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Introduction. Biorelevant dissolution media reconstitute the composition of the contents of the gastrointestinal tract. They are used as dissolution media in the evaluation of dissolution profiles of different dosage forms. Simulated biological fluids allow prediction of in vivo test results. The development of the composition of simulated salivary fluid allows the evaluation of drug properties under physiologically relevant conditions.Aim. Evaluation of the release of the drug product "glycine, sublingual tablets, 100 mg", domestically produced in Simulated Saliva 5 pH 6.8.Materials and methods. The preparations used for analysis were: «Glycine, sublingual tablets, 100 mg», domestically produced with valid expiration date. Comparative dissolution kinetics test was carried out on the dissolution test apparatus DT 6 (ERWEKA GmbH, Germany). Chromatographic separation and detection were performed on a Waters W1525 Binary HPLC Pump high-performance liquid chromatograph (Waters Corporation, USA) equipped with column and sample thermostat, degasser, autosampler and Waters 2487 Dual Absorbance Detector (Waters Corporation, USA). Detection was performed at a wavelength of 254 ± 2 nm after derivatization of the glycine molecule with 4-toluenesulfonyl chloride. A Grace Platinum C18-EPS 5 μm 4.6 × 250 mm Grace Platinum C18-EPS 5 μm 4.6 × 250 mm column (Grace, USA) and a Grace Platinum C18-EPS 5 μm 4.6 × 250 mm pre-column (Grace, USA) were used. The following software was used for the study: validated Microsoft Excel spreadsheet for calculating glycine release values.Results and discussion. The technique for quantitative determination of glycine was developed and validated under CDKT in purified water medium and Simulated Saliva 5 pH 6.8. The validated analytical range of the methodology was 10–110 % of the nominal concentration of the dosage form in 300 mL volume of medium. The developed analytical technique was validated in the biopredictive in vitro test of glycine preparations. During the study in Simulated Saliva medium for drug formulations, more discriminative data were obtained, which were expressed as: different dissolution rate, curvature of the slope of the dissolution profile and time to reach the plateau in contrast to the dissolution medium purified water.Conclusion. The quantification technique was developed and validated for biopredictive tests of tablets "Glycine, sublingual tablets, 100 mg". The analytical range of the technique was 10–110 % of the nominal concentration of the dosage form in 300 mL volume of medium. The results of the test in artificial saliva medium were more discriminatory.

Introduction. The introduction of devices – analogues of GIS (hereinafter–- Gastro-intestinal simulator) is one of the current ways to develop in-vitro assessment of the quality of solid dosage forms. Testing on a physiologically relevant test device (hereinafter referred to as PRT) makes it possible to predict pharmacokinetic profiles due to more relevant conditions, including the use of biorelevant dissolution media, physiological volumes of the gastrointestinal tract, as well as transit between them.Aim. Conduct a study of cladribine tablets on a physiologically relevant tester in order to predict the behavior of the drug in the human gastrointestinal tract.Materials and methods. The objects of the study are "Mavenclad®, tablets, 10 mg" (series 2200754, expiration date until 04.2025, NERPHARMA, S.r.L., Italy) and "Cladribine, tablets, 10 mg" of domestic production with valid expiration date. During the study, the reagents necessary for the preparation of biorelevant dissolution media and quantitative determination by HPLC. Physiologically relevant test were carried out using an apparatus of our own production, consisting of a DT-6 dissolution tester (ERWEKA GmbH, Germany), a water bath equipped with a Thermomix WB-4 heating element (B. Braun, Germany), and peristaltic pumps (Kamoer, China). The quantitative content of released cladribine was assessed using a highly efficient liquid chromatograph "Khromatek-Kristall HPLC 2014" (ZAO SKB "Khromatek", Russia) using a validated method at a wavelength of 252 nm, analysis time – 7 min, column – Grace HPLC Column Platinum C18-EPS, 250 × 4.6 mm, 5 mm (Grace, USA), temperature – 35 °C, elution mode – isocratic (A : B 80 : 20), mobile phase A – 0.1 % H3PO4 solution, phase B – acetonitrile.Results and discussion. Profiles were obtained to assess the dynamics and degree of release of the studied drugs in various parts of the human gastrointestinal tract. Despite the expected degradation of cladribine in an acidic environment (pH1.2), under physiologically relevant conditions, the drug reached the third section (small intestine model) without degradation. Complete release of cladribine from the test and reference dosage forms was observed. Also, in the future, based on the data obtained, it is possible to predict pharmacokinetic profiles using physiologically based pharmacokinetic modeling approaches.Conclusion. A PSF study was conducted for the drugs "Mavenclad®, tablets, 10 mg" and "Cladribine, tablets, 10 mg". Quantitative determination was carried out by HPLC-UV method. The test results showed complete release of both drugs and reaching the intestinal tract, indicating the absence of degradation of cladribine in the region simulating the stomach.

