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Vol 25, No 4 (2022)

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Pharmaceutical chemistry

Determination of the flavonoid content in flowers, leaves and shoots of Crataegus L

Kurkin V.A., Volkova N.A., Pravdivtseva O.E., Kurkina A.V., Trifonova P.V., Dubishchev A.V., Agapov A.I., Egorova S.N.

Abstract

Relevance. The medicinal preparations based on the raw materials of hawthorn (Crataegus sp.) are successfully used as cardiotonic agents both in the Russian Federation and abroad. However, only flowers and fruits are used as raw materials of hawthorn in our country, whereas foreign experience shows the possibility of using hawthorn cormus collected at the flowering stage of the plant as raw materials. Crataegus sanguinea Pall., Crataegus monogina Jacq. and Crataegus submollis Sarg. can serve as sources of a new type of raw materials. As studies conducted earlier have shown, the leaves of the Crataegus sanguinea Pall. have a cardiotonic effect and a diuretic effect, and the cormus of the Crataegus submollis Sarg. can be used as antidepressants. The aim of the study was a comparative study of the flavonoid composition of flowers, leaves and shoots of some species of the genus hawthorn (Crataegus L.). Material and methods. The flowers, leaves and cormus of Crataegus sanguinea Pall., Crataegus monogina Jacq. and Crataegus submollis Sarg. were collected during the flowering period of plants on the territory of the Botanical Garden of Samara University in May 2021. The content of flavonoids was determined in all types of raw materials. Qualitative analysis was carried out by thin-layer chromatography and UV-spectrophotometry. Differential spectrophotometry was chosen as the method of quantitative analysis. Results. The comparative analysis of hawthorn flowers, leaves and cormus showed that the hawthorn flowers of the studied are similar in flavonoid composition, whereas the leaves and shoots differ markedly, although within the same species their flavonoid composition is similar. In all types of raw materials, the content of chlorogenic acid and hyperoside is noted. Conclusions. The flavonoid composition of hawthorn cormus and leaves is similar in the corresponding species, therefore, quantitative analysis of hawthorn leaves and shoots should be carried out by differential spectrophotometry. In the case of leaves and cormus of Crataegus submollis Sarg., it is advisable to determine the total flavonoids calculated on hyperoside (wavelength 412 nm), and in the case of Crataegus sanguinea Pall. and Crataegus monogina Jacq, to determine the content of the total flavonoids calculated on of 211-O-rhamnoside of vitexin (wavelength 392 nm). Hawthorn leaves and cormus are promising types of medicinal plant raw materials. Crataegus submollis Sarg. is a promising source of medicinal herbal preparations.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):3-9
pages 3-9 views

Multidimensional analysis of x-ray fluorescence spectra as a possible approach for a comparative study of the elemental composition of fruit and seeds of Mangifera indica, Actinidia deliciosa, Nigella sp

Morozova M.A., Marukhlenko A.V., Maksimova T.V., Nyambose J.

Abstract

Relevance. The specificity of plant raw materials as an object of XRF is due to the heterogeneous accumulation of elements at the subcellular and tissue levels - in plants, elements are present in adsorbed, colloidal, ionic forms, as organomineral complexes and polymeric compounds. Thus, the study of plant substrates is limited by the presence of matrix effects, which require a set of standards - reference samples. The purpose of this work is to investigate the degree of interspecific and intraspecific differences in the elemental profiles of plant samples without using a standard by the means of principal component analysis for X-ray fluorescence spectra processing. Material and methods. The proposed approach was applied to study the elemental profiles of fruits genus Actinidia, Manqifera and seeds of plants genus Niqella. The work was performed on an EDX-7000 Shimadzu enerqy-dispersive X-ray fluorescence spectrometer. For data processing by the principal component method, OriginPro 2017 software (OriginLab, USA) was used. Results. As a result of the work, the efficiency of the principal component method in processing the X-ray fluorescence data is shown. Thus, the analysis of the elemental composition of dried seeds of kiwi fruits showed that the samples, regardless of the region of growth, are quite similar in chemical composition, which, apparently, is species specific. It has been established that the use of XRF to assess the calcium content in the edible part of the fruit according to stable and reproducible results obtained for seeds is promisinq. On the example of mango fruits, it was shown that the grinding of fruit tissues leads to an increase in the intensity of the fluorescence signal of all elements. It has been shown that X-ray fluorescence analysis doesn't make it possible to reveal all elements in the composition of the plant matrix, however, the ratio of fluorescence signals adds up to a characteristic and species-specific pattern. Conclusion. The results obtained indicate the possibility of using the technique for express analysis of the elemental composition of plant objects and raw materials, including for the purpose of establishing the type of producing plant or determining its geographical region of origin

