Method for producing of biologically active preparation from larch wood

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

 

The invention relates to biotechnology and agriculture, particularly to a method of obtaining biologically active drug from larch wood, which can be used in agriculture to regulate the growth and development of plants.

There are various methods of processing wood and bark of larch to obtain a wide range of biologically active products for food industry and medicine (V.A. Babkin, Ostroukhova L.A., Ivanova SZ and other Products for deep chemical processing of biomass larch. Grew up with. chem. J., 2004, .XLVIII, No.3, pp.62-69). The main active ingredient derived drugs is dihydroquercetin (DQC)is a flavonoid natural origin with a wide spectrum of biological activity.

Known, for example, the allocation method DQC by processing of larch wood with water, extraction with aqueous acetone, evaporation of the extract and crystallization DQC (RF Patent N2038094, CL AC 35/78, op. 27.06.95).

There is a method of allocating DQC by extraction by heating pre-deresinated sawdust larch wood with an organic solvent - ethyl acetate (RF Patent N2034559, CL AC 35/78, op. 10.05.95).

A method of obtaining DQC from crushed wood of larch, including pre-impregnation of the latter with water (at a temperature 2-40° C), extraction with ethyl acetate (ratio of raw material: solvent 1:4), evaporation of the extract and handling of gasoline (or hexane) without heating to remove tar. Not the extract is treated with hot water (temperature of 95-98°C), filtered from impurities. Upon cooling of the aqueous solution falls crystals DQC.

The main disadvantages of the known methods are the high consumption of organic extractants (gasoline and ethyl acetate, acetone, ethyl acetate), and correspondingly high consumption of electricity for their evaporation and heating of the extraction, as well as the complexity and the multi-stage process, involving the need of extractive removal of tar and receive DQC a high degree of purity.

Closest to the claimed method of the prototype, is a method for DQC from crushed wood of larch, comprising mixing the prepared vegetable raw materials with the extractant, which is used as the acetate in a mass ratio of 1:(5-8), heating the mixture up to the boiling point of ethyl acetate, part 77°and carrying out the extraction process at this temperature for 1-5 h, followed by filtration of the obtained hot extract by evaporation in a rotary evaporator at a temperature of 35-42°and a residual pressure of 0.9-0.12 ATM to the floor the treatment of dry residue. The obtained dry residue is mixed with hexane or petrol in a mass ratio of 1:(8-20), and the mixture was kept at a temperature of 18-24°With stirring for 15-40 min, after which the solvent is distilled off, the mixture is filtered, and the resulting extracted target product is mixed with water temperature of 90-95°With mass ratio of 1:10 and kept at this temperature for 20-40 minutes, after which the mixture is cooled (Patent RF №2082425, CL AC 35/78, op. 27.06.97). Formed during cooling, the crystals of dihydroquercetin is separated by filtration and subjected to drying at a temperature of 100-120°C for 3-5 hours

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

An object of the invention is obtaining new biologically active agent on the basis of dihydroquercetin, the reduction rate of the extractant and the reduction of waste (sewage).

The goal of the project is achieved by the proposed method lies in the following.

The wood of Siberian larch or white-naped peeled from the bark, shredded in a rotary grinder to a particle size of 3-10 mm and dried in air at 30-40°C to a residual moisture content of 10-12%. High the high raw material re-crushed to a particle size of 1-5 mm on the blade grinder and loaded into the extractor. The extraction was carried out four times each time with fresh solvent, which is used as heated to 40-60°87-88% isopropyl alcohol (azeotropic mixture of isopropyl alcohol-water), when the ratio of raw material:solvent 1:(2,5-3,0). The extracts are combined and evaporated in 70-90 times to obtain the target product as a dark brown viscous mass with low specific smell, soluble in organic solvents. All return the distilled solvent to the process, the loss of solvent during distillation and used the meal consists of 3-4 liters per 1 kg of the desired product depending on the process conditions (time of year, temperature, vacuum), and the solvent does not change its extracting ability and, therefore, allows carry out the process without changes, which dramatically reduces the flow of the extractant. The yield of the target product leaves to 18.7-21.9% of the weight used (dried) raw materials. Target product contains a mixture of resinous extractive substances of unknown structure and dihydroquercetin. The content of the DQC in the target product is 6.4-10%.