Introduction. The most important stage of pharmaceutical development of a generic drug is a clinical trial involving humans – a bioequivalence study. Considering the importance of finding rivaroxaban drugs in the list of vital and essential drugs, as part of ensuring technological sovereignty, the use of scientific robust and effective methods for determining the quality of the dosage form is required.Aim. Conduct a study of rivaroxaban tablets on a physiologically relevant tester to predict pharmacokinetic profiles.Materials and methods. The objects of the study are "Xarelto®, film-coated tablets, 10 mg" (series BXJS871, with an expiration date of October 31, 2024, Bayer AG, Germany), "Xarelto®, film-coated tablets, 20 mg" (series BXKDF32, with an expiration date of May 17, 2026, Bayer AG, Germany) and "Rivaroxaban, film-coated tablets, 10 mg" and "Rivaroxaban, film-coated tablets, 20 mg", domestically produced, with valid expiration dates. During the study, reagents were used to prepare dissolution media and perform quantitative determination. The physiologically relevant test was performed on the SC PRT-6 device (LLC "Scientific Compliance", Russia). The quantitative content of released rivaroxaban within the comparative dissolution kinetics test in a medium of 0.1 in a medium of 0.1 % sodium lauryl sulfate solution in a phosphate buffer solution pH 6.5 was carried out on a SF-2000 spectrophotometer (LLC "OKB Spektr", Russia). The quantitative content of released rivaroxaban within the comparative dissolution kinetics test in biorelevant dissolution media and physiological relevance test was assessed on a high-performance liquid chromatograph "Chromatec-Crystal HPLC 2014" (CJSC "Chromatec", Russia). Pharmacokinetic profiles were modeled in the PK-Sim® (Systems Biology Software Suite 11.2, Bayer Technology Services GmbH, Germany) program based on the data obtained within the physiologically relevant test. The clinical study of rivaroxaban tablets was a prospective, open-label, randomized, crossover, two-stage comparative study in two groups of volunteers with a single dose of drugs on the fast condition. The study randomized 30 healthy male volunteers aged 18–45 years.Results and discussion. A complex of in vitro tests was conducted, profiles were obtained that allow us to evaluate the dynamics and degree of release of the studied drugs in various parts of the human gastrointestinal tract. A comparison of the sequential and hybrid schemes for conducting the physiological relevance test was carried out. Within the framework of the set of tests, qualitative and quantitative correlation with the clinical trials data was observed only for the hybrid physiological relevance test scheme. Based on the results of physiological relevance test using different schemes, pharmacokinetic profiles for a pair of drugs were predicted and the prediction error was assessed.Conclusion. A set of scientific in vitro tests was conducted for the drugs "Xarelto®, film-coated tablets, 10 mg and 20 mg", "Rivaroxaban, film-coated tablets, 10 mg and 20 mg". Based on the physiological relevance test results, pharmacokinetic profiles for a pair of drugs were predicted with low error and high reliability. As part of the comparison of data obtained during clinical trial and modeling, the smallest prediction error was noted when performing physiological relevance test using a hybrid scheme.

