On October 29, 2021, the rector of the SPCPU Igor Anatolyevich Narkevich celebrated his 55th anniversary.

The Center for Shared Use of Scientific Equipment "Analytical Center of the Federal State Budgetary Educational Institution of Higher Education SPKhFU of the Ministry of Health of Russia" (CKU) was established on the basis of the University in 2011 with the aim of carrying out research in the field of creating highly effective and innovative medicines, taking into account modern scientific approaches and carries out scientific, research and educational functions.

Introduction. Analysis of the clinical and laboratory picture of the SARS-CoV-2 infection suggests the presence of microcirculation and oxygen transport disorders, hemolysis of erythrocytes, intra-alveolar fibrin formation and microthrombus formation in the patient’s pathogenesis. Accordingly, the search for potential anticoagulants, erythrocyte antiplatelet agents, membrane stabilizing drugs and mild thrombolytic drugs can prevent the development of life-threatening complications and reduce the mortality of COVID-19 patients.

Aim. Isolation of formononetin-7-O-β-D-glucopyranoside from the grass of Ononis arvensis L. and identification of the molecular mechanisms of its effect on platelet activation in vitro, induced by TRAP-6 (Thrombin receptor activated peptide) and ADP (adenosine diphosphate).

Materials and methods. Terrestrial parts of Ononis arvensis L. were collected in the SPCPU nursery of medicinal plants (Leningrad region, Vsevolozhsky district, Priozerskoe highway, 38 km). Isolation of formononetin-7-O-β-D-glucopyranoside was carried out by preparative high performance liquid chromatography on a Smartline device (Knauer, Germany) equipped with a spectrophotometric detector. The structure of formononetin-7-O-β-D-glucopyranoside was confirmed by one-dimensional and two-dimensional NMR spectroscopy (Bruker Avance III, 400 MHz, Germany), as well as high-resolution mass spectrometry (HR-ESI-MS) (Bruker Micromass Q-TOF, Germany). The study of the effect of formononetin- 7-O-β-D-glucopyranoside on induced platelet activation was carried out on human platelets isolated from the blood of healthy volunteers. To research the effect of formononetin-7-О-β-D-glucopyranoside on platelet aggregation flow cytofluorometry with Cyto-FLEX (Beckman-Coulter, USA) was used.

Results and discussion. According to the method of fractionation and purification of the total extract of O. arvensis developed in previous studies, formononetin-7-O-β-D-glucopyranoside was isolated in an individual form for subsequent biological studies with a total yield of 30 % in comparison with its content in the original extract. In samples with formononetin-7-O-β-D-glucopyranoside and ADP, there is a pronounced inhibition of platelet activation – the percentage of active platelets ranges from 6.3–6.6 % at doses of formononetin-7-O-β-D-glucopyranoside 1 μM, 3 μM and 30 μM. The inhibitory effect of formononetin-7-O-β-D-glucopyranoside is not dose-dependent (p ≤ 0.05). In samples with formononetin-7-O-β-D-glucopyranoside and TRAP, there is also a pronounced inhibition of platelet activation. The percentage of active platelets is 8 % at 1 μM formononetin-7-O-β-D-glucopyranoside doses, 15 % at 3 μM doses, and 16 % at 30 μM doses.

Conclusion. Administration of formononetin-7-O-β-D-glucopyranoside at doses of 1 μM, 3 μM, 30 μM strongly inhibits platelet activation induced by ADP and TRAP-6. For ADP, there is no dose-dependent effect, while for TRAP there is a weak dose-dependent effect, the greatest inhibition efficiency is achieved with the minimum investigated dose of 1 μM. In all cases, the results obtained are statistically significant.

Introduction. The use of certified reference materials (CRMs) ensures metrological traceability and comparability of analysis results performed in different laboratories, by different analysts, at different times. Genistein is a promising substance with a wide spectrum of pharmacological action. genistein is widely used in dietary supplements. Development of regulatory documents for CRM of genistein will ensure the quality of drugs and dietary supplements.

Aim. Aim of our study is to improve of the ways of synthesis and determination of spectrum characteristics of genistein for the certification of CRM.

Materials and methods. We used synthetic genistein, (Ph.D. V. Yu. Kovtun SPC "Pharmzashchita") (sample № 1) and genistein synthesized and studied at the departments of pharmaceutical chemistry and chemical technology of medicinal substances SPCPU (sample № 2). Infrared spectra of genistein samples were collected on an FSM 1201 infrared Fourier spectrometer (OOO Infraspek, Russia) via KBr pellets technique. All the spectra were collected in the 4000–500 cm−1 range. The NMR (1H and 13C) measurements were performed with a BrukerAvance III NMR spectrometer (400 and 100 MHz) (Bruker, Germany) in DMSO-d6 solvent. Raman spectra were recorded by an ORTES-785TRS-2700 analytical Raman scattering system at a laser power of 100 mW (OPTEC JSC, Russia). Laser interaction time was 5, 10, 20 and 60 seconds. The results were processed using the software "BWSpec 4.10_4", USA. GC-MS was performed on an Agilent Technologies 7890A gas chromatograph (Agilent Technologies, USA) with a 7693 autoinjector and a Hewlett Packard 5975C mass selective detector.

Results and discussion. The synthesis was carried out according to the developer's method. The stage "removal of the alkyl protection" has been improved. The spectra of the synthesis intermediate of genistein (biochanin A) correspond to the literature data. Samples of genistein were investigated by methods: MC and NMR 13С, 1Н. The structure of the investigated substance was confirmed; Raman and IR spectroscopy showed that the spectra of the samples do not differ from each other and there are no additional signals.

Conclusion. The spectrum characteristics of samples of genistein were obtained by NMR, IR and Raman spectroscopy, which will be used in the regulatory documentation for CRM of genistein. All of this will make it possible to control the quality of medicines based on it and to identify substandard dietary supplements.

Introduction. Medicinal plant raw materials are one of the most important sources of herbal remedies used both for the prevention and treatment of a number of diseases [1, 2]. Even with the development of modern science and chemistry, medicinal plant raw materials are widely used in both folk and official medicine. The advantage of medicinal plants is their wide range of biological activity, low toxicity and the possibility of long-term use without significant side effects. Human economic activity has a noticeable negative effect on the condition of wild-growing plants: their stocks are decreasing, and some species disappear altogether. Today, given the high level of development of industry and agriculture, the procurement of raw materials for wild medicinal plants is not always possible. Information on the quantitative assessment of the raw material base of wild medicinal plants in the Middle Urals, the content of biologically active substances in medicinal plant raw materials is partially outdated, which determines the need for their systemic resource study and chemical-pharmacognostic study.

Aim. Comprehensive assessment of the state of populations of wild medicinal plants in the Middle Urals.

