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Vol 22, No 5 (2019)


A novel method for extraction of essential oil compounds from Foeniculum vulgare Mill. fruits

Boyko N.N., Pisarev D.I., Zhilyakova E.T., Maljutina A.Y., Novikov O.O.


The aim of the work was to prove experimentally the possibility of extraction of main essential oil components from the Foe-niculum vulgare Mill. fruits by perfluoro organic solvents Novec 1230 and Novec 7100. Materials and methods. For study purposes, we used Foeniculum vulgare Mill. fruits with particle fraction of 0.1-0.5 mm. Quantitative and qualitative analyses of extracts were carried out by GC-MS and RP-HPLC methods. Two methods of extraction were used: simple maceration, plant raw material / solvent ratio 1:10 w/v, temperature 24±1 °С, maceration time 24 hours; circulation method: plant raw material / solvent ratio 1:5 w/v, solvent circulation time from 1 to 5 hours. Results and discussion. During maceration, the highest yield of anethole was observed in n-hexane, 96±4%, and in solvent Novec 7100, 94±4%, whereas in solvent Novec 1230, the yield of anethole was three times less, 28±2%. Solvents Novec 7100/1230 extracted identical compounds, and trans-anethole was the dominant one among them. The yield of anethole within two hours of solvent circulation was up to 82±4% for Novec 7100, and 89±4% for Novec 1230. Conclusions. The possibility of extraction of main components of essential oil from the plant raw material with perfluoro organic solvents Novec 1230/7100 by the example of Foeniculum vulgare Mill. fruits was experimentally tested. It was determined that the level of solvent molecule fluorination influenced its extraction properties with respect to anethole and fatty oil.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):3-9
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The comparison of methods for the quantitative determination of coenzyme Q10

Zozina V.I., Melnikov E.S., Krasnykh L.M., Goroshko O.A., Kukes V.G.


Background. Coenzyme Q10 is a biologically active compound that performs many important functions in the body: electron transfer in the mitochondrial cascade of adenosine triphosphate synthesis, antioxidant, preventing lipid peroxidation. Study Objective. The aim is to develop and compare methods for the quantitative determination of coenzyme Q10 in order to identify the most sensitive, simple and reproducible one. Materials and methods. Coenzyme Q10 was determined using high performance liquid chromatography with spectrophotomet-ric and mass spectrometric detectors. In developing the HPLC-MS / MS it was used the electrospray ionization (ESI) in positive mode. Detection was carried out in the mode of monitoring multiple reactions (MRM). Results. The study developed and validated two methods for the determination of coenzyme Q10 in plasma. It was shown that the most sensitive and reproducible technique was HPLC-MS / MS (the LLOQ constituted 0.10 μg / ml). One-step sample preparation involving proteins precipitation, used in the HPLC-MS / MS method was more convenient than in HPLC-UV one. The LLOQ of ubiquinone in the HPLC-UV method was 0.5 μg / ml. Using the HPLC-MS / MS technique, we analyzed the average concentration of endogenous CoQ10 in patients with cardiovascular pathologies administrating statins, p-blockers and calcium channel blockers, which constituted 0.39 μg / ml. Conclusion. It was concluded that the HPLC-UV method is unsuitable for routine practice use due to the high LLOQ (0.5 μg / ml) and its low sensitivity. While the of HPLC-MS / MS method was selective, sensitive, fast and convenient for introduction into routine practice.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):10-14
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Investigation of release kinetics of doxorubicin from drug carrier based on Fe(0) microparticles under influence of ultrasound irradiation and different values of pH

Vlasov S.S., Di Martino A., Yusubov M.S., Guryev A.M., Krivoschekov S.V., Sviridova E.V., Postnikov P.S., Belousov M.V.


Aim. To obtain new drug carrier for doxorubicin based on modified Fe(0) microparticles and evaluate its release kinetics under influence of different values of pH and ultrasound irradiation. Material and methods. Size and zeta-potential of microparticles were determined on Zetasizer Nano ZS. Surface modification (covalent binding of residues of benzoic acid) was confirmed by FTIR spectroscopy. Encapsulation efficiency (EE) and loading capacity (LC) of doxorubicin (DOX) was determined by UV spectroscopy (480 nm). Release studies were carried out in Stuart SI 500 incubator at a constant temperature (37 °C), stirring rate (100 rpm) and different pH values (3.3; 5.5; 7.4). For investigation of influence of ultrasound (US) irradiation on the release kinetics ultrasound field with frequency and power 75 kHz and 2 W/cm2 respectively was used. Ultrasonic bath Elmasonic S10H was used as a source of ultrasound irradiation. Results and discussions. Size and zeta-potential of Fe-CS-DOX conjugate were 4.43 and -9.07 respectively. Loading capacity of doxorubicin was 0.54 mg/mg. Percentage of released drug with and without US irradiation were 96 and 18% respectively (in 12 hours after starting of the experiment). Conclusion. In this study, the release of doxorubicin from drug carrier, based on Fe (0) microparticles at different pH values was investigated and the influence of ultrasound irradiation on the release kinetics was confirmed. In 12 hours after starting of the release, the amount of released drug was increased more than 4 times. So, the obtained conjugate Fe-CS-DOX leaves great promise for its further use as a drug carrier.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):15-19
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Determination of etoricoxib by high performance liquid chromatography

Kutyakov V.A., Nemikhin V.V., Kachin S.V.