Introduction. Deferasirox is a complexing drug and belongs to class II according to the biopharmaceutical classification system (BCS), has acidic properties and belongs to subclass “a” (acid). This class is characterized by high permeability and low solubility, which limits the absorption of the active substance into the blood. As a result, the development of drugs with an active substance that can be assigning BCS to this class is a difficult task, and for generic drugs it is also associated with a high risk of obtaining unproven equivalence during clinical trials. To minimize the above risks, a physiologically relevant test was carried out with further data processing and construction of putative pharmacokinetic profiles.Aim. The aim of the study is to conduct a physiologically relevant test (PRT) to predict in vitro pharmacokinetic profiles and compare them with in vivo data as part of a bioequivalence study of deferasirox.Materials and methods. The objects of the study are "Deferasirox, film-coated tablets, 360 mg" of a domestic manufacturer and "Jadenu®, film-coated tablets, 360 mg" (WTN22 series, expiration date until 10.2023, Novartis Pharma Stein AG, Switzerland). A physiologically relevant test was performed on the device SC PRT-6, Compliance. Quantitative analysis was carried out by HPLC-UV method.Pharmacokinetic profiles were modeled using PK-Sim® (Systems Biology Software Suite 11.2, Bayer Technology Services GmbH, Germany) based on data obtained from physiologically relevant test.Results and discussion. A physiologically relevant drug test for deferasirox was performed and release profiles were obtained, which formed the basis of a physiologically based pharmacokinetic model together with data on the physicochemical properties of the studied compound and literature data on the pharmacokinetics of deferasirox. The pharmacokinetic profiles obtained as part of the simulation on a virtual population were compared with data obtained during clinical trials.Conclusion. A physiologically relevant test for the drug deferasirox was carried out, quantitative determination in the samples was carried out by HPLC-UV. The test resulted in data that allowed prediction of pharmacokinetic profiles that reflected the same differences observed in the profiles of the test and reference drug in the comparative pharmacokinetics and bioequivalence study of deferasirox drugs.

Introduction. Sorafenib is an antineoplastic drug belonging to class IIc according to the biopharmaceutical classification system (BCS) due to the presence of both acidic and basic properties. In addition to low solubility, sorafenib is characterized by high variability during clinical trials, in particular bioequivalence studies (BE). To selecting batches that can be recommended for BE studies, the dissolution kinetics test is currently widely used, however, the results of this test are not always sufficient and additional tests, for example, a physiologically relevant test, are advisable. To minimize the risks of obtaining nonequivalent results during the BE study, a physiologically relevant test (PRT) was carried out with further data processing and interpretation of the results of physiologically based pharmacokinetic modeling (PBPK).Aim. The aim of the study is to conduct a physiologically relevant test (PRT) for the purpose of selecting a candidate batch for subsequent BE study of sorafenib drugs using the physiologically based pharmacokinetic model (PBPK).Materials and methods. The objects of the study are Nexavar®, film-coated tablets, 200 mg (Bayer AG, Germany) (one batch) and Sorafenib, film-coated tablets, 200 mg (two batches) (Russia). The physiologically relevant test was performed on the SC PRT-6 device (LLC "Scientific Compliance", Russia). Quantitative analysis was performed by HPLC-UV on the Chromatec-Crystal HPLC 2014 device (CJSC "Chromatec", Russia). The plasma concentration profiles were simulated using PK-Sim® software (Systems Biology Software Suite 11.2, Bayer Technology Services GmbH, Germany).Results and discussion. As part of the study, a method for the quantitative determination of sorafenib was developed and validated, a method for sample preparation was developed, and a method for conducting the PRT for sorafenib, as a representative of the IIc subclass of BCS, was developed. Based on the study results, release profiles were obtained that were used to select a candidate series for the BE study. The series were selected based on the PBPK analysis on a virtual population consisting of 36 healthy volunteers with activated enteropathic circulation, characteristic of sorafenib.Conclusion. The PRT was carried out for the drug sorafenib. Quantitative determination was carried out by HPLC-UV according to the developed and validated method. The test resulted in obtaining data that were subjected to PBPK analysis. It was shown that the studied batches have high risks of non-equivalence during the bioequivalence study.

