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Vol 25, No 1 (2022)
- Year: 2022
- Articles: 8
- URL: https://journals.eco-vector.com/1560-9596/issue/view/5737
Articles
Comparative study of the essential oil composition of pine buds and microstrobils (Pinus sylvestris L.)
Abstract
Aim of the research. Comparative study of the essential oil composition obtained from pine buds and microstrobiluses (Pinus sylvestris L.), growing in Buryatia. Material and methods. The pine buds and microstrobils samples was collected in during May-June 2019-2020 in background areas, and nearby Ulan-Ude. Essential oil was obtained by pharmacopoeial method 2 - hydrodistillation. The content of essential oil was determined using volumetric method. The qualitative composition and relative quantitative content of the components of essential oils was identified by gas-chromato-mass spectrometric method. Results. The yield of essential oil from pine buds was 0.67-0.75%, from microstrobils - 0.39-0.53% calculated to absolutely dry raw material. The main components of buds and microstrobils essential oil are: α-pinene (8.0-20.4%), β-pinene (8.1-12.5%), β-myrcene (5.5-12.4 %), 3-carene (6.5-16.8%), limonene + β-pellandrene (18.3-20.4%), terpinolene (1.0-2.1%); caryophyllene (0.8-2.0%), germacrene D (0.3-1.9%), α-murolen (0.5-1.9%), γ-cadinene (0.5-2.1 %), δ-cadinene (0.7-5.6%), T-murolol (0.1-3.4%), а-cadinol (0.1-4.1%); dehydroabietane (0.2-1.0%). It can be noted that the content of α-pinene in microstrobils is higher (2 times) than in the buds. The content of sesquiterpene and diterpene compounds such as germacrene D, cadinens, T-murolol, α-cadinol, dehydroabietane in buds essential oils of the is 1.5-2 times higher than in microstrobils, which may be due to the presence of resinous substances in the buds. Samples of essential oils from pine trees growing nearby Ulan-Ude are characterized by an increased content of oxygen-containing terpenoids due to the intensification of oxidation processes catalyzed by pollutants. Essential oils of the buds and microstrobils of Scots pine from Buryatia have a higher level of α-pinene (8.0-20.4%) compared to the populations of the Tomsk region (6.9-11.3%), and less 3-carene - 6, 5-16.8% in Buryat samples, 21.9-27.2% - in Tomsk, which may be due to climatic conditions of growth. It should be noted that 3-carene has a pronounced allergenic effect, and therefore, such an essential oil with a high component content should have limited use. Conclusion. The chemical composition of essential oils of pine microstrobils has been studied for the first time. A comparative analysis with essential oil of pine buds, which is a pharmacopoeial raw material with proven efficacy, has been carried out, its sufficient content and similarity of the component composition have been shown. The essential oil of microstrobils with a high content of α-pinene is of practical interest for use in therapeutic and prophylactic purposes. The use of microstrobils as a source of pollen, and as an independent raw material after its separation, solves the problem of rational use of plants, due to the production of not only pollen, but also other products containing biologically active substances.
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Identification of cannabimimetics MDMB(N)-022 in urine using SPE AND GG-MS
Abstract
The relevance of research. In recent years cannabimimetic MDMB(N)-022 spreads throughout the world. Appearance of new potentially dangerous psychoactive substances makes it necessary to develop methods of their determination in biological fluids in toxicological and forensic chemistry research. Purpose of this work is to examine possibility of isolation of MDMB(N)-022 metabolites from urine, using cartridges with a mixed sorbent C8/SAX and identification of received biotransformation products by gas chromatography-mass spectrometry method. Materials and methods. During sample preparation we used enzymatic hydrolysis with p-glucuronidase (Type HP-2, From Helix Pomatia, 100000 U/ml), solid-phase extraction with a mixed sorbent C8/SAX cartridges to isolate first phase metabolites in 11 urine samples and subsequent procedure of derivatization. Recieved derivatives were examined in gas chromatograph Agilent 7820 equipped with mass-spectrometric detector Agilent 5975 (Agilent, USA).). Results. The possibility to use cartridges for SPE with a mixed sorbent C8/SAX to screening tests of urine for the presence of metabolites of cannabimimetics MDMB(N)-022 was shown. The main biotransformation directions of MDMB(N)-022 where defined. They include hydrolysis of ester bonds with following conjugation with glucuronic acid, hydrolysis of amide linkage, N-dealkylation of alkyl radical at nitrogen atom of indazole cycle, oxidative carboxylation of alkyl chain, hydroxylation and dihydrodiol metabolite formation. An analytic characteristics of main MDMB(N)-022 metabolites and their derivatives received by gas chromatography-mass spectrometry were given.
