RU2707300C2 - Method of producing drug having thyrotropic and antioxidant activity - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.

SUBSTANCE: invention refers to chemical-pharmaceutical industry, namely to creation of a method for producing an agent possessing thyrotropic and antioxidant activity, which is a dry extract consisting of the following herbal raw materials: walnut leaves, hazel leaves, turnip leaves, aerial part of the turn, overhead part of prickly-bones, vertebral leaves, aerial part of duckweed, chicory roots, aerial part of the tartar. Said vegetal raw material is milled into a particle size of not more than 2 mm and extracted with 65 % ethyl alcohol in ratio of raw material: extractant equal to 1:16, at temperature 60 °C and continuous stirring. Process is repeated three times. Combined extracts are evaporated and purified by separation. Purified extract is residually evaporated and dried in vacuum drier at temperature of 60 °C and milled in a propeller type mill.

EFFECT: method under the invention widens the range of products correcting thyroid disorders.

1 cl, 1 dwg, 11 tbl, 1 ex

Description

It is known that an increase in thyroid diseases is associated with geochemical, social, environmental and other factors [Arkhipova, 2012]. The thyroid gland is not only one of the largest endocrine organs in terms of size and weight, but also one of the most significant from the standpoint of the role for the body of secreted hormones - iodothyronines. The level of iodine consumption by the population most affects the frequency of thyroid pathology. Around the world, an additional means of prevention is iodization of edible table salt, as well as the administration of iodine tablets to the most vulnerable segments of the population (children, pregnant and lactating women). In Russia, more than 30% of the territory where almost 80% of the population has a massive lesion of the thyroid gland, mainly endemic goiter. According to world statistics, at least 3% of the population suffer from various diseases of the thyroid gland (increased or decreased glandular function, nodular formations). It is known that the insufficient content of thyroid hormones in the body leads to a disorder of the structural and functional relationships of the nervous, cardiovascular, immune and other systems [Zakharia, 1997; Prikhodko 1976]. Thyroid hormones are essential for tissue growth, in the process of laying and functioning of the brain, the formation of intelligence, regulation and maintenance of metabolism. Pharmacological correction of the hypothyroid state of the body is based on the replacement of iodine deficiency and thyroid hormones. Despite the vast arsenal of modern drugs with thyrotropic action used to treat thyroid diseases, the need for effective and safe drugs remains. Given the clinical manifestations, with hypothyroidism it is advisable to use a comprehensive approach to treatment, the purpose of which is not only the normalization of thyroid hormonal status, but also the improvement of the state of the neurovegetative, cardiovascular, immune systems, correction of cognitive impairment and emotional-affective disorders. In the complex treatment of patients with the specified profile of disorders, effective sedative and metabolic therapy is prescribed. For this purpose, herbal sedatives, anabolic drugs, tranquilizers, mainly benzodiazepine series, antidepressants are used. The drugs used do not always have sufficient clinical efficacy and can cause side effects with prolonged use. In this regard, significant interest is justified in the search for new drugs that can prevent and correct hypothyroidism.

Along with this, given the dependence of mental processes and clinical manifestations of hypothyroidism on the level of thyroid hormones in the blood, the development of secondary immunodeficiency, it seems advisable to correct pathological manifestations of hypothyroidism with immunomodulatory agents (tartar, nettle, repeshka, succession) [Plant, 2008; Vegetable, 2009; Vegetable, 2012] antioxidant (walnut, chicory, hazel, duckweed) [Vegetable, 2008; Vegetable, 2009; Vegetable, 2012; Vegetable, 2013; Vegetable, 2014; Fukunda, 2003], in addition, the sequence, turnip, rosemary and cocklebur contain ecdysteroids which, in turn, have adaptogenic and anabolic effects. [Harmatha, 2007]. The object of the study was a complex herbal remedy (hereinafter the extract), consisting of walnut leaves, hazel leaves, burlap leaves, aerial parts of a string, aerial parts of a thorny thorn, nettle leaves, aerial parts of duckweed, chicory roots, aerial parts of tartarn in a ratio of 18, 16, 15, 14, 12, 8, 7, 5, 5%, respectively, and its effect on the course of experimental hypothyroidism. In the extract, the main active ingredients are walnuts, hazel, burlap, string, gingerbread, and chicory, duckweed, nettle and tatarnar enhance and modulate the action of the main components.

