Method of producing α(1,2)-l-skeleton-α(1,4)-d-galactopyranosyluronan from acorus calamus l. rhizome

FIELD: chemistry.

SUBSTANCE: method of producing α(1,2)-L-skeleton-α(1,4)-D-galactopyranosyluronan from Acorus calamus L rhizome is carried out in two steps. At the first step, crushed material is extracted with acidified water and heated on a boiling water bath while periodically mixing; after settling, the mixture is heated once more, left to cool to room temperature and filtered through a multilayer fabric filter; the filtrate is evaporated under a vacuum; the obtained solution is slowly added to 96% ethyl alcohol or to a solution of reclaimed ethanol and left in a cool place to settle the residue, after which the settled solution is drained, and the residue is filtered through a paper filter by successively washing with 96% ethyl alcohol and then with acetone. At the second step, without drying, the residue is transferred from the filter and dissolved in purified water while stirring rapidly; the obtained solution is then centrifuged; the obtained solution is purified from low-molecular weight compounds by filtering through a semi-permeable membrane; the purified solution is frozen and freeze-dried at certain conditions.

EFFECT: improved method.

3 dwg, 6 tbl

 

The invention relates to the field of pharmaceutical industry, to methods for producing an α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L., which can be used as a tool for complex therapy of malignant tumors.

The known means of enhancing antitumor and antimetastatic activity of cytotoxic drugs [RU 2471496 C1, published on 10.01.2013], which is a polysaccharide (galactosialidosis pectins), isolated from plant material the rhizomes of calamus (Acorus calamus L.). Polysaccharide α(1,2)-L-frame-α(1,4)-D-galactopyranoside has the most pronounced biological activity, described galactosaemia pectin, and can be used to develop new medicines for the treatment of malignant tumors. Question for the isolation and purification of α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L. requires the development of new technical approaches.

A method of producing from plant material of galacturonans anti-inflammatory effect [RU 2344829 C1, published 27.01.2009]. A method of producing from plant material of galacturonans includes a preliminary treatment of the raw material aqueous solution of formalin, the solution is acidified water, extraction water�th solution of ammonium oxalate, centrifugation of the extract, adding HCl to a certain value, the exposure under certain conditions, neutralization, filtration, concentration of the extract to ultrafiltration column and lyophilic drying of the target product

The closest to the proposed is a method of producing the amount of water-soluble polysaccharides from the rhizomes of calamus (Acorus calamus L.) [EN 2311918 C1, published on 10.12.2007]. The method is two-fold extraction with water of the rhizomes of calamus, the deposition of polysaccharides two-fold volume of 96% ethanol, centrifuging and drying. The tool has an immunostimulating effect. The disadvantage of this method is that there was no phase separation of α(1,2)-L-frame-α(1,4)-D-galactopyranoside obtained from the amounts of polysaccharides.

The technical result of the invention is to obtain the target product with a higher degree of purification, increase output and expand the Arsenal of methods of obtaining medicines from the rhizomes of calamus.

To achieve the technical result in the method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L., including the extraction of vegetable raw material with water under heating, followed by precipitation with 96% ethanol, filtration and drying the precipitate, in the first stage crushed to 3-7 mm raw EC�tracerout water acidified to pH of 2.0 to 3.0 with concentrated hydrochloric acid, and heated in a boiling water bath for 1 hour with occasional stirring, after a 24-hour settling, the mixture is again heated for 1 hour, leave to cool to room temperature and filtered through layers of fabric filter, the filtrate is evaporated under vacuum at a temperature not exceeding 50°C up to 100 ml, the resulting solution was slowly poured into 300 ml of 96% ethyl alcohol or the regenerated solution of ethanol with a concentration of not less than 60% when the ratio extract:ethanol 1:3, accordingly, and leave in a cool place for settling of sediment for 24 hours, after which the supernatant solution is decanted, and the residue was filtered through a filter paper by washing sequentially with 96% ethyl alcohol, then with acetone, in the second stage, the precipitate without drying, is transferred from the filter, dissolved in 100 ml of purified water with rapid stirring for 3 hours at room temperature, then the solution was centrifuged for 30 min at 5000 rpm, then the resulting solution is purified from low molecular weight compounds by filtration through a semipermeable membrane with a pore size of 5 kDa, next, the purified solution is frozen and freeze-dried.

