RU2305939C1 - Method for producing of biologically active preparation from larch wood - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: method involves removing bark from Siberian larch wood; grinding wood to 3-10 mm sized particles and drying in air at temperature of 30-40 C until residual moisture content is no more than 15%; repeatedly grinding dried raw material to 1-5 mm sized particles and charging into extractor; performing four-fold extraction of raw material using 87-88%-isopropyl alcohol at raw material to extractant ratio of 1:(2.5-3); boiling down extract by 70-90 times; measuring content of dihydroquercetin; adding five-fold emulsifier excess and mixing resultant mixture to produce homogeneous emulsion.

EFFECT: reduced consumption of organic solvent, decreased amount of production wastes, and simplified process.

4 cl, 2 tbl, 3 ex

Description

The invention relates to biotechnology and agriculture, specifically to a method for producing a biologically active preparation from larch wood, which can be used in agriculture to regulate plant growth and development.

There are various methods of processing wood and larch bark to obtain a wide range of biologically active products for the food industry and medicine (Babkin V.A., Ostroukhova L.A., Ivanova S.Z. et al. Products of deep chemical processing of larch biomass. Ros Chemical Chem., 2004, vol. XLVIII, No. 3, pp. 62-69). The main active ingredient of the preparations obtained is dihydroquercetin (DHQ), a naturally occurring flavonoid with a wide spectrum of biological activity.

A known method, for example, is the isolation of DHQ by treating larch wood with water, extraction with aqueous acetone, evaporation of the extract and crystallization of DHQ (RF Patent N2038094, class A61K 35/78, op. 27.06.95).

A known method for the allocation of DHQ by extraction with heating of the pre-tarred sawdust of larch wood with an organic solvent - ethyl acetate (RF Patent N2034559, class A61K 35/78, op. 10.05.95).

A known method of producing DHQ from crushed larch wood, including preliminary impregnation with the last water (at a temperature of 25-40 ° C), extraction with ethyl acetate (ratio of raw materials: extractant 1: 4), evaporation of the extract and processing with gasoline (or hexane) without heating to remove the resin . The tarless extract is treated with hot water (temperature 95-98 ° C), filtered from impurities. Upon cooling, crystals of DHQ precipitate from the aqueous solution.

The main disadvantages of the known methods are the high consumption of organic extractants (gasoline and ethyl acetate, acetone, ethyl acetate), and, accordingly, the high energy consumption for their evaporation and heating during extraction, as well as the complexity and multi-stage process associated with the need to remove extractive resinous substances and obtain DQF high purity.

Closest to the claimed method, the prototype is a method for producing DHQ from crushed larch wood, comprising mixing the prepared plant material with an extractant, which is used ethyl acetate in a mass ratio of 1: 5-8, respectively, heating the mixture to a boiling point of ethyl acetate of 77 ° C, and the extraction process at this temperature for 1-5 hours, followed by filtration of the obtained hot extract, evaporation in a rotary evaporator at a temperature of 35-42 ° C and the rest exact pressure of 0.9-0.12 atm to obtain a dry residue. The resulting dry residue is mixed with hexane or gasoline in a mass ratio of 1: (8-20), respectively, and the mixture is kept at a temperature of 18-24 ° C with stirring for 15-40 minutes, after which the extractant is distilled off, the mixture is filtered and the resulting extract the target product is mixed with water at a temperature of 90-95 ° C in a mass ratio of 1:10 and maintained at this temperature for 20-40 minutes, after which the mixture is cooled (RF Patent No. 2082425, class A61K 35/78, op. 27.06. 97). The crystals of dihydroquercetin formed during cooling are separated by filtration and dried at a temperature of 100-120 ° C for 3-5 hours.

The disadvantages of the method are multi-stage, the complexity of the process of separation of DHQ, high consumption of extractant and organic solvents, a large volume of drain (waste) water.

An object of the invention is to obtain a new biologically active preparation based on dihydroquercetin, reducing the consumption of extractant and reducing the amount of production waste (waste water).

The technical task is achieved by the proposed method, which consists in the following.

