Vol 21, No 5-6 (2021)

BACKGROUND: Most drugs in the form of tablets contain diosmin in the micronized flavonoid fraction. This is the reason for the low level recovery by means of solid-liquid extraction.

AIM: to determine optimal conditions for sample preparation of some drugs with active pharmaceutical ingredient diosmin with the use of mathematical experiment planning (Box-Behnken design).

MATERIALS AND METHODS: The study subjects were tablets Detralex® 1000 mg, Venarus® 900 mg, Detravenol® 1000 mg, Flebaven® 1000 mg, Phlebopha® 600 mg, Phlebodia 600 mg. Estimation of diosmin recovery by the method of solid-liquid extraction was made according to the algorithm of full factorial design with three level factors (Box-Behnken design).

RESULTS: The effect of some factors determining the extraction of diosmin from tablets Detralex® 1000 mg, Venarus® 900 mg, Detravenol® 1000 mg, Flebaven® 1000 mg, Phlebopha® 600 mg, Phlebodia 600 mg was estimated. Mathematical (regression) models of solid-liquid extraction of diosmin for each drug were developed. The most significant factor determining the diosmin recovery from drugs (tablet formulation) is the sodium hydroxide concentration. Other factors (acetonitrile concentration, extraction time, centrifugation time, addition of ammonium sulfate) fail to exert significant effect. The optimal value of sodium hydroxide concentration is 0.02 mol/l. Diosmin recovery doesn’t increase after the increase of the sodium hydroxide concentration above 0.02 mol/l.

BACKGROUND: Medicinal plant raw material “Licorice roots” contains a whole complex of biologically active compounds (triterpene saponins, flavonoids, etc.) and is widely used in medicine as raw materials for the production of anti-inflammatory and expectorant medications. However, despite its widespread use and well-studied chemical composition, the problems in the field of standardization of licorice roots remain urgent. The current pharmacopoeial method for determining the quantitative content of glycyrrhizic acid is multi-stage, it consists in the extraction of nitric acid with the use of 3% acetone solution, the addition of the concentrated ammonia solution to form a precipitate (ammonium salt of glycyrrhizic acid), its filtration and dissolution in water, followed by spectrophotometric determination. There are other methods for determining the content of this biologically active compound. They involve high performance liquid chromatography. No results for quantification of glycyrrhizic acid by this method in isocratic elution mode were found in the available scientific literature.

AIM: The aim is to develop the method for determining the quantitative content of glycyrrhizic acid in licorice roots using high performance liquid chromatography in an isocratic elution mode.

MATERIALS AND METHODS: The objects of the study were licorice roots harvested in 2018 in the Botanical Garden of Samara University, commercial samples of plant raw materials, state standard samples of monosubstituted ammonium salt of glycyrrhizic acid (glycyram, PM 42-0034-00) and the work glycyrrhizic acid standard. High performance liquid chromatography analysis was carried out by using a Milichrom-6 chromatograph.

RESULTS: As a result of the investigation, the method for determining the quantitative content of glycyrrhizic acid by reversed-phase high performance liquid chromatography with UV detection (258 nm) was developed. The mobile phase was acetonitrile: 1% acetic acid solution in water in the ratio of 40:60, the elution rate was 100 μl/min. The optimal extraction conditions were selected: extractant was 40% ethyl alcohol, extraction for 60 minutes at the ratio of “raw material-extractant” was equal to 1:30. Validation analysis showed that the developed methods demonstrated satisfactory metrological indicators.

CONCLUSIONS: The obtained results can be used to update the pharmacopoeial monograph “Licorice roots”.

BACKGROUND: Juglans nigra L. is a species of trees in the Juglandaceae family. This plant of the genus Juglans L. has not been sufficiently studied in comparison with other species, including Juglans regia L. It is a perspective medicinal plant raw material, its derivatives have an antimicrobial, general tonic effect. The known antimicrobial and general tonic activity of Juglans L preparations presented on the pharmaceutical market is probably due to substances of a flavonoid nature. These data indicate the relevance of studying the flavonoids of the Juglans nigra L. bark.

AIM: The aim of this study is to develop methods for qualitative analysis and quantitative determination of biologically active compounds for tincture of Juglans nigra L. bark with the using of spectrophotometry, thin layer chromatography, microcolumn high performance liquid chromatography.

MATERIALS AND METHODS: The object of the study was the tincture of Juglans nigra L. bark, samples of which were prepared in March-April 2020 on the territory of the Botanical Garden of Samara University, standard samples of myricitrin, myricetin. UV spectra were recorded with the use of Specord 40 spectrophotometer (Analytik Jena). Chromatographic analysis was carried out by thin-layer chromatography (qualitative analysis), reversed-phase high performance liquid chromatography on a Milikhrom-6 microcolumn liquid chromatograph (quantitative analysis).

