common synanthropic weed or ruderal plant, C. majus spreads in all temperate Eurasia and acclimates well to North America. Little is known about the natural variation of bioactive metabolites, including several aforementioned isoquinoline alkaloids. In this review, we put together older and recent literature data on phytochemistry, pharmacology, and clinical studies on C. majus aiming at a critical evaluation of state-of-the-art from the viewpoint of historical and folk indications. The controversies around this herb, the safety and drug quality issues and a prospective role in phytotherapy are discussed as well.

Polyaniline (PANI) is one of the best known conductive polymers with multiple applications. Recently, it was also used in separation techniques, mostly as a component of composites for solid-phase microextraction (SPME). In the present paper, sorbent obtained by in situ polymerization of aniline directly on silica gel particles (Si-PANI) was used for dispersive solid phase extraction (d-SPE) and matrix solid-phase extraction (MSPD). The efficiency of both techniques was evaluated with the use of high performance liquid chromatography with diode array detection (HPLC-DAD) quantitative analysis. The quality of the sorbent was verified by Raman spectroscopy and microscopy combined with automated procedure using computer image analysis. For extraction experiments, triterpenes were chosen as model compounds. The optimal conditions were as follows: protonated Si-PANI impregnated with water, 160/1 sorbent/analyte ratio, 3 min of extraction time, 4 min of desorption time and methanolic solution of ammonia for elution of analytes. The proposed procedure was successfully used for pretreatment of plant samples., Competing Interests: The authors declare no conflict of interest.

The root of Symphytum officinale L. is commonly used in folk medicine to promote the wound healing, reduce the inflammation and in the treatment of broken bones. The objective of our investigation was to analyse the extract from S. officinale in term of its antioxidant activity and the effect on cell viability and proliferation of human skin fibroblast (HSF). Moreover, the quantification of main phenolics and allantoin was conducted using HPLC-DAD method. Five compounds were found: rosmarinic, p-hydroxybenzoic, caffeic, chlorogenic and p-coumaric acid. DPPH, FRAP and TPC assay showed the high antioxidant activity of the extract. MTT test proved the stimulatory effect on cell metabolism and viability of HSF cells. Moreover, no changes in cytoskeleton structure and cells shape were observed. The obtained results indicate that non-toxic extract from S. officinale root has strong antioxidant potential and a beneficial effect on human skin fibroblasts.

Chelidonium majus L. is a rich source of isoquinoline alkaloids with confirmed anti-inflammatory, choleretic, spasmolytic, antitumor, and antimicrobial activities. However, their chromatographic analysis is difficult because they may exist both in charged and uncharged forms and may result in the irregular peak shape and the decrease in chromatographic system efficacy. In the present work, the separation of main C. majus alkaloids was optimized using a new-generation XB-C18 endcapped core-shell column dedicated for analysis of alkaline compounds. The influence of organic modifier concentration, addition of salts, and pH of eluents on chromatographic parameters such as retention, resolution, chromatographic plate numbers, and peak asymmetry was investigated. The results were applied to elaborate the optimal chromatographic system for simultaneous quantification of seven alkaloids from the root, herb, and fruit of C. majus .

Context: Viscum album L. (Loranthaceae) is a semi-parasitic plant used in pharmacy and medicine mostly for its hypotensive and anticancer activity. The effects may be related to the presence of triterpenic acids, such as betulinic (BA) and oleanolic (OA) acids., Objectives: In our investigations the content of triterpenic acids in V. album from different host trees depending on the season of harvest was determined., Material and Methods: V. album herb was dried and extracted with ethyl acetate using ultrasound energy. The reversed phase HPLC-PDA method was used for the analysis of triterpenic acids. The structure of the target components was confirmed by mass spectrometry with an electrospray ionization source., Results: Diversity in the content of both compounds was noted; however, OA was the dominant triterpenic acid and the amount thereof was ∼10 times higher than that of BA. The analysis of changes in the amount of triterpenic acids during the spring-winter period revealed the highest content of OA in summer (from 6.84 to 13.65 mg/g). In turn, in the other seasons of harvest, the content was in the range of 4.41-9.83, 6.41-9.56 and 5.59-12.16 mg/g for spring, autumn and winter, respectively. In most cases, a similar tendency was observed for BA., Discussion and Conclusion: In most cases, the highest amount of the investigated compounds was found in summer; thus, this period seems to be optimal for acquisition of plant material rich in triterpenic acids.

