RU2151139C1 - Method of preparing biologically active sum of triterpenic acids - Google Patents

Abstract

FIELD: biotechnology, more particularly preparation of biologically active sum of triterpenic acids. SUBSTANCE: method comprises steaming the original stock (wood green or fir bark) and subsequently drying and extracting it with organic solvent, treating extract with aqueous solution of alkali agent, acidifying and recovering the desired product. EFFECT: more efficient preparation method due to shorter extraction time, and increased yield of the desired product. 5 ex, 1 tbl

Description

 The invention relates to biotechnology, and in particular to methods of isolating a biologically active natural mixture (amount) of triterpene acids from green wood and fir bark.

 A known method of producing a biologically active amount of triterpenic acids by extracting dried fir needles (woody greens) fir with a mixture of petroleum ether and ethyl acetate (1: 1 by volume), treating the extract with an aqueous solution of sodium bicarbonate, acidifying with hydrochloric acid and isolating the target product with ethyl acetate (RF patent N 2025976 )

 This method is not effective enough due to its duration and low yield of the target product.

 A known method of obtaining a biologically active amount of triterpenic acids, according to which fir bark is used as the starting material, extraction is carried out with a hydrocarbon solvent and ethyl acetate sequentially or a mixture thereof in a ratio of 1: 0.8: 1.2, the target product is extracted with ethyl acetate, followed by extraction of triterpene acids alkali solution, acidification of an aqueous alkaline solution and the allocation of the target product from it (RF patent N 2061487).

 The disadvantage of this method is the low yield of the product - the biologically active amount of triterpenic acids.

 The problem to which the invention is directed, is to increase the efficiency of the method by reducing the extraction time and, in some cases, increasing the yield of the target product.

 The problem is solved as follows. Plant wood raw materials — wood greens or fir bark — are steamed to produce fir oil, then dried, the dried raw materials are extracted with an organic solvent, the amount of triterpenic acids is extracted from the extract with an alkaline agent solution, the aqueous alkaline solution is acidified, and the target product is isolated, for example, by filtration.

 Comparative analysis with the closest analogue shows that the claimed solution is characterized in that the vegetable raw materials are treated with steam before extraction with an organic solvent, which destroys the wax coating of the needles, breaks the cell membranes of the needles or bark cells, and thus facilitates the access of the solvent inside the cells to all cellular structures, due to which a more complete extraction of the target product from the raw material is achieved with a lower consumption of solvent.

 The method is as follows.

 Fresh green wood or fir bark is treated with steam, resulting in the extraction of essential oil, dried and extracted with an organic solvent, then the extract is treated with an aqueous solution of an alkaline agent, the aqueous-alkaline solution is separated, acidified with mineral acid and the desired product is isolated by filtration.

 The method is illustrated by the following examples.

 Example 1

Fresh woody greens (500 g) were treated with hot steam (t 109-124 ^o C) for 13-15 hours, while receiving 7.2 g of fir oil, dried steamed greens to a moisture content of 4-6%, pine needles were ground to 0, 1-0.4 mm. The resulting 152 g of dry needles was extracted with methyl tert-butyl ether (600 ml) by infusion for 2 hours. 500 ml of the extract was obtained, which was treated with 100 ml of a 2% sodium hydroxide solution. The separated aqueous alkaline solution was acidified with HCl to pH 2 and the desired product was isolated by filtration. Received 17.9 g of triterpenic acids, which is 11.8% by weight of the extracted raw materials.

 Example 2

 It was carried out under the conditions of example 1, but with the extraction of steamed, dried and ground wood greens with various organic solvents. The results are shown in the table.

 Example 3

 Fresh woody greens (500 g), not steamed, were dried and ground, the resulting 252 g of dried greens was extracted with methyl tert-butyl ether, further processing was carried out under the conditions of Example 1. 14.1 g of triterpenic acids were obtained. The yield of the target product was 5.6%.

 Example 4

 Fresh fir bark (500 g) was treated with hot steam and dried under the conditions of Example 1. The resulting 177 g of dried and ground bark was extracted and subjected to further processing also under the conditions of Example 1.

 Received 9.02 g of triterpenic acids, which amounted to 5.1% of the extracted raw materials and 4.2 g of fir oil.

 Example 5

 The bark of fir was dried (without steam treatment), the dried raw material was processed under the conditions of example 1. The yield of the target product was 1.7%.

 The target product is the biologically active amount of triterpenic acids obtained in all examples, analyzed by high performance liquid chromatography and corresponds to the standard mixture of triterpene acids, according to TU N 9291-001-03533895-94 for the preparation of triterpenic acids "SILK".

 The advantage of the proposed method, in comparison with the known, is to obtain the target product, triterpene acids, in particular from woody fir fir, of higher quality. Used in the known method wood greens ("fir foot") 40% consists of wood and 60% of the needles. When extracting from wood, a number of components pass into the extract, in particular dehydroabietic acid, which “pollute” the product, being its ballast and, thus, reducing the activity of the product. The separation of needles from wood is a very time-consuming process and therefore economically impractical. At the same time, after processing the green woods with steam and subsequent drying, the needles are separated from the wood by light shaking, which allows us to use needles in the proposed method with virtually no wood impurities.

 In addition to the target product - the biologically active amount of triterpenic acids, in the present method receive fir oil, distilled from woody herbs and fir needles with steam.

Claims (1)

 A method of obtaining a biologically active amount of triterpenic acids from dried plant material by extraction with an organic solvent, extracting from the extract the amount of triterpene acids with an aqueous solution of an alkaline agent, acidifying an aqueous alkaline solution and isolating the target product from it, characterized in that wood greens are used as the starting material or fir bark, pre-treated with steam.

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