RESULTS: As a result of the chromatographic study, myricitrin were revealed, it appeared to be the dominant flavonoid substance. This compound has R_f value of about 0.4 in the solvent system of chloroform-ethanol-water (25:18:2). A method for the quantitative determination of the total flavonoids calculated on myricitrin in tincture of Juglans nigra L. bark was developed by means of differential spectrophotometry. It was determined that the content of the total flavonoids calculated on myricitrin in the tincture of Juglans nigra L. bark was 0.84 ± 0.07%. A method for the quantitative determination of myricitrin in tincture of Juglans nigra L. bark by high performance liquid chromatography was developed. The content of the dominant flavonoid as myricitrin (myricetin-3-O-α-L-rhamnopyranoside) in the tincture of Juglans nigra L. bark was 0.42 ± 0.06%.

CONCLUSIONS: The methods of qualitative analysis and quantitative determination of flavonoids for tincture of Juglans nigra L. bark with the use of modern instrumental methods of analysis (spectrophotometry, thin layer chromatography, high performance liquid chromatography) has been developed. These results can be used to standardize the tincture of Juglans nigra L. bark.

The article studies the influence of hymatomelanic acids, one of the fractions of humic substances of peloids, on the course of rheumatoid arthritis. This substance was standardized by using physicochemical analysis. The elemental composition of the substance as well as a number of spectral (IR, NMR) characteristics were revealed. The list of components formed as a result of hydrolysis of the initial preparation and their relative content was determined. Fatty acids and their derivatives are the predominant constituents of peloid hymatomelanic acid hydrolyzate. The tissues of immunocompetent organs including regional lymph nodes, spleen, and also rat liver were studied. For the experiment, a model of adjuvant arthritis was selected and four groups of animals were formed. They were withdrawn from the experiment on the 17, 21 and 26 day. The histological picture indicated that when modeling rheumatoid arthritis, such pathological changes in the immunocompetent organs of rats as edema of the parenchyma and stroma, sclerosis, thickening of the walls of blood vessels, develop during the whole period of observation. A list of descriptors of the studied drug was obtained and it was found that the use of peloid himatomelanic acids in the model of adjuvant arthritis had a pronounced anti-inflammatory effect, as evidenced by the normalization of the morphofunctional state of the spleen, regional lymph nodes and liver.

The article is devoted to the development of new moisture (loss on drying) determination techniques by infrared thermogravimetric method. This method is aimed at 23 pharmacopeial types of air-dry medicinal plant raw materials of 7 morphological groups (roots, bark, leaves, fruits, seeds, herbs, flowers). These techniques can be included into the State Pharmacopoeia of the Russian Federation. The obtained results allow to develop the project proposals concerning the changes and the supplementation of the State Pharmacopoeia of the Russian Federation monographs.

The fruits of various species of the genus Crataegus L. contain flavonoids and are widely used in domestic and foreign medicine for the production of cardiotonic drugs. Crataegus submollis Sarg. is grown in our country as the edible and ornamental plant, but it is not currently used in medicine. Fruit juice of Crataegus submollis Sarg. exerts diuretic effect and an antidepressant activity. Dried fruit pulp can be used to produce extraction preparations. The aim of the work is to compare and analyze the samples of tinctures obtained from the fruits of the Crataegus submollis Sarg. and the pulp of its fruits. Tinctures based on 70% ethyl alcohol were obtained on the basis of dried fruits of Crataegus submollis Sarg. and the pulp of its fruits. The quality of the obtained tinctures was studied by determining of the total amount of flavonoids. Tinctures made of dried pulp of Crataegus submollis Sarg. fruits are as good as industrial samples of preparations in terms of flavonoid content. The fruits of the Crataegus submollis Sarg. and the pulp of its fruits can serve as medicinal plant raw materials for obtaining tinctures.

BACKGROUND: Meadowsweet [Filipendula ulmaria (L.) Maxim.], is the representative of flora of the European parts of Russia. It is widespread in Samara Region. The flowers of the meadowsweet are the pharmacopoeial type of raw materials, but the fruits of this plant are not used in medicine. The fruits of the meadowsweet and its closely related type of the dropwort are the promising source of biologically active compounds of phenolic and sterol nature. Introduction of new types of medical plant raw materials into official medical practice requires the development of regulatory documentation, including morphological and anatomical diagnostics of types.

AIM: The purpose of this work was to study and compare morphological and anatomical features of the structure of meadowsweet fruits and dropwort.

MATERIALS AND METHODS: The material of the study was the dry fruits of the meadowsweet and dropwort, collected in June-July 2019 in Samara Region. The research was carried out by means of light microscopy in transmitted and reflected light in a light field.