New in the proposed method is that the first �Tapa crushed to 3-7 mm raw material was extracted with water, acidified to pH of 2.0 to 3.0 with concentrated hydrochloric acid, and heated in a boiling water bath for 1 hour with occasional stirring, after a 24-hour settling, the mixture is again heated for 1 hour, leave to cool to room temperature and filtered through layers of fabric filter, the filtrate is evaporated under vacuum at a temperature not exceeding 50°C up to 100 ml, the resulting solution was slowly poured into 300 ml of 96% ethyl alcohol or the regenerated solution of ethanol with a concentration of not less than 60% when the ratio extract:ethanol 1:3 and leave in a cool place for settling of sediment for 24 hours, after which the supernatant solution is decanted, and the residue was filtered through a filter paper by washing sequentially with 96% ethyl alcohol, then with acetone, in the second stage, the precipitate without drying, is transferred from the filter, dissolved in 100 ml of purified water with rapid stirring for 3 hours at room temperature, then the solution was centrifuged for 30 min at 5000 rpm, then the resulting solution is purified from low molecular weight compounds by filtration through a semipermeable membrane with a pore size of 5 kDa, next, the purified solution is frozen and freeze-dried.

A new method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside out to�revis Acorus calamus L. was based on the experimental results.

A new method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L. for the expert explicitly does not follow from the prior art and description it is not detected by the authors in the patent and scientific and medical literature. The proposed method is tested in laboratory conditions. Thus, the proposed technical solution meets the criteria of the invention, namely of "novelty", "inventive step" and "industrial applicability".

The method is as follows

stage 1. Weighed raw materials rhizomes of Acorus calamus L. with a particle size of 3-7 mm (20.0 g), pour a solution of 400 ml of purified water and 2 ml of concentrated acid hydrochloric (pH 2,0-3,0) and heated in a boiling water bath for 1 hour with occasional stirring. After a 24-hour settling, the mixture is again heated for 1 hour, leave to cool to room temperature and filtered through layers of fabric filter. The filtrate is evaporated under vacuum at a temperature not exceeding 50°C to 100 ml of the resulting solution was slowly poured into 300 ml of 96% ethyl alcohol or the regenerated solution of ethanol with a concentration of 60% or more and leave in a cool place for settling of sediment for 24 hours. Supernatant solution is decanted, and sadakathullah through the paper filter washing consistently 96% ethyl alcohol, then with acetone.

stage 2. The residue, without drying, is transferred from the filter into a glass beaker and dissolved in 100 ml of purified water with rapid stirring on a magnetic stirrer for 3 hours at room temperature. Then the solution was centrifuged (5000 rpm, 30 min - the solution should be transparent and contain Muti). After centrifugation, the resulting solution is purified from low molecular weight compounds by filtration through a semipermeable membrane with a pore size of 5 kDa, and the purified solution is frozen and freeze-dried.

Chemical structure of selected polysaccharide was determined by the methods of chromatography-mass spectrometry and NMR spectroscopy (Figures 1 and 2).

The quality of the final product was determined by molecular mass distribution (exclusion chromatography), the content of uronic acids (spectrophotometric method) and the impurity content of protein (Lowry method).

Determination of the molecular weight distribution

Molecular mass distribution (MMD) characterizes the degree of homogeneity of the polymeric substance - dependence of the distribution of mass-average molecular weight and relative content in the substance. Was used refractometric detection, based on the ability �(1,2)-L-frame-α(1,4)-D-galactopyranoside to refract light passing. The chromatography was performed on a chromatographic system Ultimate 3000 ("Dionex, Germany) equipped with a plunger pump at a feed rate of eluent of 0.1-5 ml/min, the holder of columns, refractometric detector. The chromatography conditions were chosen empirically: column - TSK GMPWXL, 300*78 mm, 13 µm; flow rate of eluent (water) to 1 ml/min; column temperature - 25°C; temperature of the cell of the detector is 40°C.

The analysis of the final product α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. showed that the content of high-molecular fraction ≥5 kDa is not less than 99%.

The definition of the content of uronic acids

The content of uronic acids was determined by the carbazole-sulfuric method (modified by the addition of sulfamic acid), adapted for α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. in terms of the d-galacturonic acid by spectrophotometry. The optical density of the investigated solution was determined at a wavelength of 535 nm.

Solution comparison served as a mixture of sulfuric acid with sodium tetraborate (1500 μl), purified water (250 μl), sulfamic acid (10 µl) and carbazole (50 µl).

