RU2712094C1 - Method of producing purified inulin from vegetable raw material - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention refers to pharmacy and medical industry and can be used to produce purified inulin from elecampane roots. That is ensured by three-step extraction by purified water. Then, plant material is separated by filtration. Further, the precipitate is washed on the filter. Filter with precipitate is dried at temperature from 100 to 105 °C to constant weight. Raw material used is crushed roots of elecampane high to linear size of 0.5–1.0 mm. Extraction is carried out at raw material : extractant ratio of 1 g : 15 ml for 30 minutes. Prior to flushing, inulin is removed from impurities, where precipitate obtained after filtration under vacuum is dissolved in heated to 80 °C purified water. Then 50 % calcium chloride solution is added to it at rate of 5 drops per 1 g of initially taken vegetal raw material and 0.5 g of finely dispersed aluminum oxide powder. It is held for 20 minutes, then it is filtered under vacuum at residual pressure 0.4–0.8 atm. Produced filtrate is successively passed through ion-exchange columns with anionite in hydroxyl form AV-17-8 and cationite in hydrogen form KU-2-8 to pH of eluate 6.5–7.5 and degree of purity of inulin equal to 97 %. After that for sedimentation of inulin to eluate there added again is threefold amount of 95 % ethanol with stirring, cooling in freezing chamber at temperature of -18 °C for 1 hour. Precipitate is then filtered through a pre-dried ash-free paper filter under vacuum at residual pressure of 0.4–0.8 atm.EFFECT: invention enables to carry out deep purification of the end product from impurities with a degree of purity of 97±2 %, in the absence of use of hazardous and toxic substances.1 cl, 3 dwg, 1 ex

Description

The invention relates to pharmacy and the medical industry, and in particular to a method for producing biologically active substances from medicinal plant materials, and in particular, can be used to obtain inulin from high elecampane roots.

For medical and pharmaceutical purposes, mainly aqueous extracts from the roots of elecampane high are used, and the pharmacological effect is due to water-soluble compounds, the basis of which are water-soluble polysaccharides represented by inulin (up to 40% in the roots of elecampane high) [Kurkin V.A. Pharmacognosy // Samara: Samara State Medical University, 2004. - 249 s].

A number of methods are known for producing inulin from Jerusalem artichoke tubers [RU 2398779, A23L 1/214, 2009; RU 2148588, C08B 37/00, 1998]. However, none of them shows the yield of inulin from its content in raw materials and the degree of purity of the obtained product from related substances.

The disadvantage of this method is the large amount of impurities in the finished product, which allows it to be considered a technical or raw product, not suitable for use in medical and food purposes.

A known method of producing inulin from crushed roots of dandelion officinalis using exhaustive extraction [RU 2351166 C1, A23L 1/236, A61K 36/00, 2009]. As a raw material, the roots of dandelion are used, which are washed, dried and crushed to a particle size passing through a sieve with a hole diameter of 2 mm, poured with water and subjected to exhaustive extraction for 3-5 days. The resulting colorless solution is treated with 96% ethyl alcohol and inulin is precipitated at a temperature below minus 15 ° C.

The disadvantage of this method is the duration of the process, low yield of the target product and a large number of impurities in the finished product.

A known method of producing inulin from dry chopped Jerusalem artichoke tubers [RU 2131252, A61K 31/715, A61K 35/78, 1996], including obtaining water extracts from dry chopped Jerusalem artichoke tubers by mixing them with hot water in a ratio of 1: 6, holding for 12 hours for swelling at 70 ° C and extracting extracts at 80 ° C, the aqueous extraction is separated and another three times for 40 minutes, extraction is carried out with water in a ratio of 1: 4, followed by combining the extracts, then they are treated with calcium carbonate at 80 ° C for 60 minutes, filtered, upa filtrate They eat, crystallize crude inulin at 4 ° C in the refrigerator for 5 days after adding 1: 1 alcohol, separate the inulin precipitate, dissolve it in hot water at 75 ° C, and further purify it with anion exchange resin, aluminum oxide, calcium carbonate with further purification with cation exchange resin and anion exchange resin, followed by crystallization and reprecipitation from a water-alcohol mixture 1: 3 at 40 ° C for 24 hours in a refrigerator, the product is dried and crushed.

The disadvantage of this method is the low yield of the product (6.7-7%), a large number of technological operations (more than 30), the duration of the process (more than 7 days).

A known method of producing inulin from chicory high and elecampane, including the preparation of inulin-containing raw materials, their mechanical cleaning, washing the roots, rhizomes, stems, chopping and mixing [RU 2619758 C1, C 37B 37 / 18,2017]. The mixed and crushed pieces of raw materials are extracted twice with hot water at a temperature of 75 ° C for 2-3 days with constant stirring. The obtained extract of inulin in bulk is treated with 96% ethanol in a ratio of 1: 1 by volume, followed by precipitation of inulin at a temperature of minus 16 ° C.

The disadvantage of this method is the duration of the process, low yield of the target product and a large number of impurities in the finished product.