Materials and methods. Determination of stocks of raw materials of the studied species of medicinal plants was carried out on specific thickets according to the generally accepted method. The authenticity of the raw materials was established by a macroscopic method when collecting raw material samples. In the course of the study, samples of medicinal plant materials of 5 types were prepared. The determination and assessment of the main indicators of the good quality of medicinal plant raw materials (the content of active and extractive substances, moisture in the mass upon drying, total ash and ash insoluble in a 10% solution of hydrochloric acid) was carried out according to the methods and requirements of the State Pharmacopoeia of the Russian Federation XIV edition. In the raw materials Artemisiae absinthii herba and Leonuri herba, the amount of extractives was determined by the gravimetric method. The quantitative assessment of the content of essential oil in the samples of Origani vulgaris herba and Tanaceti vulgaris flores was carried out by the method of hydrodistillation. To determine the quantitative content of the sum of flavonoids in Hyperici herba, Artemisiae absinthii herba, Leonuri herba and the sum of flavonoids and phenolcarboxylic acids in Tanaceti vulgaris flores, a spectrophotometric method was used.

Results and discussion. In the course of resource and phytochemical studies of representatives of the medicinal flora of the Middle Urals, a comprehensive assessment of the state of populations of wild medicinal plants – sources of medicinal plant raw materials (Origani vulgaris herba, Hyperici herba, Tanaceti vulgaris flores, Artemisiae absinthii herba and Leonuri herba) was carried out. The results are included in the electronic inventory of wild medicinal plants of the Middle Urals.

Conclusion. The conducted complex of studies will allow updating information about the medicinal flora of the Middle Urals in order to use raw materials for the creation of medicines.

Introduction. Treatment and prevention of diseases of the oral mucosa is one of the priority tasks in dentistry. In practice, antibacterial agents are often used in the complex treatment of inflammatory and destructive processes. However, long-term, uncontrolled usage of such drugs leads to numerous complications: drug tolerance, weakening of the therapeutic effect, dysbiosis of the oral cavity and gastrointestinal tract, etc. Therefore, at present, the question of search for alternative to antibiotic therapy remains open. As an alternative, it is necessary considering the usage of effective and safe herbal medicines that are easy to digest, less toxic, practically do not cause side effects and allergic reactions, and have a light regulating and normalizing effect.

Aim. The aim of the present study is to develop the composition and technology of effervescent granules for the preparation of a solution for rinsing the oral cavity based on phytosubstances.

Materials and methods. Dry extracts were obtained from medicinal plant materials: medicinal sage leaves, medicinal calendula flowers, yarrow herb, medicinal rhizomes and roots and astragalus woolly herb. Sodium carbonate, citric acid, anhydrous, microcrystalline cellulose – 90 (EMCOCEL®90M), povidone (Plasdone™ K-29/32) and calcium stearate were used as auxiliary substances in the granule technology. In laboratory conditions, granules based on phytoextracts were obtained by pressing wet masses. Numerical indicators of medicinal plant raw materials, technological properties of dry extracts and granules, as well as indicators of the quality of granules were determined according to the methods described in the State Pharmacopoeia XIV.

Results and discussion. The numerical indicators of medicinal plant raw materials (grinding of raw materials and the content of impurities, total ash in medicinal plant materials and ash insoluble in hydrochloric acid, humidity, content of extractives) were determined and the good quality of the raw materials used in the subsequent stages of drug development was confirmed. Dry extracts from each type of medicinal plant raw materials have been developed and the technological properties of dry extracts have been determined. The composition and technology of effervescent granules by pressing wet masses has been developed. To improve the flowability and reduce the hygroscopicity of the granulated material, microcrystalline cellulose – 90 (EMCOCEL®90M) was used as a filler. To create an effervescent dosage form, citric acid and sodium bicarbonate were added to the granules. The mass for granulation was moistened with a 10% alcohol-water solution of Plasdone™ K-29/32. A draft specification of quality indicators for effervescent granules based on phytoextracts is proposed.

Conclusion. In the course of the research work, the numerical indicators of medicinal plant raw materials were determined and its quality was confirmed, which made it possible to use it for further production of dry extracts. The extraction conditions were selected for each type of raw material, dry extracts were developed, and quality indicators were determined in accordance with the requirements of the State Pharmacopoeia XIV. Excipients were selected taking into account the properties of dry extracts, the composition and technology of effervescent granules based on phytoextracts was developed, a draft specification for effervescent granules was proposed in accordance with the requirements of the State Pharmacopoeia XIV.

Introduction. The article presents the results of studying the technological properties of individual excipients widely used in the compositions of existing orally dispersed tablets (ODT) for subsequent planning a multifactorial experiment. Samples of excipients were analyzed according to such pharmacopoeial indicators as description, flowability, bulk density, compressibility, fractional composition, solubility in water.

Aim. The aim of the work is to create a list and study the technological properties of candidate substances for the role of auxiliary substances in the composition being developed by the ODT.

Materials and methods. The technological properties of excipient samples were studied according to the methods of the State Pharmacopoeia of the XIV edition using the flowability tester GTL (ERWEKA, Germany), the bulk density tester SVM 221 (ERWEKA, Germany), the tablet press PGR-10 (LabTools, Russia) and the tablet hardness tester TBH 125 TDP (ERWEKA, Germany).

Results and discussion. As a result of the study, experimental data on the technological properties of excipient samples were collected, and the selected samples were compared according to pharmaceutical and technological indicators.

Conclusion. In the course of the study, a list of auxiliary substances for the development of the composition of ODT was formed and studies of their technological properties were carried out. The obtained experimental data will allow to develop an optimal matrix of a multifactorial experiment for the development of the composition of ODT and justify the choice of excipients.

Introduction. Direct compression technology is one of the most common tablet technologies. As known, many active pharmaceutical ingredients are not suitable for this technology without the addition of special excipients. A useful tool for determining the suitability of powdered materials for direct compression technology is the Sediment Delivery Model (SeDeM) method, based on the concept of Quality by Design. The presented method allows not only to assess the suitability of a material for direct compression, but also helps to predict the composition of a solid dosage form in the form of a tablet, which, in turn, leads to a significant reduction in experimental work carried out in the development of a new drug.

Aim. Prediction of the compositions of matrix tablets based on sodium 4,4'-(propanediamido)dibenzoate with prolonged release, obtained by direct compression using the method of mathematical modeling SeDeM.

Materials and methods. The objects of the study were the original substance sodium 4,4'-(propanediamido)dibenzoate, as well as a number of auxiliary substances, which included polymers used for dosage forms with prolonged release, a dusting component – magnesium stearate, and a filler – lactose monohydrate. Physicochemical and technological properties of APIs, explosives, obtained tablet mixtures and tablets were studied in accordance with the requirements of the State Pharmacopoeia of the Russian Federation XIV ed. and EP 9th ed.

Results and discussion. The properties of the substance and excipients were assessed in accordance with the SeDeM method. It was found that the substance 4,4'-(propanediamido) sodium dibenzoate is not suitable for direct pressing due to poor flowability and low compressibility. Hypromellose Methocel K4M had good compressibility, but it did not have sufficient flowability. The other tested polymers had satisfactory properties for the direct compression technology. The composition of the tablet mixtures was calculated using the SeDeM method, the obtained tablet mixtures had satisfactory technological characteristics for obtaining tablets by direct compression. The tablets obtained as a result of the experiment also met the pharmacopoeial requirements.

