Using an experimentally selected extractant for balanced extraction of flavonoids, polysaccharides and polyphenols from bur-marigold herb, calendula flowers and hawthorn leaves and flowers, 3 suitable dry extracts and 1 combined extract in the selected ratio of raw materials were obtained. It was concluded that it was reasonable to standardize the combined herbal extract on the content of flavonoids in terms of luteolin-7-glucoside and content of polyphenols in terms of pyrogallol by UV method and the content of terpenoid compounds, in terms of oleanolic acid by densitometric method. The combined extract at a dose of 30 mg/kg in the model of anaphylactic shock has showed antiallergic activity at the level of tesalin and exceeds diazoline; the ability of the combined extract to reduce the permeability of skin capillaries at the level of the tesalin has established on the model of active cutaneous anaphylaxis and it significantly exceeded diazoline; the combined extract did not differ in membrane-stabilizing properties from the effect of desloratadine and was significantly superior to the reference drugs diazolin and tesalin in the test of indirect degranulation of mast cells. As a result, it was concluded that the mechanism of anti-allergic action of the combined extract is probably associated not only with antihistamine properties but also with the ability to stabilize cell membranes.
The interest to study the genus Bidens and in particular Bidens tripartita L. (bur-marigold herb) is unmitigated due to its antioxidant, anti-allergic, antimicrobial, antifungal, hepatoprotective, immunostimulating and hypotensive activity. This pharmacological activities are determined by the presence of flavonoids, polyphenolic compounds, polysaccharides, components of essential oils, polyacetylenes, etc. The aim of the research was to study the process of extracting various biologically active substances (polysaccharides, flavonoids, polyphenols) from bur-marigold herb with water-alcohol mixtures of various concentrations, to study their component composition, as well as to evaluate their effect on the oxidative effect of free radicals. Materials and methods. The object of the research was the herb of Bidens tripartita L. Raw materials were collected and procured on the territory of Kharkiv, Zhytomyr and Poltava regions of Ukraine. All used methods for the quantitative determination of biologically active substances were pharmacopeial and described in various monographs in the European Pharmacopoeia and State Pharmacopoeia of Ukraine. Results. The extracts of the bur-marigold herb were analyzed for the content of the extractable matter, flavonoids, polysaccharides and it was found that they are maximally extracted (about 80 %) in the 1st percolate with a drug/solvent ratio of 1:10 for all the extractants. It was found that with an increase of ethanol concentration in the extraction mixture, the content of polyphenols and flavonoids, is expectedly increased, as well as the value of the antioxidant activity of the corresponding extracts, but the amount of extracted polysaccharides is significantly reduced. Conclusions. With the selected evaluation criteria, it was found that 40 % ethanol is optimal from the point of view of balanced extraction of flavonoids, polysaccharides and polyphenols. A strong positive and statistically significant correlation was found between the content of polyphenols in the obtained extracts and their antioxidant activity (Pearson correlation coefficient, r=0.9998), slightly weaker for flavonoids (r=0.9886), and an inverse correlation between polysaccharides content and TEAC- value.
Aim: Literary sources on qualitative composition, quantitative content of biologically active substances, and standardization methods of pumpkin seeds have been studied. It has been established that, along with Δ7-sterols, whose action is associated with treatment of the prostate gland, pumpkin seeds contain amino acids, cucurbitin in particular, which are responsible for their antihelminthic effect. The aim is to study the qualitative and quantitative content of amino acids in domestic samples of pumpkin seeds in order to clarify the possibility of pumpkin seed standardization for this class of biologically active substances.Methods: The investigation of the possibility of amino acids’ identification in the herbal drug by the thin-layer chromatography (TLC) method was carried out on silica gel 60 F254 plates, Merck and Silica gel 60, Merck, on aluminum and glass supports in a mixture of the following solvents: butanol–acetone–acetic acid–water (35:35:10:20). The conclusions have been made based on the presence of characteristic zones of amino acids in the chromatograms after the treatment with ninhydrin solution. Quantitative determination of amino acids in the herbal drug was carried out by absorption spectrometry using HP spectrophotometer HP-8453 UV-VIS, Hewlett Packard (USA).Results: The chromatographic profile of the amino acid fraction of domestic samples of pumpkin seeds was studied using a TLC technique. In the chromatograms of test solutions from all samples, the zones were detected at the level of histidine, aspartic acid, glutamic acid, glycine, and leucine zones, as well as a zone with color (olive) that differs from the other zones of amino acids and which is possibly a zone of cucurbitin.On the basis of the results of qualitative research, the quantitative content of the sum of amino acids according to a UV method developed has been determined, which including purification from the fatty oil, further extraction with an aqueous alcohol solution, selection of aliquots for the reaction with ninhydrin solution, and subsequent determination of the absorbance of the test solution and the solution of glutamic acid at a wavelength of 400 nm.It has been established that the content of the sum of amino acids expressed as glutamic acid in domestic samples is about 2%.Conclusions: The research of the possibility of pumpkin seed standardization by the content of amino acids was carried out. The chromatographic profile of the amino acid fraction of domestic samples of pumpkin seeds was studied using a TLC method developed. The quantitative content of the amino acids sum by the absorption spectrophotometry method was investigated. The techniques developed will be recommended for inclusion in the draft of national monograph “Pumpkin seeds.”