For the calibration curve, constructed by galacturonic acid, found its contents ( % ) optical density of the sample.

The calculations were carried out according to the formula (1)

,

Cxthe amount of uronic acids was found in the calibration schedule, mg/ml; 1, 1,81 - breeding; 0,05 - volume of the test solution taken for analysis, ml; is 0.0010 - accurate mass UCS, g; 1000 - translation g in mg.

The target protein - α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. shall contain not less than 20.0% galacturonic acid.

Determination of protein impurities

Despite several purification stages in obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. in its composition may contain substances of protein nature, which has similar physico-chemical properties of the polysaccharides. In GF XII are a few methods to determine the protein content, the most widespread methods of Lowry and Bradford. The latter is less frequently used because it has several disadvantages: the specificity of positive amino acids, which leads, sometimes, to create bias and inflated indicator of the protein content; the reagent Kumasi G250 is not stable in solution, which leads to the need at the next analysis to prepare a series of standard solutions of bovine serum albumin and to calibrate. Therefore, it was chosen the method of Lowry as a working method for determining protein impurities in α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. the Analysis showed a slight with�holding protein not more than 1.8%.

A new method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L. was based on the experimental results.

The justification mode of the method

1. Study of parameters extraction.

Intermediate product to obtain the α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. is a complex of water-soluble polysaccharides (VRPS) from the rhizomes of calamus, therefore, the first phase for developing optimal technological parameters of the method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. with the purpose of increasing the yield of the final product were determined the optimal parameters of the extraction of the complex VRPS from raw materials in their relationship.

In the experiment studied the effect on the output of the complex VRPS such parameters as:

the size of particles of raw materials;

- pH of the extractant (regulated hydrochloric acid);

- during heating in a water bath.

To study the particle size of the feedstock calamus rhizomes were divided sieve method on fractions according to the particle size of raw materials. Use a set of sieves with holes of a diameter of from 1 to 7 mm. After sieving of raw materials received fractions with different particle size: 1.0 to 2.0 mm; 2,0-3,0 mm; 3,0-5,0 mm 5,0-7,0 mm.

To study the influence of pH on the extraction of complex VRPS from the rhizomes �Ira marsh used aqueous solutions, acidified with hydrochloric acid to pH=2,0; 2,5; 3,0; 3,5.

The use of acidified extractant for separation of polysaccharide complex (KTC) from the rhizomes of calamus due to the nature of the active ingredients. In plant raw materials part of the polysaccharides (PS) is in the form of salts that are not soluble in aqueous solutions. Therefore, to improve the solubility of PS in the extractant is added hydrochloric acid, low concentration of which contribute to the destruction of the PS salts and transition them into the solution.

Study of the effect of time of heating in a water bath due to the fact that the number of extracted substances from vegetable raw materials in proportion to the duration of the extraction. As the length of time the qualitative composition of the extract is deteriorating due to the amount of ballast substances. It is therefore necessary to strive to ensure that the completeness of extraction was achieved in the shortest possible time, because the quality of the extraction process right to be judged not on the amount of extracted substances, and the component composition of the product. Investigated the extraction time: 20; 40; 60 and 80 min.

To reduce the amount of research used the method of mathematical modeling on the basis of Greco-Latin square. The analysis of the experimental results was performed using the software package "Statistica for Windows 6.0". Factor�, influencing the process of entering complex VRPS presented in table 1. Plans were made so that each level of factor once present in each row (column) of the plan, and each combination of factor levels was seen only once.

The optimization criteria were: the emergence of a set VRPS and the release of acidic polysaccharides. To evaluate the significance of these factors on the plan of the experiment conducted 16 experiments in conditions that are prescribed by the planning matrix. The results of the experiment were subjected to analysis of variance. Homogeneity of variance was checked using F-Fisher's criterion with regard to the number of degrees of freedom. The matrix of experimental design and the results are presented in table 2.

Analysis of the obtained data allowed to identify factors that have a significant impact on the process of polysaccharides from raw materials (table 3).

The data obtained allowed to determine the content of the complex WRPS in the raw sum of squares of deviations from the mean caused by the impact on this indicator, and display the level of influence of factors in %:

- controlled factors SSI=SSA+SSB+SSC=13,10245;

- uncontrolled random factors and measurement errors SSII=3,21495;

- total SS=SSI+SSII=16,31740.