A known method of producing inulin from Jerusalem artichoke tubers by repeatedly extracting extractives from the aqueous extract of the feedstock by heating it with separation of the water extraction each time and subsequent combining of the water extracts [RU 2485958 C1, A61K 31/715, A61K 36/28, B01D 11/02 , 2013]. The obtained extracts are subjected to purification by precipitation of pectin substances with a calcium salt, after which the extraction mixture is filtered, the filtrate is evaporated and inulin precipitated, the inulin precipitate is separated, dissolved in hot water, purification is additionally carried out with alumina followed by precipitation of inulin, the product is dried and crushed at certain conditions. The disadvantage of this method is the low yield of the final product (14.6%), the duration of the process (more than a day), a large number of technological operations, the use of toxic and hazardous substances (concentrated sulfuric acid), insufficient degree of purification for the use of the finished product for medical purposes.

The prototype of the claimed invention is a method for producing inulin from crushed roots of dandelion medicinal (RF patent 2635996 (IPC A61K 31/715, A61K 36/288, B01D 11/02; publ. 11/17/2017) using exhaustive extraction of feedstock with purified water heated to The method involves the separation of plant materials, the precipitation of water-soluble polysaccharides, their washing, drying, and the extraction is carried out in an ultrasonic bath with a frequency of 35 kHz for 40 minutes at a temperature of 80 ° C, the plant material is separated by filtration, and water-soluble polysaccharides precipitated with 95% ethanol, cooling in a freezer for 1 hour, and subsequent drying of the filter with sediment is carried out at a temperature of 100-105 ° C to constant weight. The disadvantage of this method is the low degree of purity of the target product.

The present invention is to develop a method of extracting inulin from the roots of elecampane with a high degree of purity of the target product in the preparation of inulin in the absence of the use of dangerous and toxic substances.

The object of the invention is solved by the fact that in the method for producing inulin from elecampane roots, the biologically active substances are extracted from raw materials three times with hot water purified in an ultrasonic bath, which can significantly accelerate the extraction process, as well as increase the yield of polysaccharides from raw materials; after which the mixture of water-soluble polysaccharides is precipitated with ethyl alcohol, the precipitate is dissolved and the purification of calcium with chloride (from pectin substances), aluminum with oxide (from pigment substances) and ion exchange resins (for deeper purification from other impurities) is carried out, after which inulin is precipitated ethyl alcohol and dried to constant weight.

The technical result of the invention consists in deep purification of the target product from impurities with a degree of purity equal to 97 ± 2%, in the absence of the use of hazardous and toxic substances.

The technical result is achieved in that in a method for producing purified inulin from elecampane roots, including triple extraction with purified water heated to a boiling point, from crushed roots of elecampane high, where the flask with raw materials and extractant is placed in an ultrasonic bath with a frequency of 35 kHz at a temperature of 80 ° C; separation of plant material by filtration; precipitation of water-soluble polysaccharides with three times the amount of 95% ethanol with stirring, cooling in a freezer at a temperature of -18 ° C for 1 hour; filtering the precipitate through a previously dried ashless paper filter under vacuum at a residual pressure of 0.4-0.8 atm; washing the filter cake, carried out sequentially with a solution of 95% ethyl alcohol in purified water, taken in a volume ratio of 3: 1, with a feed: solution ratio of 1:15, and a mixture of ethyl acetate and 95% ethyl alcohol, taken in a volume ratio 1: 1, with a ratio of raw materials: solution 1:10, drying the filter with sediment at a temperature of from 100 to 105 ° C to constant weight, according to the invention, ground elecampane roots of high to linear size 0.5-1.0 mm are used as raw materials ; extraction is carried out at a ratio of raw materials: extractant 1 g: 15 ml for 30 minutes; before washing, the inulin is purified from impurities, where the precipitate obtained after the filtration step under vacuum is dissolved in purified water heated to 80 ° C, 50% calcium chloride solution is added to it at the rate of 5 drops per 1 g of the originally taken plant material and 0.5 g fine powder of aluminum oxide, incubated for 20 minutes, then filtered under vacuum at a residual pressure of 0.4-0.8 atm; the resulting filtrate is successively passed through ion-exchange columns with anion exchange resin in the hydroxyl form AB-17-8 and cateonite in the hydrogen form KU-2-8 to a pH of 6.5-7.5 and an inulin purity of 97%, after which, for precipitation three times the amount of inulin to the eluate is again added three times 95% ethanol with stirring, cooling in the freezer at -18 ° C for 1 hour, then the precipitate is filtered through a pre-dried ashless paper filter under vacuum at a residual pressure of 0.4-0 , 8 atm.

The precipitate obtained after precipitation of water-soluble polysaccharides with ethanol also contains impurities of pectin, some pigments, and some organic acids. To remove pectins, it is proposed to add calcium salts in the form of a solution of calcium chloride, to remove pigments - finely divided powder of aluminum oxide. For a deeper purification from the remaining impurities, it is also proposed to pass an additional precipitate solution through the columns of cateonite and anionite. The degree of purification of the finished product was carried out by thin-layer chromatography, comparing with a standard sample (plates — Silufol, system — 55% ethanol, developer — solutions of resorcinol and sulfuric acid diluted with subsequent heating, Rf ~ 0.80).