Carlina genus plants e.g. Carlina acanthifolia subsp. utzka have been still used in folk medicine of many European countries and its biological activity is mostly associated with root essential oils. In the present paper, Raman spectroscopy (RS) was applied for the first time for evaluation of essential oil distribution in root of C. acnthifolia subsp. utzka and identification of root structures containing the essential oil. Furthermore, RS technique was applied to assess chemical stability of oil during drying of plant material or distillation process. Gas chromatography-mass spectrometry was used for qualitative and quantitative analysis of the essential oil. The identity of compounds was confirmed using Raman, ATR-IR and NMR spectroscopy. Carlina oxide was found to be the main component of the oil (98.96% ± 0.15). The spectroscopic study showed the high stability of essential oil and Raman distribution analysis indicated that the oil reservoirs were localized mostly in the structures of outer layer of the root while the inner part showed nearly no signal assigned to the oil. Raman spectroscopy technique enabled rapid, non-destructive direct analysis of plant material with minimal sample preparation and allowed straightforward, unambiguous identification of the essential oil in the sample., (Copyright © 2017. Published by Elsevier B.V.)

Various species of the Carlina genus have been used in traditional medicine in many countries to treat numerous skin disorders, including cancer. The objective of this work was to assess the anticancer properties of root and leaf extracts from Carlina acaulis subsp. caulescens and C. acanthifolia subsp. utzka . Anti-tumor properties of the extracts were explored using a tetrazolium-based cell viability assay and flow cytometric apoptosis analysis, followed by immunodetection of phosphoactive ERK1/2 in UACC-903, C32, and UACC-647 human melanoma cell lines. Normal human fibroblasts were used as a control. Leaf extracts inhibited the viability of all tested melanoma cell lines in a dose-dependent fashion while the fibroblasts were less sensitive to such extract. The root extracts inhibited the proliferation of UACC-903 and UACC-647 cells only at the highest doses (300 μg/mL). However, the C32 and fibroblast cells exhibited an increase in the cellular proliferation rate and no caspase activity was observed in response to the root extracts (100 μg/mL). An increase in caspase activity was observed in melanoma cells treated with the leaf extracts of both Carlina species. Leaf extracts from C. acaulis subsp. caulescens (100 μg/mL) inhibited proliferatory ERK1/2 in UACC-903 and C32 cells, as demonstrated by the decrease in ERK1/2 phosphorylation. No reduction in phospho-ERK1/2 was observed in the tested cell lines treated with the root extracts, apart from UACC-647 after incubation with the C. acanthifolia subsp. utzka root extract (100 μg/mL). There was no change in ERK1/2 phosphorylation in the fibroblasts. The extracts from the leaves and roots were analyzed by HPLC and the analysis showed the presence of triterpenes and phenolic acids as the main extract components. The research demonstrated that the extracts from the leaves of the plants were cytotoxic against the human melanoma line and induced apoptosis of the cells. The triterpene fraction present in the tested extracts may be responsible for this activity.

The aim of the study was to determine the response of metallicolous and nonmetallicolous Echium vulgare L. populations to chronic multi-metal (Zn, Pb, Cd) and acute Zn (200, 400 μM) and Pb (30, 60 μM) stress. Three populations of E. vulgare, one from uncontaminated and two from metal-contaminated areas, were studied. Two types of experiments were performed - a short-term hydroponic experiment with acute Zn or Pb stress and a long-term manipulative soil experiment with the use of soils from the sites of origin of the three populations. Growth parameters, such as shoot and root fresh weight and leaf area, as well as organic acid accumulation were determined. Moreover, the concentration of selected secondary metabolites and antioxidant capacity in the three populations exposed to Pb or Zn excess were measured. Both metallicolous populations generally achieved higher biomass compared with the nonmetallicolous population cultivated under metal stress in hydroponics or on metalliferous substrates. Plants exposed to Pb or Zn excess or contaminated soil substrate exhibited higher malate and citrate concentrations compared with the reference (no metal stress) plants. It was observed that Zn or Pb stress increased accumulation of allantoin, chlorogenic and rosmarinic acids, total phenolics, and flavonoids. Moreover, it was shown that Pb sequestration in the roots or Zn translocation to the shoots may play a role in enhanced metal tolerance of metallicolous populations under acute Pb/Zn stress., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Trace analysis plays an important role in medicine for diagnosis of various disorders; however, the appropriate sample preparation is required mostly including mineralization. Although graphite furnace atomic absorption spectrometry (GF AAS) allows the investigation of biological samples such as blood, serum, and plasma without this step, it is rarely used for direct analysis because the residues of the rich organic matrix inside the furnace are difficult to remove and this may cause spectral/matrix interferences and decrease the lifetime of the graphite tube. In our work, the procedure for determination of Se, Cr, Mn, Co, Ni, Cd and Pb with the use of the high resolution continuum source GF-AAS technique in whole blood samples with minimum sample pre-treatment was elaborated. The pyrolysis and atomization temperature as well as the time of signal integration were optimized to obtain the highest intensity and repeatability of the analytical signal. Moreover, due to the apparatus modification, an additional step was added in the for graphite furnace temperature program with minimal argon flow and maximal flow of air during pyrolysis stage to increase the oxidative condition for better matrix removal. The accuracy and precision of the optimized method was verified using certified reference material (CRM) Seronorm Trace Elements Whole Blood L-1 and the developed method was applied for trace analysis of blood samples from volunteer patients of the Orthopedics Department., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The methanol extracts from three populations of Carlina vulgaris L. were examined for the chlorogenic acid content, mineral content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity. Two populations originated from natural nonmetallicolous habitats (NN (populations from Nasiłów) and NP (populations from Pińczów)), and one metallicolous population (MB) was collected from Bolesław waste heap localized at the place of former open-cast mining of Ag-Pb and Zn-Pb ores dating back to the 13th century and 18th century, respectively. The level of Zn, Pb, Cd, Fe, Ni, and Mn was significantly higher in the root and leaves of MB plants as a result of soil contaminations compared to those of the NN and NP ones. The highest antioxidant potency has been showed by the plants growing in a nonmetallicolous habitat. The flower head extracts obtained from the nonmetallicolous populations also contained the largest amount of chlorogenic acid, whereas the lowest was determined in the roots (ca. 2-3.5 mg/g and 0.2-0.4 mg/g of air-dry weight, resp.). These studies provide important information on the influence of a habitat on the quality of herbal materials and the content of the biologically active primary and secondary metabolites.