Conclusion. Prediction of the composition of sustained-release tablets based on the original substance sodium 4,4'-(propanediamido)dibenzoate was carried out using the SeDeM method. It was found that this method is suitable for the development of the composition of tablets based on sodium 4,4'-(propanediamido)dibenzoate.

Introduction. Soft gelatin capsules are a promising dosage form comprising essential oils as active agents. Joint research of the staff of the Perm State Pharmaceutical Academy, the Research Institute of Nutrition of the Ministry of Energy and Industry of the Republic of Tajikistan have proposed the composition of gelatin mass for encapsulation by the rotary matrix method. The mechanical and physical-technological parameters required to preserve the strength and elasticity of the capsule shell during the production process and storage are determined.

Aim. Study of the rheological properties of gelatin masses based on gelatin of different grades, as well as the migration of essential oils through capsule shells.

Materials and methods. Pharmaceutical active substances "Lipovitol" and "Limoneol" obtained in the Republic of Tajikistan were used as active substances introduced into the composition of soft gelatin capsules. Sunflower oil was used as the solvent. Gelatin samples were used to obtain gelatin masses: 1 – Foodchem (China), 2 – Brodnickie Zaklady Zelatyny Sp. zo.o. (Poland), 3 - Italgelatine s.p.a. (Italy), 4 – Ewald-Gelatine GmbH (Germany), 5 – Weishardt International (France); glycerol; sunflower oil. Gelatin mass for manufacturing soft capsules was prepared in a closed reactor. Capsules were prepared on an automatic encapsulation line RJWJ – 115 Soft Gelatin Encapsulator Machine (China). The structural and mechanical properties of soft gelatin masses were determined on a rotary viscometer RV type "Reotest 2" (Germany). The dynamics of the process of migration of essential oils and its components were studied by changing their amount in a capsule by chromato-mass spectrometry method on a chromatograph Varian CP 3800 with a quadrupole mass spectrometer 4000 MS as a detector (USA).

Results and discussion. When studying the rheological properties of model compositions, it was found that for all samples of gelatin masses there is a decrease in values of effective viscosity when the shear rate increases, which characterizes the tested samples as a structured dispersion system. Additional studies have shown that the gelatin masses have thixotropic properties. Samples of gelatin masses 3–5 had narrower hysteresis loops, while sample 5 the narrowest, restoration of the structure took place quite quickly. Capsules obtained from gelatin mass 3 and 4 samples had a strong seam and were well cut out of the tape. From the mass of sample 5, high strength ribbons were obtained, a high temperature was required to seal the capsules, in some capsules the seam was not glued on one side, as a result, the capsules were rigid and brittle. As a result of the study, the rheological optimum of the gelatin mass suitable for preparing capsules by a rotary matrix method was determined, which has boundaries in the ranges of shear rates of 0.556–243 s-1 and viscosity ranges of 11.46-5028.76 Pa ⋅ s and shear stress of 2788–2808 Pa developing at these rates. When studying the migration of active substances through the capsule shell, it was found that over three years of storage of capsules in a closed polymer can, the content of essential oil in Lipovitol capsules decreased by 4.88 %, in Limoneol capsules by 5 %, which indicates partial migration of oil through the gelatin shell. The content remained within the permissible deviations (±10 %). The content of essential essential oil components also remained within acceptable deviations throughout the shelf life.

Conclusion. The optimal composition of the shell for producing soft capsules by a rotary matrix method is justified. It was found that the rheological optimum of gelatin mass is characterized by viscosity ranges of 11.46-5028.76 Pa ⋅ s and shear stress of 2788–2808 Pa. According to the results of the study of the migration of essential oils through the shell, has been established the shelf life of soft gelatin capsules in glass jars made of dark glass and a temperature of 15 to 25 °C – 3 years.

Introduction. Osteoarthritis (OA) is the most common joint disease that affects more than 10 % of the world's population. More than 600 000 people are diagnosed for the first time each year, but these data do not reflect the true prevalence of the disease, since not all patients seek help from hospitals [1, 2].

Aim. Pharmaceutical development of the composition and technology of a gel based on meloxicam, a purine derivative and an immunomodulating component for the treatment of OA with pharmacological substantiation of the content of active substances.

Materials and methods. A combination of three active pharmaceutical substances was studied: a non-steroidal anti-inflammatory drug – meloxicam, a purine derivative and an original immunomodulator M. Sodium alginate, natrozole and xanthan gum were considered as gelling agents. Were identified two technological modes of obtaining a gel base. The concentrations of active substances were selected based on the results of preclinical studies. OA was modeled by the combined administration of 0.1 ml of a mixture of Freund's complete adjuvant with a 10 % talc suspension in isotonic sodium chloride solution in a ratio of 1 : 10 into the hock (tarsus) joint cavity. The criteria for choosing the optimal composition of the gel were the size of the damaged joint, exercise tolerance and the histological picture in comparison with intact and control animals. For quantitative data, sample mean values (M) and standard deviations (SD) were calculated. The results corresponded to the laws of normal distribution, statistical processing was carried out using one-way analysis of variance (One-Way ANOVA) using the GraphPad Prism 8.0.2 software, USA at the level of statistical significance of differences p < 0,05 и p < 0,001.

Results and discussion. The composition was developed and the technology of the topical dosage form based on sodium alginate was proposed. Preclinical data indicate that the highest efficacy is achieved when using a formulation containing 3 % purine derivative, 5 % immunomodulator M and 0.5 % meloxicam. The developed composition for the effectiveness of suppressing the symptoms of OA showed results that exceeded the reference drug.

Conclusion. An original combined agent for the treatment of OA has been developed. Due to the selected component composition, with greater efficiency, it was possible to reduce the dosage of meloxicam to 0.5 %, and the use of sodium alginate as a gelling agent contributed to the prolongation of the action of the gel and the subsequent reduction in the number of applications.

Introduction. Arterial hypertension is one of the main risk factors for the development of cardiovascular diseases. Drug treatment of arterial hypertension is associated with a number of difficulties: often requires combination therapy, also a possible change in either dosages or drugs during treatment during the patient's life. Three-dimensional printing allows to create individual medicines on-demand.

Aim. Study suitability of Kollidon® VA 64 as a matrix-polymer for the preparation of immediate release ramipril printing tablets.

Materials and methods. Substance: ramipril; excipients: Kollidon® VA 64, Kollidon® CL-F, Soluplus®, PEG 1500, sodium carbonate anhydrous, Poloxamer 188, sodium stearyl fumarate, mannitol; reagents: hydrochloric acid, acetonitrile for ultra-HPLC, sodium octanesulfonate for HPLC, orthophosphoric acid 85 %, sodium perchlorate analytical grade, triethylamine, standard: ramipril USP (№1598303). Ramipril filaments were prepared by hot melt extrusion on the extruder Haake™ miniCTW (Thermo Fisher Scientific). The tablets were printed on a hand-made 3D printer. The printlets were studied for friability and hardness. Uniformity and quantitative determination of ramipril and impurities in tablets and filaments were determined by high performance liquid chromatography on a Shimadzu Prominence LC liquid chromatograph. Stability of ramipril was studied on a DSC 3+ Mettler Toledo by differential scanning calorimetry. Also, the stability of ramipril was determined by the Raman spectroscopy on an analytical system ORTES-785TRS-2700.