Next target product move into the mixer, equipped with built-in filters, measuring the content of DQC add a fivefold excess of emulsifier, and the resulting mixture was stirred within 4-8 hours until a homogeneous emulsion.

To obtain a commercial preparation "Larixin" target product in the form of an emulsion is diluted with water to a content of the DQC in it, equal to 5% (50 g/l) and again stirred for 4-6 hours.

The commercial product is a 5%aqueous emulsion of a dark brown color with a specific smell. Field tests have confirmed the high efficiency of the drug, "Larixin" on the growth and development of many agricultural crops, in particular wheat.

Determining the distinctive features of the proposed method in comparison with the prototype are:

1. 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 faction resinous substances in raw materials, as well as to prevent extraction of the residual moisture of the raw material and dilution water extractant.

2. The extraction of raw materials carry out 87-88% isopropyl alcohol at a ratio of raw material:solvent = 1:(2,5-3,0), which allows extraction of mainly phenolic substance DQC not below 50% of the amount of solids in the extract. When used for the extraction of anhydrous isopropyl alcohol extracted a significant amount of non-polar substances (waxes, fats, neutral terpenoids), and the use of the propyl alcohol with humidity of about 20% starts to be extracted soluble polysaccharides (arabinogalactan).

3. In the target product measure content DQC add a fivefold excess of emulsifier, and the resulting mixture was stirred until a homogeneous emulsion that allows you to completely dissolve the target product and prevent delamination of the mixture and precipitation of the DQC.

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

Example 1.

35 kg larch wood, purified from the bark, grind on a rotary grinder 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) re-grind to particle sizes of 1 mm on the blade crusher, loaded into the extractor with a capacity of 100 liters and was extracted with 4 portions heated to 60°With 88% aqueous isopropyl alcohol at a ratio of raw material:solvent 1:2.5 to 6 hours. The combined extract (266 l) was evaporated at 60°With vacuum 160 mm Hg to obtain 3.8 l of concentrate, and measured the content of DQC by HPLC (measurement result is 6.4% or 0,243 kg). The target product was added 1.2 kg preheated to 60°emulsifier OP-7 and mixed with a mechanical stirrer for 4 hours to obtain 4.8 l uniform dark brown emulsion. The yield of the target product amounted to 18.7% by weight of the extracted raw materials. To obtain a commercial product to 4.8 l of the emulsion was added to 0.5 liter of water until the baking soda is Jania DQC in water emulsion, equal to 5%, the resulting mixture was mixed for another 6 hours and left to stand for 4 hours.

Example 2.

400 kg butt end part of larch wood, purified from the bark, split the logs to grind on a rotary grinder to a particle size of 10 mm and was dried at 40°to a moisture content of 10%. Dried raw materials (160 kg) re-grind on the blade grinder to a particle size of 5 mm, then loaded into the extractor with a capacity of 630 l and was extracted four times 87% aqueous isopropyl alcohol, heated to 40°when the ratio of raw material:solvent 1:3,0 within 8 hours. The combined extract (1756 l) was evaporated at a temperature of 50°in a vacuum of 150 mm Hg to obtain 20 liters of concentrate (target product), measured the content of DQC (8.5%), the calculated total content DQC - 1,7 kg Concentrate moved into the mixer, equipped with built-in filters, and added five times the amount of emulsifier OP 10 - 8,5 kg of the Mixture was mixed anchor stirrer for 8 hours to obtain a homogeneous dark brown emulsion. The yield of the target product 21.9% by weight of the extracted raw materials. To obtain a commercial product to the obtained emulsion was added to 7 liters of water and mixed for another 4 hours. After 6 hours of sludge received homogeneous, nerastraivaisya dark brown aqueous emulsion with a content of DQC equal to 5%.

