CN112724192B - Method for extracting and preparing aescine sodium from buckeye seeds - Google Patents
Method for extracting and preparing aescine sodium from buckeye seeds Download PDFInfo
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- CN112724192B CN112724192B CN202011619879.XA CN202011619879A CN112724192B CN 112724192 B CN112724192 B CN 112724192B CN 202011619879 A CN202011619879 A CN 202011619879A CN 112724192 B CN112724192 B CN 112724192B
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- sodium
- solution
- buckeye
- aescin
- aescine
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- 229930182493 triterpene saponin Natural products 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 230000002666 vasoprotective effect Effects 0.000 description 1
- 201000002498 viral encephalitis Diseases 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
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Abstract
The invention provides a method for extracting and preparing aescine sodium from buckeye seeds. The preparation method comprises the following steps: crushing and breaking the wall of buckeye seed raw materials; leaching the crushed and wall-broken buckeye raw material by using an acidic ethanol aqueous solution to obtain a primary extract; concentrating the primary extract, and eluting the grease by using a solvent to obtain a water-phase solution; adding high molecular polymer surfactant into the water phase solution, and performing rotary evaporation to obtain aescin concentrated solution; decolorizing the concentrated solution of aescin; carrying out sodium cation exchange resin chromatography on the decolorized aescin concentrated solution, and collecting eluate; the eluent is filtered, crystallized, recrystallized and dried to obtain the sodium aescinate. The invention has simpler process conditions and is environment-friendly; the adopted high molecular polymer surfactant can effectively remove impurities and selectively form micelles with beta-aescine, and the purity of the prepared sodium aescine is more than 95%, wherein the mass content of the beta-aescine is as high as more than 70%.
Description
Technical Field
The invention belongs to the technical field of natural pharmaceutical chemistry, and relates to a method for extracting and preparing aescine sodium from buckeye seeds.
Background
Buckeye (Aesculi, also known as perilla seed and prism) is the mature seed of Aesculus chinensis (Bge) of aesculaceae, Aesculus hippocastanum, Aesculus chestnut (Aesculus lupeostatum), Aesculus wilsonii Rehd or Aesculus chinensis Bge. As a common traditional Chinese medicine, semen aesculi is sweet and warm in nature and enters liver and stomach meridians, has the effects of regulating qi, relieving epigastric distention, harmonizing stomach and relieving pain, and is mainly used for chest and abdomen fullness and epigastric pain. The buckeye seeds mainly contain 32% of fatty oil, 36% of starch, 15% of fiber and the like, wherein the fatty oil is mainly glyceride of oleic acid and stearic acid. Buckeye seeds are widely distributed in China, aescin extracted from buckeye seeds has good curative effect and good application prospect, is one of the few pure natural plant medicines which are known at present and have stable and exact curative effect and less side effect, the clinical application range of the buckeye saponin is continuously expanded, and the buckeye saponin has wide application prospect.
The research on the efficacy of the aescin shows that the aescin has the effects of diminishing inflammation, resisting exudation, restoring capillary permeability, improving venous tension, improving blood circulation, promoting brain cell recovery and the like, can relieve and eliminate the formation of edema or hematoma, and has oxygen free radical resistance and neuroprotective effect. Can be widely used for treating epidemic encephalitis B, cerebrovascular diseases, peripheral nerve diseases, Hunt's syndrome, tumor, anal edema, facial neuritis, nephrotic syndrome I and other diseases. One significant advantage is that it has the anti-inflammatory and circulatory improving functions of hormones, but without the side effects and contraindications of hormones. The number of manufacturers engaged in the production of the drug and preparing to develop the drug is increasing at home and abroad.
Mature fruit of horse chestnut contains more than 30 saponin components, and the research on the effective components thereof is continued till now. The research in the last 50 th century found that the fruit of horse chestnut can extract two crystalline components respectively, wherein the hemolytic saponin is named aescin, and the other blood-insoluble saponin is named saponin hyposaponin. Wherein the aescin is a natural mixture of triterpene saponin, and exists in plant in the form of alpha-aescin and beta-aescin. In the last 60 s of the century, Merkel et al demonstrated that beta-aescine, an active ingredient with anti-edema, anti-exudation and vasoprotective effects, was used. At present, the medicament widely applied clinically is beta-aescine sodium which is a sodium salt compound of the beta-aescine.