As a prototype, kelp was used in tablets (Evalar, Russia) at a dose of 3 mg / kg, which compensates the daily norm of iodine by 133% (according to the annotation to the drug).

The technical result of the invention is the expansion of the range of herbal medicines that have a reliable and secure raw material base and corrective disorders of the thyroid gland, affecting the main links in the pathogenesis of the disease due to the multicomponent nature of the drug. The claimed drug is a dried shredded extract from the following plants: walnut leaves, hazel leaves, burlap leaves, aerial parts of a string, aerial parts of a thorny thorn, nettle leaves, aerial parts of duckweed, chicory roots, aerial parts of tatarn taken in the ratio of 18, 16 , 15, 14, 12, 8, 7, 5, 5%. Raw materials are crushed in a mill for grinding dry samples MRP-2 to a particle size of not more than 2 mm and extracted with 50-60% ethanol in a ratio of raw materials-extractant equal to 1:16 (taking into account the coefficient of water absorption), at a temperature of 55-65 ° C and constant stirring. The process is repeated three times. The extraction time at the 1st and 2nd phase contacts is 60 minutes, at the 3rd phase contact - 30 minutes. The extracts are filtered through sulphide cloth, combined, evaporated to 1/3 of the original volume and purified by separation. The purified extract is doubled to 1/5 of the original volume, dried in a vacuum dryer and ground in a propeller-type mill. The yield of the finished product is 27.0 ± 0.5% by weight of the plant material. The final product is a fine brown powder with a pleasant odor and a bitter astringent taste, with a moisture content of not more than 5%. The product we obtained is characterized as a means of correcting thyroid dysfunction.

Qualitative determination of BLV in dry extract

Based on the qualitative phytochemical analysis [State, 1987], the presence of:

1. Tannins

0.05 g of the extract is dissolved in 5 ml of distilled water. To 2 ml of the resulting solution add 2-3 drops of a solution of iron-ammonium alum, a black-green color (tannins) appears.

2. Phenol Carboxylic Acids

Two-dimensional upward chromatography on paper brand FN-12 in solvent systems:

A. n-Butanol - acetic acid - water (4: 1: 2)

B. 15% acetic acid

When viewing the chromatogram in UV light, after treatment with ammonia vapors, identified with standard samples:

caffeic acid - R _f = 0.80 (A), 0.42 (B);

chlorogenic acid - R _f - = 0.67 (A), 0.70 (B).

3. Flavonoids

1.1. 0.1 g of the extract is dissolved in 50 ml of 50% ethyl alcohol. To 5 ml of the resulting solution add 3 ml of a 2% alcohol solution of aluminum chloride; a yellow-green color appears (flavonoids).

1.2. Two-dimensional upward chromatography on paper brand FN-12 in solvent systems:

A. 15% acetic acid solution

B. n-Butanol - acetic acid - water (40:12:28)

When viewing the chromatogram in UV light before and after the appearance of 5% alcohol solution of aluminum chloride, nine substances of a flavonoid nature with Rf ₁ 0.03 (A) and 0.87 (B) were found; Rf ₂ 0.05 and 0.92; Rf ₃ 0.04 and 0.73; Rf ₄ 0.27 and 0.84; Rf ₅ 0.33 and 0.62; Rf ₆ 0.40 and 0.38; Rf ₇ 0.52 and 0.16; Rf ₈ 0.50 and 0.52; Rf ₉ 0.59 and 0.54. The presence of rutin, quercetin, isoramnetin, cinaroside, kempferol and their glycosides is proved. Identification was carried out with standard samples.