Objective: when conducting forensic studies in the forensic chemistry department of the Krasnoyarsk Regional Bureau of Forensic Medicine, there were many cases of identification of nonsteroidal anti-inflammatory drugs in cadaveric material, such as Ibu-profen, Diclofenac, and others. Drugs found predominantly in individuals suspected of non-medical use of psychoactive substances can serve as specific markers of the latter. Earlier, Etoricoxib was not defined in expert material, therefore the aim of the presented research was to develop a method for determining this agent in cadaver blood. Materials and methods: cadaver blood was tested, a standard solution of Etoricoxib prepared from a commercial drug Costarox (Sandoz), tested for purity and content of the active substance, the qualification of reagents complied with the requirements for examination. The analysis was carried out on a «Milichrom A-02» liquid microcolumn chromatograph with a spectrophotometric detector of the UV-range, multiwave detection, with the MultiChrom processing program. The study was carried out in compliance with the requirements of evidence-based medicine. The reliability of the study is provided by the formation of control groups and comparisons, using the correct methods of statistical processing of results. Results. A method for determining the non-steroidal anti-inflammatory agent Etoricoxib by the method of gradient high-performance liquid chromatography in cadaveric blood in relation to forensic chemical studies has been developed, ensuring a reduction in the duration and material costs of the analysis without compromising the evidence of the results. This effect is achieved by using as a mobile phase a mixture of eluents - acetonitrile (B) and lithium perchlorate (A) (gradient elution from 5 to 100% eluent B in eluent A), as well as the use of an internal standard (Anekain). The detection limit of etoricoxib was 2 ng/ml, the limit of quantitative determination was 6 ng/ml. The relative standard deviation of Sr does not exceed 0.08. The percentage of extraction of Etoricoxib from cadaveric blood ranged from 32.7 to 37.4%. Conclusions: The described method is characterized by a fairly high sensitivity, reproducibility, efficiency, express. Obtaining quantitative values of the concentration of Etoricoxib will allow to differentiate the purpose of the drug, to specify the cause of death. The described technique is used in the practice of the Krasnoyarsk Bureau of Forensic Medical Examination and can be recommended for routine forensic chemical studies in order to establish the presence and content of Etoricoxib in human cadaveric blood.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):20-24
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Physiological characteristics suspension culture of Rhodiola rosea L. at cultivation on the modified nutrient medium

Savin P.S., Savina T.A., Myasnikova S.B.


To use cell plant cultures in the production of biologically active substances, one must know their properties, including the patterns of growth and biosynthetic activity. As a result of the study of the physiological features of the Rhodiola rosea L. culture of the Rr(S) 2013VILAR strain in the dynamics of biomass accumulation under conditions of deep cultivation on a macrosolated nutrient medium, the duration of culture growth was determined to be 15-20 days. The process of culture growth under conditions of deep cultivation can be described by a growth curve having an S-shape. The factor of physiological heterogeneity of the suspension culture is noted, which can significantly influence the final results on biomass accumulation and its bioproductivity. It was found that the maximum consumption of oxygen by the cell culture is 12-10-4 mol/l-h and falls on the 8th day of growth, when the cells are in the phase of regular growth and begin to actively share. To maintain the active growth of the suspension culture of Rhodiola rosea L. strain Rr(S) 2013 VILAR in the cultivation vessels, it is necessary to create a mass transfer coefficient of 8 h-1. It is shown that the accumulation of the sum of phenylpropanoids in the cell culture of Rhodiola rosea L. of the Rr(S) 2013VILAR strain occurs in parallel with the growth of biomass.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):25-29
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Clinical and biochemical predictors in patients with obesity and newly diagnosed type 2 diabetes mellitus

Mikaelyan N.P., Skripkina P.A., Gaidina T.A., Mikaelyan A.V.


The aim of the study was to study some clinical and metabolic parameters in patients with insulin resistance in obesity and newly diagnosed type 2 diabetes mellitus. In 57 women aged 23 to 39 years with obesity 3-4 degrees and the first identified type 2 diabetes mellitus, the lipid composition of the erythrocyte membranes and the state of lipid peroxidation and antioxidant activity were studied. The control group consisted of 15 women without endocrine pathology. The study of biochemical predictors in patients with newly diagnosed diabetes mellitus type 2 and in patients with obesity, along with dyslipidemia, showed a violation of the composition of fatty acids in the blood due to the group of omega-3 and omega-6. Increased intensity lipid peroxidation on the background of reduced activity of antioxidants and the level of omega-3 fatty acids indicates the development of oxidative stress, the most pronounced in type 2 diabetes compared to obesity. The results of the study also indicate the presence of the antioxidant properties of omega-3 fatty acids, as evidenced by the direct correlation between omega-3 fatty acids and the activity of antioxidant enzymes. Conclusion: 1. In patients with diabetes mellitus type 2 and in patients with obesity, dyslipidemia, a violation of the composition of fatty acids are noted. 2. The development of oxidative stress, most pronounced in type 2 diabetes mellitus, was established. 3. Direct correlation relationships between omega-3 fatty acids and the activity of the studied enzymes indicate the presence of the antioxidant properties of omega-3 fatty acids.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):30-34
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The bioenergetic characteristics of mitochondria of the rat liver at low body temperatures

Khalilov R.A., Khizrieva S.I., Dzhafarova A.M., Abdullaev V.R.


