RU2106876C1 - Method for obtaining totality of hydroxy-cinnamic acids producing cholagolic effect - Google Patents

Abstract

FIELD: pharmacology; production of totality of dry purified hydroxy- cinnamic acids from Valeriana officinalis herb. SUBSTANCE: valerian officinalis herd is comminuted by milling and extracted with 60-70-% aqueous ethanol solution, weight ratio herb-extract is 1:5-7. Extraction is conducted by filtration method. Extract is evaporated until ethanol is removed. Resulting evaporated mass is diluted with sodium chloride to reach sodium chloride content 2-10% in eluent, followed by evaporating purified final product, eluates, and by drying. Inventive novelty of method resides in fact that sodium chloride is used, as eluent, for the first time in technology and its use leads to maximum recovery of hydroxy-cinnamic acids from all concomitant substances. Yield of final product reaches 3.0% based on weight of vegetable raw material. EFFECT: high yield of product; simplified technology. 4 tbl

Description

 The invention relates to the field of pharmacy and relates to a method for producing a dry substance of a purified amount of hydroxycinnamic acids from a medicinal valerian herb having a certain choleretic activity.

 Known methods for producing from plant sources of funds with choleretic activity.

 So, according to the method (Zubitsky D.N., Filippov Yu.A. A method of obtaining funds having choleretic activity. RF Patent N 1816216, class A 61 K 35/78. -BI N 18, 1993) receive a combined tool containing a mixture of extracts from a number of plants, including valerian root, with 50% alcohol, with the addition of various salts.

In the method (Glyzin V.I., Smirnova L.P., Obyedkova E.F. and others. A method of obtaining funds with choleretic activity. A.S. N 1361760, class A 61 K 35/78. - BI N 19, 1994), the product is obtained by extracting the flowers of two types of tansy when heated to 80 ^o C three times with ethanol at a ratio of raw materials: extractant (1: 7). The extracts are evaporated, the residue is treated with hot distilled water, then purified with dichloroethane. The target product is extracted with butanol, evaporated, the residue is dissolved in ethanol and dried at 65 - 100 ^o C with the release of the sum of 3.7 - 4.3% of the raw material.

 In the method (Gella E.V., Vavilova N.K., Grimova T.A. Method for producing flavonoids, iridoids and hydroxycinnamic acids. - A.S. USSR N 904710, class A 61 K 35/78. - BI N 6, 1982) the sum of flavonoids, iridoids and hydroxycinnamic acids is obtained from tuberous snake grass, extracting the raw material with 70% alcohol in a ratio of 1: 30, the extract is evaporated to water, cooled, the precipitate is separated and dissolved four times in 70% ethanol, separated alcohol, dissolve the residue in hot distilled water, combine with the aqueous filtrate, cool, apply on a column with polyamide. The column is washed with water, then eluted with 70% ethanol. Aqueous and alcoholic eluates are combined, evaporated and spray dried. The yield of the purified amount of flavonoids, iridoids and hydroxycinnamic acids is 8.1% of the feed.

 In the method (Matsumoto T., Kuesi T. Flavonoid glycoside. US Pat. No. 4,753,929, class A 61 K 31/70. - Chemistry chemotherapy, No. 10, 1989, 0232 P.) quercetin coumaroylglucosylramnoside is obtained from gingo leaves by extraction raw materials with lower alcohol, followed by degreasing the extract with carbon tetrachloride and treating the residue with ethyl acetate, butanol, followed by purification on a silica gel column.

 Compared with the claimed in these methods, various solvents are used, the processes occur when heated with a large consumption of solvents, long-term cleaning using various sorbents.

 The prototype of the proposed method is a method for producing oxycinnamic acids (Gella E.V., Zhukov I.M., Grimova T.A. et al. Method for producing oxycinnamic acids. - A.S. USSR N 1165405, class A 61 K 35 / 78. BI N 15, 1985), according to which the grass of the butterflies are extracted by heating in a water bath for 3 hours three times with 96% ethanol with a ratio of raw materials: extractant 1: 10 (total volume of extractant 30 l). The resulting alcoholic extracts were filtered and evaporated, the residue was diluted with hot water and cooled. The solution was filtered and applied to an alumina column. The product is eluted with distilled water, the eluates are evaporated, the precipitate is cooled, acidified with sulfuric acid to pH 2. The target product (the sum of hydroxycinnamic acids) is removed from the aqueous phase with a mixture of diethyl ether-n-butanol in a ratio of 1: 17. The organic solvent is evaporated, the residue treated with 70% ethanol and obtain a powder of the target product of 1.8% (by weight of plant materials).

