UA123718U - METHOD OF OBTAINING PLANT SUBSTANCE WITH HYPOGLYCEMIC ACTION - Google Patents

Abstract

The method of obtaining a complex of biologically active substances with hypoglycemic action includes a technological stage of extraction, and as plant raw materials use stevia leaves (Stevia rebaudiana), which is extracted with hot water purified (t - 80-90 ° C) during the day with periodic stirring With 96% ethanol, the resulting extract was filtered and evaporated in a vacuum rotary evaporator to obtain a dry extract.

Description

The useful model belongs to pharmacy and medicine, namely to methods of obtaining a complex of biologically active substances from medicinal plant raw materials, which exhibits a hypoglycemic effect, and can be used to correct the level of glucose in the blood in the treatment of diseases that are associated with metabolic disorders.

In official and traditional medicine for the treatment of diabetes mellitus, sugar-lowering agents and dietary supplements based on plant raw materials are often used, since herbal preparations are characterized by low toxicity and can be a significant addition to the pharmacotherapy of diabetes mellitus. The domestic market of herbal hypoglycemic agents is limited, which necessitates the search for new medicinal raw materials with a sugar-lowering effect and the creation of medicinal products based on them for the treatment and prevention of diabetes.

A well-known official collection for the treatment and prevention of diabetes is "Arfazetin" I2Y, which includes: rosehip fruits, St. John's wort, leaves of common bean fruits, horsetail grass, chamomile flowers, rhizomes and roots of Eleutherococcus spinosa, common blueberry shoots. The disadvantage of this remedy is the inconvenience of dosing when preparing the decoction by patients, as well as heartburn and allergic reactions may occur with long-term use.

There is also a known method of obtaining a hypoglycemic agent from dogwood leaves |Z). According to this method, the raw material is successively extracted with 50-95% ethanol, and then with purified water with additional purification of the water extract and subsequent combination of the extracts and their evaporation. The disadvantage of this method is the use of non-standard raw materials.

The purpose of the useful model is to expand the range of plant-based medicinal products with hypoglycemic action and to create a new product with a specific action from stevia leaves (Biemia heratsadiapa), which can be used as a substance in the development of products in various dosage forms for use in medicine.

The basis of a useful model is the task of developing technological techniques for obtaining a plant substance aimed at maximally complete extraction of biologically active substances from plant raw materials.

When solving the technical problem, it was taken into account that the use of aqueous extraction of plant material treated with 96 95 ethanol, as well as subsequent drying

From the obtained extract, it leads to obtaining a dry substance with a high content of biologically active substances. Degreasing of vegetable raw materials 96 96 with ethanol made it possible to free the aqueous extract from ballast substances and mucus.

In the course of previous studies, with the aim of choosing the optimal extractant for extracting a complex of biologically active substances from the leaves of stevia (Hyemia heratsaipa), we studied a number of solvents, namely: purified hot water (previously degreased raw materials with 96 95 ethanol), purified hot water (1-80 -90 "С) and 20 95 ethanol; the quantitative content of flavonoids, hydroxycinnamic acids, fructans, and polyphenolic compounds was determined in each of the extracted extracts using the spectrophotometric method, as well as extractive substances using the gravimetric method.

Through selection, we determined that for obtaining a phytosubstance from the leaves of stevia (Hyemia heratzdiapa) with the most complete extraction of extractive substances, flavonoids, hydroxycinnamic acids, fructans and polyphenolic compounds, it is optimal to use as an extractant hot water purified during the preliminary treatment of plant raw materials with 96 95 ethanol.

Also, in the course of a complex phytochemical study of stevia leaves (5yemia heratsaipa), we established the qualitative composition and determined the quantitative content of biologically active substances - amino acids, organic acids, a complex of phenolic compounds (flavonoids, hydroxycinnamic acids, tannins), polysaccharides, essential and fatty oils.

The method of obtaining a plant substance with a glycoglycemic effect, in particular from the leaves of stevia (Biemia heratsiapa), includes the technological stage of extraction with hot purified water during the day with periodic stirring, and the crushed plant material is pre-moistened with 96% ethanol. The obtained extract is drained, filtered and concentrated, followed by drying in a vacuum rotary evaporator until a dry substance is obtained.

