CN109574976B - Method for industrially extracting dihydroquercetin from larch roots - Google Patents
Method for industrially extracting dihydroquercetin from larch roots Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/32—2,3-Dihydro derivatives, e.g. flavanones
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
Abstract
The invention relates to a method for industrially extracting dihydroquercetin from larch roots in northeast regions, which takes the larch roots discarded in artificial forest selective cutting or all-fell cutting as raw materials, and prepares the pure dihydroquercetin through twelve steps of raw material selection, raw material cleaning, raw material treatment before extraction, concentration, sedimentation, secondary concentration, primary crystallization, primary centrifugal separation, recrystallization, secondary centrifugal separation, freeze drying and crushing. The method utilizes the conditions of abundant resources of the discarded larch roots (main roots and lateral roots) after cutting, relatively rich content of dihydroquercetin (1.5-1.9%), energy conservation, environmental protection and high yield of dihydroquercetin.
Description
Field of the invention
The invention relates to the field of natural product chemistry, in particular to a method for industrially extracting dihydroquercetin from larch artificial forest selective cutting or all-cutting waste tree roots serving as a raw material.
Background
The larch is a natural plant widely existing in northeast China, has straight trunk end, and is very suitable for being used as wood in various fields such as buildings, furniture and the like. The root of the plant is not valuable and is often discarded as waste.
Dihydroquercetin is a natural medicine with obvious antioxidation, can be used as main and auxiliary raw materials of medicines and bioactive additives of foods and can be widely applied to medicines and health-care functional foods. Has wide application in the fields of medicine, industry and agriculture.
Dihydroquercetin belongs to flavanone compounds, vitamin P is a strong antioxidant, the adsorption of free radicals in natural antioxidants can reach 32 mu mol/mg (the purity of dihydroquercetin is 95%), nowadays, along with the improvement of living water of people, people pay more and more attention to health preservation and long life, vitamin P is widely needed by people as a trace element which is necessary for human bodies and cannot be synthesized by the people, dihydroquercetin as a natural antioxidant has a remarkable effect on the aspect of prolonging the service life of cells by eliminating free radicals, and the dihydroquercetin can be expected to have stronger biological activity and medicinal value.
Studies have shown that it can affect lipid metabolism by modulating enzymatic activity; has certain influence on lymphocyte proliferation 9; simultaneously, the composition shows certain anticancer effect and antiviral effect; and the influence of the polypeptide on intracellular enzyme 15 and the cardiovascular system is also studied to a certain extent.
Dihydroquercetin was first extracted by Fukui, a japanese scholar, and was patented in the united states, canada, russia, et al (US2744919A) (CA532804A) (RU84561C 1); corresponding extraction methods are also provided in domestic (CN1844095) (CN1858046) (CN101333203) (CN101333204) and other patents.
However, the above methods all have the disadvantages of using highly toxic solvent in the extraction process, or having complicated extraction process or low extraction purity.
For example, U.S. Pat. No. 4, 2744919A discloses a dihydroquercetin extract which is obtained by extracting dihydroquercetin with water or an organic solvent having a relatively high polarity, and then purifying the extract by concentration, extraction, crystallization, etc., but this method can only obtain a crude dihydroquercetin product, which has many impurities and many crystallization times. Canadian patent (CA532804A) provides a method for extracting dihydroquercetin from bark with hot water, concentrating, extracting with low polar alcohol, ketone, ether, etc., removing solvent, and crystallizing with hot water. The organic solvent used in the method has certain toxicity, and is inflammable and explosive in production and has high danger.
Chinese patents (CN1844095) (CN1858046) (CN101863869A) (CN102924420B) and the like all provide methods for extracting dihydroquercetin from larch, but some of the methods have the disadvantages of toxic or flammable properties such as the use of methyl tert-butyl ether, acetonitrile or chlorine-containing solvents and the like, and have higher cost; some processes are complex, are not beneficial to industrial production and have low yield; some can only extract crude products, and the purity is lower.
In conclusion, the method for industrially extracting the high-purity product with low toxicity, high efficiency and economy has great significance for the development and utilization of the dihydroquercetin.
Disclosure of Invention
The invention aims to provide a method for industrially extracting dihydroquercetin from larch roots in northeast regions, which utilizes the conditions of abundant resources of the discarded larch roots (main roots and lateral roots) after cutting, enriched content of the dihydroquercetin (1.5-1.9 percent) and the like, saves energy, protects environment and has high yield of the dihydroquercetin.
The invention aims to realize the method for industrially extracting the dihydroquercetin from the roots of the larch in the northeast region, which comprises the following steps:
1) selecting raw materials: the larch of the Korean larch near the northwefting N41 degrees 46 '16.72' east longitude E126 degrees 56 '9.73' is 45-50 years old, the snow accumulation amount is deep in the last year before felling, the root of the dorsal slope tree is the first choice, and the content of dihydroquercetin after mixing the main root and the lateral root is 1.5-1.9%.
