RU2237488C1 - "baikalsky-6 tea" as preparation of diuretic activity and method for its obtaining - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: the present innovation deals with elaborating preparations upon plant basis of diuretic activity. The suggested preparation includes mountain cranberry leaves, prostrate knotweed grass, bearberry and dioecious nettle leaves , calendula flowers taken at certain quantitative ratio. It could be in the form of medicinal herbal species, dry extract, capsules or tablets. It is, also, suggested the method to obtain such a preparation of diuretic activity: the above-mentioned raw material should be extracted at 80-90 C with purified water at raw material : extragent ratio 1:10-12 by taking into consideration the coefficient of water absorption, moreover, the first extracting is carried out for 120 min, and the second - for 60 min, then extracts should be combined, separated and dried in vacuum dryer followed by reducing. The present method enables to obtain greater output of extractive and active substances.

EFFECT: higher efficiency.

5 cl, 3 ex, 8 tbl

Description

The invention relates to the field of pharmacy and relates to the creation of a plant-based product with diuretic activity and a method for producing the product.

The urological collection is known (RF patent No. 2103003 dated 01/27/1998), containing bearberry or lingonberry leaf, dill, rhizome and roots of Eleutherococcus or leuzea, mint leaf and calendula flowers.

It is known to use the leaves of lingonberry vulgaris as a diuretic (Complete reference of medicinal plants. - M., 2001, p. 906).

These funds have insufficient therapeutic activity.

There is a method of obtaining a decoction from the collection of diuretic No. 2, consisting of the leaves of bearberry ordinary - 40 g, the fruits of juniper ordinary - 40 g, the roots of Ural licorice - 20 g (radar, 2002, issue 4. S.876).

To prepare the broth, the crushed collection in a ratio of 1:20 is poured with room temperature water, taken into account the coefficient of water absorption, in an appropriate container and insisted on a boiling water bath with frequent stirring for 30 minutes. Filter and add water to the required volume of extraction (GF XI, issue 2. S.147-148).

A known method of obtaining the sum of extractive substances with diuretic activity, including extraction of plant materials with ethyl alcohol in the ratio of raw materials: extractant 1: (10-14) three times for 60 minutes, followed by evaporation of the combined extracts, separation and drying (RF patent No. 1737797 dated 30.11. 1994).

A disadvantage of known sources is the low yield of extractive and active substances.

The objective of the present invention is to increase the diuretic activity of the agent by creating a multicomponent plant composition and by increasing the yield of extractive and active substances (sum of phenolic compounds) by the proposed method.

A remedy having diuretic activity is proposed, including the leaves of lingonberry vulgaris, the grass of the mountaineer, the leaves of bearberry ordinary, the leaves of the nettle, the flowers of the calendula officinalis in the following ratio of components, wt.%:

lingonberry leaves 20

Highlander's Grass 30

bearberry leaves 20

dioica nettle leaves 15

calendula officinalis flowers 15,

while the tool can be made in the form of a collection, dry extract, capsules, tablets.

Also, a method for producing a agent having diuretic activity is provided, which consists in the fact that the above raw materials are extracted at a temperature of 80-90 ° C with purified water at a ratio of raw materials: extractant 1: (10-12) taking into account the coefficient of water absorption, while the first extraction is carried out in for 120 minutes, and the second for 60 minutes, then the combined extracts are evaporated, separated and dried in a vacuum dryer, followed by grinding, while the extracts are evaporated to 1/5 of the original volume and dried at a temperature of 55 ± 5 ° C in 5 hours

The proposed method allows to obtain the final product, which is an amorphous powder from brown to dark brown in color with a specific smell and bitter taste. The loss in mass upon drying is 3.7-4.2%.

The determination of the authenticity and quantitative content of active ingredients of Baikalsky-6 dry tea extract has been studied by modern physicochemical methods.

The phenolic compounds were separated by two-dimensional chromatography on Filtrak FN-16 paper in systems: 1. 15% acetic acid, 2. n-butanol-acetic acid-water (BUV) 4: 1: 2.

Flavonoid compounds were detected by their own fluorescence in UV light (adsorption zones from pale yellow to brown). After chromatograms developed with a 3% alcohol solution of aluminum chloride and heated in an oven at a temperature of 100 ° С for 2-3 min, the fluorescence of the zones intensified and the flavonoid zones acquired a bright yellow, greenish-yellow color. In UV light, the phenol carboxylic acid adsorption zones exhibit blue fluorescence. After processing chromatograms in ammonia vapors, the color of the zones becomes bright blue.