According to the national standard of China, the main 4 components of the aescin are named as aescin A, aescin B, aescin C and aescin D in sequence according to the sequence of peaks appearing by liquid chromatography. Aescin A and aescin B belong to beta-aescin, and aescin C and aescin D belong to alpha-aescin. At present, an effective means for separating different effective components in the aescin is lacked, and total aescin (Totalescin) sodium is mainly used in clinical application. Recent researches have considered that the toxic reaction of the aescin is mainly caused by alpha-aescin, and the reduction of the content of the alpha-aescin in the extract can reduce the toxic and side effects of the aescin extract.
The extraction process of aescin in the prior art comprises two steps of primary extraction and purification. The ethanol or methanol reflux method is usually adopted for the initial extraction, and the main problem is that the heat resistance of the aescin is poor, and the glycosidic bond is easy to break when the temperature is higher than 60 ℃, so that the extraction temperature cannot be too high, and the extraction time is longer. Microwave or ultrasonic assisted extraction has also been reported, but the actual effect of microwave assisted extraction is difficult to verify due to the easy cleavage of glycosidic bonds. In addition, researches try to carry out initial extraction on aescin by adopting a supercritical extraction method, but the process conditions are complicated, so that the production cost is obviously improved, and the practical application value is lacked. In addition, the initial extraction of the aescin by an ultrahigh pressure extraction method has the advantages of high total extraction rate of the aescin and less impurities in the extract, and has the disadvantages that the safety hazard of the production process is increased by adopting a high-pressure reaction vessel, and the alpha-aescin and the beta-aescin are difficult to selectively extract.
For the purification of the primary extraction liquid, ion exchange columns are mostly adopted in the prior art, and ultrafiltration or repeated crystallization is also adopted.
Micelle (Micelle) refers to an ordered collection of molecules that begins to form in large numbers after a certain concentration of surfactant in an aqueous solution is reached. Surfactants are dissolved in water and when their concentration is low they are monomolecularly dispersed or adsorbed on the surface of the solution to reduce surface tension. When the concentration of the surfactant is increased to the point that the surface of the solution is saturated and can not be adsorbed any more, molecules of the surfactant begin to transfer into the solution, and as the hydrophobic parts of the molecules of the surfactant have low affinity with water and the hydrophilic parts have high attraction, when the concentration reaches a certain value, the hydrophobic parts of a plurality of molecules of the surfactant attract each other and associate with each other to form an association, namely a micelle. In micelles, the hydrophobic groups of the surfactant molecules aggregate to form the inner core of the micelle, and the hydrophilic polar groups form the outer layer of the micelle. The water-insoluble substance molecules can be wrapped in the micelle cores by the hydrophobic groups of the micelles to form a water-soluble micelle solution. When the surfactant concentration of the aqueous solution of the micelle is reduced or the pH value or surface tension of the solution is changed, the micelle is broken, and molecules encapsulated in the inner core are released. Recent studies have shown that the molecular structure of the surfactant is selective to the molecules of the substance to be coated. Taking plant-extracted terpenoid paclitaxel as an example, only a few surfactants such as mPEG-PLLA can effectively wrap paclitaxel to form micelles, while docetaxel which is a terpenoid cannot be effectively wrapped by mPEG-PLLA to form micelles.
In conclusion, because the components contained in the raw materials of aescin are very complex and difficult to separate, the prior art faces two difficulties in selecting the extraction rate and the purity of the effective components. If the extraction rate is pursued, various biological impurities in the raw materials are mixed into the primary extraction product, and the difficulty of the purification process is increased. To take care of the simplicity of the purification process, the extraction rate in the initial extraction stage cannot be too high. At present, no report of adopting micelle technology in the process of purifying aescine is found.
Disclosure of Invention
Based on the problems in the prior art, the first purpose of the invention is to provide a method for preparing sodium aescinate by extracting buckeye seeds; the second purpose of the invention is to provide sodium aescinate obtained by the method.
In order to solve the technical problems, on one hand, the invention provides a method for extracting and preparing sodium aescinate by using buckeye seeds, which comprises the following steps:
crushing and breaking the wall of buckeye seed raw materials; leaching the crushed and wall-broken buckeye raw material by using an acidic ethanol aqueous solution to obtain a primary extract; concentrating the primary extract, and eluting the grease by using a solvent to obtain a water-phase solution; adding high molecular polymer surfactant into the water phase solution, and performing rotary evaporation to obtain aescin concentrated solution; decolorizing the concentrated solution of aescin; carrying out sodium cation exchange resin chromatography on the decolorized aescin concentrated solution, and collecting eluate; the eluent is filtered, crystallized, recrystallized and dried to obtain the sodium aescinate.
The buckeye raw materials adopted by the invention are selected from horse chestnut, European horse chestnut or Japanese chestnut, the production place is not limited, and the picking period is 9-10 months.