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):10-14
pages 10-14 views

Chemical and toxicological examination of zaleplon and its metabolites in urine

Volkova A.A., Kalekin R.A., Orlova A.M., Moskaleva N.E., Markin P.A., Astashkina O.G.

Abstract

Relevance. Benzodiazepine derivatives are psychoactive substances, modulators of gamma-aminobutyric acid (GABA) receptors, exhibiting hypnotic, sedative, anxiolytic and anticonvulsant effects. Based on this mechanism of action, the use of benzodiazepine receptor agonists of the non-benzodiazepine series is widely used in medical practice. Non-benzodiazepine anxiolytics include the so-called "Z-drugs", after the name of the first drugs from this group (zopiclone, zolpidem, zaleplon). According to the mechanism of action, they are selective agonists of the omega-1 subtype (zopiclone, zolpidem) and the omega-2 subtype (zaleplon) of benzodiazepine receptors of the macromolecular GABA receptor complex. Currently, zaleplon is the most affordable of this group of drugs in Russia. The aim of the study is to develop a technique for detecting zaleplon and its metabolites in urine by HPLC-MS with BP using Orbitrap technology for the purposes and tasks of forensic medical examination. Material and methods. Morning urine was taken from patients who took therapeutic concentrations of zaleplon as prescribed by a doctor. The active substance zaleplon was studied as an alcohol solution with a content of 1 mg/ml of the substance under study. For the study, high-resolution HPLC-MS was used using Orbitrap technology on the instrument. Since the symptoms of side effects in zaleplon are similar to benzodiazepine derivatives when sent for forensic examination for chemical and toxicological examination, the sample preparation was carried out according to the generally accepted scheme for compounds of this group. Results. A technique for identifying zaleplon in urine extracts by HPLC-MS with BP using Orbitrap technology as a confirmatory method during forensic chemical and chemical-toxicological examination of urine has been developed. Retention times of the 5-oxosaleplone metabolite after oral administration were detected and established, and characteristic ions for use in identification were determined. Conclusion. A validation evaluation of the developed methodology was carried out, which indicates the suitability of the proposed methodology for forensic chemical and chemical-toxicological analysis. The relative standard deviation (RSD,%) in assessing the convergence and intra-laboratory reproducibility of retention times did not exceed 5% for a biological object - urine.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):15-22
pages 15-22 views

Medical chemistry

Changes in the activity of lysosomal cysteine proteases of spermoplasm in patients with impaired reproductive function

Ishtulin A.F., Korotkova N.V., Matveeva I.V., Isakov S.A., Minaev I.V., Polyakova P.M.

Abstract

Relevance. One of the serious problems of modern society is male infertility, which accounts for 7% of the total structure of infertile marriages. The reasons leading to it are not always clear. All researches of ejaculate are mainly based on chemical microscopic explorations. The biochemical indicators of sperm are currently being determined in insufficient volume, therefore the introduction of new biochemical markers for the diagnosis of decreased fertility in men is very relevant. Object. To examine the activity of cathepsins B, L and H contained in the acrosome of spermatozoa among men with impaired reproductive function. Materials and methods. The material for the investigation was spermoplasm. Spermoplasm was obtained from 30 patients diagnosed with chronic prostatitis and 30 patients diagnosed with varicocele with complaints of reproductive health. The control group consisted of 30 patients without diseases of the genitourinary system during physical examination and without complaints in anamnesis about reproductive health. Spermoplasm was obtained by centrifugation of a sperm sample for 10 minutes at a speed of 1000 rpm. The activity of cathepsins B, L, H in the spermoplasm was determined by the spectrofluorimetric method according to A.J. Barrett and H. Kirschke. Results. The activity of cathepsin B among patients diagnosed with chronic prostatitis was reduced by 3.4 times, in patients diagnosed with varicocele by 18.5 times. The activity of cathepsin L among patients with chronic prostatitis is reduced by 3.5 times, with varicocele by 20 times. The activity of cathepsin H in chronic prostatitis decreases by 3.3 times, in patients with varicocele by 25.5 times. Conclusions. The activity of lysosomal cysteine proteases B, L, H among patients with impaired reproductive function was explored. A decrease in the activity of cathepsins B, L, H was found among these patients, as distinct from the patients without signs of fertility disorders. In this way, changing the activity of cathepsins B, L and H can be considered as one of the possible markers for the diagnostics of male infertility.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):23-28
pages 23-28 views

