Research of the chemical composition of Artemisia tschernieviana Bess. by gas chromatography method with mass detection

The aim of the work was to study chromato-mass spectroscopic of Artemisia tschernieviana Bess. grass and to establish possible prospects for the use of this plant in medical practice. Materials and methods. Artemisia tschernieviana Bess. grass was chosen as the object of research. Raw materials were collected in July 2021 in the flowering phase (Zaporizhzhia). The tincture was obtained by maceration and the raw material was extracted with methyl alcohol at room temperature for 10 days according to the method of making tinctures. The study of the chemical composition of Artemisia tschernieviana Bess. Grass was carried out using gas chromatograph Agilent 7890B GC System (Agilent, Santa Clara, CA, USA) with mass spectrometric detector Agilent 5977 BGC/MSD (Agilent, Santa Clara, CA, USA) and chromatographic column DB-5ms (30 m × 250 μm × 0.25 μm). Results. During the study it was found that the composition of Artemisia tschernieviana Bess. grass includes 21 compounds (2 in the isomeric state). β-myrcene (17.22 %), sabinene (14.54 %), 2-(4a,8-dimethyl-7-oxo-1,2,3,4,4a,7-hexahydronaphthalen-2-yl) propionic acid (11.57 %), 2-dodecen-1-yl-succinic anhydride (10.84 %) and chamazulene (9.02 %) accounted for the largest percentage. According to the literature review, myrcene has antibacterial, anesthetic and analgesic effects. Sabinene is characterized by antibacterial action. 2-(4a,8-dimethyl-7-oxo-1,2,3,4,4a,7-hexahydronaphthalen-2-yl)-propionic acid is a fatty acid with possible antibacterial and anti-inflammatory activity. 2-dodecen-1-yl-succinic anhydride is used to produce anhydride-modified form of insulin that can be used in the treatment of diabetes. Hamazulen has anti-inflammatory activity. Conclusions. In view of all the above, we can conclude that Artemisia tschernieviana Bess. grass can be recommended for further research as a source of potential anti-inflammatory and antibacterial agents.

Comparative chromato-mass spectrometric study of tinctures based on Echinacea purpurea (L.) Moench

An urgent problem today is the spread of secondary immunodeficiencies associated with increased stress, urbanization, and negative changes in the environment that lead to impaired immune function. Herbal remedies are milder, less likely to become addictive and have side effects, and are long-lasting. A promising plant with immunomodulatory action is Echinacea purpurea (L.) Moench, a member of the family Asteraceae. The main active ingredients of echinacea are phenolic compounds, carbohydrates, and alkylamides. The chemical composition of echinacea is also represented by flavonoids, tannins, saponins, higher fatty acids, amino acids, betaine, essential oil, macro- and microelements (Se, Co, Ag, Mo, Zn, Mg, K, Na, Fe, etc.). Drugs have immunostimulatory, antioxidant, membrane-stabilizing effect, promote healing of wounds, burns, ulcers, are used in infectious and viral diseases, especially HF. They are traditionally used for furunculosis, septicemia (infection of the blood), pyorrhea, tonsillitis, especially for the treatment of boils, carbuncles, and abscesses. It is also important that in modern pharmaceutical science the leading place is occupied by research related to the introduction into medical practice of herbal medicines, the study of their chemical composition, standardization, development of optimal technologies for the manufacture of phytopreparations. That is why the objects of our research were tinctures based on Echinacea purpurea. The aim of the work is to study and compare the component composition of tinctures based on Echinacea purpurea (L.) Moench using gas chromatography (GC). Materials and methods. Tinctures of rhizomes with roots of Echinacea purpurea were used for the study: experimental – made according to industrial technological recipe (1:5, extractant – ethanol 70 %) from raw materials harvested in July 2019 at the research site of Zaporizhzhia State Medical University and control – finished products LLC “Zhytomyr Pharmaceutical Factory”, series 20319. Tinctures were investigated on a gas chromatograph Agilent 7890B with mass spectrometric detector 5977B. The NIST14 mass spectrum library was used to identify the components. Results. 31 (experimental) and 23 (control) components were identified by GC in rhizomes with rhizomes of Echinacea purpurea. The results of the study indicate that the qualitative and quantitative chemical composition of rhizomes with roots of Echinacea purpurea was characterized by complexity and variability. This fact complicates the process of standardization of phytopreparations and can affect their effectiveness and safety. Therefore, members of the genus Echinacea should be the subject of further in-depth chemical study. Conclusions. A comparative analysis of the component composition of tinctures based on Echinacea purpurea (L.) Moench was studied and performed with the help of GC. The GC method is suitable for the determination of natural BAS in the composition of phytopreparations from echinacea and can be used in the development of methods for their standardization. The results of the study were of value for further development and improvement of analytical regulations for raw materials and phytopreparations from Echinacea purpurea. They can also be the basis for the creation of new substances with immunomodulatory activity based on the underground organs of echinacea.