Siberian or Daurian larch wood is cleaned of bark, crushed on a rotary crusher to particles of 3-10 mm in size and dried in air at 30-40 ° C to a residual moisture content of 10-12%. The dried raw materials are re-crushed to particles with a size of 1-5 mm on a knife crusher and loaded into the extractor. Extraction is carried out four times each time with a fresh solvent, which is heated to 40-60 ° С 87-88% isopropyl alcohol (azeotropic mixture of isopropyl alcohol-water), with the ratio of raw material: extractant 1: (2.5-3.0) . The extracts are combined and evaporated 70-90 times to obtain the target product in the form of a dark brown viscous mass with a weak specific odor, readily soluble in organic solvents. All distilled solvent is returned to the process, the loss of solvent during the distillation process and with the meal used is 3-4 liters per 1 kg of the target product, depending on the technological conditions (season, temperature, vacuum), while the solvent does not change its extraction ability and, therefore, allows you to conduct the process without changes, which dramatically reduces the consumption of extractant. The yield of the target product leaves 18.7-21.9% of the weight of the used (dried) raw materials. The target product contains a mixture of extractive resinous substances of unknown structure and dihydroquercetin. The content of DHQ in the target product is 6.4-10%.

Next, the target product is transferred to a mixer equipped with built-in filters, the content of DHQ is measured, a five-fold excess of emulsifier is added, and the resulting mixture is stirred for 4-8 hours until a homogeneous emulsion.

To obtain the commercial drug Larixin, the target product in the form of an emulsion is diluted with water to the content of DHQ in it equal to 5% (50 g / l) and re-mixed for 4-6 hours.

A commercial preparation is a 5% dark brown aqueous emulsion with a specific odor. Field tests have confirmed the high effectiveness of the drug "Larixin" on the growth and development of many agricultural plants, in particular wheat.

The defining hallmarks of the proposed method, compared with the prototype, are:

1. The crushed larch wood is dried in air at a temperature of 30-40 ° C to a residual moisture content of 10-12%, which allows to preserve the natural content of dihydroquercetin and the fraction of resinous substances in the feedstock, as well as to prevent the extraction of residual moisture from the raw material and dilution with water of the extractant.

2. The extraction of raw materials is carried out with 87-88% isopropyl alcohol at a ratio of raw materials: extractant equal to 1: (2.5-3.0), which allows you to extract mainly phenolic substances with a DCV content of at least 50% of the total dry matter in the extract. When anhydrous isopropyl alcohol is used for extraction, a significant amount of non-polar substances (waxes, fats, neutral terpenoids) are extracted, and when using isopropyl alcohol with a moisture content of more than 20%, soluble polysaccharides (arabinogalactans) begin to be extracted.

3. The content of DHQ is measured in the target product, a five-fold excess of emulsifier is added, and the resulting mixture is mixed until a homogeneous emulsion is obtained, which allows the target product to completely dissolve and exclude delamination of the mixture and precipitation of DHQ.

The invention is illustrated by the following examples of specific performance of the method.

Example 1

35 kg of larch wood, purified from bark, was crushed on a rotary crusher to a particle size of 3 mm, dried at a temperature of 30 ° C for 48 hours to a residual moisture content of 12%. Dried raw materials (26 kg) were re-crushed to a particle size of 1 mm on a knife crusher, loaded into an extractor with a capacity of 100 l and extracted with 4 portions of 88% aqueous isopropyl alcohol heated to 60 ° C at a ratio of raw material: extractant 1: 2.5 for 6 hours. The combined extract (266 L) was evaporated at 60 ° C in a vacuum of 160 mm Hg. to obtain 3.8 l of concentrate, and the content of DHQ was measured by HPLC (measurement result - 6.4% or 0.243 kg). To the target product was added 1.2 kg of OP-7 emulsifier heated to 60 ° C and mixed with a mechanical stirrer for 4 hours to obtain 4.8 L of a uniform dark brown emulsion. The yield of the target product was 18.7% of the weight of the extracted raw materials. To obtain a commercial preparation, 0.5 L was added to 4.8 L of the emulsion. water to the content of DHQ in the aqueous emulsion equal to 5%, the resulting mixture was stirred for another 6 hours and left to stand for 4 hours.

Example 2

400 kg of the butt of the larch wood, peeled from the bark, chopped into logs, crushed on a rotary grinder to 10 mm particles and dried at 40 ° C to a moisture content of 10%. Dried raw materials (160 kg) were re-crushed on a knife crusher to 5 mm particles, then they were loaded into an extractor with a capacity of 630 L and extracted four times with 87% aqueous isopropyl alcohol heated to 40 ° C at a ratio of raw materials: extractant 1: 3.0 v within 8 hours. The combined extract (1756 L) was evaporated at a temperature of 50 ° C in a vacuum of 150 mm. Hg to obtain 20 l of concentrate (target product), the content of DHQ (8.5%) was measured, the total content of DHQ was calculated - 1.7 kg. The concentrate was transferred to a mixer equipped with built-in filters, and a five-fold amount of an emulsifier OP 10 - 8.5 kg was added. The mixture was stirred with an anchor stirrer for 8 hours until a homogeneous dark brown emulsion was obtained. The yield of the target product was 21.9% of the weight of the extracted raw materials. To obtain a commercial preparation, 7 L of water was added to the resulting emulsion and mixed for another 4 hours. After a 6-hour sludge, a homogeneous, non-stratified dark brown aqueous emulsion was obtained with a DHQ content of 5%.