When studying the absorption spectra of dry alcohol extracts, it was found that the maxima of the absorption spectra of alcohol solutions of the polyextract are close to those of GSO cinaroside. When a solution of aluminum chloride I was added to the polyextract solution, the maximum of the absorption spectrum shifted to 405 nm and was close to the maximum of the absorption spectrum of a standard cynaroside sample taken under similar conditions (Fig. 1).

Quantitative determination of BLV in dry extract

About 0.05 g (accurately weighed) of the extract is dissolved in a 25 ml volumetric flask in 10 ml of warm water (40-60 ° C), 5.2 ml of 96% alcohol are added, the solution volume is adjusted to the mark with water (solution A). 3 ml of solution A is placed in a volumetric flask with a capacity of 25 ml, 2 ml of a 2% solution of aluminum chloride in 96% alcohol are added, the volume of the solution is adjusted with 20% alcohol to the mark. After 40 minutes, the optical density of the solution was measured on a spectrophotometer at a wavelength of 405 nm in a cuvette with a layer thickness of 10 mm. As a comparison solution, use a solution consisting of 3 ml of solution A, 1 drop of diluted acetic acid and adjusted with 20% alcohol to the mark in a 25 ml volumetric flask. In parallel, the optical density of the state standard sample (GSO) of cinaroside is measured. 1 ml of the prepared GSO cinaroside solution is placed in a 25 ml volumetric flask, 1 ml of a 2% solution of aluminum chloride in 95% alcohol is added and the volume of the solution is adjusted with 20% alcohol to the mark. The comparison solution is a solution consisting of 1 ml of GSO cinaroside, 1 drop of diluted acetic acid and brought to 20% alcohol to the mark in a 25 ml volumetric flask.

The content of the sum of flavonoids in terms of cinaroside absolutely dry extract in percent (X) is calculated by the formula:

where D is the optical density of the test solution;

D _{o the} optical density of the GSO solution of cinaroside;

m is the mass of extract in grams;

m _o is the mass of GSO cinaroside in grams;

W is the mass loss upon drying of the extract in percent.

Note. Preparation of a GSO cinaroside solution: about 0.05 g (accurately weighed) GSO cinaroside, previously dried at a temperature of 130-135 ° C for 3 hours, dissolved in 85 ml of 95% alcohol in a 100 ml volumetric flask when heated in a water bath, cooled, quantitatively transferred to a volumetric flask with a capacity of 100 ml, bring the volume of the solution with the same alcohol to the mark and mix.

Found: 10.72 ± 0.18%.

The proposed method of obtaining is quite simple, does not require a complex purification scheme, allows you to get a constant product. The technology can be implemented in enterprises producing drugs.

Example

180 grams of walnut leaves, 160 grams of hazel leaf, 150 grams of burlap leaves, 140 grams of the aerial parts of a string, 120 grams of aerial parts of a prickly thorn, 80 grams of nettle leaves, 70 grams of aerial parts of duckweed, 50 grams of chicory roots, 50 grams of aerial parts of tatarnik meeting the requirements of the Global Fund XI edition 2, Art. crushed in a mill for grinding dry samples MRP-2 to a particle size of not more than 2 mm (sieve No. 20 GOST 214-83). The crushed raw materials are loaded into an extraction apparatus with a stirrer and external steam heating. Pour 16 liters of 65% ethyl alcohol in a ratio of raw materials-extractant equal to 1:16, taking into account the coefficient of water absorption of raw materials. Extracted at a temperature of 60 ° C with constant stirring for 60 minutes The extract is filtered through a tin cloth in the collection. Two more extractions are carried out for 60 and 30 minutes, each time feeding 65% to the extractor) alcohol equal to the volume of the drained extraction (1st discharge - 11.85 L, 2nd discharge - 11.65 L, 3rd discharge - 10.00 L) .). The combined extracts are evaporated to 1/3 of the original volume and purified by separation. The purified extract is doubled to 1/5 of the original volume, dried in a vacuum dryer at a temperature of 60 ° C for 8 hours and ground in a propeller-type mill. Get 26.6. g of the finished product, which is 26.6% of the weight of the feedstock. The dry extract is a fine brown powder with a pleasant odor and a bitter astringent taste; crumple, moisture content - 4.55%.