The value of the degree of influence each�wow factor was K A=8,49%; KB=64,24%; KC=7,56%; overall control factors - KI=80,3%; uncontrollable random factors and measurement errors - KII=19,7%.

Consequently, the greatest influence on the output of the PSC supported the factor B - pH of the extractant, the factors A (particle size) and C (heating time) did not demonstrate a significant effect on the output of the PSC.

The sum of squares of deviations in the analysis of factors on the yield of acidic polysaccharides (CP) was equal SSI- 0,23795, SSII- 0,12096 and SS - 0,35891.

The greatest influence on the process of allocating KP had factor B - pH of the extractant (KB=36,53%). The particle size and heating time to a small extent influenced the release of KP from plant material (KA=22,22 and KC=7,55%). The share of uncontrolled random factors accounted for 33.7 per cent.

Thus, it is established that the most intensively efficiency of extraction of polysaccharides increases with decreasing pH of the medium.

To estimate the parameters inside factors were calculated the average values and variances for each parameter (table 4).

The data obtained show that with the increase of particle size of raw materials from 1 mm to 7 mm output complex VRPS increased. For vegetable raw materials with a particle size of 2-3 mm, 3-5 mm and 5-7 mm these figures differed slightly, within error. When you change the pH of the extractant from 3.0 to 2.0 output comp�of EXA VRPS was significantly increased. The extraction time had no significant influence on the yield of polysaccharides and the data obtained cannot be reliably judge what time of extraction is optimal.

The definition of KP shows that the yield increases with increasing the particle size of the raw material and the pH of the extractant in the acidic side. The highest content of CP was observed at pH 2.0 and the particle size of 3-7 mm. the extraction Time has no significant effect on the yield of acid polysaccharides. None of the factors have no significant effect on the output of the CP.

Analysis results indicate that the degree of grinding of raw materials and the extraction time had little impact on the extraction of polysaccharides from the rhizomes of calamus, and pH of the extractant is a significant factor for the output of the complex WRPS.

Thus were defined:

the size of particles of raw materials - 3-7 mm;

- pH of the extractant (regulated hydrochloric acid) to pH=2,0;

- during heating in a water bath for 1 hour.

2. Study of the conditions of evaporation of the extraction.

To select the operating conditions of evaporation of the extract on a rotary evaporator was given a series of experiments on the variation of technological parameters of processing. The data obtained are presented in table 5.

As seen from table 5, the minimum time of mpariwa�Oia is achieved by the following parameters of the rotary evaporator: temperature of the water bath 50°C, the rotation speed of the evaporator flask 40 rpm, the coolant temperature in the refrigerator at 2-5°C and a system pressure of 30 mbar. These conditions were chosen as the operating parameters at the stage of evaporation of water extraction.

3. Study of the effect of the concentration of ethanol in the precipitation of the polysaccharide complex with ethanol.

At the stage of deposition of the polysaccharide complex ethyl alcohol remains a large number of water-ethanol solution containing 45-70% ethyl alcohol. This waste may be regenerated by distillation in a rotary evaporator until the concentration of ethanol 60-80% and re-used for the deposition of the polysaccharide complex from the aqueous extract, which will lead to the cheapening of the product and reduce the amount of hazardous waste production. The use of regenerates with ethanol content of 60-80% requested additional experimental studies of the effect of different concentrations of ethyl alcohol(60, 65, 70, 75, 80 and 96%) on the yield and quality indices of α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L.

Data on the concentration of ethanol, ratio of the extract and precipitating agent, received a Deposit, the content of galacturonic acid and yield is shown in table 6.

The table shows that concentration� ethyl alcohol affect the density of the sediment, the precipitate obtained in all cases was homogeneous, but different volume, mass, color and texture. When the concentration of ethanol, the precipitate became more and more thick, more strong dehydration of sludge while reducing the weight.

The ratio of extract and ethyl alcohol affect product yield, at a ratio of 1:2 product yield decreased.

Neither the concentration of ethanol or the ratio of the extract and precipitating agent did not affect the content of galacturonic acid in the final product.

Thus, as the optimal mode of deposition, it is advisable to use reclaimed ethyl alcohol with a strength of 60% or more in the ratio 1:3.

