CN107602641B - Method for extracting aescine from buckeye seeds under ultrahigh pressure based on freeze drying - Google Patents
Method for extracting aescine from buckeye seeds under ultrahigh pressure based on freeze drying Download PDFInfo
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Abstract
the invention discloses a method for extracting aescine from buckeye seeds by ultrahigh pressure based on freeze drying. According to the invention, through carrying out freeze drying treatment on buckeye seeds, the internal structure of medicinal material tissues is changed, cell walls are easier to break, active ingredients are easier to exude, the pressure and the times of ultrahigh pressure extraction are reduced, and the energy consumption and the cost are reduced.
Description
Technical Field
The invention belongs to the field of pharmacy, relates to an extraction method of aescin, and particularly relates to a method for extracting aescin from buckeye seeds under ultrahigh pressure based on freeze drying.
background
aescin, also called aescin acid, is the general name of total saponin, beta-aescin or isoaescin extracted from seeds of aesculus plant of aesculaceae, and belongs to triterpenoid saponin. The aescin has the functions of resisting inflammation, resisting edema, etc. and is used clinically in treating cerebral edema, wound and other diseases. Most of the current methods for extracting aescin are methods of ethanol thermal reflux or leakage, and for example, patent documents such as CN 104402963A, CN 102659897A, CN 106589045A, CN 104804060A and the like adopt the methods.
With the continuous progress of the extraction technology of traditional Chinese medicines, more and more new methods are used for extracting active ingredients of aescin. CN 106491672A discloses a method for extracting aescin by supercritical carbon dioxide extraction, CN 106554384A discloses a method for extracting aescin by ultrasonic assistance, CN 102309529 a discloses a method for extracting aescin by microwave, and CN 101967175 a discloses a method for extracting aescin by ultrasonic-microwave synergy. However, until now, no report of extracting aescin by using an ultrahigh pressure method is found.
The ultrahigh pressure extraction is also called ultrahigh cold isostatic pressure extraction, which is characterized in that hydrostatic pressure of 100-1000MPa is applied to mixed solution of an extraction solvent and the traditional Chinese medicine, and the mixed solution is kept for a period of time under preset pressure, so that the pressure inside and outside plant cells is quickly released after the pressure is balanced. Because the osmotic pressure inside and outside the cell suddenly increases, the structure of the cell membrane changes, so that the effective components inside the cell can pass through the cell membrane and be transferred to the extracting solution outside the cell, and the aim of extracting the effective components of the traditional Chinese medicine is fulfilled. The method can effectively extract multiple effective components such as flavone, saponin, polysaccharide and alkaloid. Compared with other extraction modes, the ultrahigh pressure extraction has the advantages of low temperature, short time, high efficiency and the like.
Buckeye seed is hard in texture, cell walls are difficult to break, a solvent is difficult to permeate into medicinal material tissues, high pressure (generally over 500 Mpa) and more times (generally over 3 times of repeated extraction) are needed during ultrahigh-pressure extraction, and therefore energy consumption and cost are increased.
disclosure of Invention
The invention aims to provide a method for extracting aescine under ultrahigh pressure, which comprises the step of carrying out freeze drying treatment on buckeye firstly, so that the internal structure of a medicinal material tissue is changed, cell walls are easier to break, active ingredients are easier to seep out, the pressure and the times of ultrahigh pressure extraction are reduced, and the energy consumption and the cost are reduced.
The method provided by the invention comprises the following steps:
(1) crushing buckeye seeds into powder with the granularity of 5-100 meshes, soaking the powder in water for 0.5-3 hours, then fishing out the powder, draining water on the surface, and quickly freezing the powder in a freezer;
(2) Vacuum drying the frozen buckeye seeds, controlling the vacuum degree in the drying process within the range of 10-20pa, and controlling the temperature within the range of 10-50 ℃;
(3) Adding 5-20 times weight of extraction solvent into dried buckeye, sealing in a pressure-resistant flexible soft bag, placing in an extraction container of an ultrahigh pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: raising the pressure in the extraction container to 100-400MPa within 5-30 min;
2) And (3) pressure maintaining stage: keeping the pressure for 5-20 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 5-30 s;
4) repeating the steps 1) -3)1-3 times;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
Preferably, the extraction solvent is water or 10-50% ethanol. According to the principle of similar compatibility, the content of active ingredients extracted by the solvent is the highest.