2) Raw material cleaning and washing: sawing off and removing fibrous roots and tips of lateral roots with the diameter less than 60mm, peeling off the rest parts of barks, washing away silt by using high-pressure water, determining that no stone blocks are carried between the lateral roots, then air-drying, and storing in a warehouse conforming to GAP standard for later use.
3) And (3) treating raw materials before extraction: crushing the raw material stored in step 2) into wood chips with a particle size of 2x2x4 mm.
4) And (3) extracting and concentrating: adding the wood chips treated in the step 3) into an extraction tank by adopting a cold leaching method, then adding 95% high-grade n-hexane and purified water, and adjusting the pH value of the mixture by using analytically pure glacial acetic acid, wherein the wood chips: purifying water: n-hexane: the mass ratio of the glacial acetic acid is 1:1:4:0.001, the cold soaking time is 14 hours, after the extraction process is finished, the extracting solution is pumped into a single-effect external circulation evaporator by vacuum for concentration, the paste is collected when the vacuum degree is 0.06MPa, the temperature is 65 ℃ and the density of the paste is 1.15-1.20, and meanwhile, the aim of recovering the normal hexane is also achieved.
5) And settling: in the extraction process, a certain amount of turpentine is leached out, so that the turpentine needs to be removed under certain conditions, the concentrated extract is added into a mixing tank, then purified water is added, the ratio of the extract to the purified water is 1:3, the mixture is uniformly placed into 6 transfer barrels when the temperature in the tank is about 95 ℃, the volume of the barrels is 50L, the barrels are placed into a medium water bath, the temperature of water in the tank is 44-46 ℃, when the temperature of the water is reduced to 28-30 ℃, the separation of liquid and solid is completed, and the sedimentation process is completed.
6) And secondary concentration: after the turpentine is removed, the liquid is concentrated to the density of 1.20 by using a single-effect evaporator under the conditions that the vacuum degree is 0.08Mpa and the temperature is 65-68 ℃, and then the concentrated solution is put into a primary crystallization barrel.
7) And primary crystallization: adding 95% ethanol into the primary crystallization barrel, stirring for 10min, cooling to 20-25 deg.C, placing 8-10 titanium rods with diameter in the liquid, standing for 96-108 hr to obtain primary crystals, taking out the titanium rods after the primary crystallization process is completed, and cleaning for the next time.
8) And primary centrifugal separation: and uniformly placing the primary crystals in 4 filter cloth bags, performing solid-liquid separation by using a three-leg centrifuge, wherein the rotation speed of the centrifuge is 1500rpm, operating for 20min, and transferring the liquid to an ethanol recovery device.
9) And (3) recrystallizing: weighing the crystals in the cloth bag in the centrifugal process, pouring the crystals into a reaction kettle, simultaneously adding purified water, sealing, heating, putting the liquid into a recrystallization barrel when the temperature is 95-97 ℃, and repeating the primary crystallization process in the step 7).
10) And secondary centrifugal separation: repeating the process 8).
11) And (3) freeze drying: and (3) dividing the crystal in the cloth bag after centrifugal separation into four discs, and putting the four discs in a BLK-FDYY-3 vacuum freeze dryer at the temperature of-15 ℃ and the vacuum degree of 1.0pa for 16h to obtain a dried product after 16 h.
12) And crushing: pulverizing and sieving the dried powder to obtain light white powder, namely the product dihydroquercetin, which is qualified by 200 meshes.
The invention has the following advantages and positive effects:
1. the invention is a set of energy-saving and environment-friendly process which is suitable for industrial mass production. The raw materials adopted in the invention are the discarded larch roots after the artificial forest is cut, the roots of the larch artificial forest with the gradient of less than or equal to 25 degrees are inevitable products of ginseng planting and 45-50 years of rotation of the artificial forest, and the extracted sawdust is a raw material of a new energy material, namely biomass particles, so that the method is green, economic and environment-friendly and conforms to sustainable development. Such as cold soaking and concentrating under closed and vacuum condition; and refining and recycling the recovered organic solvent.
2. The technology of the invention can realize the preparation of the dihydroquercetin product with high purity and high yield. The solvent adopted in the extraction process is relatively strong in pertinence, such as: when water or alcohol is used as an extracting agent in the traditional process, more impurities such as arabinogalactan, rosin, lignin and the like are generated, the process of separating the substances is complicated, and the purity of a final product is easily reduced. The purity of the final product is up to more than 95 percent, and the yield is up to 5 per mill
3. The invention uses ethanol with proper proportion to participate in the crystallization process, uses a titanium rod (nucleus for crystallization) to place a critical point for breaking the material balance, accelerates the speed of tending to crystallization and improves the crystallization quality, has high product purity, and can recycle the titanium rod, which is the main innovation point of the invention.