Features of the effect of antioxidant status on the production of cytokines and pro-inflammatory molecules under stimulation of human whole blood cells with mitogen and lipopolysaccharide

Logatkina A.V., Bondar S.S., Nikiforov V.S., Bondar N.V., Terekhov I.V., Parfenyuk V.K.

Abstract

Relevance. Antioxidants play an important role in the prevention of various pathological conditions, including radiation damage, infections, immune response, etc. Antioxidant deficiency against the background of increased peroxidation processes is associated with the progression of various pathological processes, including infectious inflammation. The relationship between the activity of the antioxidant system and the state of lipid peroxidation is interrelated with the phosphorylation of individual intracellular enzymes involved in the functioning of receptor-associated signaling pathways. It has been shown that antioxidants have an immunomodulatory effect in infectious processes, contributing to a change cytokine production, phagocytosis activity, antibody production, etc. The aim of the study was to characterize the effect of antioxidants on the production of cytokines stimulated by mitogen and lipopolysaccharide and the content of terminal MAPK/SAPK protein kinases in mononuclear leukocytes. Material and Methods. The material for the study was venous blood taken in the morning from the cubital vein from practically healthy individuals from among blood donors of both sexes aged 20-25 years. In whole blood mononuclear cell (MNC) lysates, the content of stress-activated protein kinase JNK 1 and 2 isoforms (JNK1/2), mitogen-activated protein kinase p38, and the level of the p65 subunit of the nuclear transcription factor NF-kB were assessed by ELISA. In the supernatants, the level of interleukins (IL) was studied: IL-1ß, -2, -4, -6, -8, -10, -12, -13, -17A, -19, -21, -22, -23, - 28, TNFa, IFNy, RAIL-1, TRGF, GM-CSF, lipopolysaccharidebinding protein, cAMP, concentration of antioxidants and peroxides. Results. The analysis showed that a high level of antioxidant activity in the blood against the background of mitogenic stimulation and exposure to lipopolysaccharide cells is associated with an increase in the production of IL-10, IL-12, GM-CGF, IL-4, COX-2, cAMP, SOD, as well as a decrease in levels of IL-1, IL-17A, lipopolysaccharide-binding protein, a decrease in the content of JNK and p38 protein kinases in MNCs. Conclusions. A high level of antioxidants contributes to a partial weakening of the pro-inflammatory activity of whole blood cells subjected to mitogenic stimulation and activation by lipopolysaccharide. The immunoregulatory effects of antioxidants are determined by changes in the MNC activity of terminal protein kinases of the MAPK/SAPK signaling pathway, in particular, JNK and p38 is possible due to cAMP activation of the PKA/CREB signaling pathway.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):29-39
pages 29-39 views

Problems of experimental biology and medicine

Some aspects of the preclinical study of the safety of the dry extract of Potentilla alba L

Krepkova L.V., Babenko A.N., Lemyaseva S.V., Borovkova M.V., Kuzina O.S.