Results and discussion. Ramipril filaments with a diameter of 1.75 mm were obtained by melt extrusion at a temperature of 105 °C. They were homogeneous in quantitative content of the active substance. From the resulting filaments, tablets were printed in five configurations with three filling densities: 30 %, 50 % and 100 %. Degradation of ramipril in filaments and tablets is not observed. The melting point of the selected mixture is lower than the melting point of matrix-polymer. It makes possible to lower the processing temperature. Tablets with 100 % filling provide an immediate release of ramipril.

Conclusion. Kollidon® VA 64 is suitable as a matrix-polymer for the development of immediate release ramipril printlets. Kollidon® VA 64 provides the necessary physical and processing properties of the filament required for FDM printing.

Introduction. Cerebrovascular disease (CVD) is the most important medical and social problem of modern neurology because they have the highest rates of morbidity, mortality and disablement in the population. The growing incidence of CVD as a result of an aging population worldwide requires the emergent development of therapeutics, diagnostic and preventive tools. However, the development of drugs for the treatment of brain diseases has limitations due to the presence of the blood-brain barrier, which protects the brain against most molecules from the bloodstream entering the central nervous system. At the St. Petersburg State Chemical Pharmaceutical University of the Ministry of Health of Russia the alpha-2 adrenergic agonist mafedine was synthesized, which has mild psychostimulant and anxiogenic effects and which may be used in the treatment of traumatic brain injury as a neuroprotective agent.

Aim. The development of a dosage form of mafedine in order to improve its penetration into the central nervous system.

Materials and methods. Mafedine (pharmaceutical substance) [6-oxo-1-phenyl-2-(phenylamino)-1,6-dihydropyrimidin-4-olate sodium] (St. Petersburg State Chemical-Pharmaceutical University of the Ministry of Health of Russia); lecithin, span-60, Tween-80, Poloxamer 188, mannitol, vitamin E, ascorbic acid, methylene chloride, dimethyl sulfoxide, acetonitrile, trifluoroacetic acid. The fine emulsion of mafedine was obtained by ultrasound. The dosage form of mafedine was obtained by freeze drying. Residual solvents were determined by gas chromatography. Quantitative analysis of mafedine was performed by high-performance liquid chromatography. Particle size and zeta potential of emulsion were determined on a Zetasizer Nano ZS.

Results and discussion. Lyophilizate of mafedine was obtained and presenting as a light yellow porous, odorless tablet. The average mass of dry tablet was (0,17 ± 0,01) g with mafedine content is (26 ± 1) mg. The water content in the lyophilizate was 3,85 %. The quantity of methylene chloride in the lyophilizate correspond to the requirements for residual solvent content. The reconstitution time of lyophilizate into a primary emulsion was 3–5 seconds. The reconstituted dispersion was yellow, odorless, and did not break within 2 days during storage. The pH of the reconstituted emulsion was 7,34. The average particle size was (164,7 ± 6,4) nm, the zeta potential was –32 mV.

Conclusion. The developed dosage form is stable according to its physicochemical and pharmaceutical characteristics and is suitable for experimental study on models as a neuroprotective and neurorehabilitation agent.

Introduction. Understanding the mechanisms of accumulation of individual groups of biologically active substances in promising types of plant raw materials and the possibility of predicting them is important for solving fundamental and applied problems of pharmaceuticals. To date, differences have been revealed in the qualitative and quantitative composition of secondary metabolites in the aboveground and underground of Comarum palustre L., however, the issue remains unstudied.

Aim. Comparative metabolomic study of the composition of the primary metabolites of the aboveground and underground parts of Comarum palustre L.

Materials and methods. The object of the study was the aboveground and underground parts of Comarum palustre L., harvested in the vicinity of the nursery of medicinal plants of the St. Petersburg State Chemical and Pharmaceutical University (Leningrad Region, Vsevolozhsky District, Priozerskoe Highway, 38 km) in 2019 and dried. Metabolomic studies based on GC-MS method was perfomed. A statistical analysis based on the MetaboAnalyst 5.0 platform was used.

Results and discussion. Analysis of the chromatograms obtained using the GC-MS method revealed the content of 933 primary metabolites in the aboveground and underground parts of Comarum palustre L., 120 of which were identified. Using a number of statistical methods, 10 metabolites from monosaccharides, acids and alcohols, making the greatest contribution to the manifestation of differences between the studied samples, were identified.

Conclusion. The study revealed the relationship between the composition of primary and secondary metabolites in medicinal plant raw materials.

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.

Introduction. The article presents the development of solutions based on the comenic acid substance. The criteria of the studied compositions that affect their stability during storage are evaluated within the framework of the Quality-by-Design concept. The optimal compositions of comenic acid solutions have been established.

Aim. The purpose of the study is to develop solutions based on the comenic acid substance and determine the most stable variants of execution.

Materials and methods. The study of comenic acid solutions was carried out by using a laboratory pH meter PB-11-P11 (SARTORIUS, Germany) and a liquid/ion chromatograph "Stayer" ("Akvilon" JSC, Russia).

Results and discussion. The study made it possible to determine the most stable compositions of solutions based on the comenic acid substance and to establish optimal indicators of their stability criteria. It was found that solutions of comenic acid are the most stable in the pH range: from 4.0 to 6.0. At the same time, regardless of the studied methods of neutralization of comenic acid, solutions are unstable at concentrations of 25 mg/ml or more.

Conclusion. As a result of the study, the optimal compositions of solutions based on the comenic acid substance were determined. A comparative analysis of excipients that increase the solubility of comenic acid in aqueous solvents is performed. The stability criteria of the studied solutions are established and their values for ensuring the stability of the developed drug are determined.

Introduction. The standard samples (SS) use is a necessary condition for the medicines' quality control implementation. Their development is an urgent problem for the pharmaceutical industry, especially for new biologically active compounds that can be further used as pharmaceuticals.

Aim. This work aim is to establish the 5-butyl-1,2-diphenyl-6-oxo-1,6-dihydro pyrimidone-4-olate sodium quantitative content, for which anti-inflammatory and analgesic activity was previously proven, in a standard sample.

Materials and methods. This work aim is to establish the 5-butyl-1,2-diphenyl-6-oxo-1,6-dihydro pyrimidone-4-olate sodium quantitative content, for which anti-inflammatory and analgesic activity was previously proven, in a standard sample. The main method for establishing a substance quantitative content in the SS is the material balance method. The water determination was carried out according to K. Fisher's method (semimicro method). Sulphated ash was determined according to the XIV edition Russian Federation State Pharmacopoeia General Pharmacopoeia Monograph "Sulphated ash". Related impurities and their content were assessed using the HPLC method on a Flexar liquid chromatograph equipped with a diode array detector (Perkin Elmer, USA). The residual solvents' determination was carried out by the headspace method using a gas chromatograph GC-2010Plus Shimadzu with a flame ionization detector. As an additional method for establishing the main component quantitative content, acidimetric titration with the equivalence point potentiometric indication was carried out.