Drawings
FIG. 1 is a liquid chromatogram of dihydroquercetin prepared in example 1 of the present invention.
FIG. 2 is a liquid chromatogram of dihydroquercetin prepared in example 2 of the present invention.
FIG. 3 is a liquid chromatogram of dihydroquercetin prepared in example 3 of the present invention.
FIG. 4 is a liquid chromatogram of dihydroquercetin prepared in example 4 of the present invention.
FIG. 5 is a liquid chromatogram of dihydroquercetin prepared in example 5 of the present invention.
Fig. 6 and 7 are nuclear magnetic spectrograms of dihydroquercetin prepared by the invention.
FIG. 8 is a mass spectrum of dihydroquercetin prepared by the present invention.
Detailed Description
The following further details the practice of the invention:
a method for industrially extracting dihydroquercetin from larch (northeast China) roots is characterized by comprising the following steps:
a method for extracting dihydroquercetin from tree root by extracting, settling, and crystallizing is provided. The method comprises the steps of selecting certain tree roots as raw materials, cleaning, pretreating, extracting, concentrating, settling, concentrating for the second time, crystallizing for the first time, centrifugally separating, recrystallizing, separating for the second time, and freeze-drying and crushing to obtain the pure dihydroquercetin.
The present invention will be further described with reference to examples, but it is not limited to any one of these examples or the like.
Example 1: experimental batch 201804002
The main equipment is as follows: 6m3 multifunctional extraction tank, 500L/h single-effect external circulation evaporator; 300L/h single-effect external circulation evaporator, 500L batching tank, 100L reaction kettle and 100L crystallization barrel; 2 BV-5131 water circulation vacuum pump; BLK-FDYY-3 vacuum freeze drier, BFY-1000 pulverizer.
A detection instrument: agilent1260
And (3) standard substance: sigma 03890585-10MG batches: 201703011
The operation process is as follows:
1. weighing 1000kg of qualified raw materials, putting the raw materials into a 6m & lt 3 & gt multifunctional extraction tank, adding 1000kg of purified water and 1000mg of glacial ethanol, then adding 4000kg of n-hexane (95%) to close a feeding port and a pipeline valve, carrying out cold soaking for 14h, then pumping the extracting solution into a 500L/h single-effect external circulation evaporator by utilizing vacuum, concentrating, wherein the vacuum degree is 0.06mpa, the temperature is 65 ℃, adding 3000kg of purified water into the extraction tank, and then heating to recover the n-hexane in the wood chips. When the concentrated solution is discharged at a sampling density of 1.18, 114kg of the concentrated solution is obtained by weighing.
2. Pouring the concentrated solution into a blending tank, adding 432kg of purified water into the tank, heating to 95 ℃, uniformly placing the mixture into 6 transfer barrels, putting the 6 transfer barrels into a water bath tank, keeping the water temperature at 45 ℃, keeping the water temperature at 29 ℃ when the water temperature is 13h, taking out the transfer barrels, and removing turpentine on the surface of the liquid.
3. And (3) pumping the liquid in the barrel into a 300L/h single-effect external circulation evaporator for secondary concentration by utilizing vacuum, wherein the vacuum degree is 0.08Mpa, the temperature is 65 ℃, after 2 hours and 45 minutes, the concentration operation is stopped after the density of the sampled concentrated solution is 1.20, and the concentrated solution is placed into a primary crystallization barrel and weighed as 45 kg.
4. Adding 11.26kg of 95% ethanol into the primary crystallization barrel, stirring for 10min, sealing, and cooling in a water bath. The temperature of water in the tank is room temperature, the temperature of liquid in the barrel is measured to be 22 ℃ after 6 hours and 20 minutes, 9 titanium rods are placed, the titanium rods are taken out after 96 hours, and the formation of crystals is confirmed.
5. Uniformly placing the crystals into 4 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running for 10min at a low-gear rotating speed of 1000 revolutions per minute, automatically starting running for 10min at a high-gear rotating speed of 1500 revolutions per minute, taking out the filter cloth bags, and weighing to obtain 15.66kg of solids.
6. Putting 15.66kg of solid into a reaction kettle, adding 15.66kg of purified water, sealing and heating, stopping heating after a safety valve at the top of the kettle is opened, putting the materials in the kettle into a recrystallization barrel, weighing 7.84kg of ethanol (95%), adding the ethanol into the barrel, stirring for 10min, sealing the barrel cover, putting the barrel into a water bath, measuring the temperature in the barrel to be 20 ℃ for 4 hours and 20 minutes, placing 6 titanium rods, taking out the titanium rods when 100 hours, and confirming the formation of recrystallization.
7. Uniformly distributing the crystals into 2 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running at a low speed for 10min, automatically starting a high speed for 10min, taking out the cloth bag, distributing the solids into 3 drying trays, and conveying the solids into a drying room.