Many of the pathological effects of hypothermia are directly or indirectly associated with changes in the functioning of mitochondria, the sensitive indicators of which are their bioenergetics characteristics. In this paper, the bioenergetic characteristics of isolated rat liver mitochondria in normal and hypothermia of different depths were investigated. Moderate (30 °C) hypothermia was found to significantly increase the respiratory rate of mitochondria. The deepening of the hypothermic state to 20 °C continues to stimulate respiration, however, increase in its rate becomes less pronounced relatively moderate hypothermia. In both moderate and deep hypothermia, the rate of oxidative phosphorylation is increased, and respiratory controls, P/O, and sensitivity to 2.4 DNP of mitochondrion are reduced. Many of these changes in mitochondrial respiratory parameters are more pronounced with moderate hypothermia. A comparative analysis of the bioenergetic characteristics of mitochondria obtained in the study of glutamate- and succinate-dependent respiration suggests that the stimulation of mitochondrial respiration during hypothermia occurs mainly due to changes in the functioning of Complex I of the respiratory chain.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):35-41
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Creation of zinc-enriched medicine and food plants

Syroeshkin A.V., Makarova M.P., Maksimova T.V., Pleteneva T.V.


Objective. To develop the basics of technology for obtaining new elements modified plant products that can be used for hypo-elementoses correction. Materials and methods. Water samples: deionized highohmic water (BD, D/H=140 ppm); deuterium depleted water (DDW, D/H=12 ppm); medicinal and nonoficinal plants (leaves and seeds). Atomic absorption spectrometry with electrothermal atomization and Zeeman effect; Xray fluorescence energy dispersive spectrometr; Low-Angle Laser Light Scattering, LALLS); Spirotox-test equipment. OriginPro®9, OriginLab Corporation, USA. Results. For the first time, the integrated approach was used to develop the technology fundamentals for plant accumulation by essential microelements. The approach includes isotopic control of plant development; new laser methods for quality control of water solutions for plants irrigation and hydroponics; the methods for the control of microelements enrichment of plant materials; online control of biotoxicity (Spirotox test) of the original plant materials and modified plants. For example, it has been demonstrated the ability to create zinc (II) modified plants with a high content of essential trace elements, up to 1.4 mg / g dry weight, which corresponds to zinc content increase by three orders of magnitude. Conclusion. Plant materials enriched with essential microelements can be recommended as the objects for the creation of the medicinal products that do not cause undesirable side effects, in contrast to the synthetic preparations used for the correction of hypo-elementoses.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):42-47
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Localization of phenolic compounds in cells and tissues of medicinal plants (Dioscorea caucasia Lypsky, Euonymus nana Bieb., Aristolochia manshuriensis Kom.), cultivated under in vitro conditions

Kalashnikova E.A., Zaytseva S.M., Doan T.T., Kirakosyan R.N.


The aim of the work is to study the localization of phenolic compounds in callus cultures of medicinal plants (Dioscorea caucasia Lypsky, Euonymus nana Bieb., Aristolochia manshuriensis Kom), used as raw material sources in pharmacology. Methods and materials. The object of the study was callus cultures obtained from intact plants and regenerative plants (Dioscorea caucasia Lypsky., Euonymus nana Bieb and Aristolochia manshuriensis Kom.). Callus tissue was cultured on a nutrient medium containing Мurasiga and Skuga mineral salts and growth regulators at a temperature of 250 C and a 16-hour photoperiod, In alcohol extracts of callus cultures by spectrophotometric method, the content of the sum of soluble phenolic compounds (with Folin-Denis reagent), flavans (with vanillin reagent) and flavonols (with aluminum chloride) was determined. The localization of polyphenols was determined by histochemical methods (0.08% raster of Fast Blue reagent, reaction with vanillin reagent in hydrochloric acid vapor). Results. The resulting long-passable, well-proliferating callus cultures are characterized by a high ability to form polyphenols (flavans and flavanols). Flavanols were major components of the phenolic complex. In the process of culturing callus cultures, the level of accumulation of all studied classes of polyphenols is reduced in comparison with intact plant tissues. Summary. Under in vitro conditions, the species-specific ability to synthesize phenolic compounds is preserved, which is confirmed not only by quantitative determination of phenolic compounds, but also by histochemical studies. The polyphenols in cells of the calli were localized in micro- and macrovascular, in the intercellular spaces and cell walls. Among callus cells, epiblates with a central vacuole containing a large number of polyphenols in the form of an amorphous substance, as well as small and large-faceted inclusions, were occasionally encountered.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(5):48-54
pages 48-54 views

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