Identification of substances was carried out with standard samples. Used 0.05% solutions of state standard samples (GSO): rutin, quercetin, myricetin, hyperoside, luteolin, as well as arbutin, gallic, chlorogenic, caffeic acids. The presence of quercetin was established - Rf ~ 0.05 (I), Rf ~ 0.71 (II); routine - Rf ~ 0.57 (I), Rf ~ 0.32 (II); luteolin - Rf ~ 0.50 (I), Rf ~ 0.65 (II); myricetin - Rf ~ 0.05 (I), Rf ~ 0.35 (II); hyperoside - Rf ~ 0.45 (I), Rf ~ 0.55 (II); caffeic acid - Rf ~ 0.34 (I), Rf ~ 0.79 (II); gallic acid - Rf ~ 0.63 (I), Rf ~ 0.57 (II); chlorogenic acid - Rf ~ 0.48 (I), Rf ~ 0.75 (II).

To characterize the authenticity of Baikalsky-6 dry tea extract, the TLC method was used on Kieselgel 60 F-254 plates (Merck), in the ethyl acetate-acetic acid-water system (5: 1: 1). Solvent passage 10 cm. UV observation -light (365 nm) before and after treatment with a 3% alcohol solution of aluminum chloride, followed by exposure in an oven at a temperature of (100-105) ° C for 2-3 minutes by comparing the color and chromatographic mobility of the detected zones with that standards identified: rutin - Rf ~ 0.27; hyperoside - Rf ~ 0.41; luteolin - Rf ~ 0.93; k Vercetin - Rf ~ 0.89.

By TLC on a Silufol plate in a chloroform-ethanol system (7: 3) followed by processing of the chromatogram with a 10% aqueous-alcoholic sodium hydroxide solution, after 5-7 minutes with freshly prepared diazoreaktiv, arbutin was identified: Rf ~ 0.23, manifested in the form of a bright red spot at the witness level.

The study of phenolic compounds by HPLC. The analysis was carried out on a Gilson instrument followed by computer processing of the research results using the Multichrom for Windows program.

Elution conditions: a Platinum EPS C-18 metal column of 4.6 × 250 mm LUNA C-18 size was used as the stationary phase, methanol-water-phosphoric acid (40: 60: 0.5) was used as the mobile phase. The analysis was performed by gradient elution at room temperature. The flow rate of the eluent is 1 ml / min. Analysis time 50.75 minutes. Detection was carried out using a UV detector at a wavelength of 254 nm.

The proposed method of obtaining funds is quite simple, does not require laborious costs, allows you to get a product of constant composition.

To prove the materiality of the proposed features of the method, namely the type of extractant, the ratio of raw materials-extractant, the duration and frequency of the extraction process, a series of experiments were carried out, the data of which are presented below.

The choice of optimal extraction parameters was controlled by the yield of extractive substances (HF XI, issue 1. P.295) and the content of active substances (sum of flavonoids and phenological glycosides).

To select the type of extractant, the processes of extraction of raw materials with water and ethyl alcohol of various concentrations were studied: 20, 30, 40, 50, 60, 70, 80, 96%. From the analysis results (table 1), we can conclude that the optimal extractant is 30% alcohol and hot water, which allows you to extract the maximum amount of extractive and active substances. A pharmacological screening study of extracts obtained by various extractants was also carried out. The extract obtained by extraction with hot water has a more pronounced diuretic effect. In this regard, it was proposed to carry out extraction with hot water, which is also economically advantageous.

The plant composition includes plants of various morphological and anatomical structures. The particle size of the crushed raw materials plays an important role in the intensification of the process. The degree of grinding of each type of raw material was studied.

A quantitative assessment was carried out according to the yield of extractives. The results of the analysis are shown in table 2.

Grinding of each type of raw material should be carried out separately, to the particle size: for bearberry and lingonberry leaves - 2 mm: bird grass, grass nettle and marigold flowers - 5 mm.

The dependence of the yield of the amount of extracted substances in various proportions to the amount of raw material was studied. The experimental data presented in table 3 indicate that the optimal ratio of raw materials-extractant 1: (10-12) for a single bay of raw materials was selected. A further increase in the volume of extractant is impractical, since the extraction efficiency is not achieved, since there is no increase in the yield of active substances.