Results and discussion. The percentage was determined for the following indicators: water, residual organic solvents, related impurities, sulphated ash. Using the material balance method, it was found that the 5-butyl-1,2-diphenyl-6-oxo-1,6-dihydropyrimidin-4-olate sodium percentage in a standard sample is 96.01 ± 0.50 %. It was found by acidimetric titration that the 5-butyl-1,2-diphenyl-6-oxo 1,6-dihydropyrimidin- 4-olate sodium quantitative content in SS is 95.12 ± 0.02 %. The difference in the certified value can be explained by the fact that during titration, the SS aciform is released, which precipitates in an aqueous medium and contributes to a shift in the equilibrium and pH value. Consequently, the equivalence point is reached somewhat earlier. However, the data are practically comparable, but it is necessary to use the value obtained by the material balance method.

Conclusion. A standard sample certification parameters were determined: water content, residual organic solvents, sulphated ash, related impurities. The main component quantitative content was determined using the material balance method and titrimetry (acidimetry with the equivalence point potentiometric indication).

Introduction. Orthilia secunda (L.) House is a perennial herb that grows in Europe, Siberia, Asia Minor and Central Asia. The herb of Orthilia secunda is actively used in folk medicine as a diuretic, wound-healing and anti-inflammatory agent. From literary sources it is known that this medicinal plant raw material (PRM) contains flavonoids, tannins, organic acids, vitamins, as well as simple phenols and their derivatives (arbutin and hydroquinone). The presence of arbutin is responsible for the plant's high antioxidant and anti-inflammatory properties. But the use of Orthilia secunda in official medicine is limited due to the lack of complete information on the chemical composition and criteria for standardization of this type of medicinal product.

Aim. Identification and quantification of arbutin by chromatographic methods in Orthilia secunda (L.) House, harvested in various phytocenotic zones.

Materials and methods. The investigated medicinal plant material – the herb of Orthilia secunda – was harvested in various phytocenotic zones: in July 2018, harvesting was carried out in the northern part of Kazakhstan (Kokshetau district), in July-August 2019 in the Perm Territory and in the Tyumen Region. Preliminary identification of arbutin and related phenols – gallic acid and hydroquinone – was carried out by high performance thin layer chromatography (HPTLC) on a CAMAG instrument with a UV cabinet (Merck HPTLC silica gel 60 F154 plates, 20 × 10), semi-automatic Linomat 5 applicator (sample application). Elution of the plates was performed in a CAMAG Automatic Developing Chamber (ADC2). Image fixation was performed on a CAMAG Scanner 3 spectrodensitometer. The quantitative determination of arbutin was carried out by the method of highperformance liquid chromatography, which was carried out on a Prominence LC-20 device (Shimadzu, Japan) according to the validated method described in the European Pharmacopoeia 10.0. Diode array detector SPD-M20A, column Intersil C18 column (250–4.6 mm, 5 μm) (Phenomenex, USA). The results were processed using the LabSolution software. The identification and quantification of arbutin was carried out in comparison with a standard solution containing a reference sample (RS) of arbutin (C = 0,025 mg/ml) and RS of hydroquinone (C = 0,0125 mg/ml).

Results and discussion. HPTLC analysis made it possible to detect arbutin and gallic acid – the main product of hydrolytic degradation/ precursor of the biosynthesis of tannins of the hydrolysable group – in the herb of Orthilia secunda from different places of growth. HPLC analysis demonstrates a different chromatographic profile of Orthilia herb harvested in different phytocenotic zones. However, in all studied objects, the absence of hydroquinone and the presence of substances that can presumably be attributed to its derivatives were confirmed, which is confirmed by the visual similarity of the spectra of these compounds and the proximity of the extrema. It was found that arbutin does not belong to the marker (majority) compounds of Orthilia. Its content is low and reaches a maximum (about 0,021 %) in the herb of Orthilia secunda growing on the territory of Kazakhstan, while in the herb of Orthilia harvested in the Perm Territory arbutin was not identified. From the data obtained, it follows that the greatest accumulation of arbutin occurs in areas with a warmer and drier climate (northern part of Kazakhstan).

Conclusion. HPTLC analysis of the herb Orthilia secunda allowed the identification of arbutin and gallic acid (the main precursor of tannins of the hydrolysable group). The results of HPLC analysis of Orthilia herb harvested in various phytocenotic zones suggest quantitative differences in the content of arbutin depending on the region of growth. From the experimental data, it follows that Orthilia growing in the northern part of Kazakhstan accumulates the maximum (0,021%) amount of arbutin, in comparison with the samples harvested in the Tyumen region and the Perm region. At the same time, Orthilia harvested in the Perm Territory does not accumulate arbutin. The presence of hydroquinone has not been confirmed (by HPTLC and HPLC methods); therefore, it is not justified to talk about the hydrolytic cleavage of arbutin in the process of biosynthesis or drying. However, in all studied objects there are peaks of substances with spectral characteristics like hydroquinone, which makes it possible to assume the presence of its derivatives. Therefore, it is not advisable to position arbutin as a marker compound of Orthilia secunda harvested on the territory of the Russian Federation, and to standardize raw materials for this compound.

Introduction. The article presents the results of the detection of pharmacologically active secondary metabolites in black crowberry Empetrum nigrum L. using the method of high performance thin layer chromatography (HPTLC).

Aim. To show the efficiency of HPTLC for conducting preliminary phytochemical analysis to determine the main groups of metabolites in promising medicinal plant species.

Materials and methods. HPTLC analysis was carried out on a CAMAG device (Switzerland), using MERCK HPTLC silica gel 60 F154, 20 × 10 cm plates. For the evaporation of the samples, a Heidolph vacuum rotary evaporator (Germany) was used. The aerial parts (shoots) of Empetrum nigrum were harvested next to St. Petersburg State Chemical and Pharmaceutical University (SPCPU) nursery garden of medicinal plants (Leningrad Region, Vsevolozhsky District, Priozerskoe Highway, 38 km) in August 2019.

Results and discussion. In the course of the research, four fractions from the aerial parts of Empetrum nigrum were obtained: hexane, dichloromethane, butanol, and water. Then, these fractions were investigated by HPTLC in two solvent systems – n-butanol : acetic acid : water (BAW) (4 : 1 : 2) and hexane : dichloromethane : methanol (HDM) (1 : 2 : 0.5). After scanning densitometric analysis of the plates eluted in the HDM system, it was revealed, that the hexane and dichloromethane fractions have a similar composition and contain the greatest amount of compounds, compared to the butanol and water fractions, and in the BAW system, it was found, that the butanol fraction contains the greatest variety of metabolites. As a result of UV spectroscopy, it was found, that the main groups of compounds contained in the hexane and dichloromethane fractions are derivatives of chalcones, dihydrochalcones, bibenzyls and 9,10-dihydrophenanthrenes. While in the butanol fraction, the main groups of secondary metabolites were derivatives of flavonoids and tannins

Conclusion. The data obtained allow us to note the efficiency, speed and simplicity of HPTLC for conducting preliminary phytochemical analysis to determine the main groups of metabolites of promising medicinal plant species.

