Determination of individual coumarin in a dry extract of Melilotus officinalis L.

Introduction. Substances of plant origin exhibit a variety of pharmacological activity in combination with a relatively low toxicity and frequency of side effects. In this regard, a promising direction is the standardization of substances of plant origin in the development of new herbal medicinal products.Aim. Qualitative and quantitative determination of individual coumarin in a dry extract of Melilotus оfficinalis L.Materials and methods. A dry extract of the Melilotus officinalis L. was used as an object of research. Determination of individual coumarin was carried out by high-performance liquid chromatography.Results and discussion. As a result of the research, coumarin was identified by high performance liquid chromatography based on comparison of retention times of the sample with retention times of a standard sample of coumarin in the sample of dry extract under research. The coumarin contents in the dry extract was 0.642 ± 0.007 %.Conclusion. The data obtained on the qualitative and quantitative content of coumarin in the dry extract of Melilotus оfficinalis L. were used in the development of the draft version specification of quality indicators.

Antioxidant and Antiinflammatory Effects of Epilobium parviflorum, Melilotus officinalis and Cardiospermum halicacabum Plant Extracts in Macrophage and Microglial Cells

Background: We investigated the phenolic content characterizing different plant extracts from Epilobium parviflorum, Cardiospermum halicacabum, and Melilotus officinalis, their antioxidant, antiinflammatory effects, and their mechanism of action. Methods: plant samples were macerated in 40% ethanol or hot/ cold glycerate and assessed for polyphenols content. The antioxidant activity was investigated by DPPH radical scavenging assay and H2DCFDA test in LPS-stimulated RAW264.7 macrophages and N9 microglial cells. MTS experiments and antiinflammatory properties verified cellular toxicity through NO assay. Interaction with A2A adenosine receptors was evaluated through binding assays using [3H]ZM241385 radioligand. Results: Polyphenols were present in 40% ethanol plant extract, which at 0.1–10 µg/µL achieved good antioxidant effects, with a DPPH radical scavenging rate of about 90%. In LPS-stimulated cells, these plant extracts, at 1μg/μL, did not affect cell vitality, displayed significant inhibition of H2DCFDA and NO production, and inhibited ZM 241385 binding in CHO cells transfected with A2A receptors. RAW 264.7 and N9 cells presented a density of them quantified in 60 ± 9 and 45 ± 5 fmol/mg of protein, respectively. Conclusion: Epilobium parviflorum, Cardiospermum halicacabum, and Melilotus officinalis extracts may be considered a source of agents for treating disorders related to oxidative stress and inflammation.

Ensiling Reduces Seed Viability: Implications for Weed Management

Ensiling, a lactic acid fermentation process, is mainly used to preserve biomass. In addition, it has been shown to affect seed viability of some plant species. The extent to which this makes ensiling suitable as a weed control measure, however, has not yet been determined. Both the range of controllable species and the parameters of an ensiling process that safely kills seeds are still undefined. We aimed to determine the effect of varying substrate and ensiling conditions on the seed viability of 10 species selected to represent a wide range of different seed traits. Five different types of silages were made from maize or mixtures of wildflower and maize biomass and ensiled in lab-scale silos for 8 months. The pure maize silages were prepared under conditions either ideal or suboptimal for ensiling forage. Seeds of important weeds (Chenopodium album, Abutilon theophrasti) and of species from a wildflower mixture suitable for ensiling and biogas production (Cichorium intybus, Daucus carota, Echium vulgare, Malva alcea, Malva sylvestris, Melilotus albus, Melilotus officinalis) were tested. Seed viability was determined using a combination of tetrazolium and germination tests. Ensiling reduced seed viability across all 10 species significantly. Seed-killing efficacies of ensiling, however, differed widely among the species studied, largely related to whether the species could produce hard (physically dormant) seeds. Seeds from species without hardseededness were completely inactivated by ensiling, while the seed-killing efficacies for hardseeded species ranged from 5 to 60%. Variation in ensiled substrate and ensiling conditions had no consistent effect on seed survival. We concluded that ensiling has the potential to sustainably reduce seed viability of a wide range of species and therefore should be adopted as a component of integrated weed management in organic agriculture.

Bio-inspired silver selenide nano-chalcogens using aqueous extract of Melilotus officinalis with biological activities

For the first time, an aqueous extract of Melilotus officinalis was used to synthesize bimetallic silver selenide chalcogenide nanostructures (Ag2Se-NCs). The formation of NCs was confirmed and characterized by UV–visible and FTIR spectroscopy, SEM and TEM imaging, XRD and EDX crystallography, zeta potential (ZP) and size distribution (DLS). The bioactivities of biosynthesized Ag2Se-NCs, such as antibacterial, antibiofilm, antioxidant and cytotoxicity potentials, were then examined. Bio-based Ag2Se-NCs were successfully synthesized with mostly spherical shape in the size range of 20–40 nm. Additionally, the MIC and MBC values of Ag2Se-NCs against β-lactam-resistant Pseudomonas aeruginosa (ATCC 27853) were 3.12 and 50 µg/ml, respectively. The DPPH scavenging potential of Ag2Se-NCs in terms of IC50 was estimated to be 58.52. Green-synthesized Ag2Se-NCs have been shown to have promising benefits and could be used for biomedical applications. Although the findings indicate promising bioactivity of Ag2Se-NCs synthesized by M. officinalis extract (MO), more studies are required to clarify the comprehensive mechanistic biological activities.