Experimental studies were performed on 56 female rats with an initial weight of 180-200 g. The animals were kept in accordance with the "Rules of Laboratory Practice" (GLP) and Order of the Ministry of Health of the Russian Federation No. 708N of 08/23/2010 "On Approving the Rules of Laboratory Practice". Before the start of the experiments, animals meeting the criteria for inclusion in the experiment were divided into groups taking into account the principle of randomization. The experimental work was carried out in accordance with the "Rules for the use of experimental animals" (Appendix to the order of the Ministry of Health of the USSR No. 755 of 08/12/07), "the Rules adopted in the European Convention for the Protection of Vertebrate Animals (Strasbourg, 1986). The research protocol was agreed with the ethics committee of the IOEB SB RAS (protocol No. 3 of February 7, 2013).

Animals were divided into six groups: an intact group, a control group, and four experimental groups that received dry complex herbal extract extract and a comparison drug. Hypothyroidism in animals of the control and experimental groups was modeled by intragastric administration of mercazolil (AKRIKHIN, Russia) at a dose of 10 mg / kg for 28 days [Arkhipova, 2012]. Then, animals of the I-III experimental groups were intragastrically injected with an aqueous solution of the extract of the dry studied complex herbal remedy in doses of 100, 200 and 300 mg / kg once a day for 20 days; animals of the experimental group IV — an aqueous solution of kelp powder in a dose of 6.2 mg / kg (in terms of iodine 7.5 μg / kg) (EVALAR, Russia). Animals of the control and intact groups received an equivolume amount of water purified according to a similar scheme.

The influence of the tested means on the psycho-emotional state of animals was studied in the tests “open field” and “elevated cruciform maze” (PCL) according to the Guidelines for the study of the tranquilizing (anxiolytic) effect of pharmacological substances [Guide ..., 2012]. In the PCL test, the number of entries into open and closed arms, the residence time of animals in open and closed arms and at the central site were recorded. The time for testing animals was 5 minutes. In the “open field” test, the following were recorded for 3 minutes: horizontal activity (the number of central and peripheral squares crossed), vertical activity (the number of climbs on the hind legs), mink reflex (number of peeping into minks), number of grooming acts (cleaning) and acts bowel movements. The total motor activity was judged by the sum of the vertical, horizontal components and the mink reflex.

For biochemical studies, animals were decapitated under ether anesthesia. Thyroid mass was measured. An enzyme-linked immunosorbent assay determined serum levels of thyroid-stimulating hormone (TTG), free triiodothyronine (fT3), free thyroxin (fT4) using the Tyroid ELISA kit (Russia) using the STAT-FAX 2100 (AWARENESS TECHNOLOGY INC, USA). Based on these indicators, the integral thyroid index (ITI) and the peripheral conversion index (IPC) were calculated [Clinical ..., 2015].

ITI is the ratio of the thyroid hormones themselves to their pituitary regulator: ITI = (fT3 + fT4) / TTG

An increase in this index is the earliest sign of hyperthyroidism, while a decrease in ITI even reflects the initial stages of hypothyroidism.

IPC - an indicator of tissue conversion of thyroxine into its biologically more active metabolite triiodothyronine - and is calculated by the formula: IPC = fN4 / fT3

The intensity of lipid peroxidation (lipid peroxidation) was evaluated by the increase in serum peroxidation product - malondialdehyde (MDA) in blood serum [Stalnaya, Garishvili, 1977]. The state of the endogenous antioxidant system was characterized by the activity of catalase in blood serum [Korolyuk, 1988], superoxide dismutase (SOD) in red blood cells [Makarenko, 1988] and the content of reduced glutathione (HB) in the blood [Akerboom, Sies N., 1981].