RESULTS: Morphological and anatomical characteristics of the fruits of two types of meadowsweet were revealed. They hadn’t been described in the literature before. So the fruits of the meadowsweet have the following diagnostic features: the stele of the pedicels is of transitional type, lysigenic receptacles located in them are uneven, pubescence of the epidermis occurs due to simple unicellular trichomes; they also have the three-layer structure of the pericarp, a significant quantity of single crystals in the parenchyma of the parmelia, the cone-shaped form of the conducting bundle in the parmelia seam. The fruits of the dropwort are characterised by retort-shaped hairs that make up the thick pubescence of the multiple fruits, peculiar conducting system of the parmelia in the form of one large collateral bundle of the cone-shaped form, located on the side of the seam.

CONCLUSIONS: The obtained results allow to develop the chapter “Microscopic signs” as a part of the pharmacopoeial monograph devoted to a new perspective medicinal plant raw materials of “Fruits of the meadowsweet” and “Fruits of the dropwort”.

The food spirulina [Spirulina platensis (Nordst.) Geitl.] is a blue-green multicellular filamentous microscopic alga widely cultivated in many countries of the world, including the Russian Federation. The aim of this study is to develop approaches to the standardization of biomass of Spirulina platensis. As a result of the conducted research, approaches to the standardization of biomass of Spirulina platensis have been developed. It is aimed to determine the content of β-carotene, phycocyanin and proteins by using TLC and spectrophotometry. It was determined that the content of β-carotene in the samples of biomass of Spirulina platensis calculated on absolutely dry raw material varies from 63.0 mg% to 94.0 mg%, phycocyanin from 3.50% to 4.30% and protein from 53.50% to 61.09%. It was established that β-carotene, C-phycocyanin and protein were the most important diagnostically significant biologically active compounds of the biomass of Spirulina platensis. The analytical aspects of the study of Spirulina platensis biomass considered in this paper are the scientific basis for the substantiation of the creation of pharmaceuticals based on it.

BACKGROUND: Currently, fundamental and experimental data confirming the role of free radicals in physiological and pathological processes have been accumulated. The literature describes conflicting information about the effect of antioxidants on the organism: they are able to protect not only normal cells from the alterative effect of free radicals, but also tumor cells, thereby supporting the survival and growth of the latter. Therefore, the study of the effect of Thymus marchallianus extract on the processes of lipoperoxidation in vivo against the background of the tumor process is relevant.

AIM: To study the effect of Thymus marchallianus extract on the quantity of lipid peroxidation intermediates in the blood plasma of mongrel white rats with liver cancer PC-1 and compare with in vivo antioxidant activity.

MATERIALS AND METHODS: The object of research was the dried Thymus marchallianus grass. Water and alcohol extracts were used in the experiment. Fifteen male white mongrel rats with a tumor were included in the experiment: the first group was intact, the second group received the water extract of Thymus marchallianus, and the third group received the alcohol extract of Thymus marchallianus. The activity of lipoperoxidation processes was evaluated, the content of intermediate lipid peroxidation intermediates – malon dialdehyde, lipid hydroperoxides and medium-mass molecules in the blood plasma of experimental rats were estimated by means of conventional spectrophotometric methods. Determination of the antioxidant activity of the infusion and alcohol extraction in the in vitro experiment was carried out by the titrimetric method.

RESULTS: In the control groups and the group of animals that received intraperitoneal water and alcohol extracts of Thymus marchallianus, increase in the intermediate products of lipid peroxidation was not revealed. The amount of malon dialdehyde, lipid hydroperoxides and medium-mass molecules in the blood plasma of experimental rats did not differ significantly from the level of similar indicators of intact animals. In the in vitro experiment, water and alcohol extracts from raw Thymus marchallianus showed antioxidant activity.

CONCLUSIONS: For the first time, the experiment was conducted to study the effect of water and water-alcohol extracts of Thymus marchallianus on lipoperoxidation processes in mongrel white rats with alveolar liver cancer PC-1. The activity of formation of intermediate products of lipid peroxidation did not change, since their content in the blood plasma of experimental animals did not differ from the level of similar indicators of intact animals. In vitro the infusion and alcohol extract demonstrated antioxidant activity.