Introduction. Search for new plant species containing biologically active substances (hereinafter – BAS) is one of the leading tasks of pharmacognosy as a science. The search for flavonoid glycosides in plant raw materials is especially relevant, since they have anti-inflammatory, antioxidant, immunostimulating, as well as weak fungicidal and bacteriostatic action. Staphylea pinnata L. is an endemic plant of the Caucasus, cultivated not only in Georgia, but also in the Russian Federation in the Northern and Northwestern Caucasus. In foreign literature there are studies of antioxidant, antiproliferative and cytotoxic activity of leaf extracts of several species of S. pinnata L., as well as inhibitory activity of COX-1, COX-2 and LTB4 formation. Meanwhile, no serious Russian-language scientific studies on either the chemical composition or pharmacological action of generative organs of S. pinnata were found in the literature. This work is part of a comprehensive phytochemical study of S. pinnata. The aim of the work is to study the qualitative and quantitative composition of flavonoids in the studied object.Aim. To isolate, identify and quantify flavonoids in flowers and buds of Staphylea pinnata L.Materials and methods. Alcohol-water extracts from dried generative organs of the studied plant were used as analyzed solutions. Solutions were analyzed on a spectrophotometer SF-2000 (LLC "OKB Spectr", Russia) after sample preparation with aluminum chloride and on an HPLC Nexera-i LC-2040 (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, degasser and autosampler using an individually selected elution gradient of the mobile phase (0.1 % orthophosphoric acid/acetonitrile solution). The primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan). Compounds from the flavonoid group were identified by retention times. Detection was carried out using a UV detector with an absorption wavelength of 365 ± 2 nm.Result and discussion. Alcohol-water extracts from flowers and buds of S. pinnata L. were obtained. Quantitative evaluation by spectrophotometry for flavonoid content was carried out. A gradient elution mode for HPLC was selected for simultaneous determination of 7 flavonoid glycosides. These chromatographic conditions allowed the identification and quantification of astragaline, cynaroside, cosmosiin, narcissin and rutin in flowers and buds of Staphylea pinnata L. Flavonoid glycosides: raponticin and kaempferol were not detected.Conclusion. Flavonoid glycosides were isolated from the generative organs of S. pinnata L., a technique for quantitative determination of flavonoid glycosides in alcohol-water extracts was developed, astragalin, cynaroside, cosmosiin, narcissin and rutin were detected and quantified.

Introduction. Amino acids (AA), the primary metabolites in plants, play a crucial role in various physiological processes, including the synthesis of phenolic compounds. Drug products and dietary supplements made from medicinal plants can become a rich source of both nonessential and essential amino acids. High levels of free amino acids found in herbal raw material often indicate the presence of biotic and abiotic stress in the plants. Therefore, understanding the dynamics of bioactive compound accumulation in plants throughout their phenological phases of development is critical to optimizing their potential health benefits.Aim. To compare qualitative composition and dynamics of AA accumulation in the underground organs of four representatives of the Rumex genus: R. confertus, R. crispus L., R. obtusifolius L., R. aquaticus L. of three different vegetative phases.Materials and methods. Water extracts from underground organs of the studied plants were analyzed using two different methods for qualitative and quantitative analysis. Extracts were applied to the chromatographic plates TLS Silica gel 60 F254 (Merk, Germany) 20 × 20 cm with a micro-syringe (LLC "Tsvet", Russia). After elution, the plates were treated with a 2 % ninhydrin solution. Quantitative analysis was carried out using the SF-2000 spectrophotometer (LLC "OKB Spectr", Russia).Results and discussion. The amino acid profile of the underground organs of R. confertus, R. crispus L., R. obtusifolius L., R. aquaticus L. during three vegetative phases was determined using the TLC method; the quantitative analysis performed using spectrophotometry.Conclusion. The most diverse amino acid (AA) profiles were found in the withering phase in all Rumex species, with up to 9 amino adsorption zones. In contrast, the least diverse AA profiles were observed in the flowering phase, ranging from 2 to 4 adsorption zones. The quantitative content of AA was lowest in the flowering phase, increased during the regrowth phase, and peaked in the withering phase. However, R. crispus L. showed an unusual pattern, with the highest quantitative content of AA detected in the regrowth phase.