Example 3.

The field is s trials of the drug, "Larixin conducted on the experimental field of the Altai Gosagrouniversiteta (ochos "suburban" Pervomaisky district) on the varieties of spring wheat Omsk 24 and Altai 92.

The effectiveness of pre-sowing treatment of seeds of spring wheat preparation "Larixin" tested on the grade of the Omsk 24. The seeds were processed by dry etching before sowing the following drugs: drug SILK (standard), the rate of flow of 50 ml/t, the flow rate of the working solution of 10 l/t and the drug, "Larixin", rate: 100 ml/t, the flow rate of the working solution of 10 l/T. the Control seeds were treated water. The effectiveness of pre-sowing treatment of seeds of spring wheat are presented in table 1, which shows that the seed treatment with the drug, "Larixin allowed to receive a yield increase of 8.1 kg/ha or by 24.5% more than in the control. Decreased damage to the plants of wheat root rot by 25%.

The processing efficiency of vegetative plants of spring wheat preparation "Larixin" tested on the grade of the Altai 92. For this vegetative plants were sprayed at the tillering stage, the preparation of silk with a consumption rate of 50 mg/ha, and the drug, "Larixin" with a consumption rate of 100 ml/ha When this solution consumption 300 l/ha of the Control plants were treated with water. The effectiveness of foliar treatment crops of spring wheat are presented in table 2, which shows that under the influence of the drug, "Larixin" the grain yield increased by 8.7 t/ha or by 35.7% compared with the control and 4.0 t/ha or 13.8% of the battle is to the standard (silk).

Using the proposed method will allow you to:

- get a new biologically active drug, which is also DQC contains a fraction of resinous substances, and has growth-stimulating and fungicidal action;

- 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 sewage, because the target product is the sum of the extract containing a mixture of resinous extractive substances of unknown structure and dihydroquercetin, and liquid waste is generated at the stage of purification of the DQC from resinous and related substances and deposition DQC from the cooled aqueous solution (prototype).

Table 1
OptionsYieldThe mass of 1000 grains, gGluten contentClass grainBankable. 1 RUR cost, rubles
kg/hagain%%group
Control (10 l/t)33,0 -10032,330III-
Silk, 50 ml/t40,17,1121,536,030,4III8,4
Larixin, 100 ml/t41,48,1124,534,632.8II9,6
Table 2
OptionYieldThe mass of 1000 grains, gGluten contentClass grainBankable. 1 RUR cost, rubles
kg/hagain%%group
Control (water)24,2-10033,528,8IIIII-
Silk, 50 ml/ha28,94,7 19,435,6528,9IIII6,65
Larixin, 100 ml/ha32,98,735,735,6930,1IIa 7.62
R %2,94
NDS0,952,45

1. The method of obtaining biologically active drug from larch wood, containing dihydroquercetin, including the grinding of raw materials, drying the last in the air to a moisture content of not more than 15%, extraction with an organic solvent, evaporation of the extract, wherein the crushed wood dried at 30-40°as extractant used 87-88%isopropyl alcohol, extraction is carried out at a ratio of raw material:solvent 1:(2,5-3), the extract evaporated in 70-90 times, measure the content of dihydroquercetin and add a fivefold excess of emulsifier, followed by stirring the mixture until a homogeneous emulsion.

2. The method according to claim 1, characterized in that the raw materials are crushed first to a particle size of 3-10 mm, and then to a particle size of 1-5 mm

Cab according to claim 1, characterized in that the emulsifier used OP-7 or OP-10.

4. The method according to claim 1, characterized in that before using the drug in the form of an emulsion is diluted with water to a content of dihydroquercetin equal to 50 g/l



 

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12 cl, 10 tbl, 7 ex, 1 dwg

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24 cl, 3 ex, 2 dwg

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3 tbl, 3 ex

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3 cl, 2 tbl, 8 ex

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6 cl, 107 tbl, 1 ex

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1 tbl

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1 tbl

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32 cl, 11 ex

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