Introduction. A number of studies have shown that various genetic and environmental factors can affect the biosynthesis and accumulation of secondary metabolites. In particular, it is known that the local geoclimate, seasonal changes, external conditions such as light, temperature, moisture and soil fertility can affect the chemical composition and, as a result, the therapeutic properties of plants used in the pharmaceutical and food industries. Empetrum nigrum L. is a rich source of various pharmacologically active secondary metabolites – chalcones, dihydrochalcones, bibenzyls, 9,10-dihydrophenanthrenes, flavonoids, and proanthocyanidins. In the scientific literature, there is no data on the variation in the chemical composition of E. nigrum depending on the growing area. The obtained data are necessary for a reasonable choice of the collecting location for the plant, with the aim of its further chemical and pharmacological research for the development of promising drug candidates.

Aim. To carry out a comparative analysis of secondary metabolites composition in the aerial parts of Empetrum nigrum growing in different regions of the Russian Federation.

Materials and methods. Samples collected in three different areas were used to compare HPLC profiles: sample 1 was collected next to SPCPU nursery garden of medicinal plants (Leningrad region, Vsevolozhsky district, Priozerskoe highway, 38 km) in July 2020, sample 2 was collected on the Kola Peninsula, around the Khibiny mountains in July 2020, sample 3 was collected on the Kamchatka Peninsula, next to Khalaktyrsky beach (Pacific Ocean coast) in July 2020. Extracts were analyzed by analytical high performance liquid chromatography (HPLC) using a Prominence LC-20 device (Shimadzu, Japan) equipped with a diode array detector.

Results and discussion. As a result of the research, for the first time, a significant variation in the qualitative chemical composition in the aerial parts of Empetrum nigrum growing in different regions of Russian Federation was established. Sample 3, collected on the Kamchatka Peninsula, in comparison with samples 1 and 2, contain the greatest variety of polar secondary metabolites belonging to the classes of flavonoids, tannins, and phenol carboxylic acids, while in the shoots collected in the Leningrad region, the major metabolites were weakly polar compounds belonging to the classes of chalcones, dihydrochalcones, bibenzyls, and 9,10-dihydrophenanthrenes, and in sample 2, collected in the Khibiny mountains, the lowest qualitative content of secondary metabolites was found. This variation may be caused by various environmental factors (biotic and abiotic).

Conclusion. For the first time, the comparison of HPLC profiles of aerial part samples of E. nigrum, collected in different regions of the Russia Federation has been carried out. As a result, significant variations in the secondary metabolites composition of the studied samples were established, depending on the regions and growing conditions of the plants. The data obtained can be used for a reasonable choice of the collection location for the plant, with the aim of its further chemical and pharmacological research for the development of promising drug candidates.

Introduction. Non-alcoholic fatty liver disease is one of the most common chronic diseases of this parenchymal organ among the adult population. The search and creation of supporting drugs is an urgent task of modern pharmacy. The malonic acid derivative, sodium 4,4'-(propanediamido) dibenzoate, synthesized by the employees of the Department of Organic Chemistry of the SPСPU, has antisteatous activity, is a potential agent for the treatment of liver diseases. Sustained release tablets were prepared based on sodium 4,4'-(propanediamido)dibenzoate. An integral part of the pharmaceutical development of a medicinal product is the development of a method for conducting the Dissolution test and the selection of optimal conditions, which became the purpose of this study.

Aim. To develop the "Dissolution" test method for the sustained-release tablets based on sodium 4,4'-(propanediamido)dibenzoate.

Materials and methods. The objects of research are the active pharmaceutical ingredient sodium 4,4'-(propanediamido)dibenzoate, as well as sustained-release tablets based on this substance. Equilibrium biopharmaceutical solubility was determined by UV-spectrophotometry. To establish the conditions for the "Dissolution" test, an ERWEKA DT-620 dissolution tester (ERWEKA GmbH, Germany) was used.

Results and discussion. The suitability of the UV-spectrophotometry method for the quantitative determination of sodium 4,4'-(propanediamido) dibenzoate in solutions was determined. The established high biopharmaceutical solubility of sodium 4,4'-(propanediamido)dibenzoate in a buffer solution with a pH of 6,8, as well as in a 0,01 M solution of hydrochloric acid with a pH of 2,6, determined the choice of these media for the "Dissolution" test of the dosage form. The apparatus "Rotating basket" (rotation speed of 100 rpm in a dissolution medium with a volume of 1000 ml) was reasonably chosen for the test on the basis of the obtained linear dependence of the rate of release of the substance on time, as well as the best test results by the end of the experiment.

Conclusion. A study of the biopharmaceutical properties of the original substance with antisteatous activity has been carried out. High biopharmaceutical solubility was established in media with pH 2,6 and pH 6,8. The conditions of the "Dissolution" test for sustained-release tablets based on the original sodium 4,4'-(propanediamido)dibenzoate were experimentally substantiated.

Introduction. Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the world, and non-alcoholic steatohepatitis is the second most common cause of liver transplantation in the adult population. An urgent task is to find and develop an optimal model of NAFLD in laboratory animals, which would reproduce all the features of this disease in the clinic.

Aim. Modification of the NAFLD model in laboratory animals (rats), which allows the obtained data to be transmitted to humans as fully as possible.

Materials and methods. The study was conducted on 52 outbred white male rats of the same age. As the basis of the model, a hypercaloric high-fat diet was used with the addition of food appeal enhancers (sodium glutamate and liquid shrimp extract) and for the first-time conditions of hypodynamia were used – restriction of the motor activity of animals using specially designed cells, in which an individual 11 × 18 cm cell was allocated for each individual. The duration of the study was 12 months. In the course of the experiment, body weight, physical performance, biochemical parameters of blood serum and urine in dynamics were assessed, and lethality was recorded. After the end of the study, the mass of internal organs, visceral and epididymal fat was analyzed, and a histological examination of the liver was performed.

Results and discussion. In the course of the experimental study, the development of NAFLD in rats of the control group of animals was histologically confirmed. A high mortality rate was revealed in the group of animals with pathology. Compared with animals of the intact group, a statistically significant increase in their body weight, liver weight, visceral and epididymal fat, a decrease in physical performance, disturbances in lipid, carbohydrate and protein metabolism were revealed, as well as signs of deterioration of the protein synthesis and excretory functions of the liver.

Conclusion. A number of advantages of the NAFLD model with a combination of a hypercaloric diet and hypodynamic conditions were revealed, including the similarity of the conditions for the formation and pathogenesis of the disease in experimental animals and humans, which ensures the adequacy of data translation from preclinical practice to clinical practice.

Introduction. Selenopyran is an organic selenium compound with sharply hydrophobic properties. An increase in solubility in water (and as a consequence – and bioavailability) is possible due to the formation of inclusion complexes with cyclodextrins.