preferably, the granularity of the buckeye powder is 10-30 meshes. The particle size is too large, the solvent is not easy to permeate the medicinal materials to permeate into tissue cells, and the contact between the active ingredients and the solvent is reduced, so that the extraction efficiency is reduced; on the other hand, if the particle size is too small, not only the dissolution of the active ingredient increases, but also the dissolution of impurity components such as tissue, protein and tannin of the medicinal material also increases, which may lead to a decrease in the content of the active ingredient in the extract.
Preferably, the extraction solvent is 10-15 times of the weight of buckeye seeds. If the amount of the extraction solvent is too small, the active ingredient is difficult to be dissolved out after reaching the dissolution balance, resulting in low extraction efficiency, and if the amount of the extraction solvent is too large, the cost is increased.
preferably, the pressure raising stage is to raise the pressure in the extraction vessel to 200-300MPa within 10-15 min. The higher the pressure is, the faster the solvent infiltration and the transfer speed of the active ingredients are, and the density, the activity and the structure of the medicinal material basic block can be influenced, so that the dissolution speed of the active ingredients is accelerated, and the dissolution amount is increased; however, if the pressure is too high, the elution of the impurity components is also increased, which is disadvantageous in extraction and increases the cost. And the faster the pressure is raised, the more easily the cell wall is broken.
preferably, the time of the pressure holding stage is 10-15 min. Within this range, the solute is dissolved in a better equilibrium.
Preferably, the time of the pressure relief stage is 10-15 s. The pressure of the medicinal material tissue cells is quickly reduced from ultrahigh pressure of hundreds of megapascals to normal pressure, the explosive expansion of fluid and medicinal material matrix volume is generated under the action of reverse pressure, the medicinal material tissues are strongly impacted to cause deformation, and meanwhile, the cell structures are loosened, perforated and cracked, so that the medicinal material components are fully contacted with the solvent and quickly diffused outwards. Therefore, it is theoretically preferable that the pressure release time is as short as possible, but if the time is too short, a large amount of ineffective components may be dissolved.
preferably, the steps 1) -3) are repeated for 1 time, the extracted active ingredients are not only sufficient, but also the period is shortened, and the energy is saved.
the ethanol concentration referred to in the present invention refers to volume concentration, for example, 10-50% ethanol refers to 10-50ml ethanol per 100ml ethanol water solution.
the invention has the beneficial effects that:
1) compared with the existing extraction method, the method disclosed by the invention can improve the content of the aescin in the extract, reduce impurities and improve the transfer rate of the aescin, so that the utilization rate of medicinal materials is higher, the extraction is more sufficient, and the extraction cost is reduced.
2) Compared with the extraction methods such as thermal reflux, decoction, microwave, ultrasound and the like, the method is carried out at lower temperature (room temperature), so that the energy consumption and the damage of active ingredients are reduced, and the obtained extract has better medicinal activity.
3) The pretreatment and extraction time of the method is not more than 5 hours in total, wherein the ultrahigh pressure extraction time is not more than 3 hours in total, and compared with the existing method, the method greatly shortens the extraction time and improves the extraction efficiency.
4) According to the method, before ultrahigh pressure extraction, the buckeye medicinal material is subjected to freeze drying pretreatment, the medicinal material is soaked, a large amount of moisture permeates into the medicinal material tissue, then the frozen moisture is directly sublimated into steam from ice solid without melting in a high-vacuum environment by utilizing the ice crystal sublimation principle, a large number of holes are reserved in the medicinal material tissue during the sublimation and overflow process of the moisture, a porous spongy structure is formed in the medicinal material tissue, and an ultrahigh pressure solvent can permeate into histiocytes through the medicinal material more easily to extract active ingredients, so that the ultrahigh pressure intensity is reduced, the ultrahigh pressure extraction frequency is reduced, and the energy consumption and the cost are reduced.