8. And (3) conveying the drying tray into a freeze dryer, setting the working program of the freezer, and drying for 16 hours at the operating temperature of-15 ℃ and the vacuum degree of 1.0 pa.
9. After the drying procedure is finished, the drying tray is taken out, the materials are collected, and are crushed and sieved to obtain dihydroquercetin powder with the granularity of 200 meshes, the weighing is 5.75kg, the yield is 5.75 thousandths, and the purity of the dihydroquercetin is detected to be 94.45%.
Detection of dihydroquercetin:
and (3) qualitatively and quantitatively detecting the dihydroquercetin prepared by the step (1) by taking liquid chromatography as a detection means and taking a sigma 03890585-10MG dihydroquercetin standard substance as a standard substance.
FIG. 1 is a liquid phase chromatogram of dihydroquercetin (201804002) prepared by the above method: the purity of dihydroquercetin is 94.45% as calculated by the peak integral area ratio in the figure
Example 2: experimental batch 201805006
The main equipment is as follows: 6m3 multifunctional extraction tank, 500L/h single-effect external circulation evaporator; 300L/h single-effect external circulation evaporator, 500L batching tank, 100L reaction kettle and 100L crystallization barrel; 2 BV-5131 water circulation vacuum pump; BLK-FDYY-3 vacuum freeze drier, BFY-1000 pulverizer.
A detection instrument: agilent1260
And (3) standard substance: sigma 03890585-10MG batches: 201703011
The operation process is as follows:
1. weighing 1000kg of qualified raw materials, putting the raw materials into a 6m3 multifunctional extraction tank, adding 1000kg of purified water and 1000mg of glacial ethanol, then adding 4000kg of n-hexane (95%) to close a feeding port and close a pipeline valve, carrying out cold soaking for 14h, then pumping an extracting solution into a 500L/h single-effect external circulation evaporator by utilizing vacuum, concentrating, wherein the vacuum degree is 0.06Mpa, the temperature is 65 ℃, adding 3000kg of purified water into the extraction tank, and then heating to recover the n-hexane in the wood chips. When the concentrated solution is discharged at a sampling density of 1.18, 150kg of concentrated solution is obtained by weighing.
2. Pouring the concentrated solution into a blending tank, adding 450kg of purified water into the tank, heating to 95 ℃, uniformly putting the mixture into 6 transfer barrels, putting the 6 transfer barrels into a water bath, keeping the water temperature at 45 ℃, keeping the water temperature at 30 ℃ for 13h, taking out the transfer barrels, and removing turpentine on the surface of the liquid.
3. And (3) pumping the liquid in the barrel into a 300L/h single-effect external circulation evaporator for three times of concentration, wherein the vacuum degree is 0.08Mpa, the temperature is 65 ℃, after 2 hours and 55 minutes, the concentration operation is stopped after the density of the sampled concentrated solution is 1.18, and the concentrated solution is placed into a primary crystallization barrel and weighed to be 47.6 kg.
4. Adding 11.9kg of 95% ethanol into the primary crystallization barrel, stirring for 10min, sealing, and cooling in a water bath barrel. The temperature of water in the tank is room temperature, the temperature of liquid in the barrel is measured to be 23 ℃ after 6 hours and 28 minutes, 9 titanium rods are placed, the titanium rods are taken out after 99 hours, and the formation of crystals is confirmed.
5. Uniformly placing the crystals into 4 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running for 10min at a low-gear rotating speed of 1000 revolutions per minute, automatically starting running for 10min at a high-gear rotating speed of 1500 revolutions per minute, taking out the filter cloth bags, and weighing to obtain 16.3kg of solids.
6. Putting 16.3kg of solid into a reaction kettle, adding 16.3kg of purified water, sealing and heating, stopping heating after a safety valve at the top of the kettle is opened, putting the materials in the kettle into a recrystallization barrel, weighing 8.2kg of ethanol (95%) and adding into the barrel, stirring for 10min, sealing the barrel cover, putting into a water bath, measuring the temperature in the barrel to be 20 ℃ for 4 hours and 20 minutes, placing 6 titanium rods, taking out the titanium rods when 100 hours, and confirming the formation of recrystallization.
7. Uniformly distributing the crystals into 2 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running at a low speed for 10min, automatically starting a high speed for 10min, taking out the cloth bag, distributing the solids into 3 drying trays, and conveying the solids into a drying room.
8. And (3) conveying the drying tray into a freeze dryer, setting the working program of the freezer, and drying for 16 hours at the operating temperature of-15 ℃ and the vacuum degree of 1.0 pa.
9. After the drying procedure is finished, the drying tray is taken out, the materials are collected, and the dihydroquercetin powder with the granularity of 200 meshes is obtained through crushing and sieving, the weighing is 7.92kg, the yield is 7.92 thousandths, and the purity of the dihydroquercetin is detected to be 94.32%.