The ratio "raw material: extractant" is 1:86, which is necessary and sufficient for the extraction process.

The method is carried out as follows. Chopped to the size of particles that pass through a sieve with a hole diameter of Mo 11200 (with a maximum tolerance for the hole of 0.77 mm), plant raw materials are treated with ethanol 96 95, left for 15 minutes at room temperature. Then the raw materials are poured with hot purified water (80-90 "С) in a ratio of 1:8, infused for 24 hours at room temperature, stirring periodically. The aqueous extract is drained and filtered through a paper filter. It is condensed and dried on a rotary vacuum evaporator to obtain a dry substance (mass loss during drying is 3.26 95).

Example 1 100 g of air-dried stevia leaves (Pyemia heratsaipa), crushed to the size of particles that pass through a sieve with a hole diameter of 11200, were moistened with 96 95 ethanol and left for 15 minutes at room temperature to degrease the raw material. Then the processed raw materials were transferred to a percolator and poured with 800 ml of hot purified water (80-90 "С), infused for 24 hours at room temperature with periodic stirring. The obtained aqueous extract was filtered through a paper filter, thickened to 1:5 vol. to it by evaporation at a temperature of 80-90 "С. The condensed aqueous extract was dried on a rotary vacuum evaporator to a dry powder (mass loss during drying was 3.26 Ob).

The obtained finished product - dry extract - is a crystalline powder of dark brown color with a greenish tint, with a specific smell, sweet to the taste, soluble in cold water, sparingly soluble in 96 95 ethanol.

Example 2

The study of the hypoglycemic effect of a dry extract from the leaves of stevia (Biemia heratsaipa) was conducted on 35 white non-lily female rats weighing 250-295 g. The animals for the experiment were divided into 5 groups of 7 animals each (n - 7). The first group of animals was a control (intact control - IR), the second group consisted of animals that were injected subcutaneously with a solution of dexamethasone (KAKA, Slovenia) at a dose of 0.125 mg/kg once a day (control pathology - KP); the third group consisted of animals that received an aqueous solution of stevia leaf extract at a dose of 250 mg/kg intragastrically once a day against the background of KP; the fourth and fifth groups consisted of animals that, on the background of KP, received comparison drugs: metformin in the form of tablets "Siofor" BERLIN-

CHEMICAL AG (MENARINI GROUP) and official plant collection "Arfazetin" (PJSC "Viola",

Ukraine) in doses of 100 mg/kg and 9 ml/kg, respectively, which were administered in similar regimes. The blood glucose level was measured with a VIOMIME glucometer of the Widpiezi CM 550 series (Switzerland). All animals were kept in standard vivarium conditions (natural alternation of light and darkness, air temperature 21-42 "С, relative humidity 55-70 95) and received complete ration granulated food for rats 4 times a day and drinking water without restrictions (feeding was stopped after 12 hours before blood sampling to determine glycemia).

The content of glucose in the blood was determined for 1 day (initial level, before the introduction of dexamethasone), and

Also on the 7th and 14th day of the experiment.

The results of determining the glycemia of rats of all groups and the analysis of the obtained data are shown in Table 1.

Table 1

Hypoglycemic activity of dry extract of stevia leaves on the model of dexamethasone hyperglycemia in rats (M--t, p--7)

A group of animals. 2, Difference 2, Difference level, Glycemia relative Glycemia relative mmol/l | mmol/l | initial level, | mmol/l | initial level,

Cho Cho

Notes: 1. "- statistically significant differences (p<0.05) relative to the initial level in each group 2. " statistically significant differences (p<0.05) relative to control pathology (dexamethasone without correction)

Z. and - statistically significant differences (p<0.05) relative to metformin

The results of the research showed a reliable increase in the basal (after 12-hour fasting) blood glucose level of KP rats on the 7th day (6.04:2-0.21 mmol/l) and on the 14th day (6.43:20, 17 mmol/l) relative to the normal initial level (4.59240.13 mmol/l).

IR animals were characterized by a normal level of glycemia both at the beginning of the experiment and during the study.