Introduction. The search for new promising sources of biologically active substances is one of the tasks of modern pharmacognosy. Among the huge variety of species of the genus Astragalus L., only a few of them have been studied. In addition, none of them is included in the current edition of the State Pharmacopoeia of the Russian Federation. In our opinion, the study of four species of this genus of Astragalus L. (A. dasyanthus, A. varius, A. testiculatus, A. henningii) growing in the Volga region is of interest.Aim. Study of the glycoside composition of flavonoids in the astragalus herb of four species (A. dasyanthus, A. varius, A. testiculatus, A. henningii) by HPLC-UV using external standards.Materials and methods. The material was samples of astragalus grass (A. dasyanthus, A. varius, A. testiculatus, A. henningii), collected on the territory of the Saratov region in accordance with the basic rules of collection and preparation and dried to an air-dry state. Aqueous-alcoholic extracts (1 : 50) were analyzed. Qualitative and quantitative analysis of flavonoid glycosides was carried out on an Agilent 1260 chromatograph (Agilent Technologies, USA) equipped with a diode array detector, manual sample injection (Agilent G1328C manual injector, Agilent Technologies USA) and an Agilent Open Lab CDS system for collecting and processing chromatographic data. The elution gradient was selected individually using 0.1 % phosphoric acid/acetonitrile solution as solvents. Compounds were identified by retention times and UV spectra of reference standards (RS). The quantitative content of each detected component was expressed in mg/g of dry raw material. All studies were repeated five times, the results were statistically processed using MS Exсel 2010.Results and discussion. The chromatographic profiles of aqueous-alcoholic extracts from the herbs of four types of astragalus were studied. Differences were found in both the qualitative composition and quantitative content of flavonoid glycosides in the grass of the analyzed species. The largest number of compounds was found in the herb of A. varius (6 compounds), 4 compounds were identified in the herb of A. dasyanthus and A. henningii, and 3 compounds were identified in the herb of A. testiculatus. The dominant components in the herb of A. henningii are orientin and rutin, in the herb of A. testiculatus and A. varius it is cynaroside, and in A. dasyanthus it is rutin.Conclusion. A HPLC-UV method was developed for determining the quantitative content of flavonoid glycosides in extracts from the herbs of four species of astragalus (A. dasyanthus, A. varius, A. testiculatus, A. henningii) and the chromatographic profiles of the analyzed samples were studied.

Introduction. The study of the dynamics of accumulation of biologically active substances (hereinafter – BAS) in relation to the phonological phases of plant development is of great scientific and practical interest. The quantitative content of aglycones and glycosides of flavonoids depends on the life cycle of the plant and its vegetation phase.Aim. To identify and quantify flavonoids in the underground organs of Rumex confertus Willd., Rumex aquaticus L., Rumex crispus L. and Rumex obtusifolius L. harvested during different phases of vegetation.Materials and methods. Extracts from the underground organs of the studied plants, obtained according to the method from the pharmacopoeial article on R. confertus, were analyzed. Chromatographic separation and detection were performed on a Nexera-i LC-2040 high-performance liquid chromatograph (Shimadzu Corporation, Japan). The chromatograph was equipped with a column thermostat, chromatography column Grace HPLC-COLUMN 250 × 4.6mm platinum C8-EPS 5 mm (Grace, США) and Guard column Phenomenex SecurityGuardTM Cartridges Widepore C18 4 × 3,0 mm, a degasser, an autosampler (injection volume: 10 µl), and an ultraviolet detector. Detection was carried out at a wavelength λ = 365 ± 2 nm. Mobile phase contains 0.1 % phosphoric acid in water (v/v) (eluent A); acetonitrile (eluent B) with flow rate: 0.9 ml/min.Results and discussion. All studied objects were analyzed. The authenticity of the substances contained were confirmed using the external standard, and their quantitative content was determined. The discovered and quantified substances were: 3-O-rutinoside of quercetin (rutin), 3-O-rutinoside of isorhamnetin (narcissin), 3-O-glucoside of kaempferol (astragalin), luteolin, kaempferol and isorhamnetin. 7-O-glucoside of luteolin (cynaroside) and 7-O-beta-D-glucoside apigenin (cosmosiin) were not found. The aglycone luteolin had the biggest share in the total quantitative content of flavonoids. It is noted that this aglycone is contained in larger quantities in relation to other flavonoids in all studied objects, regardless of the phase of vegetation.Conclusion. In the process of the research, a method for the quantitative determination of flavonoids in alcohol-water extracts was developed. Aglycones and glycosides of flavonoids were identified and quantified in the underground organs of R. confertus Willd., R. aquaticus L., R. crispus L. and R. obtusifolius L. of three different vegetations.