Разработка методики определения флудиоксонила в растениводческой продукции методом газовой хроматографии

In the publication the methods of fludioxonyl determination in vegetable pea, potato and winter wheat by the method of gas chromatography with the mass-spectrometric detector is presented. The ex-traction method of soil preparation differs by the simplicity of execution, small use of reagents, expres-sion and provides with the purity of extracts necessary for getting the reproduced quantitative results.

A new functional food ingredient enriched by Phytoecdisteroids and Polyphenols from quinoa grains (Chenopodium quinoa Willd.)

Objective: Quinoa grains (Chenopodium quinoa Willd.) contain a large number of secondary metabolites. Phytoecdysteroids and polyphenols, secondary metabolites, are of great interest for research today. The aim of the study is a method development for the extraction of biologically active substances (BAS) from quinoa grains to obtain a functional food ingredient (FFI) enriched with phytoecdysteroids and polyphenols. Materials and methods: The content of phytoecdysteroids at each stage of quinoa extract production was determined by two methods: HPLC with a spectrophotometric detector and HPLC with a mass spectrometer. The content of total polyphenols was determined by the Folin-Ciocalteu method. The flavonoid profile was determined using a liquid chromatography system with a diode array spectrophotometric detector and a triple quadrupole mass spectrometric detector. Results: The chosen technological scheme for the production of FFI can concentrate more than 200 times the major phytoecdysteroid 20-hydroxyecdysone major in quinoa in the final product relative to the raw material. The maximum yield of flavonoids in the final product was observed in cases of 20 and 40% ethanol extraction. Conclusion: These results lead to the conclusion that highly enriched sources of BAS can be obtained using fairly simple technological approaches and commercially available reagents from quinoa grains.

The study of compounds distilled with water vapor of Populus laurifolia Ledeb. leaves

Poplar plants (Populus L.) are one of the promising species for inclusion in the State Pharmacopoeia of Ukraine. To date, extensive experience has been accumulated in the use of extracts from buds, leaves and bark of various types of poplars in folk and official medicine as effective anti-inflammatory, antipyretic, analgetic, antibacterial agents. According to literature data only buds are pharmacopoeial raw materials in some countries of the world. Poplar leaves are of interest as a new promising type of medicinal plant raw material, since they have a chemical composition close to the buds, moreover, they are much easier to harvest and the possible volume of raw materials procurement is much larger. The laurel-leaf poplar (Populus laurifolia Ledeb.) is assigned to the section of balsamic poplars (Tacamahaca). It is grown in forest culture, used for landscaping and creation of field-protective plantations in Ukraine. Continuing a comprehensive pharmacognostic study of raw materials of plants of the genus poplar, purpose of this work was to study composition and content of compounds distilled with water vapor of leaves of laurel-leaf poplar cultivated in Ukraine. Leaves for research were harvested in June 2019, from trees growing on the shores of the Murom reservoir in the village of Russkiye Tishki, Kharkiv region (50°09′30″N 36°29′10″E). The composition of compounds distilled in water vapor was investigated on Agilent Technologies 6890N (USA) chromatograph with a 5973N mass spectrometric detector. The total content of compounds distilled with water vapor was 2 126.1 mg/kg. 46 compounds were identified. Terpenoids were presented by 20 compounds with a total content 1 444 mg/kg, the remaining 26 compounds were aromatic substances (256.2 mg/kg) and compounds of various groups (216.8 mg/kg). The dominant compounds in content were β-eudesmol (776.4 mg/kg), γ-eudesmol (298 mg/kg), α-bisabolol (125.9 mg/kg), eugenol (70.9 mg/kg). The data obtained indicate the rich chemical composition of leaves of laurel-leaf poplar and are the basis for further in-depth study of this raw material for assessment the possibility of using it as a medicine.