The amount of secondary metabolites in plants can be enhanced or reduced by various external factors. In this study, the effect of strontium ions on the production of phytoestrogens in soybeans was investigated. The plants were treated with Hoagland's solution, modified with Sr(2+) with concentrations ranging from 0.5 to 3.0 mM, and were grown for 14 days in hydroponic cultivation. After harvest, soybean plants were separated into roots and shoots, dried, and pulverized. The plant material was extracted with methanol and hydrolyzed. Phytoestrogens were quantified by HPLC. The significant increase in the concentration of the compounds of interest was observed for all tested concentrations of strontium ions when compared to control. Sr(2+) at a concentration of 2 mM was the strongest elicitor, and the amount of phytoestrogens in plant increased ca. 2.70, 1.92, 3.77 and 2.88-fold, for daidzein, coumestrol, genistein and formononetin, respectively. Moreover, no cytotoxic effects were observed in HepG2 liver cell models after treatment with extracts from 2 mM Sr(2+)-stressed soybean plants when compared to extracts from non-stressed plants. Our results indicate that the addition of strontium ions to the culture media may be used to functionalize soybean plants with enhanced phytoestrogen content.

Actaea racemosa L. is used as a component of drugs or dietary supplements to alleviate the menopause symptoms. Its biological activity is associated with the presence of phenolic compounds. In our work, the analysis of isoflavones and phenolic acids - caffeic acid (CA), ferulic acid and isoferulic acid (iFA) - both free and bonded in two species of Actaea, was conducted using HPLC-PAD technique. Moreover, the antioxidant effect of extracts from different parts of the investigated plants was determined on the basis of DPPH assay. Significant variation of CA and iFA content was observed. The highest content of CA was found in A. racemosa, while Actaea cordifolia contained the highest amount of iFA. Isoflavones were not found in the investigated plants. The antioxidant activity assay showed the high free radical-scavenging ability of the extracts obtained from different parts of the plant.

The paper describes the fabrication and application of a novel sensor (a boron-doped diamond electrode modified with Nafion and lead films) for the simultaneous determination of paracetamol and ascorbic acid by differential pulse voltammetry. The main advantage of the lead film and polymer covered boron-doped diamond electrode is that the sensitivity of the stripping responses is increased and the separation of paracetamol and ascorbic acid signals is improved due to the modification of the boron-doped diamond surface by the lead layer. Additionally, the repeatability of paracetamol and ascorbic acid signals is improved by the application of the Nafion film coating. In the presence of oxygen, linear calibration curves were obtained in a wide concentration range from 5×10(-7) to 2×10(-4) mol L(-1) for paracetamol and from 1×10(-6) to 5×10(-4) mol L(-1) for ascorbic acid. The analytical utility of the differential pulse voltammetric method elaborated was tested in the assay of paracetamol and ascorbic acid in commercially available pharmaceutical formulations and the method was validated by high performance liquid chromatography coupled with diode array detector., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Soy (Glycine max (L.) Merr.) is an annual plant cultivated worldwide mostly for food. Moreover, due to its pharmacological properties it is widely used in pharmacy for alleviating the symptoms of osteoporosis. The aim of the present study was to investigate the biofortification of soy treated with various concentrations of strontium. Soy was found to have a strong capacity to absorb Sr(2+) (bioconcentration factor higher than 1). A positive linear correlation (R(2) > 0.98) between the amount of strontium in the growth medium and its content in the plant was also observed. Moreover, at a concentration of 1.5 mM, strontium appeared to be nontoxic and even stimulated plant growth by approximately 19.4% and 22.6% of fresh weight for shoots and roots, respectively. Our research may be useful to obtain vegetable products or herbal preparations containing both phytoestrogens and strontium to prevent postmenopausal osteoporosis.