In rats receiving an aqueous solution of a dry extract of stevia leaves at a dose of 250 mg/kg orally against the background of subcutaneous administration of dexamethasone, the level of glycemia by the end of the experiment was lower than the parameters of KP both on the 7th and on the 14th day, which characterizes the positive effect of the substance under study on carbohydrate metabolism under the conditions of simulation of a pathological condition, namely, impaired glucose tolerance under the influence of chronic administration of large doses of a glucocorticoid drug. Thus, on the 7th day of the experiment in the animals of this group, the average level of glycemia increased by 13.57 95 relative to the initial level and was (5.26:20.22) mmol/l, which is 18.21 95 lower than the dynamics of the increase in the level blood glucose in the group of KP rats, where the glucose level increased by 31.78 95 relative to the baseline. On the 14th day of the experiment, the average value of glycemia in rats that received the studied extract on the background of KP was (6.13:20.13) mmol/l; the increase in the level of blood glucose relative to the baseline in the rats of this group was 32.41 95 and was 7.72 95 less compared to KP rats (in this group, the glucose level at the end of the experiment increased by 40.19 95 compared to the baseline).

On the 7th day of the experiment, the average level of glycemia increased by 27.88 95 compared to the initial one and amounted to (5.7020.22) mmol/l, which was lower by only 3.90 95 , than on the 7th day of the experiment in the animals of this group, the average level of glycemia increased by 13.57 95 relative to the initial level and was (5.26:20.22) mmol/l, which is 18.21 95 lower than the dynamics of the increase in the level blood glucose in the group of KP rats, where the glucose level increased by 31.78 Fo compared to the initial one. Stevia leaf extract on the background of KP compared to the reference "Arfazetin" reduced the level of glycemia by 14.31 95 on the 7th day and by 7.01 95 on the 14th day.

On the 7th day of the experiment, the average value of glycemia in rats receiving the reference drug metformin on the background of KP was (5.83:50.18) mmol/l, and the increase in blood glucose level relative to the initial level in rats of this group was 14.93 95, which was 16.85 95 less compared to KP rats and only 1.36 95 compared to rats given stevia extract on a KP background. On the 14th day, the increase in the level of blood glucose relative to the baseline in metformin-treated rats was 121.13 95, which was 19.06 95 less compared to KP rats.

An increase in the level of blood glucose relative to the baseline in rats administered stevia extract,

Zo was 32.41 95, which was 11.28 95 higher than in metformin-treated animals.

Therefore, the use of an aqueous solution of a dry extract of stevia leaves contributed to a significant increase in glucose tolerance in an experiment on rats in the simulation of chronic dexamethasone hyperglycemia.

Thus, the plant substance obtained by the proposed method in the form of a dry extract of stevia leaves is promising for the creation and use of medicines with a sufficiently pronounced hypoglycemic effect.

Sources of information: 1. Efimov A.S. Phytotherapy of obesity and diabetes // A.S. Efimov, A.V. Shcherbak -

K.: Naukova dumka, 1993. - 96 p. 2. Meretska I.V. The effect of a new hypoglycemic plant collection on the immune status in diabetes / I.V. Meretska // Tavricheskiy medical and biological journal. - 2012. - T. 15, Me4 (60). - P. 251-253.

Z. Pat. 89735). Ukraine, IPC AbETK 36/40, AbIK 3/10. Therapeutic and preventive agent with hypoglycemic effect from dogwood leaves / V.A. Rybak, L.M. Maloshtan, O.V. Kryvoruchko, V.M.

Kovalev. - Mo c201314662; Application December 16, 2013; Publ. 04/25/2014, Bul. Mob.-4 p.

Claims (1)

USEFUL MODEL FORMULA The method of obtaining a complex of biologically active substances with a hypoglycemic effect, which includes a technological stage of extraction, which is distinguished by the fact that stevia leaves (5yemia heratsaipa) are used as plant raw materials, which are extracted with hot purified water (Her - 80-90 C) during the day with periodic stirring, and the raw materials are pre-treated with 9695 ethanol, the resulting extract is filtered and evaporated in a vacuum rotary evaporator to obtain a dry extract.