In FIG. 1 shows the results of the isolation of inulin (%) from the roots of Elecampane high with varying degrees of grinding of raw materials and the temperature of the ultrasonic bath.

In FIG. Figure 2 presents the results of the isolation of inulin (%) from the roots of Elecampane high with varying multiplicity and duration of extraction.

In FIG. Figure 3 presents the results of the isolation of inulin (%) from the roots of Elecampane high with varying ultrasound frequencies and the ratio of raw materials to extractant

Example 1. To obtain inulin, an analytical sample of the raw material is crushed to particles with a size of 0.5-1.0 mm About 1 g (accurately weighed) of the crushed raw material is placed in a flask with a capacity of 50 ml, 15 ml of purified water is added, heated to boiling point, placed in an ultrasonic bath with a frequency of 35 KHz at a temperature of 80 ° C, extracted for 30 minutes. The extraction is repeated 2 more times, adding 10 ml of water.

Water extracts are combined and filtered through 3 layers of gauze with a cotton swab placed in a 5 cm diameter glass funnel. Precipitation is carried out in triplicate with 95% ethyl alcohol, stirred, cooled in a freezer at -18 ° C for 1 hour. Then the contents of the flask are filtered through a pre-dried and suspended ashless paper filter, laid in a glass filter POR 16 with a diameter of 40 mm, under vacuum at a residual pressure of 0.4-0.8 atm. The resulting precipitate is dissolved in 10 ml of purified water heated to 80 ° C, add 5 drops of a 50% solution of calcium chloride and 0.5 g of fine aluminum oxide powder, incubated for 20 minutes, then filtered under vacuum at a residual pressure of 0.4-0.8 atm. The obtained filtrate is successively passed through ion-exchange columns with anion exchange resin in the hydroxyl form AB-17-8 and cation exchange resin in the hydrogen form KU-2-8 to a pH of 6.5-7.5 and an inulin purity of 97%. To precipitate inulin, a triple amount of 95% ethanol is again added to the eluate with stirring, cooling in a freezer at a temperature of -18 ° C for 1 hour, the precipitate is filtered through a pre-dried ashless paper filter under vacuum at a residual pressure of 0.4- 0.8 atm. The filter cake was washed successively with 15 ml of a solution of 95% ethyl alcohol in purified water (3: 1), 10 ml of a mixture of ethyl acetate and 95% ethyl alcohol (1: 1). The filter cake is dried first in air, then at a temperature of 100-105 ° C to constant weight.

The proposed method allows to obtain inulin from the roots of elecampane high with a degree of purity of up to 97 ± 2% when the yield of the target product is up to 20.75% in terms of absolutely dry raw materials.

Claims (1)

A method of obtaining purified inulin from the roots of Elecampane high, including triple extraction with purified water heated to a boiling point, from the crushed roots of Elecampane high, where a flask with raw materials and extractant is placed in an ultrasonic bath with a frequency of 35 kHz at a temperature of 80 ° C; separation of plant material by filtration; precipitation of water-soluble polysaccharides with three times the amount of 95% ethanol with stirring, cooling in a freezer at a temperature of -18 ° C for 1 hour; filtering the precipitate through a previously dried ashless paper filter under vacuum at a residual pressure of 0.4-0.8 atm; washing the filter cake, carried out sequentially with a solution of 95% ethyl alcohol in purified water, taken in a volume ratio of 3: 1, with a feed: solution ratio of 1:15, and a mixture of ethyl acetate and 95% ethyl alcohol, taken in a volume ratio 1: 1, with the ratio of raw materials: solution 1:10, drying the filter with sediment at a temperature of from 100 to 105 ° C to constant weight, characterized in that the crushed elecampane roots of high to linear size 0.5-1 are used as raw materials, 0 mm; extraction is carried out at a ratio of raw materials: extractant 1 g: 15 ml for 30 minutes; before washing, the inulin is cleaned of impurities, where the precipitate obtained after the filtration step under vacuum is dissolved in purified water heated to 80 ° C, 50% calcium chloride solution is added to it at the rate of 5 drops per 1 g of the originally taken plant material and 0.5 g fine powder of aluminum oxide, incubated for 20 minutes, then filtered under vacuum at a residual pressure of 0.4-0.8 atm; the obtained filtrate is successively passed through ion-exchange columns with anion exchange resin in the hydroxyl form AB-17-8 and cation exchange resin in the hydrogen form KU-2-8 to a pH of 6.5-7.5 and an inulin purity of 97%, after which, for precipitation three times the amount of inulin to the eluate is again added three times 95% ethanol with stirring, cooling in the freezer at -18 ° C for 1 hour, then the precipitate is filtered through a pre-dried ashless paper filter under vacuum at a residual pressure of 0.4-0 , 8 atm.

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