Aim. The aim of this work was to study the effect of a gel containing a clathrate complex of selenopyran with β-cyclodextrin on the rate of wound healing on a model of a conditionally aseptic full-thickness planar wound in rats.

Materials and methods. The object of the study was a gel containing a clathrate complex of selenopyran with β-cyclodextrin (the content of selenopyran in the gel was 0.1 %). A model of a full-thickness planar wound in sexually mature male rats was used. 20 individuals were divided into 2 groups – intact (without treatment) and experimental (received gel treatment). Efficacy was assessed by the change in wound area at 3, 5, 7, 9, 11 and 14 days after application of wound.

Results and discussion. The results of the study showed that the relative area of the wounds in the treated animals by the 3rd day of the experiment was less than in the intact ones. On the fifth day of the experiment, the differences were statistically significant (57.49 ± 12.51 % in treated animals versus 85.27 ± 26.61 % in intact animals). By the 14th day of the experiment, there were practically no differences in the groups of animals.

Conclusion. The results obtained indicate that when using a gel containing selenopyran in combination with β-cyclodextrin, it accelerates the transition from the inflammation phase to the proliferation phase. This is most likely due to the antioxidant properties of selenopyran. Considering the lower concentration of selenopyran in comparison with the therapeutic concentrations of other antioxidants (taurine, allantoin), it can be considered as a promising wound healing agent for further study.

Introduction. Alopecia is a polyetiological disorder characterized by hair loss and reducing their number per unit area. Baldness causes psychological and social discomfort to patients, in connection with what an important task is to develop formulations that are more effective than the reference agents.

Aim. Investigate the possibility of applying the original substance Y in several dosage forms for the treatment of alopecia in comparison with reference drugs: minoxidil and burdock oil.

Materials and methods. The research subject was the original substance Y, for which several dosage forms were made: gel, alcohol and oil compositions. The study on the effectiveness and safety of the developed formulations was carried out on 9 groups of male C57BL/6 mice. Depilation with further assessment of the percentage of hair follicles in the growth and resting phases was tested as a pre-clinical model of alopecia. In the study of the mechanism of action of substance Y, chemiluminescent assay was performed compared with natural antioxidant quercetin in the system luminol – 2,2'-azo-bis(2-amidinopropane)dihydrochloride, in potassium-phosphate buffer medium (pH = 7.4). Statistical processing of the results was carried out using two-way ANOVA using GraphPad Prism 8.0.2, USA software at the level of statistical significance of differences p < 0.05 and p < 0.005.

Results and discussion. Based on the results of histological analysis and visual changes, it was found that the effectiveness of the topical forms of substance Y decreases in the following order: gel, alcohol form, oil composition. The use of a combination of the gel base with the test substance Y resulted to the appearance of a larger number of hair follicles in the growth phase than when using the reference preparation – 2 % minoxidil solution (the differences are statistically significant). Chemiluminescent assessment of antioxidant activity showed the lack of antioxidant effect in substance Y.

Conclusion. The study combines two pharmaceutical profiles: technological and pharmacological. In the course of the experiments, the prospects of the gel form of the original substance Y for topical therapy of alopecia were shown. In the near future, it is planned to study the mechanism of action of substance Y, as well as registration of patent protection for a new drug.

Introduction. The search for and development of new drugs capable of reducing the severity of neurological deficit in traumatic brain injury are a critical task for investigational pharmacology. Chromone-containing allylmorpholines are a new group of neuroprotective drug candidates that have been shown to inhibit acetylcholinesterase and butyrylcholinesterase, and block N-methyl-D-aspartate receptors in vitro.

Aim. This study aimed to evaluate the neuroprotective activity of the allylmorpholine derivative (E)-4-[3-(8-bromo-6-methyl-4-oxo-4H-chromen- 3-yl)-1-cyclohexylallyl]morpholin-4-ium chloride (33b) in vivo using a rat model of traumatic brain injury.

Materials and methods. Traumatic brain injury was induced using the controlled cortical impact model. The allylmorpholine derivative was administered intraperitoneally at 1, 10, or 50 mg × kg-1 b.w. at 1 h after trauma induction, and then daily for the next 6 d. The neurological deficit was assessed using the Limb Placing, Open Field, Elevated Plus Maze, Beam Walking, and Cylinder tests.

Results and discussion. At all doses administered, the allylmorpholine derivative had no positive effect on the motor function or exploratory behavior following traumatic brain injury. In the Elevated Plus Maze, 10 mg × kg-1 b.w. of the compound further suppressed exploratory behaviour in the injured animals, which appears to be consistent with its sedative properties observed previously in zebrafish.

Conclusion. Despite the previously described in vitro affinity of allylmorpholines towards several molecular targets crucial for the pathogenesis of brain trauma and posttraumatic functional recovery, an allylmorpholine derivative had no neuroprotective effect in a rat model of traumatic brain injury in this study. These results further emphasize the importance of in vivo evaluation of potential neuroprotective drug candidates.

Introduction. Lately, medical services have reported a lot of cases caused by taking Tropicamide alone or with other drugs together. Moreover, it has been declared that the increase in the number of resistance cases to Tropicamide consumption has. Due to those facts, Tropicamide was included in the List of Drugs for Medical Use that should be served by the prescriptions in 2015. However, nowadays in Russia there are many combinations of medicines, for instance, Tropicamide and α-adrenergic agonist (phenylephrine) (Midrimax, Fenikamid, Appamide plus) that are not under that regulation. As a result, those medicines are served in pharmacies without any prescriptions. Thus, method developing for Tropicamide determination in the hair samples to establish his consumption period has become a perspective one.

Aim. The research aimed to develop a method for the isolation and determination of Tropicamide in the hair samples.

Materials and method. Reference standard of Tropicamide was used in this research. The following enzymes – papain, chymopsin, chymotrypsin, and hyaluronidase – were applied in the experiment. To design the long-term consumption of Tropicamide, laboratory animals (Guinea pigs, average masses about 200 – 250 g) with fair and brown nature colour hair were used in this research. The hair of laboratory animals was dyed by professional hair-dye "Estel Professional De Luxe". The following equipment was applied: balance "Sartorius СР224S", pH-meter " FiveEasy ", ball mill Retsch MM-200. The hair samples extracts were analyzed by gas chromatography with mass selective detection (Gas chromatograph model 7890А with mass selective detector model 5977 and MassHunter GC/MS software by Agilent Technologies).

Results and discussion. All developed methods of enzymatic hydrolysis (by papain, chymopsin, chymotrypsin, and hyaluronidase) revealed comparable results for the Tropicamide determination in the hair samples. The research showed that the amount of the analyte isolated from the pigmented hair was a bit higher in comparison with the other hair samples (fair hair), despite the melanin gives chemical steadiness property to hair stuff. Moreover, the amount of Tropicamide extracted from the dyed hair samples increased by 30 %. The degradation products of the analyte of interest were not found in the extracts obtained for the dyed hair samples. Thus, the colorant does not destroy the xenobiotic during the hair dying procedure and does not impact the enzymatic hydrolysis process. The values of the validation parameters (precision and accuracy) met the required criteria for bioanalytical methods. Therefore, the enzymatic hydrolysis method can be recommended for application in laboratory practice.