Combined herbal medicines for the treatment and prevention of allergic diseases are perspective substitutes for the «classic» drugs of H-1 receptor blockers of the 1st and 2nd generation. Literary-logical methods are quite effectively used for scientifically justified design of these drugs, namely phytopharmacological drug design (PPD) and phytochemical drug design (PCD). The aim of the work – the search for ways of the scientifically justified design of a combined multifunctional medicine based on the bur-marigold, which normalizes the functioning of the body's defence systems, increases the body's nonspecific resistance, normalizes the immune status using literary-logical and statistical methods of analysis. The design of the preparation was carried out using literature data for PPD and experimental and literature data for PCD. Statistical processing method – Spearman's rank correlation coefficient was used. Design aspects of the combined herbal medicine, which includes a bur-marigold herb, calendula flowers, hawthorn leaves and flowers were considered using of a system-comparative analysis of information and its processing by the method of the Spearman's rank correlation coefficient determination. When PCD development of various combinations of herbal drugs, information about the content of such biologically active substances (BAS) as flavonoids, oxycinnamic acids, polyphenols, triterpenes, proteins/amino acids, polysaccharides was used. When PPD development the spectra of pharmacological activity were determined and a quantitative assessment to them in the system of marks for each of the herbal drug of the combined medicine was given. The PCD and PPD data into the matrix for calculating the Spearman rank correlation coefficient of the «action–sum of BAS» was entered. As a result, a strong, direct and statistically significant correlation relation (p from 0.924 to 0.997) between the content of oxycinnamic acids, polyphenols, triterpenes, proteins/amino acids, polysaccharides and such pharmacological actions as capillary-strengthening, anti-inflammatory, immunostimulating; choleretic, hepatoprotective, improving metabolism; diuretic, hypotensive, sedative, tonic, desensitizing, antipruritic, reparative for the studied herbal combinations was found. The obtained results of the PCD and PPD development as well as the study of the correlation «action–sum of BAS» by calculating the Spearman's rank correlation coefficient, this suggests the developed combined herbal medicinal product with multifunctional action is perspective in the complex therapy of allergic diseases accompanied by a decrease in the body's resistance and immunostatus.