Detailed description of the invention
the present invention will be described in detail below with reference to specific examples.
Example 1
(1) crushing buckeye seeds into powder with the granularity of 15 meshes, soaking the powder in water for 2 hours, then fishing out the powder, draining off the water on the surface, and quickly freezing the powder in a freezer at the temperature of-25 ℃;
(2) vacuum drying the frozen buckeye seeds, controlling the vacuum degree of the drying process to be 10pa and the temperature to be 40 ℃;
(3) Adding 20% ethanol 12 times the weight of buckeye seeds into the dried buckeye seeds, putting the buckeye seeds and the buckeye seeds into a pressure-resistant flexible soft bag, sealing the mixture, then putting the mixture into an extraction container of an ultrahigh-pressure extraction device, and extracting the mixture at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: increasing the pressure in the extraction container to 200MPa within 15 min;
2) And (3) pressure maintaining stage: keeping the pressure for 15 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 10 s;
4) repeating steps 1) -3)1 time;
(4) Taking out the soft bag from the extraction container, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain aescin extract.
Through measurement and calculation, the yield of the extract is 16.3%, the purity of the aescin is 49.6%, and the transfer rate of the aescin is 99.6%.
Example 2
(1) crushing buckeye seeds into powder with the granularity of 30 meshes, soaking the powder in water for 3 hours, taking out the powder, draining the water on the surface, and quickly freezing the powder in a freezer at the temperature of-30 ℃;
(2) vacuum drying the frozen buckeye seeds, controlling the vacuum degree of the drying process to be 20pa and the temperature to be 50 ℃;
(3) Adding 15 times of water by weight into the dried buckeye, putting the buckeye and the buckeye into a pressure-resistant flexible soft bag together for sealing, then putting the buckeye into an ultrahigh pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: increasing the pressure in the extraction container to 300MPa within 10 min;
2) and (3) pressure maintaining stage: keeping the pressure for 10 min;
3) A pressure relief stage: quickly releasing the pressure to normal pressure within 15 s;
4) Repeating the steps 1) -3) for 2 times;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
through measurement and calculation, the yield of the extract is 17.1%, the purity of the aescin is 47.0%, and the transfer rate of the aescin is 99.0%.
example 3
(1) Crushing buckeye seeds into powder with the granularity of 10 meshes, soaking the powder in water for 1 hour, taking out the powder, draining the water on the surface, and quickly freezing the powder in a freezer at the temperature of-20 ℃;
(2) vacuum drying the frozen buckeye seeds, controlling the vacuum degree of the drying process to be 15pa and the temperature to be 30 ℃;
(3) adding 30% ethanol 10 times the weight of the buckeye seeds into the dried buckeye seeds, putting the buckeye seeds and the buckeye seeds into a pressure-resistant flexible soft bag, sealing the mixture, then putting the mixture into an extraction container of an ultrahigh-pressure extraction device, and extracting the mixture at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: increasing the pressure in the extraction container to 250MPa within 15 min;
2) And (3) pressure maintaining stage: keeping the pressure for 20 min;
3) A pressure relief stage: quickly releasing the pressure to normal pressure within 5 s;
4) repeating steps 1) -3)3 times;
(4) taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
through measurement and calculation, the yield of the extract is 17.3%, the purity of the aescin is 46.3%, and the transfer rate of the aescin is 98.7%.
Example 4
(1) Crushing buckeye seeds into powder with the granularity of 5 meshes, soaking the powder in water for 3 hours, then fishing out the powder, draining off the water on the surface, and quickly freezing the powder in a freezer at the temperature of-30 ℃;
(2) Vacuum drying the frozen buckeye seeds, wherein the vacuum degree in the drying process is controlled to be 10pa, and the temperature is controlled to be 25 ℃;
(3) Adding 50% ethanol 20 times the weight of the buckeye seeds into the dried buckeye seeds, putting the buckeye seeds and the buckeye seeds into a pressure-resistant flexible soft bag, sealing the mixture, then putting the mixture into an extraction container of an ultrahigh pressure extraction device, and extracting the mixture at normal temperature, wherein the extraction steps are as follows:
1) A boosting stage: increasing the pressure in the extraction container to 400MPa within 20 min;
2) And (3) pressure maintaining stage: keeping the pressure for 10 min;
3) A pressure relief stage: quickly releasing the pressure to normal pressure within 30 s;
4) Repeating the steps 1) -3) for 2 times;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
Through measurement and calculation, the yield of the extract is 16.8%, the purity of the aescin is 47.2%, and the transfer rate of the aescin is 97.6%.