Research of fatty and organic acids in Rhododendron luteum Sweet leaves

Rhododendron luteum Sweet of the Heath family Ericaceae is found on the territory of Ukraine both in the wild and is widely cultivated in botanical gardens, parks, squares. Leaves and inflorescences are used to treat heart disease, rheumatism, gout, and disorders of the nervous system. The literature data on the chemical composition relate to the presence of aminoacids, organic acids, essential oils, phenolic and triterpene compounds, andromedol derivatives. The aim of the work was to determine the composition and content of fatty and organic acids in the Rhododendron luteum Sweet leaves by gas chromatography-mass spectrometry. The object of the study was the leaves of Rhododendron luteum Sweet, collected on the territory of the botanical garden of H. S. Skovoroda Kharkiv National Pedagogical University in 2019. Gas chromatography - mass spectrometric study of the component composition and content of fatty and organic acids in the leaves of the studied plant on an Agilent Technologies 6890 chromatograph with a 5973 mass spectrometric detector was carried out. In Rhododendron luteum Sweet leaves 37 substances were found and identified. Fatty acids were represented by 17 compounds. Among saturated fatty acids, palmitic acid (3 276.2 mg/kg) was prevailed, among monounsaturated – oleic (736.78 mg/kg), among polyunsaturated – linolenic (1 617.65 mg/kg). The smallest amounts contained heneukocylic (46.33 mg/kg), lauric (58.17 mg/kg) and caproic (68.17 mg/kg) acids, which belong to saturated fatty acids. The presence of 20 organic acids in the raw material was also established: citric (8 680.30 mg/kg) and iso-citric (4 106.85 mg/kg) acids dominated in terms of quantitative content; oxalic (1 685.65 mg/kg) and malic (1 310.82 mg/kg) acids were contained in significant amounts. Among the derivatives of hydroxybenzoic acid, the greatest amount was presented by gentisic (187.76 mg/kg) acid, among the derivatives of hydroxycinnamic acids – p-coumaric (312.62 mg/kg). For the first time, the qualitative composition and quantitative content of fatty and organic acids in Rhododendron luteum Sweet leaves was studied by gas chromatography with a mass spectrometric detector. The obtained results indicate the prospects for further study of raw materials to develop drugs with membrane stabilizing and anti-inflammatory activity.

Ecological Aspect of Antibiotic Batumin Synthesis by Pseudomonas batumici

The species Pseudomonas batumici, isolated from the rhizosphere of eucalyptus in the humid subtropical zone, is a producer of the polyketide antibiotic batumin with highly selective activity against staphylococci. Batumin biosynthesis operon includes 28 genes or 74 151 bp. According to modern notions, the biosynthesis of energy-intensive metabolites, which probably includes batumin, is justified in the case of its multifunctionality for producers. The species P. batumici, as a representative of rhizosphere bacteria, must interact with plants and compete with the surrounding microbiota. Aim. To determine the role of batumin in the ecology of the rhizosphere producer strain P. batumici UCM B-321. Methods. The batumin producing strain P. batumici UCM B-321T was obtained from the Ukrainian Collection of Microorganisms. Antibiotic batumin was obtained by fermentation of P. batumici UCM B-321. Extraction was carried out from acidified P. batumici fermentation broth by chloroform (1:2). Chromatographic analysis of fermentation broth obtained after centrifugation was carried out by HPLC using liquid chromatograph Agilent 1200 with mass spectrometric detector Agilent G1956B. Batumin derivatives were obtained after the extraction of the fermentation broth of P. batumici using thin layer chromatography (TLC) on silica gel plates (Merck, USA) in the benzene-isopropanol system (5:1). Disc-diffusion method on phytopathogenic test-strains was used for bioautography. Biofilm formation by P. batumici strain was studied according to O’Toole by growing strain B-321 at 25 0C for 48 hours in 96-well plates on LB medium. Batumin effect upon bacterial mobility was studied using Volf and Berg method in Petri dishes with 0.5% semisolid bacterial agar. To research chemotaxis the soil strain Bacillus subtilis IMV B-7023 and the following concentrations of batumin were used: 20, 50, and 150 μg/mL. The studies were performed using Tso and Adler method. Results. Growth inhibition zones for phytopathogenic bacteria strains were the following (in mm): Pseudomonas syringae pv. syringae UCM B-1027T – 19±3, P. fluorescens IMV 8573 – 22±3, Pectobacterium carotovorum UCM B-1075T – 17±2. Activity against Xantomonas campestris pv. campestris UCM B-1049, Clavibacter michiganensis subsp. michiganensis IMV 102, Agrobacterium tumefaciens UCM B-1000 was not detected. Minimum inhibitory concentrations (MIC) in the range from 8 to 64 μg/mL for P. carotovorum UCM B-1075T, Erwinia aroidea IMV 1058, Proteus vulgaris UCM B-905 and P. fluorescens IMV 8573 are hardly comparable with the discovered against staphylococci. TLC analysis of its broth extract revealed five separate compounds with different values of retention factors: Rf1=0.42; Rf2=0.38; Rf3=0.31; Rf4=0.28; Rf5=0.25. The main component of extract was batumin, other four substances were present in minor quantities. All found substances had similar absorption maxima with the minimum differences between isomeric forms: descarbamoyl batumin-enol (Mr=505, λ=226 nm), descarbamoyl batumin-keto (Mr=505, λ=231 nm), batumin (Mr=548, λ=231 nm), batumin-enol (Mr=548, λ=228 nm) and 17-hydroxy-batumin (Mr=550, λ=229 nm). The largest inhibition zone (P. carotovorum UCM B-1075T) was on the third compound placement which represents of batumin, tiny inhibition zones were found around keto and enol form of descarbamoyl batumin. Observation of live bacterial cells in light microscope confirmed a serious disruption of motility in all these bacteria by batumin in the concentration far lower than the MIC for these organisms. Proteus actively moved in the control, but in presence of 10 μg/mL of batumin was almost no growth. The biofilm formation by P. batumici UCM B-321 was stimulated by supplementing batumin into the medium. The stimulation effect by batumin on the biofilm formation was equally strong when the compound was applied in the concentrations of 1 and 10 μg/mL. Batumin was not an attractant of the producer strain. However, in one of our experiments batumin has shown the properties of positive effector (attractant) for B. subtilis UCM B-7023 strain. Conclusion. The discovered features allow to consider the antibiotic batumin synthesized by P. batumici UCM B-321 as the essential tool for survival and competition of the producer strain in a natural habitat.