The data obtained during the experiment were statistically processed to determine the average value and the arithmetic mean and its standard error (M ± m) according to the generally accepted method using the Student t-test using the Microsoft Exel 2006 statistical software package. The differences were considered statistically significant at p <0 , 05.

The research results showed that in animals of the control group receiving mercazolil, against the background of a decrease in serum levels of fT4 and fT3 by 27% and 29%, respectively, TTG activity increased 2.5 times in comparison with the data of animals of the intact group (Table 1) . As a result, the ITI index in the control group decreased by 67%) and amounted to 37.6 ± 6.26 versus 113.0 ± 7.1 in intact, which indicates the development of hypothyroidism. Also, in animals of this group there was a tendency to disrupt the processes of conversion of thyroxine into a more active metabolite - triiodothyronine, which confirms an increase in the IPC index (by 12%) compared with intact.

The development in control animals of hypothyroidism against the background of course administration of mercazolil is also evidenced by a significant increase in thyroid mass by 48% compared with this indicator of intact animals (Table 2).

Note: * - the differences are statistically significant compared with the data of the control group of animals at p≤0.05; n is the number of animals in the group; ITI - integral thyroid index; IPC is a peripheral conversion index.

Note: * - the differences are statistically significant compared with the data of the control group of animals at p≤0.05; n is the number of animals in the group.

In animals of the first and second experimental groups receiving the test extract at doses of 100 and 200 mg / kg, despite the fact that the level of thyroid hormones did not significantly differ from the control indicator (Table 1), the serum TTG content was 52% and 55% higher than control animals. As a result, the ITI in animals of these groups was 85.2 ± 7.92 and 94.4 ± 8.42, which is 2.7 and 2.5 times, respectively, higher than that of the control animals.

The most pronounced thyrotropic effect was observed against the background of the administration of the test extract at a dose of 300 mg / kg and kelp. So, in animals of the third and fourth experimental groups, the level of fT4 increased by 27% and 25%, the level of fT3 - by 35% and 32%), respectively, compared with the indices of control animals. Against the background of the use of the studied agent, TTG activity in the blood of animals of the third and fourth experimental groups was 55% and 52% lower than the control animals, and almost reached the intact level, as a result, the ITI was higher than that in the control of 2.7 and 2, 6 times respectively.

The thyroid gland weight index also testified to the limitation of the development of hypothyroidism against the background of the test extract (Table 2). So, in the animals of the experimental group, the mass of the thyroid gland corresponded to that of intact animals and was 27-34% lower than the indicator of the control animals.

The results of animal testing in the tests “open field” and “elevated cruciform labyrinth” (PCL) showed that prolonged administration of mercazolilum reduces the motor and orientational-research activity of animals and increases their anxiety level (Table 3, 4).

It was found that in animals of the control group, when testing in the "open field", the index of total motor activity was reduced compared to that in animals of the intact group by 60% due to horizontal (by 68%) and vertical (by 20%) activities (Table 3 ) Not a single animal of the control group visited the central squares for the entire testing period, and, as a result, the mink reflex was absent. The emotional instability of control animals during experimental hypothyroidism was indicated by an increase in grooming and bowel movements by 1.6 and 2.4 times, respectively, compared with data from intact animals (Table 3).

The use of the extract of the dry complex herbal remedy normalized the behavioral indicators of animals in the open field test (Table 3). The most pronounced effect on the motor activity of animals, comparable with that of the comparison drug, the test extract showed at doses of 200 and 300 mg / kg So, the number of peripheral squares visited by animals of the second and fourth experimental groups was on average 3.3 times higher than the index of control animals; vertical activity in the second experimental group - by 11%, and in the third and fourth experimental groups - by 26%. As a result, the total motor activity in animals of these groups was 2.6; 2.8 and 2.4 times higher than that in control animals. The influence of the test extract in doses of 200 and 300 mg / kg on the orientational research behavior of animals exceeded that of the comparison drug - kelp. The number of central squares visited by animals of the second and third experimental groups was 3.6 and 5.0 times higher than that of animals treated with kelp, and, as a result, the mink reflex was 4.0 and 4.6 times, respectively.