In order to determine the duration and multiplicity of the number of extractions, the time of the onset of equilibrium concentration in the “feed-extractant” system was studied. According to the method: the mass of a sample of plant material was poured with purified water in a ratio of 1:10 and insisted at room temperature and stirring continuously for 2 hours. The process was repeated three times, filling the raw material with a new portion of extractant in an amount equal to the drained extraction. At certain time intervals, each contact of the extraction phases was poured and analyzed. The analysis results presented in table 4 show that the equilibrium state at two phase contacts is reached after 3 hours. With 1 phase contact after 120 minutes, at 2 after 60 minutes. Double extraction provides a fairly complete depletion of raw materials - up to 94% of active ingredients are recovered.

The technological scheme for producing diuretic agents has been tested on the equipment of the experimental plant NPO VILAR: n / a extractor with a capacity of 50 l, with a paddle-type mixer at 60 rpm, with heating and cooling (Volga-Vyatka CHX); vacuum evaporation apparatus (Czechoslovakia), with a capacity of 20 l / h (evaporation is carried out at a temperature not exceeding 40-70 ° C and a residual pressure of 0.1 kgf / cm ² or 9.8 kPa); separator; Vacuum drying cabinet (Poland) with a working pressure of 1.0-1.5 kgf / cm ² .

Before starting work, inspect and check the valves. On the false bottom of the extractor, a gray-tin cloth and two layers of calico are placed (when draining, the extraction is simultaneously filtered) and fixed with a spacer ring. Raw material is loaded into the extractor through the side hatch, and extractant is fed from the measuring device using a vacuum from above. Everything is carefully checked for leaks.

Examples of the method of obtaining the final product.

Example 1

300 g of bird grass, 150 g of dioica nettle leaves, 150 g of calendula officinalis flowers are ground in a mill to a particle size of 5 mm in diameter (sieve No. 50 GOST 214-83), 200 g of bearberry leaves and 200 g of lingonberry leaves are ground to size particles with a diameter of 2 mm (sieve No. 20 GOST 214-83). Then they are packaged in bags and, if appointed, brewed like tea with boiling water.

Example 2

The crushed raw materials of example 1 are loaded into an extractor with a stirrer and external steam heating. Pour 14 liters of purified water in a ratio of raw materials-extractant 1:10 taking into account the coefficient of water absorption 1: 3 and the false bottom of the extractor. Extracted at a temperature of (80-90) ° C for 2 hours with constant stirring. The extract is filtered through a filter in the collector. In a similar way, a second extraction is carried out for 1 h, feeding purified water to the extractor in an amount equal to the volume of the drained extract. 1 drain - 10.3 liters, 2 drain - 10.1 liters. The combined extracts are evaporated to 1/5 of the original volume and purified by separation. The concentrated extract is evaporated to a densely viscous state of the mass, dried in a vacuum oven at a temperature of (55 ± 5) ° C for 5 hours and crushed.

Get 276 g of the finished product, which is 27.6% by weight of the feedstock.

The diuretic activity of the finished product was evaluated in accordance with the “Guidelines ...” (1989). The experiments were carried out on white rats of both sexes of the Wistar line weighing 190-200 g. Baikalsky-6 dry tea extract in the form of an aqueous solution was administered once intragastrically at a dose of 40 mg / kg (1/15 LD ₅₀ ) - the optimal conditionally therapeutic dose was determined in preliminary experiments, in a volume of 1 ml / 100 g 30 minutes before the water load, which is 2.5% of body weight. Animals of the control group received an equivolume amount of distilled water according to a similar scheme. As a comparison drug, we used “diuretic collection No. 2” (multicomponent collection) prepared according to GF XI in a volume of 1 ml / 100 g, which is a decoction of a mixture of bearberry leaves, licorice roots, and juniper fruits. Urine collection was carried out in metabolic cells within 4 hours after water loading. The volume of urine excreted was measured, and the content of potassium and sodium ions was determined by flame photometry on a Flapho-4 instrument (Germany).

The results are presented in table 5.

As follows from the data given in table 5, a single administration of a dry extract in the specified dose had a pronounced stimulating effect on the excretion of water and sodium ions in white rats. Moreover, the diuretic activity of the test agent exceeded that by 10% in the comparison drug.