Introduction. Taurine is a non-proteinogenic amino acid. The molecule is involved in lipid metabolism, adsorbs fat-soluble vitamins, and its conjugates with bile acids contribute to the emulsification of fats in the intestine. Drugs, which include a taurine molecule, have anti-cataract, cardiotonic, metabolic effects, and also stimulate regeneration. Among the dosage forms, where taurine acts as an active substance, there is a solid dosage form – film-coated tablets. One of the methods for assessing the quality of solid dosage forms is a comparative dissolution kinetics test. High-performance chromatography with ultraviolet detection is a widely used method for quantification within the dissolution test, however, for taurine, which does not contain chromophore groups in its structure, this method is not directly applicable. To solve this problem, one can apply the method of pre-column derivatization, because of which an fragment is introduced into the structure, providing a bathochromic shift in the UV spectrum of the starting compound.Aim. Development, validation and approbation analytical method for the quantitative determination of taurine by high-performance chromatography with ultraviolet detection as part of a test comparative kinetics dissolution of taurine tablets with a dosage of 250 and 500 mg.Materials and methods. The following preparations were used for the analysis: taurine tablets, film-coated 250 mg and 500 mg, domestic production with a valid expiration date. The comparative dissolution kinetics test was carried out on a DT 126 Light instrument for the "Dissolution" test (ERWEKA GmbH, Germany). Chromatographic separation and detection were performed on a Nexera-i LC-2040 high-performance liquid chromatograph (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, a degasser, an autosampler, and an ultraviolet detector. Detection was carried out at a wavelength of 254 nm after derivatization of the taurine molecule with 4-toluenesulfonyl chloride. Were used a Shim-pack Velox C18 5 μm 4.6 × 150 mm column (Shimadzu Corporation, Japan) and a Shim-pack Velox C18 EXP Guard Column Cartridge 5 μm 4.6 × 5 mm (Shimadzu Corporation, Japan). Primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan).Results and discussion. The optimal conditions for taurine derivatization have been selected, and a method for the quantitative determination of taurine by HPLC-UV in test comparative kinetics dissolution in three dissolution media: 0.1 M hydrochloric acid solution with pH 1.2, acetate buffer solution with pH 4.5, phosphate buffer solution with pH 6.8, as well as in the quality control medium – purified water has been developed and validated. During the validation of the developed methodology, it was found that the validation characteristics are within the acceptance criteria in 4 dissolution media. The analytical range of the method was 0.05–1.2 mg/mL and allows the developed method to be used for the quantitative determination of tablets with a dosage of 250 mg and 500 mg as part of the test comparative kinetics dissolution. The method was tested in 4 dissolution media, in all media, there was a complete release in both dosages (more than 85 % by 30 minutes).Conclusion. The method was tested in three dissolution media: 0.1 M hydrochloric acid solution with pH 1.2, acetate buffer solution with pH 4.5, phosphate buffer solution with pH 6.8, as well as in the quality control medium – purified water. In all media, there was a complete release in both dosages (more than 85 % by 30 minutes).

Drug development & registration. 2023;12(2):62–72. (In Russ.) https://doi.org/10.33380/2305-2066-2023-12-2-62-72. Published: 25.05.2023.