Nowadays, diseases of the respiratory system are a common problem for national and global healthcare. The drugs of synthetic and natural origin, intended for the prevention and treatment of these diseases, are available on the pharmaceutical market. The latter are mainly represented by herbal medicinal products, among which mixture herbal products occupy a special place. Pectoral species No. 2 is a multicomponent herbal medicinal products used for diseases of the upper respiratory tract, it includes coltsfoot leaves (40%), plantain leaves (30%), and licorice roots (30%). It is produced in the form of powder and is dispensed in packs and filter sachet. On the basis of information and analytical research, some approaches to the standardization of pectoral species No. 2 are developed in this study. Currently, the infusion is the main dosage form of pectoral species No. 2; water-soluble biologically active compounds are fully extracted by water and are responsible for the manifested pharmacological effects of pectoral species No. 2 (polysaccharides and triterpene compounds). The development of modern regulatory documentation for complex herbal preparations (mixture herbal products) should be carried out taking into account scientifically based data and harmonized requirements. Additional experimental studies are required to substantiate the indicators and standards of identity and good quality of pectoral species No. 2. Modern regulatory documentation for pharmaceutical substances of plant origin and mixture herbal products made of them (including pectoral species No. 2) should include sections that take into account the principle of “raw material-to-drug standardization” and methods that can be used in research laboratories with various material and technical support.

BACKGROUND: To identify the features of coronavirus infection (COVID-19) treatment in older working age individuals the analysis of the drugs assortment and treatment schemes used in COVID hospitals in Samara was carried out.

AIM: The purpose of the research was to study the drugs used to treat senior patients in the hospitals of Samara Region, to identify the main drugs, to form a range of medical preparations and to predict the volume of their consumption.

MATERIALS AND METHODS: The study carried out a retrospective analysis of 218 cases of patients with coronavirus infection treated in Samara medical institutions from May till July 2020.

RESULTS: The main symptoms, comorbidities, the average duration of symptom manifestation prior to admission in both groups of patients, as well as the essential drugs used to treat the infection have been identified.

CONCLUSIONS: The analysis of the studied drugs used for the treatment of coronavirus infections should form the basis for the formation of an optimal range of drugs, forecasting the need for drugs in the light of scientifical pharmacoeconomic approaches.

BACKGROUND: The new coronavirus infection (SARS-CoV-2), better known as COVID-19, quickly evolved into a worldwide pandemic with a significant public health burden. Currently, there are no approved drugs or preventive therapeutic strategies to combat infection. However, there are hundreds of clinical studies around the world to discover effective treatments for COVID-19.

AIM: The purpose of the research is to conduct a content analysis of the published clinical trials devoted to the treatment of COVID-19, as well as to the synthesis of clinical experience in order to gain insight into potentially effective drugs.

MATERIALS AND METHODS: Clinical results published in PubMed, Medline, eLIBRARY, Cochrane Library.

RESULTS: The review summarized the results of clinical studies of potential therapeutic drugs used in COVID-19 therapy.

CONCLUSIONS: By the 15 of August 2021 no medications had been registered to treat the infection.

AIM: To assess the level of expression of TLR2, TLR4, TLR7 genes in the placenta and fetal membranes during full-term physiological pregnancy, without signs of onset of labor.

MATERIALS AND METHODS: The study was carried out at the premises of City Clinical Hospital No. 1 named after N.I. Pirogov (Samara), and included 15 pregnant women at 37-41 weeks of gestation without somatic pathology, exacerbation of chronic and acute inflammatory diseases, obstetric complications (placental insufficiency, preeclampsia). All patients were delivered by elective caesarean section due to fetal malposition. The level of expression of TLR2, TLR4, TLR7 genes was determined by reverse transcription and real-time PCR (RT-PCR) on the basis of the laboratory of “NPO DNA-Technology” using a set of reagents “Proba NK”.

RESULTS: The data on the features of TLR2, TLR4, TLR7 expression in the placenta and membranes during full-term pregnancy were obtained. Direct correlations were found between the production of TLR2, TLR4, TLR7 of the fetal membranes and TLR2 of the placenta. The expression of placental TLR4 and TLR7 was not associated with the production of the TLRs. Significant differences were revealed in pregnant women with inflammatory and involutive-dystrophic changes in the placenta and the level of TLR4 expression.

CONCLUSIONS: The expression of the Toll-like receptors (TLRs) in the tissues of the fetoplacental complex occurs uniformly during physiological pregnancy. There are positive correlations between the expression of TLR2, TLR4, TLR7 of the fetal membranes and TLR2 of the placenta, as well as its mass. The most significant indicator of the inflammatory process is the placental expression of TLR4.

Nowadays it is important to improve the techniques to diagnose malocclusion class II which is very common in children. 24 patients with various maxillofacial anomalies were examined. The developed diagnostic tactics in children with various maxillofacial anomalies involves a set of sequential studies: teleroentgenography, cone-beam computed tomography, ultrasound, kinesiography and electromyography. The obtained results allow assessing the morphological and functional state of the temporomandibular joint in children with distal malocclusion.