Combinations of medicinal plants ensure the collective work of several active components, which can lead to an additive or synergistic effect. We have chosen the bur-marigold-calendula-hawthorn combination to obtain an antiallergic herbal medicine, in which the bur-marigold is the main plant, calendula is a helper plant that helps in the fight against accompanying symptoms, and hawthorn harmonizes the action of the first 2 plants. The aim of the work – the study of possible synergistic interactions in a multicomponent herbal antiallergic preparation by calculating the correlation between the content of polyphenols in various combinations of plant extracts and their antioxidant activity. Both individual extracts of a bur-marigold herb, calendula flowers and hawthorn leaves and flowers and a combined target extract with the ratio of herbal drugs (6:3:1) were obtained. Two and three component ompositions were obtained by mixing extracts in appropriate ratios. The content of polyphenols was determined by SPhU 2.0, 2.8.14. The antioxidant activity was determined by the neocuproin method. The Pearson Correlation was used to calculate the relationship between the content of polyphenols in extracts and their antioxidant activity. The maximum amount of polyphenols was found in the hawthorn extract (7.2%), in the bur-marigolg extract – 5.5%, and in the calendula extract – 3.4%. When adding calendula extract to the bur-marigold extract, the antioxidant activity was expected to decrease, thus an additive effect is observed; for the combination of bur-marigold /hawthorn 1:0.5, an increase in antioxidant activity (110.6%) was observed compared to the expected value, which may be synergism; for the combination of calendula/hawthorn, an increase in activity was observed. For 3-component combinations, it was found that only the combination of bur-marigold/calendula/hawthorn 1:0.5:0.5 showed greater activity (112%) than the target extract, all others combinations showed activity within the limits of possible variations (± 10%) of the components. When calculating the correlation between the content of polyphenols in the combinations and their activity using the Pearson coefficient, a strong positive correlation was found. The correlation between the content of polyphenols in various combinations of plant extracts and their antioxidant activity of a multicomponent herbal antiallergic preparation was studied. In 2-component combinations an additive effect was found when determining antioxidant activity, and an assumption was made about the ability of a synergism only for a combination of bur-marigold/hawthorn 1:0.5. All analyzed 3-component compositions showed antioxidant activity within the limits of possible variations (± 10%) of the components. A strong positive correlation was found between the content of polyphenols in the combinations and their activity.
Aim. To improve the national monograph of burr marigold (Bidens tripartita L.) herb on macro- and microscopic characteristics of the raw material. To achieve this goal, a comparative macroscopic analysis of various species of burr marigold and an extended microscopic analysis of all parts of the plant that are part of the raw material were carried out. Materials and methods. The study objects were 7 samples of the medicinal plant raw material of Bidens tripartita L. identified and registered at the premises of the State Enterprise “Pharmacopoeia Center”. They were provided by various domestic manufacturers; a sample of the whole raw material of B. frondosa L. collected in the Akhtyrsky district ofthe Sumy region; a sample of the whole raw material of B. cernua L. provided by a domestic pharmaceutical enterprise. The raw material was harvested in the period of budding and the beginning of flowering. The dry raw material wassubject to the macroscopic analysis. The microscopic analysis was performed on the powder of the raw material sifted through a sieve with a hole diameter of 355 since according to the requirements of the European Pharmacopoeia and the State Pharmacopoeia of Ukraine this was a standardized procedure for determining the diagnostic signs of the medicinal plant raw material by microscopy. Results and discussion. As a result of the macroscopic analysis, it has been found that the raw material of Bidens tripartita L. is a mixture of the whole or fragmented stems with leaves, leaves, anthodia, and achenes. Stems are green or greenish-purple, round-oval, longitudinally grooved. Leaves are green or brownish-green, with a dentateserrated edge, opposite, with a 3-5-parted lamina with lanceolate lobes narrowed into short, slightly winged petioles. Inflorescences are erect anthodia, their width is almost equal to the height. Fruits are flattened achenes, at the apex with 2 or 3 awns, twice as short as the achene. The awns and lateral ribs of the achenes are covered with bristles, which tips are directed downwards. As a result of the microscopic analysis of Bidens tripartita L. powder, diagnosticsigns of all parts of the raw material have been found, namely: fragments of the lamina with veins, along which secretory channels with reddish-brown contents pass; fragments of the lamina from the upper or lower epidermis with the basic cells with sinuous walls, stomata anomocytic type, covering caterpillar form trichomes of 9–18 thin-walled cells, rarely thick-walled covering trichomes of 2-13 cells; fragments of the stem epidermis of rectangular elongated cells with thickened walls; fragments of the epidermis of the inner leaves of the involucres of elongated rectangular thin-walled cells and articulated branched secretory channels; fragments of epidermis of tubular flowers from polygonal cells with rounded papillary outgrowths; rounded pollen grains with spinous exine; fragments of the testa with pigmented darkbrown cells of the hypodermis and the adjacent lower layer of elongated yellowish sclereids. Conclusions. The sections “Identification A and B” (macro- and microscopic signs of the raw material) of the national monograph “Burr marigold herbN” of the SPhU 2.2 have been improved, and it allows to more accurately identify all parts of the medicinal plant raw material. The additions developed are included to the SPhU 2.5 in the text of the monograph “Burr marigold herbN”.
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