 The novelty of the proposed solution lies in the fact that the proposed set of technical methods allowed the use of a new type of raw material - grass (Technical conditions 64-4-044-83. Medicinal valerian grass. - M., 1983. - 109 pp.), To ensure a high yield of the target product and simplify the technology, get a purified amount of hydroxycinnamic acids with a certain choleretic activity.

 The novelty of the proposed method indicates the creative nature of the solution, that is, it indicates the conformity of the proposed solution to the requirement of "non-obviousness".

 The reasons that impede the achievement of the required technical result in the prototype include the following: in this case, 96% ethanol as an extractant does not ensure the complete extraction of the target product from raw materials, acidification of the substrate leads to partial hydrolysis of the native esters of hydroxycinnamic acids, which leads to the loss of part biologically active complexes, reducing the quality and yield of the target product.

 The technical result obtained when solving the problem is expressed in the use of a new type of raw material (grass), the simplification of the technology, the use of a new eluent (10 - 2% sodium chloride solution), achieving a high yield and in obtaining the choleretic effect of the target product.

Information confirming the possibility of carrying out the invention is as follows: finely ground (rolled) herb of Valerian officinalis (Ammosov A.S., Popova T.P., Litvinenko V.I. Rolling - a promising method for grinding medicinal plant materials // Abstract. scientific and practical conference dedicated to the 25th anniversary of the pharmaceutical faculty Kursk medical institute - Kursk - 1991. - P. 170 - 171) is extracted with 60 - 70% ethyl alcohol at a ratio of raw materials : extractant (1: 5 - 7) by the filtration method (Filtration extraction and its hardware design / T.P. Popova, A.S. Ammosov, V.I. Litvinenko, V.M. Mishev // Farmakom, N 2-3 (19-20 1994) .- Kharkov. - P. 43 - 49). The extraction process is carried out at a temperature of 20 - 25 ^o C. Alcohol extraction is evaporated in vacuo to remove alcohol and obtain about 1/3 of the original volume. The resulting concentrate is kept at a temperature not exceeding 10 ^o C for 5 hours. After sedimentation, the sediment of lipophilic substances is separated. The filtrate is diluted in half (1/2) with a 10% aqueous solution of sodium chloride. The resulting solution is introduced into a column filled with a polyamide sorbent in a wet way (Maksyutina N.P., Litvinenko V.I. Methods for the isolation and study of flavonoid compounds // Phenolic compounds and their physiological functions. - M., 1968. - P. 7-26 ) The elution of the target product is first carried out with a 10% sodium chloride solution, with a decrease in the concentration gradient of sodium chloride from 10 to 2%. The resulting eluates containing the target product are evaporated to 1/5 of the original volume, cooled and re-applied to a small polyamide column, washing with distilled water until a negative reaction to chlorides. The purified amount of the target product is eluted with 20% aqueous ethanol, evaporated and dried. The yield of the target product 2.5 - 3.0% by weight of plant materials.

Example 1. The feedstock, dried medicinal valerian grass, is crushed by rolling into a powder with a particle size of not more than 1 mm. 30.0 kg of the crushed raw material is loaded into a filtration-type extractor with a uniform dense layer. The raw materials in the extractor are treated with extractant - 60% aqueous ethanol. The extraction process is carried out at a temperature of 20 - 25 ^o C by filtration method with the extraction of the extract using vacuum at an average speed of about 20 l / h and with the extraction, which spend 150 l of extractant (at a ratio of 1: 5).

The resulting extract is evaporated in a vacuum evaporation apparatus at a temperature of 70 - 80 ^o C and a residual pressure of 0.1 - 0.2 kg / cm ² until complete removal of ethanol and to obtain 30 l of an aqueous concentrate.