example 5
(1) crushing buckeye seeds into powder with the granularity of 50 meshes, adding water, soaking for 1 hour, taking out, draining water on the surface, and quickly freezing in a freezer;
(2) Vacuum drying the frozen buckeye seeds, controlling the vacuum degree in the drying process within the range of 10-20pa, and controlling the temperature within the range of 10-50 ℃;
(3) Adding 5 times of 40% ethanol by weight into dried buckeye, sealing in a pressure-resistant flexible soft bag, placing in an extraction container of an ultrahigh pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: increasing the pressure in the extraction container to 100Mpa within 5-30 min;
2) And (3) pressure maintaining stage: keeping the pressure for 20 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 5 s;
4) Repeating steps 1) -3)3 times;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
Through measurement and calculation, the yield of the extract is 16.9%, the purity of the aescin is 46.6%, and the transfer rate of the aescin is 97.0%.
example 6
(1) crushing buckeye seeds into powder with the granularity of 100 meshes, soaking the powder in water for 0.5 hour, taking out the powder, draining water on the surface of the powder, and quickly freezing the powder in a freezer;
(2) Vacuum drying the frozen buckeye seeds, controlling the vacuum degree in the drying process within the range of 10-20pa, and controlling the temperature within the range of 10-50 ℃;
(3) adding 10 weight times of 10% ethanol into dried buckeye, sealing in a pressure-resistant flexible soft bag, placing in an extraction container of an ultrahigh pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) A boosting stage: increasing the pressure in the extraction container to 300MPa within 20 min;
2) And (3) pressure maintaining stage: keeping the pressure for 15 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 15 s;
4) Repeating steps 1) -3)1 time;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
Through measurement and calculation, the yield of the extract is 17.3%, the purity of the aescin is 46.1%, and the transfer rate of the aescin is 98.2%.
Test examples
The medicinal materials belonging to the same batch as examples 1-6 (the content of aescin in the medicinal materials is 8.12%) are extracted according to different modes, extraction solvents except water decoction are all 20% ethanol, and the extraction methods are all conventional methods well known in the art.
the following comparative examples were also prepared:
crushing buckeye seeds into powder with the granularity of 15 meshes, adding 20% ethanol with the weight being 12 times of the buckeye seeds, soaking for 1 hour, then putting the mixture into a pressure-resistant flexible soft bag, sealing the bag, then putting the bag into an extraction container of an ultrahigh-pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) A boosting stage: increasing the pressure in the extraction container to 200MPa within 15 min;
2) and (3) pressure maintaining stage: keeping the pressure for 15 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 10 s;
4) Repeating steps 1) -3)1 time;
5) taking out the soft bag from the extraction container, pouring out the materials, filtering, centrifuging at high speed, concentrating under reduced pressure, and vacuum drying to obtain aescin extract.
the extract is concentrated and dried, the extract yield is calculated, the aescin content is detected, the aescin transfer rate (the ratio of the total mass of the aescin in the extract to the total mass of the aescin in the extract) is calculated according to the medicinal material content, and the result is shown in the following table.