FIG. 2 is a liquid phase chromatogram of dihydroquercetin (201805006) prepared by the above method: the purity of dihydroquercetin is 94.32% as calculated by the integral area ratio in the figure
Example 3: experimental batch 201806001
The main equipment is as follows: 6m3 multifunctional extraction tank, 500L/h single-effect external circulation evaporator; 300L/h single-effect external circulation evaporator, 500L batching tank, 100L reaction kettle and 100L crystallization barrel; 2 BV-5131 water circulation vacuum pump; BLK-FDYY-3 vacuum freeze drier, BFY-1000 pulverizer.
A detection instrument: agilent1260
And (3) standard substance: sigma 03890585-10MG batches: 201703011
The operation process is as follows:
1. weighing 1000kg of qualified raw materials, putting the raw materials into a 6m3 multifunctional extraction tank, adding 1000kg of purified water and 1000mg of glacial ethanol, then adding 4000kg of n-hexane (95%) to close a feeding port and close a pipeline valve, carrying out cold soaking for 14h, then pumping the extracting solution into a 500L/h single-effect external circulation evaporator by utilizing vacuum, concentrating, adding 3000kg of purified water into the extraction tank at the temperature of 65 ℃ under the vacuum degree of 0.06 and Mpa, and then heating to recover the n-hexane in the wood chips. When the concentrated solution is discharged at a sampling density of 1.19, 138kg of concentrated solution is obtained by weighing.
2. Pouring the concentrated solution into a dosing tank, adding 414kg of purified water into the tank, heating to 95 ℃, uniformly placing the mixture into 6 transfer barrels, placing the 6 transfer barrels into a water bath, keeping the water temperature at 45 ℃ and the water temperature at 30 ℃ for 12h, taking out the transfer barrels, and removing turpentine on the surface of the liquid.
3. And (3) pumping the liquid in the barrel into a 300L/h single-effect external circulation evaporator for three times of concentration, wherein the vacuum degree is 0.08Mpa, the temperature is 65 ℃, after 2 hours and 35 minutes, the concentration operation is stopped after the density of the sampled concentrated solution is 1.15, and the concentrated solution is placed into a primary crystallization barrel and weighed as 42.6 kg.
4. Adding 10.66kg of 95% ethanol into the primary crystallization barrel, stirring for 10min, sealing, and cooling in a water bath barrel. The temperature of water in the tank is room temperature, the temperature of liquid in the barrel is measured to be 22 ℃ after 6 hours and 20 minutes, 9 titanium rods are placed, the titanium rods are taken out after 96 hours, and the formation of crystals is confirmed.
5. Uniformly placing the crystals into 4 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running for 10min at a low-gear rotating speed of 1000 revolutions per minute, automatically starting running for 10min at a high-gear rotating speed of 1500 revolutions per minute, taking out the filter cloth bags, and weighing to obtain 15.36kg of solids.
6. Putting 15.36kg of solid into a reaction kettle, adding 15.36kg of purified water, sealing and heating, stopping heating after a safety valve at the top of the kettle is opened, putting the materials in the kettle into a recrystallization barrel, weighing 7.68kg of ethanol (95%) and adding into the barrel, stirring for 10min, sealing the barrel cover, putting into a water bath, measuring the temperature in the barrel to be 20 ℃ for 4 hours and 20 minutes, placing 6 titanium rods, taking out the titanium rods when 100 hours, and confirming the formation of recrystallization.
7. Uniformly distributing the crystals into 2 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running at a low speed for 10min, automatically starting a high speed for 10min, taking out the cloth bag, distributing the solids into 3 drying trays, and conveying the solids into a drying room.
8. And (3) conveying the drying tray into a freeze dryer, setting the working program of the freezer, and drying for 16 hours at the operating temperature of-15 ℃ and the vacuum degree of 1.0 pa.
9. After the drying procedure is finished, taking out the drying tray, collecting the materials, pulverizing, and sieving to obtain dihydroquercetin powder with particle size of 200 mesh, weight of 7.112kg, yield of 7.112 ‰, and purity of 94.98%.
FIG. 3 is a liquid phase chromatogram of dihydroquercetin (201806001) prepared by the above method: the purity of dihydroquercetin was 94.98%, as calculated from the ratio of the peak-to-area in the figure.
Example 4: experimental batch 201808014
The main equipment is as follows: 6m3 multifunctional extraction tank, 500L/h single-effect external circulation evaporator; 300L/h single-effect external circulation evaporator, 500L batching tank, 100L reaction kettle and 100L crystallization barrel; 2 BV-5131 water circulation vacuum pump; BLK-FDYY-3 vacuum freeze drier, BFY-1000 pulverizer.