Chromato-mass-spectrometric identification of asymmetric dimethylhydrazine and N-nitrosodimethylamine

Introduction. Environmental safety is one of the main priorities of state policy. It ensures the legal regulation of relations in space activities to strengthen the defense and security of the Russian Federation and further extension of the international cooperation of the Russian Federation. Material and methods. In the fall areas of the stages of the booster rockets, screening studies were carried out to identify the propellant component 1.1-unsymmetrical dimethylhydrazine in atmospheric air (n=14) and drinking water (n=23), determine its metabolite N-nitrosodimethylamine quantitatively in the residents’ blood (n = 90) living in the surveyed areas before the rocket launch (n = 45) and after the launch (n = 45). Also quantitative determination of the N-nitrosodimethylamine metabolite in the residents’ urine in the observation group (n = 108) was performed. For comparison, there was selected a group of residents not related to rocket and space activities (n = 13). Identification and analysis of samples of atmospheric air, drinking water, and biological media (blood, urine) was performed using Agilent 7890A gas chromatograph (USA) with a 5975C quadrupole mass spectrometric detector (MCD) and a capillary column of the HP-FFAP 30m • 0.25mm • 0.25µm series. Results. The study revealed the absence of asymmetric 1.1-dimethylhydrazine in 100% of analyzed atmospheric air samples. During the observation period, the concentrations of N-nitrosodimethylamine in the range of 0.00039 to 0.001 mg/dm3 were found in drinking water samples that did not exceed the hygienic standard (LOCNDMA 0.01 mg/dm3). N-nitrosodimethylamine in a concentration range of 0.00095-0.346 mg/dm3 was determined in the blood samples of the population. The studies revealed that after the rocket launch, the N-NDMA concentration in the blood was 1.8 times higher than the concentration registered before the rocket launch. In the urine sample of the resident living in the surveyed area, N-nitrosodimethylamine was detected with a high degree of reliability according to the essential ion with mass 74 m/z and confirmatory ion of 42 m/z, and the concentration was quantitatively calculated at a level of C N-DMA = 0.23 μg/ml. The detection of N-nitrosodimethylamine in blood and urine, even in trace amounts, indicates the possibility of exposure. Conclusion. Performed comprehensive studies made it possible to prove the relative safety of the ecological situation in the fall areas of the booster rockets that are located close to settlements when considering the environmental pollution with unsymmetrical dimethylhydrazine and can be used for systematic monitoring.