Note: * - the differences are statistically significant compared with the data of the control group of animals at p≤0.05; n is the number of animals in the group.

In the group of animals receiving the test extract at a dose of 100 mg / kg, the total motor activity was 47% higher than that of the control animals, mainly due to horizontal activity. So, the average number of peripheral squares visited by animals of the first experimental group was 68% higher than in the control; 50% of the animals of this group entered the central squares of the installation, while in the control group no animals visited the central area of the installation.

The increase in animals of the experimental groups of motor and orientation-research activity can be explained by a decrease in their anxiety level (Table 3). The number of grooming acts in animals treated with phytoextract was 27-36% lower than that of the control animals; the number of acts of defecation was 53-82%. The use of kelp reduced these indicators relative to control by 32% and 24%, respectively.

The results of testing animals in PCL showed that the course administration of animals to the mercazolil caused a decrease in the number of entries into the open arms of the maze and the time spent in them by 54% and 65%, respectively, compared with those of intact animals (Table 4). The control animals also showed a decrease (by 60%) in the number of entries into the closed arms of the labyrinth, which indicates a decrease in motor activity in animals.

The use of the extract of the complex agent neutralized the anxiogenic effect, which develops against the background of experimental hypothyroidism. The number of entries into the open labyrinth arms in animals receiving dry extract in doses of 100-300 mg / kg increased by 1.8; 1.9 and 2.2 times, the time spent in them - 1.9; 2.0 and 2.8 times, respectively, compared with those of control animals. In animals of these experimental groups, the number of entries into the closed arms of the maze was 2.2; 2.0 and 2.6 times respectively higher than the animals of the control group.

Against the background of the introduction of kelp to animals, the number of entries into the open and closed sleeves of PKL, as well as the duration of their stay in the open sleeves, increased on average 1.5 times in comparison with the corresponding data of control animals.

Note: * - the differences are statistically significant compared with the data of the control group of animals at p≤0.05; n is the number of animals in the group.

The experiment found that experimental hypothyroidism causes an increase in the processes of free radical oxidation and a decrease in the antioxidant activity of enzymes in serum (Table 5). So, in the control group of animals there was a 1.8-fold increase in the content of MDA in blood serum compared to the intact group. One of the factors enhancing LPO processes is changes in the antioxidant system (AOS) of the body of animals of the control group, which are expressed in a decrease in serum catalase and SOD in red blood cells by 49% and 65%, as well as in a decrease in the level of GV in the blood by 22%. compared to intact.

It was established that the course administration of the studied phytochemical to animals reduces the severity of oxidative stress, thereby reducing the intensity of lipid peroxidation and increasing the activity of the antioxidant system of the body (Table 5). So, with the introduction of the test drug in doses of 100, 200 and 300 mg / kg, a decrease in the content of MDA in the blood serum was observed by 35%, 41% and 34%, respectively, compared with the control animals. A significant decrease in MDA is due to the fact that, against the background of the use of the studied herbal remedy, normalization of the antioxidant status of the organism is observed. So, in animals of the first experimental group, the activity of catalase was 1.4 times higher than that of the control animals, in animals of the second and third experimental groups - 1.6 times. The use of the test extract in doses of 100 and 300 mg / kg increased the activity of SOD in red blood cells by 1.7 times in relation to the control; higher activity of this enzyme, twice exceeding the control animals, was observed in animals of the third experimental group. The content of hepatitis B in the blood of animals of the second and third experimental groups was on average 19% higher than that of control animals. In animals treated with the comparison drug - kelp, catalase activity, SOD and the content of GV were respectively 23%, 56% and 23% higher than those of animals in the control group.

Note: * - the differences are statistically significant compared with the data of the control group of animals at p≤0,05; n is the number of animals in the group.