The concentrate is placed in a sump, closed with a lid and placed in a refrigerator, where it is kept for 5 hours at a temperature not exceeding + 10 ^o C. After settling, the lipophilic resinous part containing chlorophyll, carotenoids, resins, oils is collected in the form of a precipitate. The aqueous solution is decanted and filtered on a suction filter.

 An aqueous filtrate of about 30 L is diluted with 30 L of a 10% aqueous solution of sodium chloride.

 The resulting solution with a feed rate of 5 l / h is applied to a column with a wet powder of polyamide sorbent (column height 150 cm, diameter 25 cm, sorbent mass 5 kg). The elution of oxycinnamic acids is carried out first with a 10% sodium chloride solution and, with every 5 L, the concentration of sodium chloride solution is reduced by 2%, bringing it to 2% to obtain about 25 L of eluates.

 The resulting solution of eluates, including hydroxycinnamic acids, was evaporated in vacuo to 5 L of concentrate. The lipophilic and phenolic (flavonoid) components remain on the polyamide.

The concentrate is re-applied to the column with polyamide powder (height 50 cm, diameter 15 cm, weight 2 kg). The sorbent is washed with distilled water at a rate of 5 l / h until a negative reaction in the last portions of the eluates to the chlorides. Oxycinnamic acids are eluted with 20% aqueous ethanol until a negative reaction is obtained for hydroxycinnamic acids (chromatographic method) to give about 25 L of eluates. The eluates are evaporated in vacuum to obtain about 1.5 kg of a thick mass, which is placed on a baking sheet in a small layer and dried in a vacuum oven at a temperature not exceeding 80 ^o C for 2.5-3 hours and a residual moisture content of not more than 5%. The product is removed, crushed in a ball mill, sieved to obtain 0.75 kg of the target product, i.e. 2.55% of the mass of raw materials.

 Example 2. 30.0 kg of finely ground raw materials are loaded into the filter type extractor in a uniform layer. The raw materials in the extractor are treated with an extractant - 70% ethanol. The extraction process is carried out at room temperature with an average speed of about 20 l / h to obtain an extraction, which consumes 210 l of extractant (at a ratio of 1: 7).

The resulting extract is evaporated in a vacuum evaporator at a temperature of 70 - 80 ^o C and a residual pressure of 0.1 - 0.2 kgf / cm ² until complete removal of ethanol and to obtain 30 l of an aqueous concentrate.

The concentrate in the sump is kept for 5 hours at a temperature not exceeding plus 10 ^o C. After sedimentation, the lipophilic part is separated by decantation on a suction filter.

 An aqueous filtrate of about 30 L was diluted with 30 L of a 10% aqueous solution of sodium chloride.

 The resulting solution with a feed rate (outflow) of about 5 l / h is applied to a column of wet polyamide sorbent powder (column height 150 cm, diameter 25 cm, sorbent mass 5 kg). The elution of the amount of hydroxycinnamic acids is first carried out with a 10% aqueous solution of sodium chloride and, with every 5 L, the concentration of the sodium chloride solution is reduced by 2%, bringing it to 2%, to obtain about 25 L of total eluates.

 The resulting solution of eluates containing the sum of hydroxycinnamic acids in saline was evaporated in vacuo to 5 L of concentrate.

The concentrate at room temperature is re-applied to the column with wet polyamide powder (column height 50 cm, diameter 15 cm, sorbent mass 2 kg). The sorbent is washed with distilled water at a rate of 5 l / h until a negative reaction to chlorides in the last portions of the wash water. Oxycinnamic acids are eluted with a 20% solution until a negative reaction to oxycinnamic acids (chromatographic method) is obtained and about 1.5 kg of a thick mass is obtained, which is placed on a baking sheet in a small layer and dried in a vacuum oven at a temperature not exceeding 80 ^o C during 2.5 - 3 hours and negative humidity in the product no more than 5%. The product is removed from the baking sheet, crushed in a ball mill, sieved to obtain 0.875 kg of the target product, i.e. 2.75% of the mass of raw materials.