Chromato-mass-spectroscopic research of chemical composition of Elaeagnus angustifolia L.

The aim of the work was chromato-mass-spectroscopic research of chemical composition of Elaeagnus angustifolia L. fruits and leaves and identifying further prospects for the use of this plant in medicine. Materials and methods. Raw materials of Elaeagnus angustifolia L. were selected as objects of study. The tincture was obtained by maceration and the raw material was extracted with methyl alcohol at room temperature for 10 days according to the method of making tinctures. The study of the chemical composition of Elaeagnus angustifolia L. was carried out using gas chromatograph Agilent 7890B GC System (Agilent, Santa Clara, CA, USA) with mass spectrometric detector Agilent 5977 BGC/MSD (Agilent, Santa Clara, CA, USA) and chromatographic column DB-5ms (30 m × 250 mkm × 0,25 mkm). Results. 23 compounds (1 in the isomeric state) in fruits and 20 compounds (2 in the isomeric state) in the leaf of Elaeagnus angustifolia L. were identified. The main components of fruits were sitosterol (phytosterols) – 12.53 %, propyl acetate (esters of carboxylic acids) – 12.60 %, chamazulene (terpenes) – 11.97 % and palmitic acid (fatty acids) – 8.28 %. The main component of leaves were sitosterol (phytosterols) – 17.57 %, 1-(2-hydroxy-5-methylphenyl)-ethanone (ketone) – 8.35 %, phytol (terpenes) – 6.10 %. It is known from the literature that chamazulene has antioxidant, antinociceptive, cytotoxic activity. Sitosterol has anti-inflammatory and antidiabetic activity. Hexadecanoic (palmitic) acid has antimicrobial, antidiabetic and antioxidant properties. Phytol is characterized by a wide range of biological action – antimicrobial, antinociceptive, anti-inflammatory, antioxidant and cytotoxic. Conclusions. Based on the above, the olive can be considered as a source of antimicrobial, antinociceptive, anti-inflammatory, antioxidant, antidiabetic and cytotoxic drugs.

Research in biologically active substances of the volatile fraction from Highbush blueberry vegetative organs

Ukraine ranked second in the world after Peru in the pace of laying new Highbush blueberry plantations. Experts estimate that in 2018, in Ukraine, from 0.7 thousand hectares to 1.0 thousand hectares of new Highbush blueberry plantations were laid. Over the past 12 years, previously little-known product, Highbush blueberries have become one of the main berry crops. At present, commercial plantations blueberries take the 3rd place after black currants and strawberries. Zhytomyr region boasts the biggest areas planted with Highbush blueberries in Ukraine, followed by Volyn and Kyiv. According to estimates, the export of blueberries from Ukraine may increase to 20 thousand tons in 5 years. In addition to the use of Highbush blueberries in the food industry, they are a promising source of BAS for pharmaceutical and medical use, so their study is a promising area of pharmaceutical science. In Ukraine, there are no domestic standardized medicines manufactured from Highbush blueberries, and there are only foreign dietary and functional supplements. Abroad, Highbush blueberry medicines are used to improve vision and as astringents for colitis, enterocolitis and diarrhoea. In this regard, the development of domestic standardized medicines based on Highbush blueberries raw materials is an urgent task for modern pharmacy. The aim of the research was to study the phytochemical profile of the volatile fraction of Highbush blueberry vegetative organs (leaves, stems and fruits). Materials and methods. The objects of research were the leaves, fruits and stems of Highbush blueberry (Vaccinium corymbosum L.), in which the composition of the the volatile fraction components was studied by chromato-mass spectrometry on a gas chromatograph Agilent Technologies 6890 with a mass spectrometric detector 5973. Results. Chromato-mass spectrometry in the composition of volatile fractions of leaves, fruits and stems of Highbush blueberries revealed 65 substances. 49 substances were identified in the Highbush blueberry leaves, of which 36 substances of terpene nature, 13 organic acids, and 2 compounds were not identified. In the Highbush blueberries fruits, 47 substances were found, of which 36 substances are of terpene nature, 14 are organic acids and 3 were not identified. In the Highbush blueberries stems, 50 substances were found, of which 33 substances are of terpene nature, 14 are organic acids and one was not identified. Conclusions. As a result of phytochemical studies, it is for the first time that the component composition of volatile fractions of leaves, stems and fruits of Highbush blueberries from the flora of Ukraine was established. The content of 65 substances was identified and established, including 14 organic acids and 39 terpene compounds.