Comparative assessment of the amino acids content of some legumes species in Southern Ukraine

The data on the composition and amount of amino acids have been analyzed in the raw materials of five legume species. All of them grow in Southern Ukrainian flora (Securigera varia (L.) Lassen, Vicia cracca L., Lupinus luteus L., Melilotus officinalis (L.) Pall., Melilotus albus Medic.) and may be used as a source of amino acids, especially essential, whose resource has to be replenished from the outside. Aim. We have studied and compared the amino acid profile of some species of the Ukrainian South Legumes, and used the multidimensional statistical cluster analysis to construction of histograms based on the amino acids content and composition of such plants as: Securigera varia (L.) Lassen, Vicia cracca L., Lupinus luteus L., Melilotus officinalis (L.) Pall., Melilotus albus Medic. Materials and methods. The raw materials were harvested at the South of Ukraine and were investigated by gas-liquid chromatography. The amino acid analyzer has been used after hydrochloric acid hydrolysis at elevated temperature. Results. 19 amino acids have been identified, of which nine are essential or partially interchangeable. The non-polar amino acid proline is in the lead in terms of quantity among the essential amino acids. Its amount was 6932 mg/100 g and the ability to accumulate it was noted in Melilotus officinalis (2276 mg/100 g). The smallest proline amount was found in Lupinus luteus (388 mg/100 g). The sulfur-containing non-polar amino acid methionine is in the smallest amount in the selected plants (506 mg/100 g). Our attention was drawn to the absence of the polar amino acid glutamine among the non-essential amino acids in some plants. Securigera varia, Vicia cracca, and Melilotus officinalis did not contain glutamine. In this subgroup, the polar aspartic acid was found in the highest amount (6824 mg/100 g) with the highest content in Vicia cracca and the lowest – in Melilotus albus (2660 mg/100 g and 385 mg/100 g, respectively). Conclusions. The analysis of the presence and number of amino acids was the basis for our multidimensional statistical cluster analysis and histograms of the presentation of the amino acid profile of the studied plant members of the family Fabaceae L. In constructing the dendrogram, three clusters were identified, and representatives of one genus (Melilotus L.) were attributed to different clusters which is significant for further chemosystematic studies.

Assessment of the Suitability of Melilotus officinalis for Phytoremediation of Soil Contaminated with Petroleum Hydrocarbons (TPH and PAH), Zn, Pb and Cd Based on Toxicological Tests

The article presents issues related to the possibility of using toxicological tests as a tool to monitor the progress of soil treatment contaminated with petroleum substances (TPH, PAH), Zn, Pb and Cd in bio-phytoremediation processes. In order to reduce the high content of petroleum pollutants (TPH = 56,371 mg kg−1 dry mass, PAH = 139.3 mg kg−1 dry mass), the technology of stepwise soil treatment was applied, including basic bioremediation and inoculation with biopreparations based of indigenous non-pathogenic species of bacteria, fungi and yeasts. As a result of basic bioremediation in laboratory conditions (ex-situ method), the reduction of petroleum pollutants TPH by 33.9% and PAH by 9.5% was achieved. The introduction of inoculation with biopraparation-1 prepared on the basis of non-pathogenic species of indigenous bacteria made it possible to reduce the TPH content by 86.3%, PAH by 40.3%. The use of a biopreparation-1 enriched with indigenous non-pathogenic species of fungi and yeasts in the third series of inoculation increased to an increase in the degree of biodegradation of aliphatic hydrocarbons with long carbon chains and PAH by a further 28.9%. In the next stage of soil treatment after biodegradation processes, which was characterized by an increased content of heavy metals (Zn, Pb, Cd) and naphthalene, chrysene, benzo(a)anthracene and benzo(ghi)perylene belonging to polycyclic aromatic hydrocarbons, phytoremediation with the use of Melilotus officinalis was applied. After the six-month phytoremediation process, the following was achieved: Zn content by 25.1%, Pb by 27.9%, Cd by 23.2% and TPH by 42.2% and PAH by 49.9%. The rate of removal of individual groups of hydrocarbons was in the decreasing order: C12–C18 > C6–C12 > C18–C25 > C25–C36. PAHs tended to be removed in the following order: chrysene > naphthalene > benzo(a)anthracene > benzo(ghi)perylene. The TF and BCF coefficients were calculated to assess the capacity of M. officinalis to accumulate metal in tissues, uptake from soil and transfer from roots to shoots. The values of TF translocation coefficients were, respectively, for Zn (0.44), Pb (0.12), Cd (0.40). The calculated BCF concentration factors (BCFroots > BCFshoots) show that heavy metals taken up by M. officinalis are mainly accumulated in the root tissues in the following order Zn > Pb > Cd, revealing a poor metal translocation from the root to the shoots. This process was carried out in laboratory conditions for a period of 6 months. The process of phytoremediation of contaminated soil using M. officinalis assisted with fertilization was monitored by means of toxicological tests: Microtox, Ostracodtoxkit FTM, MARA and PhytotoxkitTM. The performed phytotoxicity tests have indicated variable sensitivity of the tested plants on contaminants occurring in the studied soils, following the sequence: Lepidium sativum < Sorghum saccharatum < Sinapis alba. The sensitivity of toxicological tests was comparable and increased in the order: MARA < Ostracodtoxkit FTM < Microtox. The results of the toxicological monitoring as a function of the time of soil treatment, together with chemical analyses determining the content of toxicants in soil and biomass M. officinalis, clearly confirmed the effectiveness of the applied concept of bioremediation of soils contaminated with zinc, lead and cadmium in the presence of petroleum hydrocarbons.