In the above method, preferably, the high molecular polymer surfactant is methoxy polyethylene glycol-polycaprolactone block copolymer (mPEG-PCL), and the molecular weight thereof is 6000 to 10000 g/mol; wherein the molecular weight of the methoxy polyethylene glycol section is 2000g/mol, and the molecular weight of the polycaprolactone section is 4000-8000 g/mol. In the method, the crushed buckeye raw material preferably has a particle size of 8 to 20 meshes.
In the above method, preferably, the method of the wall-breaking treatment comprises: adding acidic water into the crushed buckeye raw material, freezing, and then performing microwave thawing to realize wall breaking.
In the above method, preferably, the pH of the acidic moisture is 5.5 to 6.9; the weight ratio of the crushed buckeye raw material to the acidic moisture is 1: (3-4). Too much or too little of the acidic moisture addition may affect the final product yield.
In the above method, preferably, the method of leaching with an acidic aqueous ethanol solution comprises: and adding an acidic ethanol aqueous solution into the raw material of the buckeye after wall breaking, and leaching for 3-5 hours at the temperature of 40-45 ℃ to obtain a primary extract. Theoretically, the longer the leaching time, the higher the product yield. However, the leaching time of the buckeye raw material after wall breaking is too long, which can cause the purity of the final product to be reduced, and therefore, the leaching time is preferably controlled to be 3-5 hours.
In the above method, preferably, the pH of the acidic ethanol aqueous solution is 5.0 to 6.9, and the concentration of ethanol is 30% to 50%.
In the above method, preferably, the weight ratio of the buckeye raw material after wall breaking to the acidic ethanol aqueous solution is 1: (8-10).
In the above method, preferably, the method for eluting the oil and fat with the solvent after concentrating the primary extract comprises: and adding an organic solvent into the concentrated solution obtained after the primary extraction liquid is concentrated to elute the grease, and separating to obtain a water phase solution and an oil phase solvent, wherein the water phase solution is subjected to subsequent rotary evaporation treatment, and the oil phase solvent is recycled and purified for reuse. The degreasing link selects different organic solvents, and the process parameters such as the use amount of the organic solvents, the degreasing times and the like are mainly based on the consideration of production cost, so that the technical effect provided by the invention is not obviously influenced.
In the method, preferably, the volume ratio of the concentrated solution after the primary extraction solution is concentrated to the organic solvent is 1 (0.2-1); the number of times of grease elution is 1-3.
In the above method, the primary extract is preferably concentrated to 1/4 or less of the original volume to obtain a concentrated solution.
In the above method, preferably, the organic solvent includes dichloromethane and/or chloroform.
In the above method, preferably, the method of adding the high molecular polymer surfactant to the aqueous phase solution for rotary evaporation comprises:
adding a high molecular polymer surfactant into the aqueous phase solution, performing rotary evaporation at the temperature of 40-60 ℃, and concentrating the reaction system to 1/3-1/4 of the original volume to obtain the aescin concentrated solution.
In the above method, preferably, the weight ratio of the aqueous phase solution to the high molecular polymer surfactant is 100: (5-20).
In the above method, preferably, the method for decoloring the concentrated solution of aescin comprises: adding activated carbon into the aescin concentrated solution for decolorization, and then filtering and removing impurities by adopting an ultrafiltration membrane to obtain the decolorized aescin concentrated solution.
In the above method, preferably, the weight ratio of the concentrated solution of aescin to the activated carbon is 1000: (1-5).
In the above method, preferably, the pore size of the ultrafiltration membrane is 50nm (nanometer).
In the above method, preferably, the sodium cation exchange resin chromatography of the decolorized aescin concentrate includes:
adding equal volume of anhydrous ethanol (ethanol with water content less than 5%) into the decolorized concentrated solution of aescin; then, the mixture is loaded on a column sodium type cation exchange resin to be exchanged into sodium salt; then eluting by adopting 2 times of 40% ethanol solution, and cleaning the residual high molecular polymer surfactant in the column bed; then using 70% -90% ethanol solution with 2 times of column volume to elute, and collecting eluent.
In the method, the sodium cation exchange resin is obtained by treating cation exchange resin with acid and NaOH. The cation exchange resin comprises a strong acid resin and a weak acid resin, and different resin types have no significant influence on the technical effect of the invention, and the invention is not limited to the above.
In the above method, preferably, the method for obtaining sodium aescinate by drying after filtering, crystallizing and recrystallizing the eluate comprises: filtering the eluent by a filter membrane; concentrating the eluent to 1/10-1/8 of the original volume after filtration, then adding anhydrous ethanol (ethanol with the water content less than 5%) of 3-5 times of the volume, placing the mixture at the temperature of between 10 ℃ below zero and 20 ℃ below zero after dissolution, and filtering again after crystallization to obtain crystals; repeating the crystallization step for 2 times to obtain recrystallization; and (4) drying the recrystallization in vacuum to obtain sodium aescinate.