In the next series of experiments, the diuretic effect of the dry extract was evaluated in a model of toxic damage to the kidneys, which was caused by a single intraperitoneal injection of a solution of mercuric chloride (HgCl ₂ ) at a dose of 2 mg / kg (Sokolova et al., 1975). The experiments were carried out on 24 white rats of both sexes of the Wistar line weighing 160-180 g. Animals of the experimental group were intragastrically injected with an aqueous solution of dry extract at a dose of 40 mg / kg (1/15 LD ₅₀ ) in a volume of 1 ml / 100 g - determination of the optimal conditionally therapeutic dose carried out in preliminary experiments. The first administration of the test substance was carried out 1 hour before the introduction of the mercuric chloride, and then 1 time per day for 7 days. The reference preparation was “diuretic collection No. 2” prepared according to GF XI, which was administered in a volume of 1 ml / 100 g of mass, which was administered according to a similar scheme. Animals of the control group received an equivolume amount of distilled water.

It was established that the course administration of a dry extract at a dose of 40 mg / kg increases diuresis in white rats by 1.5-2 times against the background of renal damage by sublimate (table 6). Moreover, the activity of the test agent exceeded that of the comparison drug.

The acute toxicity of the dry extract was investigated according to the generally accepted Kerber method (Requirements ..., 1984). The experiments were carried out on 36 white rats of both sexes of the Wistar line weighing 160-180 g. The test product in the form of an aqueous solution was administered once intraperitoneally at doses of 300-800 mg / kg. Observation was carried out for 14 days. Studies have shown that the LD _{50 of the} dry extract is 550 mg / kg (470.1-629.9). The results obtained allow us to classify the test drug as low toxic according to the classification of K. Sidorov. (1973).

Example 3

300 g of bird grass, 150 g of dioica nettle leaves, 150 g of calendula officinalis flowers are ground in a mill to a particle size of 5 mm in diameter (sieve No. 50 GOST 214-83), 200 g of bearberry leaves and 200 g of lingonberry leaves are ground to size particles with a diameter of 2 mm (sieve No. 20 GOST 214-83). The crushed raw materials are loaded into an extractor with a stirrer and external steam heating. Pour 16 liters of purified water in a ratio of raw materials-extractant 1:12, taking into account a ratio of 1: 3 and the false bottom of the extractor. Extracted at a temperature of (80-90) ° C for 2 hours with constant stirring. The extract is filtered through a filter in the collector. In a similar way, a second extraction is carried out for 1 h, feeding purified water to the extractor in an amount equal to the volume of the drained extract. 1 drain - 11.7 liters, 2 drain - 11.4 liters. The combined extracts are evaporated to 1/5 of the original volume and purified by separation. The concentrated extract is added to a densely viscous state of the mass, dried in a vacuum oven at a temperature of (55 ± 5) ° C for 4 hours and ground.

Get 284 g of the finished product, which is 28.4% by weight of the feedstock.

The diuretic activity of the dry extract was evaluated according to the generally accepted method with 2.5% water load on 18 white rats of both sexes of the Wistar strain, weighing 190-200 g. For a description of the experimental design, see the previous section. The test product was administered once intragastrically at a dose of 40 mg / kg. The results are shown in table 7.

As follows from the data given in table 7, a change in the extraction conditions practically does not affect the diuretic activity of the test drug.

In the next series of experiments, the diuretic effect of the dry extract was evaluated in a model of toxic damage to the kidneys, which was caused by a single intraperitoneal injection of a solution of mercuric chloride (HgCl ₂ ) at a dose of 2 mg / kg (Sokolova et al., 1975). The experiments were carried out on 24 white rats of both sexes of the Wistar line weighing 160-180 g. For a description of the experimental design, see the previous section. The test agent was administered once intragastrically at a dose of 40 mg / kg. The results are shown in table 8.

Claims (5)

1. A tool with diuretic activity, including the leaves of lingonberry vulgaris, characterized in that it additionally contains bird grass, leaves of bearberry, leaves of the nettle, flowers of calendula officinalis in the following ratio, wt.%: Lingonberry leaves 20 Highlander's Grass 30 Bearberry leaves 20 Stinging nettle leaves 15 Calendula officinalis flowers 15 2. The tool according to claim 1, characterized in that it is made in the form of a collection, dry extract, capsules, tablets. 3. The method of obtaining funds with diuretic activity, including extraction of plant materials, evaporation of the combined extracts, separation and drying, characterized in that the raw material according to claim 1 is extracted at a temperature of 80-90 ° C with purified water at a ratio of raw material: extractant 1:10 -12 taking into account the coefficient of water absorption, while the first extraction is carried out for 120 minutes, and the second for 60 minutes, the drying is carried out in a vacuum dryer, followed by grinding. 4. The method according to claim 3, characterized in that the extraction is evaporated to 1/5 of the original volume. 5. The method according to claim 3, characterized in that it is dried at a temperature of 55 + 5 ° C for 5 hours