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Study of the stability of 2,6-di-tert-butyl-4-methylhydroxybenzene in biological material
Abstract
Aim purpose. Study of stability of 2,6-di-ferf-butyl-4-methylhydroxybenzene in biological material. Methods. The studies were carried out by GC-MS, TLC and UV spectrophotometry.2,6-di-ferf-butyl-4-methylhydroxybenzene was isolated from the biomatrix (liver tissue) by infusion with a mixture of ethyl acetate-acetone (7:3). The analyte was purified by combining extraction (water-ethyl acetate system) and semi-preparative column chromatography (sorbent - silica gel L 40/100 pm, eluent -hexane-acetone (9.5:0.5)), Results. It was found that at -22oC, 2oC, 10oC, 20oC and 30oC. 2,6-di-ferf-butyl-4-methylhydroxybenzene remains in the biomaterial for 469, 371, 315, 301 and 259 days, respectively. The possibility of mathematical description of the dynamics of analyte decomposition in biomaterial at the indicated temperatures using the hyperbole equation has been studied. Coefficients in the equation of hyperbola (km), Calculated from the results of the experiment, for temperatures of -22oC, 2oC, 10oC, 20oC and 30oC were respectively 11415, 7508, 4983, 3696 and 2156. A linear dependence of km was found. from the storage temperature tü, which is described by the equation: km = 182,24(50-f°) -1702,36. Conclusion. The dynamics of decomposition of 2,6-di-ferf-butyl-4-methylhydroxybenzene in the liver tissue at -22oC - + 30oC, which can be described by the hyperbole equation, is shown. The possibility of predicting the nature of analyte stability in biomaterial in the specified temperature range is shown.
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Characteristics of endophytic bacteria, isolated from the roots of Ipomoea pes-caprae (L.)
Abstract
Relevance. Endophytic bacteria live in the internal tissues of healthy plants and do not cause any harm to the host plant. They positively affect plants' growth and development and help them absorb nutrients from the environment. Endophytic bacteria protect plants from various phytopathogens due to the synthesis of phytohormones and contribute to converting insoluble mineral salts into a form accessible to plants. Studies on endophytic bacteria isolated from I.pes-caprae in Vietnam are not yet numerous. Purpose of the study. Isolate and characterize endophytic bacteria from the roots of Ipomoea pes-caprae. Identify promising strains. Material and methods. The objects of the study were I. pes-caprae roots isolated from plants collected in Thang Hai village, Tinh Hai commune, Tinh Gia district, Thanh Hoa province. Before isolating endophytic bacteria strains, root samples were thoroughly washed with distilled water and sterilized with mercuric chloride solution for 5 minutes. The homogenate was cultivated on Luria-Bertani nutrient medium. The IAA concentration from bacterial strains was determined by the method of Glickmann and Dessaux (1995). A quantitative determination of phosphate-dissolving activity was carried out by measuring the phosphate accumulation in the solution in which the strains under study were cultivated. Results. Fifteen strains of endophytic IAA-producing bacteria were isolated on LB nutrient medium supplemented with L-tryptophan. Three (TH10R, TH11T, and TH13T) strains with high IAA synthesizing ability (from 2.53 to 4.19 pg / ml) were selected. The phosphate-dissolving activity of these isolates ranged from 1.06 mg / L to 3.64 mg / L. The TH10R strain was superior to other strains in its ability to synthesize IAA and phosphate solubility and was identified as Bacilus mycoides and named Bacilus mycoides TH10R.
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Analysis of the size of protein molecules obtained from a legume plant material
Abstract
Obesity is a problem in the present-day world. Excess weight leads to the cardiovascular diseases, type 2 diabetes mellitus. As an alternative to a low glycemic index diet, there are foods that slow down the absorption of carbohydrates by inhibiting the enzymes responsible for their fixation. These products include α-amylase and glucosidase inhibitors. Protein compounds with an inhibitory activity are contained in large quantities in assorted of beans. Depending on the method of extraction, concentration, isolation, many inhibitor preparations can lose its activity during the use. Therefore, production of purified inhibitors with high biological activity is an urgent task. The modern methods for extracting substances from plant materials, for example, fractionation, supercritical extraction, lead to an increase in the activity of bioactive substances. In this regard, the purpose of the work is to study the size of the particles of protein substances extracted from the seeds of red beans using the light-scattering method and to determine their biological activity. Bovine serum albumin was used as a model object. When studying the particle size, it was shown that detection at 15° reveals only large particles, and an angle of 90° allows simultaneously small (6-20 nm) and large (up to 6000 nm) protein fractions to be fixed. In phosphate buffer, serum albumin particles form agglomerates (~ 6000 nm), and in water, particles can be present in monomeric and dimeric forms (~ 1000 nm). Protein substances obtained by extraction from red beans into a buffer solution of pH 8 are enlarged aggregates. The size of protein particles decreases from 620 nm to 380 nm when the ultrasonic irradiation was used. Studies of the biological activity of protein substances in relation to pancreatic amylase have shown that the activity of the enzyme decreases from 28 U/g to 15 U/g and to 5 U/g when using protein substances obtained by maceration and in a cavitation field, respectively.