A detection instrument: agilent1260
And (3) standard substance: sigma 03890585-10MG batches: 201703011
The operation process is as follows:
1. weighing 1000kg of qualified raw materials, putting the raw materials into a 6m3 multifunctional extraction tank, adding 1000kg of purified water and 1000mg of glacial ethanol, then adding 4000kg of n-hexane (95%) to close a feeding port and close a pipeline valve, carrying out cold soaking for 14h, then pumping the extracting solution into a 500L/h single-effect external circulation evaporator by utilizing vacuum, concentrating, adding 3000kg of purified water into the extraction tank at the temperature of 65 ℃ under the vacuum degree of 0.06 and Mpa, and then heating to recover the n-hexane in the wood chips. When the concentrated solution is discharged at a sampling density of 1.15, 152kg of concentrated solution is obtained by weighing.
2. Pouring the concentrated solution into a dosing tank, adding 456kg of purified water into the tank, heating to 95 ℃, uniformly putting the mixture into 6 transfer barrels, putting the 6 transfer barrels into a water bath, keeping the water temperature at 45 ℃, taking the water temperature at 28 ℃ at 14h, taking out the transfer barrels, and removing turpentine on the surface of the liquid.
3. And (3) pumping the liquid in the barrel into a 300L/h single-effect external circulation evaporator for three times of concentration, wherein the vacuum degree is 0.08Mpa, the temperature is 68 ℃, after 2 hours and 25 minutes, the concentration operation is stopped after the density of the sampled concentrated solution is 1.16, and the concentrated solution is placed into a primary crystallization barrel and weighed as 45.6 kg.
4. Adding 11.4kg of 95% ethanol into the primary crystallization barrel, stirring for 10min, sealing, and cooling in a water bath barrel. The temperature of water in the tank is room temperature, the temperature of liquid in the barrel is measured to be 23 ℃ after 6 hours and 30 minutes, 9 titanium rods are placed, the titanium rods are taken out after 96 hours, and the formation of crystals is confirmed.
5. Uniformly placing the crystals into 4 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running for 10min at a low-gear rotating speed of 1000 revolutions per minute, automatically starting running for 10min at a high-gear rotating speed of 1500 revolutions per minute, taking out the filter cloth bags, and weighing to obtain 16.24kg of solids.
6. Putting 16.24kg of solid into a reaction kettle, adding 16.24kg of purified water, sealing and heating, stopping heating after a safety valve at the top of the kettle is opened, putting the materials in the kettle into a recrystallization barrel, weighing 8.12kg of ethanol (95%) and adding into the barrel, stirring for 10min, sealing the barrel cover, putting into a water bath, measuring the temperature in the barrel to be 24 ℃ for 4 hours and 30 minutes, placing 6 titanium rods, taking out the titanium rods when 100 hours, and confirming the formation of recrystallization.
7. Uniformly distributing the crystals into 2 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running at a low speed for 10min, automatically starting a high speed for 10min, taking out the cloth bag, distributing the solids into 3 drying trays, and conveying the solids into a drying room.
8. And (3) conveying the drying tray into a freeze dryer, setting the working program of the freezer, and drying for 16 hours at the operating temperature of-15 ℃ and the vacuum degree of 1.0 pa.
9. After the drying procedure is finished, the drying tray is taken out, the materials are collected, and the dihydroquercetin powder with the granularity of 200 meshes is obtained through crushing and sieving, the weighing is 8.34kg, the yield is 8.34 per mill, and the purity of the dihydroquercetin is detected to be 94.56%.
FIG. 4 is a liquid phase chromatogram of dihydroquercetin (201808014) prepared by the above method: the purity of dihydroquercetin was 94.56%, as calculated from the ratio of the peak-to-area in the figure.
Example 5: experimental batch 201809007
The main equipment is as follows: 6m3 multifunctional extraction tank, 500L/h single-effect external circulation evaporator; 300L/h single-effect external circulation evaporator, 500L batching tank, 100L reaction kettle and 100L crystallization barrel; 2 BV-5131 water circulation vacuum pump; BLK-FDYY-3 vacuum freeze drier, BFY-1000 pulverizer.
A detection instrument: agilent1260
And (3) standard substance: sigma 03890585-10MG batches: 201703011
The operation process is as follows:
1. weighing 1000kg of qualified raw materials, putting the raw materials into a 6m3 multifunctional extraction tank, adding 1000kg of purified water and 1000mg of glacial ethanol, then adding 4000kg of n-hexane (95%) to close a feeding port and close a pipeline valve, carrying out cold soaking for 14h, then pumping the extracting solution into a 500L/h single-effect external circulation evaporator by utilizing vacuum, concentrating, adding 3000kg of purified water into the extraction tank at the temperature of 65 ℃ under the vacuum degree of 0.06 and Mpa, and then heating to recover the n-hexane in the wood chips. When the concentrated solution is discharged at a sampling density of 1.18, 116kg of concentrated solution is obtained by weighing.