Introduction. Promising sources of biologically active compounds (BAS) are extracts obtained from various morphological parts of plants of the numerous genus Astragalus L. One of the main groups of BAS isolated from plants of this genus are flavonoids, saponins, and polysaccharides. Sufficiently studied are Astragalus membranaceus (Fisch.) Bunge) and Astragalus mongholicus Bge., whose extracts have a wide range of pharmacological activity. Expansion of the nomenclature of medicinal plant materials and the study of BAS groups that determine the main pharmacological effects are topical studies in pharmacognosy. From this point of view, little-studied species of Astragalus (Astragalus henningii (Stev.) Klok., Astragalus testiculatus Pall., Astragalus varius S.G. Gmel., Astragalus dasyanthus Pall.), massively growing in the Volga region, are of interest.Aim. Isolation and identification of 5 aglycones of flavonoids in hydrolysates of water-alcohol extracts from Astragalus herb: A. henningii, A. testiculatus, A.varius, A. dasyanthus by HPLC/UV.Materials and methods. The objects of the study were the herb of four types of Astragalus (A. henningii, A. testiculatus, A.varius, A. dasyanthus), harvested during the period of mass flowering in the Saratov region and dried to an air-dry state. To study the composition of phenolic compounds, extraction was carried out with 70 % ethanol in the ratio of raw material : extracting agent 1 : 10 by infusion for 7 days. Acid hydrolysis was carried out with hydrochloric acid 37 % with heating for 40 min. The resulting hydrolysates were analyzed on an Agilent 1260 chromatograph (Agilent Technologies, USA) with a diode array detector, manual sample injection (Agilent G1328C manual injector, Agilent Technologies, USA) and an Agilent OpenLab CDS chromatographic data collection and processing system using an individually selected elution gradient of the mobile phase (0.1 % solution of orthophosphoric acids/acetonitrile). Compounds in the hydrolysates were identified by the retention times and UV spectra of a mixture of reference standards (RS) of quercetin, isorhamnetin, luteolin, kaempferol, and apigenin.Results and discussion. An elution gradient was selected for one time determination of 5 aglycones of flavonoids with a single analysis time of 13 minutes. These conditions made it possible to identify compounds in the hydrolyzates of water-alcohol extracts of four types of Astragalus. Luteolin, quercetin and apigenin were found in hydrolyzates of A. henningii, A. testiculatus and A. varius; isorhamnetin and kaempferol were found in A. henningii and A. varius. None of the analyzed aglycones was found in the A. dasyanthus hydrolyzate.Conclusion. After preliminary selection of the optimal conditions for the chromatographic separation of a mixture of 5 aglycones of flavonoids, hydrolysates of four types of Astragalus were analyzed. The data obtained indicate the prospects for further study of the chemical composition of Astragalus and confirm the expediency of their use for obtaining of new herbal remedies.

Introduction. The creation of new highly effective drugs requires a thorough study of the metabolome of plant raw materials and a comparative phytochemical study of the underground organs of closely related species of Rumex, such as: R. crispus, R. obtusifolius and R. aquaticus, ubiquitous in Russia. It was noted that they have a metabolome like the official R. confertus, which in turn confirms the potential for studying these species. Of scientific and practical interest is the study of the dynamics of accumulation of the leading group of biologically active substances – anthracene derivatives, depending on the phenological phases of plant development.Aim. Identify and quantify anthracene derivatives in the underground organs of R. confertus, R. crispus, R. obtusifolius and R. aquaticus harvested in three different phases of vegetation.Materials and methods. Extracts from the underground organs of the studied plants obtained according to the method from the pharmacopoeial article on R. confertus were used as the analyzed solutions. The solutions were analyzed on a Nexera-i LC-2040 chromatograph (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, a degasser, and an autosampler using an individually selected mobile phase elution gradient (0.1 % phosphoric acid/acetonitrile solution). Primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan). Compounds from the group of anthracene derivatives were identified by retention times. Detection was carried out using a UV detector with a dynamic change in the absorption wavelength during analysis from 365 ± 2 nm to 254 ± 2 nm.Results and discussion. Alcohol-water extracts were obtained from the underground organs of Rumex. An elution gradient was selected for the simultaneous determination of 5 anthracene derivatives with a single analysis time of 40 minutes. These chromatographic conditions made it possible to identify and quantify the content of emodin, 8-O-β-D-glucoside of emodin, and chrysophanol in the underground organs of R. confertus, R. crispus, R. obtusifolius and R. aquaticus in three different vegetations. Glycosides of anthracene derivatives: glucofrangulin A and frangulin A were not found in the studied objects.Conclusion. Anthracene derivatives were isolated from the underground organs of different vegetations, a method for the quantitative determination of anthracene derivatives in alcohol-water extracts was developed, emodin, 8-O-β-D-glucoside of emodin and chrysophanol were found and quantified.