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Study of the involvement of TrkB receptors in the antihypoxic effect of the neuropeptide cyclo-prolyl-glycine
Abstract
Relevance. Cycloprolylglycine (CPG) was identified as a neuropeptide in 1996. It was found to have nootropic, antihypoxic, anxiolytic, neuroprotective and analgesic activity. This complex of effects is beneficial for the development of CPG-based drugs. In this regard, the study of the mechanism of action of CPG is relevant. We have previously revealed that AMPA and TrkB receptors are involved in the anxiolytic action of CPG. Objective. Purpose of the study is to reveal the involvement of TrkB receptors in the antihypoxic effect of the neuropeptide CPG. Material and methods. The method of inhibitory pharmacological analysis using the Trk receptor blocker K252A was used. Antihypoxic activity was determined in the test of "canned" hypoxia in mice. Results. The antihypoxic effect of CPG was completely removed by a specific blocker of Trk receptors K252A. Conclusions. For the first time it was shown that TrkB receptors are involved in the antihypoxic effect of the neuropeptide cyclo-prolyl-glycine.
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Effect of the dry Rhaponticum uniflorum (L.) DC extract on the state of the immune and antioxidant systems of the body in experimental immunodeficiency
Abstract
The purpose of the study was to evaluate the immunomodulatory effect and antioxidant activity of the dry extract from the herb of Rhaponticum uniflorum (L.) DC. Materials and methods. Experiments were carried out on F1 (СВАхС57В1/6) mice and Wistar rats. Immune deficiency was simulated by a sinqle intraperitoneal introduction of cyclophosphamide in the dose 250 mq/kq. The first and second experimental qroups administered the dry extract from R. uniflorum (100 mq/kq) and a comparative druq «Echinacea P» (200 mq/kq) respectively intraqastrically once a day for 14 days aqainst the backqround of cyclophosphamide immune suppression. The third and fourth intact qroups received the R. uniflorum extract and «Echinacea P» respectively in the same doses for 14 days. The intact qroup administered the purified water accordinq to the analoqous scheme. The effect of the tested remedy on the cellular immunity was evaluated in the reaction of the delayed-type hypersensitivity (DTH), the humoral immunity was estimated by the number of antibody-forminq cells (AFC). The antioxidant status of the body was assessed by the content of malondialdehyde, the activity of SOD, catalase, and GPx in the spleen homoq-enate. The antiradical activity of R. uniflorum extract was studied in vitro by its ability to reduce 2,2'-diphenyl-1-picrylhydrazyl (DPPH') and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonate (ABTS•+), neutralize O2- and Fe2+. Results. The extract increased the DTH-index by 1.5 times and the value of absolute and relative numbers of AFC by 1.5 and 1.4 times as compared with the control qroup; it was comparable to the action of «Echinacea P» and did not chanqe the immune indicators in intact mice. The extract reduced the concentration of malondialdehyde by 1.8 times, increased SOD and catalase activity by 1.6 times; GPx - 1.4 times; in vitro the extract exhibited antiradical effect. Conclusion. The R. uniflorum extract has an immunomodulatory effect on cellular and humoral immunity in cyclophosphamide immunosuppression. Its action is based on membrane-stabilizinq and antioxidant activity.
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Determination of the free radical reactions in whole blood of Wistar rats under stress-induced physical exercise
Abstract
Purpose. Determination of the free radical reactions in whole blood of Wistar rats under stress-induced physical exercise. Materials and methods. The object of the research is whole blood of male Wistar rats. Rats in the experimental group were forced to swim in an aquarium (swimming time 40 minutes at the water temperature of 25 °C with using of Porsolt test). The swimming time was determined experimentally after the stress modeling. The values of maximum luminescence intensity (Imax) and its growth rate (L tg) were determined with spontaneous luminol-dependent chemiluminescence on BCLM 3606M biochemiluminometer. The rate of the free radical reaction was calculated by using the formula described by Yu.A. Vladimirov et al. (2011). Results. During stress-induced physical exercise Imax value significantly increases by 1.09 times (p < 0.05) and L tg value significantly decreases by 4.65 times (p < 0.05). Between the above-noted parameters the linear dependence described with the equation Imax = 128.411L tg where 128.411 is the coefficient characterizing BCLM 3606M analyzer's sensitivity to radiation of a photomultiplier tube is observed. The rate constants of the studied free radical reactions were established with this equation: the constant of the first reaction k1 = 11010 (mol/dm3)-1·s-1 is the constant of peroxynitrite formation rate, the constant of the second reaction k2 = 3.2107 (mol/dm3)-1·s-1 is the constant of the rate of reduction of glutathione-thiyl radical to reduced form of glutathione. Conclusions. As a result of our study, it was found that there is the reaction of formation of peroxynitrite from radical of nitrogen monoxide and the reaction of reduction of glutathione-thiyl radical to reduced form of glutathione during stress-induced physical exercise. For the first time free radical reactions during stress-induced physical exercise were determined with the dependence of luminescence rate on ratio of maximum chemiluminescence intensity to the coefficient characterizing BCLM 3606M analyzer sensitivity to radiation of a photomultiplier tube. For the first time the equation described by Yu.A. Vladimirov et al. (2011) used in study of free radical reactions using BCLM 360* analyzers. In the process of chemiluminescence substances are not destroyed, therefore a reaction path doesn't change. In addition, chemiluminescence is highly sensitive, which is very important when registering highly reactive radicals.
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