4. The use of centrifugation for the purification of polysaccharide complex.

In the further development of the method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L. it was observed the deposition of sediment in the semis when stored in the refrigerator (5±3°C). Sediment loose white, turbid supernatant. In the study of technological parameters used in the receipt, it was found that the residues associated with the duration of stages of filtration polysaccharide complex. During filtration the solution was heated to room temperature, the resulting solution was transferred to a water-soluble fraction of cu�chmela and other polysaccharides, which upon further cooling, the precipitate.

The problem is solved by introducing a stage of centrifugation, enabling to reduce the time stage and be cleaned in the cooling mode using a refrigerated centrifuge. To explore the possibility of using centrifugation were investigated.

To solve this technological problem, it is advisable to use speed centrifugation at 5000 rpm, the cooling of the working chamber of the centrifuge to +5°C.

5. The use of filtration through a semipermeable membrane for purification of polysaccharide complex.

At the stage of purification of the solution VRPS subjected to washing with organic solvents, but even small residual amounts of solvents may adversely affect the quality of the product. To purify the resulting product from organic solvents and low molecular weight impurities using prefabricated dialysis through a semipermeable membrane with a pore diameter of 5000 daltons.

To confirm the effectiveness of filtration, purification method, determined the composition of substances by using HPLC method.

The molecular weight of the polysaccharides included in the composition of the samples was established by retention time in accordance with the calibration� values identified by standard samples dekstranov.

From the chromatograms (Fig.3) shows that the proposed technology allows to extract the polysaccharides with the desired molecular weight. Presents peaks A and B with retention times 4,580 6,205 minutes and min correspond to the molecular mass of 380 and 720 kDa. Also on the chromatogram, we see the absence of low molecular weight impurities, which confirms the efficiency of the purification method used.

Thus, the experimentally determined parameters of the method of obtaining α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L.:

the size of particles of raw materials - 3-7 mm;

- pH of the extractant (regulated hydrochloric acid) to pH 2,0;

- during heating in a water bath for 1 hour;

- evaporation conditions: the temperature of the water bath 50°C, the rotation speed of the evaporator flask 40 rpm, the coolant temperature in the refrigerator at 2-5°C and a system pressure of 30 mbar;

for deposition, it is advisable to use reclaimed ethyl alcohol with a strength of 60% or more in the ratio of extract:astrigent 1:3;

- centrifugation for 30 min at a speed of 5000 rpm, the cooling of the working chamber of the centrifuge to +5°C;

for purification of the target product - filtering semi-finished product through a semi-permeable membrane with a pore diameter of 5000 daltons.

Annex 1

Figure 1 - Spectrum of13C-NMR �(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L.

Figure 2 - Spectrum1H-NMR of α(1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L.

Figure 3 - Chromatogram of the complex VRPS from the rhizomes of calamus.

Table 1 - Factors affecting the extraction process of polysaccharides from the rhizomes of calamus.

Table 2 - symmetric three-Factor fractional plan of the second order based on the Latin square and the results of the study of polysaccharides from the rhizomes of calamus.

Table 3 - ANOVA test results polysaccharide complex obtained from the rhizomes of calamus.

Table 4 - Average values, deviations from the average and the average quadratic deviation of the average values of the fractional plan based on the Latin square.

Table 5 - the Dependence of the rate of evaporation of 1 liter of water extraction on the technological parameters of the rotary evaporator.

Table 6 - the Effect of different concentrations of ethanol in the precipitation of the polysaccharide complex of evaporated water extract of rhizomes of Acorus calamus L. on the yield and quality indices of α(1,2)-L-frame-α(1,4)-D-galactopyranoside Acorus calamus L.

A method of producing α1,2)-L-frame-α(1,4)-D-galactopyranoside from the rhizomes of Acorus calamus L., including the extraction of vegetable raw material with water under heating, followed by precipitation with 96% ethanol, filtration and drying the precipitate, characterized in that the first stage crushed to 3-7 mm raw material was extracted with water, acidified to pH 2.0 with concentrated hydrochloric acid, in the ratio of vegetable raw material and extractant 1:20 and heated in a boiling water bath for 1 hour with occasional stirring, after a 24-hour settling, the mixture is again heated for 1 hour, leave to cool to room temperature and filtered through layers of fabric filter, the filtrate is evaporated under vacuum at a temperature not exceeding 50°C up to 100 ml, the resulting solution is slowly poured in 96% ethyl alcohol or in a solution of recovered ethanol concentration of at least 60% when the ratio of extract and ethanol 1:3 and leave in a cool place for settling of sediment for 24 hours, after which the supernatant solution is decanted, and the residue was filtered through a filter paper by washing sequentially with 96% ethyl alcohol, then with acetone, in the second stage, the precipitate without drying, is transferred from the filter dissolved in 100 ml of purified water with rapid stirring for 3 hours at room temperature, then the solution was centrifuged for 30 min at 5000 Rev/mi� and cooling of the working chamber of the centrifuge to +5°C, then the resulting solution is purified from low molecular weight compounds by filtration through a semipermeable membrane with a pore size of 5 kDa, then the purified solution is frozen and freeze-dried.