Example 3. 30.0 kg of finely ground raw materials - herbs of Valerian officinalis - are loaded into the filter type extractor with a uniform layer. Extraction is carried out at a temperature of 20 - 25 ^o C with 65% aqueous ethyl alcohol with an average speed of about 20 l / h and obtaining extract, which spend 180 l of extractant (at a ratio of 1: 6).

The resulting extract is evaporated in a vacuum evaporator at a temperature of 70 - 80 ^o C and a residual pressure of 0.1 - 0.2 kgf / cm ² until complete removal of ethanol and to obtain 30 l of an aqueous concentrate.

The concentrate is placed in a sump, closed with a lid and placed in a refrigerator, where it is kept for 5 hours at a temperature not exceeding + 10 ^° C. After settling of the lipophilic resinous part containing chlorophyll, carotenoids, resins, oils, the aqueous solution is decanted and filtered on filter.

 A filtrate of about 30 L is diluted with 30 L of a 10% sodium chloride solution. The resulting solution with an eluate exit speed of about 5 l / h is applied to a column with a wet polyamide sorbent powder (column height 150 cm, diameter 25 cm, sorbent mass 5 kg).

 The elution of the amount of hydroxycinnamic acids is carried out first with a 10% aqueous solution of sodium chloride and, with each 5 L of eluate, the concentration of sodium chloride in the eluent is reduced by 2%, adjusted to 2% in the last portions to obtain about 25 L of eluates.

 The resulting eluate solution containing the sum of hydroxycinnamic acids in saline was evaporated in vacuo to 5 L of concentrate.

The concentrate at room temperature is re-applied to the column with wet polyamide powder (column height 50 cm, diameter 15 cm, sorbent mass 2 kg). The sorbent is washed with distilled water at a rate of 5 l / h until a negative reaction to chlorides in the last portions of the wash water. Oxycinnamic acids are eluted with 20% aqueous ethanol until a negative reaction in the last portions of the eluate to hydroxycinnamic acids (chromatographic method) is obtained and about 25 L of eluates are obtained. The eluates are evaporated in vacuum to obtain about 1.5 kg of a thick mass, which is placed on a baking sheet in a small layer and dried in a vacuum oven at a temperature not exceeding 80 ^o C for 2.5 - 3 hours to a residual moisture content in the product of not more than 5 %

 The dry product is removed from the baking sheet, crushed in a ball mill, sieved through a sieve to obtain 0.900 kg of the target product, i.e. 3.00% yield by weight of plant material.

 The resulting target product, the purified amount of hydroxycinnamic acids, is standardized spectrophotometrically according to the content of hydroxycinnamic acids in terms of chlorogenic acid and other qualitative indicators (Oxycinnamic acids of Valerian officinalis / Talashova S.V., Turcan An.A., Fursa N.S. et al. / / Complex use of biomedical health problems of the population of the Tyumen region: Sat scientific work, dedicated to the 30th anniversary of the Tyumen state medical institute. Part 3. - Tyumen, 1993. - P. 32-33).

 The biological activity was checked by comparing 1% aqueous solutions obtained from the target product and alcohol tincture of valerian grass, according to test indicators of their choleretic activity. Comparative materials are given in table. 4.

 As a result of the implementation of the proposed method, a new type of plant material is used - medicinal valerian grass, milled by rolling, which is extracted with 60-70% aqueous ethanol by the filtration method, purification is carried out on polyamide, for the first time using a 10-2% chloride solution as eluent sodium to obtain the target product with choleretic activity. Comparative data on the claimed method are presented in table. 13.

Claims (1)

 The method of obtaining the amount of hydroxycinnamic acids with a choleretic effect by treating plant materials with an organic solvent, evaporation, separating the lipophilic precipitate, cleaning with a sorbent, evaporation and drying, characterized in that the valerian officinalis chopped by rolling is extracted with 60-70% aqueous ethanol by filtration by the method with the ratio of raw materials: extractant (1: 5–7), purification is carried out on polyamide, first eluted with a 10–2% sodium chloride solution, and then eluted with the eluted product 20% ethanol to give the desired product.

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