Conclusion. In the course of the study, a method for laboratory diagnostics of non-drug use of tropicamide was developed, the reproducibility of which meets the acceptance criteria for bioanalytical methods, which makes it possible to recommend it for work in laboratory practice.

Introduction. Type 2 diabetes mellitus is currently considered one of the most common non-communicable diseases. For the prevention and concomitant treatment of this pathology, various herbal remedies are successfully used, such as, for example, blueberry shoots. The plant contains phenolic compounds (anthocyanins, flavonoids, phenolcarboxylic and organic acids), which have antioxidant and hypoglycemic effects, and also accumulates macro- and microelements (Ca, Mg, Zn, Mn), which in turn can affect the course of diabetes mellitus. Complexes of elements with phenolic biological active substances (BAS) can affect the formation of a pharmacological response or change its severity. Therefore, it is possible to put forward a hypothesis about the potentiation of the antidiabetic action of phenolic compounds when they exist in the form of mineral complexes.

Aim. To carry out a comparative assessment of the antidiabetic activity of the mineral complex rutin with zinc in comparison with precursor substances and extraction from blueberry shoots to predict the effect of elements on the course of this pathology.

Materials and methods. The objects of the study were an aqueous solution (C = 0.18 mg/ml) of a model complex of rutin with zinc with a molar ratio of components of 1 : 1 and blueberry shoots purchased from a pharmacy in St. Petersburg. According to the information on the packaging, the region of raw material procurement is Altai Territory, Barnaul, the period for harvesting blueberries is July 2020. The complex of rutin with zinc was obtained according to the method described in the literature from the pharmaceutical substance rutin (Rutin, batch 332, valid until 26.03.2023, Sichuan Guangsong Pharmaceutical Co., Ltd., China, FS 000569-060514) and an aqueous solution (С = 0.13 mg/ml) zinc chloride (Zinc chloride, batch 39/G 4, valid until 09.10.2021, Neva Reaktiv, Russia, STP TU COMP 1-533-2012). The optimal ratio of components 1 : 1 for the formation of a mineral complex was established by us earlier experimentally using the Job's method. The mass of zinc chloride, which must be added to the extraction, and the mass of the complex for the preparation of its aqueous solution were calculated on the basis of the quantitative content of biologically active substances in blueberry shoots and the molar ratio of the components involved in the formation of the complex compound determined by the spectral method. The quantitative content of the main groups of biologically active substances (flavonoids, hydroxycinnamic acids, organic acids) was determined spectrophotometrically on SF-2000 instrument (Russia) and titrimetrically using the methods presented in Russian Pharmacopoeia XIV FS.2.5.0093.18 and FS.2.5.0012.15. The antidiabetic effect of the complex of rutin with zinc was evaluated in comparison with an aqueous extract from the shoots of common blueberries (the ratio of raw materials: extractant – 1 : 80), an aqueous solution of zinc chloride (concentration – 0.36 mg/ml) and their mixture (ratio 1 : 1) on a model of dexamethasone-induced type 2 diabetes mellitus in laboratory animals. Determination of blood glucose concentration was carried out using a portable glucometer "AccuChek Active" (Roche Diabetes Care, Germany). The presence of glucosuria and ketonuria was established using Ketoglyuk-1 test strips (Biosensor AN, Russia). Statistical processing of the results was carried out using the Microsoft Excel program according to OFS.1.1.0013.15.

Results and discussion. The results of the content of glucose and ketones in urine showed that in the process of modeling diabetes mellitus, glucosuria and ketonuria are characteristic for all groups of animals, which confirmed the formation of a pathological process in rats. A lower content of glucose and ketones in urine was found in a group of animals that received a complex of rutin with zinc (glucose concentration – less than 2.3 mmol/l, ketone concentration – less than 0.2 mmol/l). Measurement of the concentration of glucose in the blood showed that the complex of rutin with zinc has the most pronounced hypoglycemic effect, while the water extract from blueberry shoots and an aqueous solution of zinc chloride have a minimal antidiabetic effect (blood glucose is 6.9 mmol/l versus 8,1 mmol/l and 7.9 mmol/l, respectively).

Conclusion. The study of the influence of elements in the composition of phenolic complexes on the course of diabetes mellitus showed that the introduction of minerals has a positive effect on the severity of the pharmacological effect, which can serve as confirmation of the hypothesis about the potentiation of the antidiabetic effect of phenolic compounds when they exist in the form of mineral complexes. Thus, the complex of rutin with zinc showed the maximum activity in comparison with the solution of zinc chloride, extraction from blueberry shoots and their mixture, which suggests that the form of existence of natural phenolic compounds in the composition of mineral complexes is optimal both from the point of view of accumulation and from the side manifestations of a pharmacological response. The results of the study create the preconditions for further study of the effect of various elements in combination with marker phenolic components of antidiabetic herbal preparations on the course of diabetes mellitus, and also make it possible to conclude that natural mineral complexes have a prophylactic effect against this pathology.

Introduction. Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the world. Non-alcoholic steatohepatitis (NASH), a clinically progressive morphological form of NAFLD, ranks second in the list of reasons for liver transplantation in the adult population. In the pathogenesis of this disease, metabolism and distribution of free fatty acids (FFA) play an important role. A large number of studies have established that the level of FFA in peripheral blood directly correlates with the severity of NASH, but it is still unclear what effect fluctuations in the profile of fatty acids (FA) in the liver have in steatohepatitis.

Aim. Study of changes in the profile of fatty acids in the liver of laboratory animals with experimental non-alcoholic steatohepatitis.

Materials and methods. The study was carried out on 17 white outbred male rats, which were randomized into two groups – intact (n = 6) and control (steatohepatitis) (n = 11). Steatohepatitis was modeled by 12-month use of a hypercaloric high-fat diet against the background of hypodynamia. The content of fatty acids in the liver was determined in the reaction of methanolysis and extraction with a hexane mixture of their methyl esters. The LC was separated by gas chromatography-mass spectrometry. Calibration for quantitative calculation was carried out with deuterated tridecanoic acid. The content of saturated and monounsaturated higher FAs, their aldehydes and hydroxy derivatives, as well as sterols were studied.

Results and discussion. A total decrease in the content of FFA in the liver of animals with steatohepatitis was revealed. The most significant decrease occurred mainly in the class of monounsaturated fatty acids and cholesterol. Also, a significant decrease in the activity of Δ9-desaturase, a key enzyme in the synthesis of monounsaturated FAs from their precursor with the same carbon chain length, was revealed, which was manifested by a significant decrease in their amount in the liver. There were no statistically significant changes in the levels of aldehydes and hydroxy acids between the study groups, as well as in the level of sterols (except for cholesterol, the content of which decreased significantly).

Conclusion. Thus, in the liver of rats with steatohepatitis caused by a combination of a hypercaloric diet and hypodynamia, statistically significant changes in the profile and concentration of fatty acids were found in comparison with healthy animals. The demonstrated shifts in FA composition may reflect both adaptive and pathological changes in the liver of animals with NAFLD and require further study.