 

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FIELD: chemistry.

SUBSTANCE: invention represents a cosmetic preparation for skin in the form of a "water-in-oil" type emulsion, which contains water, ethanol, volatile oily component, carboxydecyltrisiloxane of formula (1): 0.1-5 wt % and one, two or more components, selected from the group, consisting of hydrophibised titanium dioxide, hydrophibised zinc dioxide and hydrophibised iron dioxide.

EFFECT: removal of powder squeakiness on the skin in the course of time after application and simultaneous improvement of absorption into the skin during application and absence of stickiness after application on the skin.

3 cl, 2 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical composition for treating insomnia. Said composition includes root of Polygonum multiflorum and/or its extracts, seeds of Ziziphus spinosa and/or its extracts, mulberry fruit and/or its extracts, Ganoderma and/or its extracts, lily bulb and/or its extracts, Anemarrhena rhizome and/or its extracts, root of Salvia miltiorrhiza and/or its extracts, chrysanthemum flower and/or its extracts, Poria and/or its extracts and Albizia flower and/or its extracts. Invention also relates to application of said composition.

EFFECT: claimed invention provides sedative and soporific effect, contributes to memory stimulation.

17 cl, 6 tbl, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to oral hygiene preparations. An oral care composition contains a) an aqueous phase; b) stannous tin ions dissolved in the aqueous phase; c) nitrates in the aqueous phase; wherein total nitrates is that a molar amount of nitrogen measured as a nitrate in the aqueous phase makes 1.8-0.1 of the molar amount of stannous tin ions; and d) a flavouring agent. What is also presented is a method for storage and a storage container for this composition; using nitrate for stabilising oxidised stannous tin ions dissolved in the aqueous phase. What is also presented is also a version of the composition, wherein the composition is aqueous.

EFFECT: using the group of inventions stabilises oxidised stannous tin ions dissolved in the aqueous phase by the above molar ratio of nitrate and stannous tin ions.

17 cl, 2 tbl, 15 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the pharmaceutical industry, namely to a medication for the complex therapy of patients with type 2 diabetes mellitus and a method of the complex therapy of patients with type 2 diabetes mellitus. The medication for the complex therapy of patients with type 2 diabetes mellitus contains a cleaned and dried rhizome of Curcuma longa in the form of powder with the specified size of particles. The method of the complex therapy of patients with type 2 diabetes mellitus consists of the daily intake of the cleaned and dried rhizome of Curcuma longa in the form of powder with the specified size of particles twice per day after dinner and supper at the background of the intake of pelleted hypoglycaemic agents without changing the dose and scheme of the drug therapy.

EFFECT: medication produces a direct stimulating impact on β-cells of the pancreas and ensures the development of a hypoglycaemic effect, including early stages of the disease development.

3 cl, 1 dwg, 5 tbl

FIELD: chemistry.

SUBSTANCE: invention refers to using an oral composition containing steviol, which improves the human hair or animal fur/feather/scale look. Using steviol in preparing the oral composition of nutriceuticals or a nutrient composition for improving the human hair or animal fur/feather/scale look.

EFFECT: invention provides improving the human hair or animal fur/feather/scale look more effectively.

7 cl, 4 dwg, 5 tbl, 13 ex

FIELD: veterinary medicine.

SUBSTANCE: method comprises administering a homeopathic agent. The agent "Liarsin" is used, which is administered subcutaneously before calving into the biologically active points in the centre of the front and rear udder parts.

EFFECT: invention is highly effective for prevention of parturient paresis in cows.

4 cl, 2 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and represents method of obtaining microcapsules of medication, which possess supermolecular properties, by method of precipitation with non-solvent, characterized by the fact that as medication applied is powder of unabi berries, preliminarily dissolved in butanol, as envelope - carrageenan, which is precipitated from solution in acetone by addition as non-solvent of ethanol and water, with the following drying at room temperature.