| extract yield (%) | Content of escin (%) | transfer rate of escin (%) | |
| Ultrasonic extraction | 16.7 | 38.7 | 79.6 |
| microwave extraction | 17.2 | 42.3 | 89.6 |
| Leak extraction | 13.5 | 45.2 | 75.1 |
| Hot reflux extraction | 17.2 | 40.3 | 85.4 |
| Supercritical carbon dioxide extraction | 11.3 | 47.0 | 65.4 |
| decocting in water to extract | 16.0 | 36.4 | 71.7 |
| comparative example | 17.2 | 45.0 | 95.3 |
from the data, the content and the transfer rate of the aescin in the obtained extract are obviously higher than those of the existing extraction methods such as ultrasonic extraction, microwave extraction, leakage extraction and the like by adopting ultrahigh pressure extraction; meanwhile, after the buckeye medicinal material is subjected to freeze drying treatment, the content and transfer rate of aescine in the extracted product are higher than those of the comparative example without the freeze drying treatment. In order to further improve the extraction effect, the pressure is increased, the pressure relief time is shortened, and the extraction times are increased, so that the energy consumption and the cost are increased.
Claims (7)
1. A method for extracting aescine from buckeye seeds under ultrahigh pressure based on freeze drying is characterized by comprising the following steps:
(1) crushing buckeye seeds into powder with the granularity of 5-100 meshes, soaking the powder in water for 0.5-3 hours, then fishing out the powder, draining water on the surface, and quickly freezing the powder in a freezer;
(2) Vacuum drying the frozen buckeye seeds, controlling the vacuum degree in the drying process within the range of 10-20pa, and controlling the temperature within the range of 10-50 ℃;
(3) Adding 5-20 times of weight of extraction solvent into dried buckeye, wherein the extraction solvent is water or 10-50% ethanol, putting into a pressure-resistant flexible soft bag, sealing, putting into an extraction container of an ultrahigh pressure extraction device, and extracting at normal temperature, wherein the extraction steps are as follows:
1) a boosting stage: raising the pressure in the extraction container to 100-400MPa within 5-30 min;
2) And (3) pressure maintaining stage: keeping the pressure for 5-20 min;
3) a pressure relief stage: quickly releasing the pressure to normal pressure within 5-30 s;
4) repeating the steps 1) -3)1-3 times;
(4) Taking out the soft bag from the ultrahigh pressure extraction device, pouring out the materials, filtering, centrifuging, concentrating, and drying to obtain the aescin extract.
2. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: the granularity of the buckeye powder is 10-30 meshes.
3. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: the extraction solvent is 10-15 times of the weight of buckeye seeds.
4. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: the pressure raising stage is to raise the pressure inside the extracting container to 200-300MPa within 10-15 min.
5. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: the pressure maintaining period is 10-15 min.
6. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: the time of the pressure relief stage is 10-15 s.
7. method for extracting aescine from buckeye seeds based on freeze drying according to claim 1, wherein the method comprises the following steps: repeating the steps 1) -3) for 1 time.
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| CN101974061A (en) * | 2010-10-19 | 2011-02-16 | 南京泽朗医药科技有限公司 | Method for extracting purified aescine from horse chestnut |
| CN102516011A (en) * | 2011-12-16 | 2012-06-27 | 西藏天成农牧产业股份有限公司 | Extraction method for lycopene |
| CN103550951A (en) * | 2013-11-20 | 2014-02-05 | 青岛农业大学 | Novel method for extracting or purifying effective components of traditional Chinese medicine |
| CN104402963A (en) * | 2014-10-29 | 2015-03-11 | 武汉爱民制药有限公司 | Sodium aescinate preparation method |
| CN106692703A (en) * | 2017-03-17 | 2017-05-24 | 贵州健瑞安药业有限公司 | Preparation method of dendrobium freeze-dried powder |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101974061A (en) * | 2010-10-19 | 2011-02-16 | 南京泽朗医药科技有限公司 | Method for extracting purified aescine from horse chestnut |
| CN102516011A (en) * | 2011-12-16 | 2012-06-27 | 西藏天成农牧产业股份有限公司 | Extraction method for lycopene |
| CN103550951A (en) * | 2013-11-20 | 2014-02-05 | 青岛农业大学 | Novel method for extracting or purifying effective components of traditional Chinese medicine |
| CN104402963A (en) * | 2014-10-29 | 2015-03-11 | 武汉爱民制药有限公司 | Sodium aescinate preparation method |
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