2. Pouring the concentrated solution into a blending tank, adding 438kg of purified water into the tank, heating to 95 ℃, uniformly putting the mixture into 6 transfer barrels, putting the 6 transfer barrels into a water bath, taking the transfer barrels out, and removing turpentine on the surface of the liquid, wherein the water temperature is 45 ℃ and is 29 ℃ when the water temperature is 13 h.
3. And (3) pumping the liquid in the barrel into a 300L/h single-effect external circulation evaporator for three times of concentration, wherein the vacuum degree is 0.08Mpa, the temperature is 65 ℃, after 2 hours and 45 minutes, the concentration operation is stopped after the density of the sampled concentrated solution is 1.20, and the concentrated solution is placed into a primary crystallization barrel and weighed as 45.8 kg.
4. Adding 11.48kg of 95% ethanol into the primary crystallization barrel, stirring for 10min, sealing, and cooling in a water bath barrel. The temperature of water in the tank is room temperature, the temperature of liquid in the barrel is measured to be 22 ℃ after 6 hours and 20 minutes, 9 titanium rods are placed, the titanium rods are taken out after 96 hours, and the formation of crystals is confirmed.
5. Uniformly placing the crystals into 4 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running for 10min at a low-gear rotating speed of 1000 revolutions per minute, automatically starting running for 10min at a high-gear rotating speed of 1500 revolutions per minute, taking out the filter cloth bags, and weighing to obtain 15.76kg of solids.
6. Putting 15.76kg of solid into a reaction kettle, adding 15.76kg of purified water, sealing and heating, stopping heating after a safety valve at the top of the kettle is opened, putting the materials in the kettle into a recrystallization barrel, weighing 7.88kg of ethanol (95%) and adding into the barrel, stirring for 10min, sealing the barrel cover, putting into a water bath, measuring the temperature in the barrel to be 20 ℃ for 4 hours and 20 minutes, placing 6 titanium rods, taking out the titanium rods when 100 hours, and confirming the formation of recrystallization.
7. Uniformly distributing the crystals into 2 filter cloth bags, symmetrically placing the filter cloth bags into a centrifuge, starting the centrifuge, running at a low speed for 10min, automatically starting a high speed for 10min, taking out the cloth bag, distributing the solids into 3 drying trays, and conveying the solids into a drying room.
8. And (3) conveying the drying tray into a freeze dryer, setting the working program of the freezer, and drying for 16 hours at the operating temperature of-15 ℃ and the vacuum degree of 1.0 pa.
9. After the drying procedure is finished, the drying tray is taken out, the materials are collected, and the dihydroquercetin powder with the granularity of 200 meshes is obtained through crushing and sieving, the weighing is 7.62kg, the yield is 7.62 per mill, and the purity of the dihydroquercetin is detected to be 94.85 percent.
FIG. 5 is a liquid phase chromatogram of dihydroquercetin (201809007) prepared by the above method: the purity of dihydroquercetin can be calculated from the proportion of the peak integral area in the figure, and is 94.85%.
In summary, waste larch root sawdust is used as a raw material, used extracting solution is water and low-toxicity easily-recycled n-hexane widely used in soybean oil extraction, only nontoxic and harmless ethanol and water are used in a subsequent separation process, and finally, a titanium rod is used for heterogeneous crystallization. The obtained product is verified to be dihydroquercetin (figures 6, 7 and 8) by nuclear magnetic and mass spectrometry, and is verified to be high-purity dihydroquercetin by high performance liquid chromatography. In summary, the process is an effective process for industrially providing high-yield dihydroquercetin production on the premise of high purity.
Claims (1)
1. A method for industrially extracting dihydroquercetin from larch roots is characterized by comprising the following steps: the method takes larch roots discarded in selective cutting or all-cutting of artificial forests as raw materials, and prepares the pure dihydroquercetin by twelve steps of raw material selection, raw material cleaning and washing, raw material treatment before extraction, concentration, sedimentation, secondary concentration, primary crystallization, primary centrifugal separation, recrystallization, secondary centrifugal separation, freeze drying and crushing; the method comprises the following specific steps:
1) selecting raw materials: the larch of the southern white larch near the northwefting N41 degrees 46 '16.72' east longitude E126 degrees 56 '9.73', the tree age is 45-50 years, the snow accumulation amount is deep in the last year before felling, the root of the dorsal slope tree is the first choice, and the content of dihydroquercetin after mixing the main root and the lateral root is 1.5-1.9%;
2) raw material cleaning and washing: sawing off and removing fibrous roots and tips of lateral roots with the diameter less than 60mm, peeling off the rest parts of barks, washing away silt by using high-pressure water, determining that no stone blocks are carried between the lateral roots, then air-drying, and storing in a warehouse conforming to the GAP standard for later use;