EFFECT: invention ensures simplification and acceleration of process of obtaining microcapsules, reduction of loss in the process of obtaining microcapsules (increase of output by weight).

2 ex, 18 dwg

FIELD: medicine.

SUBSTANCE: method includes the preparation of a carious cavity, opening the tooth cavity, creation of an access to root canals, extension of their orifice. Removal of a decayed material from the root canals and their drug treatment are carried out. After that, performed are: wide opening of the apical tooth orifice, mechanical and drug removal of periapical pathological exudative formations in the focus of periapical inflammation through the root canal. Before filling the canal with a filling material the gel "Lamifaren" is introduced into the focus of periapical destruction. Such introduction is performed three times after a day under temporary stopping. The gel "Lamifaren" is introduced inside in a dose of 50 g 2 times per day for 30 days.

EFFECT: application of the invention accelerates bone tissue regeneration due to the active release of an active substance calcium alginate, provides stable remission due to local and systemic detoxifying effect.

2 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to methods of purification and health improvement of an organism. For this purpose therapeutic starvation for not fewer than 5 days in case of a 7-day programme and for not fewer than 7 days in case of a 9-day programme is carried out. The duration of a recovery period constitutes two days. Food intake is realised nine times per day each day both in the process of therapeutic starvation and in the recovery period. In the period of therapeutic starvation the first food intake includes bee products "APIGRANULES 2" in a dose of one teaspoon, "KHINAZI balm" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "APITOK" in a dose of one teaspoon, "Antihelm phyto" - one capsule and still mineral water - one glass. The second food intake includes a drink with ginger, lemon juice, garlic and mint, honey - one teaspoon and one glass of apple-carrot juice. The third food intake supposes an intake of bee products "APIGRANULES 2" in a dose of one teaspoon, "KHINAZI balm" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "Antihelm phyto" - one capsule and still mineral water - one glass. The fourth food intake includes a drink with ginger, lemon juice, garlic and mint, honey - one teaspoon and one glass of apple-carrot juice. The fifth food intake consists of tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The sixth food intake consists of bee products "APIGRANULES 2" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "Antihelm phyto" - one capsule and one glass of still mineral water. The seventh food intake includes tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The eighth food intake supposes an intake of tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The ninth intake of food includes depressant tea, including mint grass, valerian root, fennel seeds, cumin seeds, epilobium or strawberry leaves and one glass of apple-carrot juice. In the recovery period in 1-st, 2-nd, 3-rd, 4-th and 6-th intakes of food the composition of products remains the same as in the process of starvation. For the fifth intake of food the patients take tea black or green with ginger, one teaspoon of honey and vegetable soup. For the seventh intake of food the patients take tea black or green with ginger, "MILK COCKTAIL WITH CHITOSAN". The eighth intake of food includes tea black or green with ginger, one teaspoon of honey, "APICAMPA" cereal. The ninth intake of food for the recovery period includes a baked apple, depressant tea. In addition, a number of procedures are carried out after each food intake during the period of therapeutic starvation and the recovery period. After the first food intake gymnastics "5 Tibetan pearls" is realised. After the second food intake exercises on training apparatuses, procedures of press-therapy, electrolypolysis and myostimulation are realised. After the third food intake exercises of therapeutic physical training or aerobics are performed. After the fourth food intake a rest in form of a walk, a halotherapy session, combined with a session of relaxation therapy are realised. After the fifth food intake a course of strip-plastic is carried out. After the sixth food intake massage by manual application with a peloid-based mixture or manual massage with honey is carried out. After the seventh food intake an infra-red sauna, shower, phytobath with medicinal herbs, honey, ginger, lemon juice is taken. After the eighth food intake a shower is taken.

EFFECT: method ensures effective health improvement of the organism with the preservation of the full value life style and a sense of a comfortable state in the starvation period, reduction of recovery period term after starvation, purification of the organism without loading on the gastrointestinal tract, weight loss, and improvement of the general state and workability of the patients.

3 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a novel compound - N,O-(2,3-dihydroxypropyl)chitosanyl-borate, having formula , where m=500-3000.

EFFECT: compound has antibacterial, immunomodulating and antitoxic effect.

1 tbl, 6 ex

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