Abstract

Relevance. Recently for the treatment of thyroid diseases it has been suggested to use herbal remedies, which are able to influence the functional activity and structure of the thyroid gland. One of such perspective objects is considered to be Potentilla alba L., Rosacea family, which is an effective regulator of thyroid hormones level and is used in a complex treatment of the mentioned pathology of endocrine system. The purpose. To estimate general toxic effect of the dry extract of Potentilla alba L. in conditions of chronic experience to determine a possibility of development on its base of new medicines recommended to treat thyroid diseases. Materials and methods. A chronic experiment was carried out on male and female Wistar rats. The studied extract was administered intragastrically for 90 days in doses of 25, 125 and 375 mg/kg. The control animals were given water. Integral health indexes of experimental animals were registered, peripheral blood samples were taken to determine hematological and biochemical indexes, functional state of cardiovascular system (ECG study), central nervous system by orientation reactions in "open field" test, excretory func tion of kidneys against 3% water load were studied. At the end of the experiment pathohistological examination of the internal organs of the rats was carried out. Results. Long-term administrations of the dry extract of Potentilla alba L. into stomach of clinically healthy rats of both sexes in 2,5, 12,5-37,5 times therapeutic doses caused hypothyroidism and hypolipidemic action. The dry extract in all tested doses showed no toxic effect on blood, cardiovascular and nervous system of rats. Prolonged administration of studied extract at the maximum tested dose (375 mg/kg) resulted in a moderate damaging effect on the liver and kidneys of rats of both sexes as well as the testes. A threshold dose of 25 mg/kg was established. Conclusions. Results of the investigation were grounds for development of new thyrotropic drugs of systemic action (0,050 g capsules, 0,150 g tablets) based on the studied extract and to recommend them for further investigation.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):40-47
pages 40-47 views

Choleretic activity of solvent fractions of Calendula officinalis L

Nikolaev S.M., Badmaev N.S., Sambueva Z.G., Olennikov D.N., Kashchenko N.I.

Abstract

The purpose: to study choleretic activity of solvent fractions (butanol, ethyl acetate, hexane, and chloroform) of Calendula officinalis flowers cultivated in Russia. Materials and methods: The studies were carried out on white Wistar rats. Bile was obtained using a polyethylene cannula inserted into the general bile duct according under the thiopental anesthetized animals. Results. The most pronounced choleretic property was shown for the butanol fraction which increased bile secretion at 27-47% compared to the control followed by the hexane fraction (20%), chloroform fraction (12%) and ethyl acetate fraction (10%). The choleretic reaction remained at a high level throughout the 4 hours after the introduction of butanol and hexane fractions. The stimulating effect on the synthesis and release of cholates is most pronounced for the ethyl acetate fraction, followed by butanol, chloroform and hexane fractions. Conclusions. Pronounced choleretic effect of butanol fraction of Calendula officinalis flowers was due to the high content of flavonoids (35-40%), which are the main class of compounds of this fraction.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):48-51
pages 48-51 views

Plant protection and biotechnology

Flavonoids and essential oils containt in peppermint (Mentha x piperita L.) raw materials grown in organic farming

Malankina E.L., Tkatchova E.N., Terechova V.I., Zujkova E.Y.

Abstract

The study of the medicinal plant quality indicators is an urgent task in the organic raw materials production. Peppermint (Mentha x piperita L.) is one of the most popular medicinal crops in the world, which is used to obtain mint leaves, essential oils, to produce herbal teas, where organic products occupy a separate segment. The pharmacologically significant compounds in mint are essential oil and phenolic compounds (including flavonoids and tannins). The aim of the work was to estimate the content of essential oil, the sum of phenolic compounds, flavonoids, and tannins in the leaf of 5 varieties of peppermint, which were grown under organic culture condition. Materials and Methods. The varieties of Mentha x piperita Mitcham, Apelsinovaya, Kubanskaya 6, Tik-Tak, and Multimentha were selected as objects. Productivity was determined in the flowering phase. The yield was determined in the flowering phase. The essential oil content was determined by the Clevenger hydro distillation method, component composition - by gas chromatography, the sum of phenolic compounds and tannins - according to the Folin-Ciocalteu method in a water-alcohol extract in terms of gallic acid. Determination of flavonoids was carried out by the aluminum chloride method in terms of rutin. Results. The content of essential oil, depending on the variety, ranged from 0.89 to 2.91% over 2 years, while the menthol content of the menthol varieties varied from 25, 99 to 50, 36%. The mint raw material is characterized by a high content of the total phenolic compounds (from 9.33% for the Mitcham variety to 10.77% for the variety Multimentha). The flavonoids content, depending on the variety, ranged from 2.25% by the variety Kubanskaya 6 to 4,22% by the variety Apelsinovaya. The average content of tannins for two years did not exceed 1,5%. Conclusion. Growing peppermint in organic culture allows to obtain raw materials with a high content of phenolic compounds, including flavonoids, and essential oil.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(4):52-60
pages 52-60 views

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