3) and (3) treating raw materials before extraction: crushing the raw material stored in the step 2) into wood chips with the granularity of 2x2x4 mm;
4) and (3) extracting and concentrating: adding the wood chips treated in the step 3) into an extraction tank by adopting a cold leaching method, then adding 95% high-grade n-hexane and purified water, and adjusting the pH value of the mixture by using analytically pure glacial acetic acid, wherein the wood chips: purifying water: n-hexane: the mass ratio of the glacial acetic acid is 1:1:4:0.001, the cold soaking time is 14 hours, after the extraction process is finished, the extracting solution is pumped into a single-effect external circulation evaporator by vacuum for concentration, the paste is collected when the vacuum degree is 0.06MPa, the temperature is 65 ℃ and the density of the paste is 1.15-1.20, and meanwhile, the aim of recovering normal hexane is also achieved;
5) and settling: in the extraction process, a certain amount of turpentine is leached out, so that the turpentine needs to be removed under certain conditions, the concentrated extract is added into a mixing tank, then purified water is added, the ratio of the extract to the purified water is 1:3, the mixture is uniformly placed into 6 transfer barrels when the temperature in the tank is about 95 ℃, the volume of the barrels is 50L, the barrels are placed into a reclaimed water bath, the temperature of water in the bath is 44-46 ℃, when the temperature of the water is reduced to 28-30 ℃, the separation of liquid and solid is completed, and the sedimentation process is completed;
6) and secondary concentration: after the turpentine is removed, the liquid is concentrated to the density of 1.20 by using a single-effect evaporator under the conditions that the vacuum degree is 0.08Mpa and the temperature is 65-68 ℃, and then the concentrated solution is put into a primary crystallization barrel;
7) and primary crystallization: adding 95% ethanol into a primary crystallization barrel, stirring for 10min, placing 8-10 titanium rods with the diameter in the liquid when the temperature is reduced to 20-25 ℃, standing for 96-108 h to obtain primary crystals, taking out the titanium rods after the primary crystallization process is finished, and washing for the next time;
8) and primary centrifugal separation: uniformly placing the primary crystals in 4 filter cloth bags, performing solid-liquid separation by using a three-leg centrifuge, wherein the rotation speed of the centrifuge is 1500rpm, running for 20min, and transferring the liquid to an ethanol recovery device;
9) and (3) recrystallizing: weighing the crystals in the cloth bag in the centrifugal process, pouring the crystals into a reaction kettle, adding purified water at the same time, sealing, heating, putting the liquid into a recrystallization barrel when the temperature is 95-97 ℃, and repeating the primary crystallization process in the step 7);
10) and secondary centrifugal separation: a step of repeating the process 8);
11) and (3) freeze drying: putting the crystal in the cloth bag after centrifugal separation into four discs and putting the four discs in a BLK-FDYY-3 vacuum freeze dryer, wherein the temperature is-15 ℃, the vacuum degree is 1.0pa, the setting time is 16h, and the dried product is obtained after 16 h;
12) and crushing: pulverizing and sieving the dried powder to obtain light white powder, namely the product dihydroquercetin, which is qualified by 200 meshes.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2318528C2 (en) * | 2006-04-03 | 2008-03-10 | Евгений Викентьевич Лифанов | Method for preparing dihydroqurcetin |
| CN101993429A (en) * | 2010-12-08 | 2011-03-30 | 天津市尖峰天然产物研究开发有限公司 | Method for extracting dihydroquercetin from larch sawdust |
| CN102924420A (en) * | 2012-10-25 | 2013-02-13 | 沈立乾 | Method for extracting dihydroquercetin from root of dahurian larch |
| CN103951645A (en) * | 2013-07-18 | 2014-07-30 | 郑毅男 | Preparation method and medicinal purpose of Larix olgensis extractive |
| CN108285440A (en) * | 2017-05-20 | 2018-07-17 | 郑毅男 | A kind of preparation method for extracting dihydroquercetin extract in larch using ultra-high pressure apparatus |
-
2018
- 2018-12-10 CN CN201811502285.3A patent/CN109574976B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2318528C2 (en) * | 2006-04-03 | 2008-03-10 | Евгений Викентьевич Лифанов | Method for preparing dihydroqurcetin |
| CN101993429A (en) * | 2010-12-08 | 2011-03-30 | 天津市尖峰天然产物研究开发有限公司 | Method for extracting dihydroquercetin from larch sawdust |
| CN102924420A (en) * | 2012-10-25 | 2013-02-13 | 沈立乾 | Method for extracting dihydroquercetin from root of dahurian larch |
| CN103951645A (en) * | 2013-07-18 | 2014-07-30 | 郑毅男 | Preparation method and medicinal purpose of Larix olgensis extractive |
| CN108285440A (en) * | 2017-05-20 | 2018-07-17 | 郑毅男 | A kind of preparation method for extracting dihydroquercetin extract in larch using ultra-high pressure apparatus |
Non-Patent Citations (1)
| Title |
|---|
| 落叶松中二氢槲皮素的提取及鉴定;金建忠，等;《浙江林业科技》;20040930;第24卷(第5期);第15-17页 * |
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