Example 3

Field tests of the Lariksin preparation were carried out on the experimental field of the Altai State Agrarian University (Prigorodnoye farm in the Pervomaisky district) on spring wheat varieties Omskaya 24 and Altayskaya 92.

The effectiveness of pre-sowing treatment of spring wheat seeds with Larixin was tested on Omskaya 24. The seeds were treated by semi-dry dressing the day before sowing with the following drugs: SILK (standard), flow rate - 50 ml / t, working solution consumption 10 l / t and the preparation " Larixin ", the consumption rate: 100 ml / t, the flow rate of the working solution is 10 l / t. Control seeds were treated with water. The effectiveness of pre-sowing treatment of spring wheat seeds is presented in Table 1, from which it can be seen that the treatment of seeds with the Lariksin preparation allowed to obtain a yield increase of 8.1 c / ha or 24.5% more than in the control. A 25% decrease in damage to wheat plants by root rot was noted.

The efficacy of treating vegetative spring wheat plants with Larixin was tested on Altai 92 variety. For this, vegetative plants were sprayed during tillering with Silk at a rate of 50 mg / ha and Larixin at a flow rate of 100 ml / ha. The flow rate of the working solution was 300 l / ha. Control plants were treated with water. The effectiveness of foliar treatment of spring wheat crops is presented in table 2, which shows that under the influence of the drug "Lariksin" grain yield increased by 8.7 kg / ha or 35.7% compared with the control and 4.0 kg / ha or 13.8% compared to the standard (Silk).

Using the proposed method will allow:

- receive a new biologically active drug, which in addition to DHQ contains a fraction of resinous substances, and has a growth-promoting and fungicidal effect;

- significantly reduce the consumption of organic solvents due to the continuous return to the stage of extraction of the same extractant;

to improve the environmental safety of the method by eliminating the discharge of wastewater, since the target product is a total extract containing a mixture of extractive resinous substances of an unspecified structure and dihydroquercetin, and liquid waste is formed at the stage of purification of DHQ from tar and related substances and precipitation of DHQ from a cooled aqueous solution (prototype).

Table 1 Options Productivity Mass of 1000 grains, g Gluten content Grain Class We pay back. 1 rub costs RUB c / ha increase % % Group Control (10 l / t) 33.0 - one hundred 32,3 thirty I II - Silk, 50 ml / t 40.1 7.1 121.5 36.0 30,4 I II 8.4 Larixin, 100 ml / t 41,4 8.1 124.5 34.6 32.8 I I 9.6 table 2 Option Productivity Mass of 1000 grains, g Gluten content Grain Class We pay back. 1 rub costs RUB c / ha increase % % Group Control (water) 24.2 - one hundred 33.5 28.8 II III - Silk, 50 ml / ha 28.9 4.7 19,4 35.65 28.9 II II 6.65 Larixin, 100 ml / ha 32.9 8.7 35.7 35.69 30.1 I I 7.62 R, % 2.94 NDS ^0.95 2.45

Claims (4)

1. A method of obtaining a biologically active preparation from larch wood containing dihydroquercetin, including grinding the raw material, drying the latter in air to a moisture content of not more than 15%, extraction with an organic solvent, evaporation of the extract, characterized in that the chopped wood is dried at 30-40 ° C , 87-88% isopropyl alcohol is used as the extractant, extraction is carried out at a ratio of raw materials: extractant 1: (2.5-3), the extract is evaporated 70-90 times, the content of dihydroquercetin is measured in it and five times are added th excess emulsifier, followed by stirring to obtain a homogeneous emulsion. 2. The method according to claim 1, characterized in that the feedstock is ground first to particles of 3-10 mm in size, and then to particles of 1-5 mm in size. 3. The method according to claim 1, characterized in that as the emulsifier use OP-7 or OP-10. 4. The method according to claim 1, characterized in that before use, the preparation in the form of an emulsion is diluted with water to the content of dihydroquercetin equal to 50 g / l.