Thus, the studied complex herbal remedy shows pronounced pharmacotherapeutic effectiveness in experimental hypothyroidism, normalizing the level of thyroid-stimulating and thyroid hormones, due to which there is a decrease in the level of anxiety in animals, an increase in motor and orientational research activity. The pronounced pharmacotherapeutic activity of the test phytochemical is due to its inhibitory effect on the free radical oxidation of biomacromolecules and the ability to activate the antioxidant system of the body. The technological part.

To identify the optimal technological conditions, we studied the main factors affecting the extraction process of plant materials: the nature of the extractant, its ratio with the raw materials, temperature conditions, the degree of grinding of the raw materials, duration and number of extractions.

Quantitative assessment was carried out according to the yield of extractives [1, 2] and the sum of flavonoids. Metrological characteristics are presented in table 6.

Water and ethyl alcohol of various concentrations (20, 30, 40, 50, 60, 70, 80, 90, 96%) were used as extractants.

As can be seen from table 7, 50-55%) alcohol is the optimal extractant, as it allows to achieve the maximum yield of the sum of flavonoids, taking into account the yield of extractives.

Identification of the optimal degree of grinding of raw materials was carried out jointly for all types of raw materials. A quantitative assessment was carried out according to the yield of extractives. Based on the data obtained (table 8) and taking into account the fact that too fine grinding of plant material leads to contamination of the extraction with difficult-to-filter high molecular weight compounds, grinding of raw materials should be carried out to a particle size of not more than 2 mm.

Due to the fact that the source of plant materials contains thermolabile substances (vitamins, iridoids), the influence of temperature on the extraction process was studied in the temperature range from 20 to 60 ° C. The data are presented in table 9, the optimum temperature is 60 ± 5 ° C.

When studying the influence of the ratio of raw material to extractant on the extraction process, it was established (table 10) that with an increase in the amount of extractant, the yield of extractives and the amount of flavonoids increase, but this increase for ratios of 1:16 and higher is insignificant, so an equal ratio 1: (14-16) (taking into account the coefficient of water absorption).

To determine the duration and the required number of extractions, the time was established for reaching equilibrium concentrations of extractive substances and flavonoids in the raw material-extractant system during a three-time extraction. For this, extraction of the crushed raw material with 40% ethyl alcohol in a ratio of 1:14 was carried out in a water bath at a temperature of 60 ° C with constant stirring in a flask with a reflux condenser. The extracts obtained at certain intervals (15, 30, 45, 60, 75, 90 min) during three extractions were analyzed for the content of extractives and flavonoids. It was established that the equilibrium concentrations are established at the 1st and 2nd phase contacts after 60 minutes, at the 3rd phase contact after 30 minutes (table 11). Threefold extraction provides a fairly complete depletion of raw materials - up to 96% of active substances are extracted.

The proposed method allows to obtain a drug of constant composition with higher thyrotropic and antioxidant activity due to the higher content of active substances (flavonoids).

It is currently not possible to calculate the economic efficiency of the proposed method, however, the above advantages, combined with a simple production scheme, contribute to the rational use of medicinal plant materials and open up the prospect of introducing this method into the pharmaceutical industry.

Used Books

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2. The State Pharmacopoeia of the USSR: issue. 2. General methods of analysis. Medicinal plant material. 11th ed. - M., 1987 .-- 400 p.

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Claims (1)

A method of obtaining a medicinal product with thyrotropic and antioxidant activity, which is a dry extract, consisting of the following plant materials in the ratio: walnut leaves 18%, hazel leaves 16%, burlap leaves 15%, aerial parts of a series of 14%, aerial parts of a thorny thorn 12%, nettle leaves 8%, aerial parts of duckweed 7%, chicory roots 5%, aerial parts of tatarnar 5%, which are ground in a mill to a particle size of no more than 2 mm and extracted with 65% ethyl alcohol three times in the ratio of raw-ext a reagent equal to 1:16 at a temperature of 60 ° C and constant stirring, after which the combined extracts are evaporated, purified by separation, dried in a vacuum dryer at a temperature of 60 ° C and ground in a propeller-type mill.

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