In the above method, preferably, the pore size of the filter is 0.22 μm (μm).
The method has the advantages that the process conditions are simple, the pollution of a large amount of exchange resin to the environment is avoided, all solvents can be recycled, and the method is environment-friendly; the wall breaking effect is good by adopting freezing and microwave thawing methods, the yield is improved, and the using amount of a solvent is reduced; the high molecular polymer surfactant mPEG-PCL can effectively remove impurities, can selectively form micelles with beta-aescin, and improves the proportion of the beta-aescin in the product; and then the high-purity sodium aescinate (the purity is more than 95 percent) can be prepared by simple purification, wherein the mass content of the beta-sodium aescinate is more than 70 percent.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in detail below. However, it will be appreciated by those of ordinary skill in the art that numerous technical details are set forth in order to provide a better understanding of the present invention in its various embodiments. However, the technical solutions claimed in the claims of the present invention can be implemented without these technical details and with various changes and modifications based on the following embodiments.
Example 1.
The embodiment provides a method for preparing aescine sodium by using buckeye seeds, which comprises the following steps:
(1) 100g of buckeye raw material (horse chestnut seeds in Hubei of the production area) is crushed to have the particle size of 8-20 meshes.
(2) Adding 300-400 g of acidic water with the pH value of 6.0-6.9 into the crushed buckeye raw materials, uniformly mixing, spreading the mixture in a plastic tray, sending the mixture into a refrigerator, thawing the mixture by using microwaves after freezing, wherein the thawing temperature is not higher than 40 ℃, and breaking the wall.
(3) Adding 800-1000 g of acidic ethanol aqueous solution (the concentration of ethanol is 30-50%, and the pH value is 5.0-6.9) into the raw material of buckeye seeds subjected to wall breaking, and leaching at the temperature of 40-45 ℃ for more than 3 hours to obtain primary extract.
(4) Concentrating the primary extract to below 1/4 of the original volume to obtain a concentrated solution, adding dichloromethane with the same volume into the concentrated solution, fully stirring to elute the grease, then separating to obtain a water phase solution and an oil phase solvent, and recycling and purifying the oil phase solvent.
(5) Adding a high molecular polymer surfactant methoxy polyethylene glycol-polycaprolactone segmented copolymer (mPEG-PCL; molecular weight is 6000-10000g/mol, wherein the molecular weight of the mPEG section is 2000g/mol, the molecular weight of the PCL section is 4000-8000g/mol, and the supplier is Shanghai Xinbo biotechnology limited), adding 5-20 parts of mPEG-PCL into each 100 parts by weight of the aqueous solution, performing rotary evaporation at 40-60 ℃, and concentrating the reaction system to 1/3-1/4 of the original volume to obtain the aescin concentrated solution.
(6) Adding activated carbon into the aescin concentrated solution for decolorization, adding 1-5 parts of activated carbon into every 1000 parts of the aescin concentrated solution, and filtering and removing impurities by adopting a 50nm (nanometer) ultrafiltration membrane to obtain the decolorized aescin concentrated solution.
(7) Adding equal volume of absolute ethyl alcohol into the concentrated solution of the aescin after the decoloration treatment; then, the mixture is loaded on a column sodium type cation exchange resin to be exchanged into sodium salt; then eluting by using 40 percent ethanol solution with 2 times volume, and cleaning the residual mPEG-PCL in the column bed; then using 70% -90% ethanol solution with 2 times of column volume to elute, and collecting eluent.
(8) Filtering the eluate with 0.22 μm (micrometer) filter membrane; concentrating the eluent to 1/10-1/8 of the original volume after filtration, then adding anhydrous ethanol with the volume of 3-5 times, placing at the temperature of-10 ℃ to-20 ℃ after dissolution, and filtering again after crystallization to obtain a crystal; repeating the crystallization step for 2 times to obtain recrystallization; and (4) drying the recrystallization in vacuum to obtain sodium aescinate.
Comparative example 1.
The comparative example provides a method for preparing aescine sodium by adopting buckeye seeds, and the preparation method is the same as that of example 1, and is characterized in that:
adding trichloromethane with the volume of 0.2 time into the concentrated solution, fully stirring, standing, and removing the trichloromethane in an oil phase; repeating the above steps for 2 times to elute the oil.
In the rotary evaporation process in the step (5), the added surfactant is methoxy polyethylene glycol-polylactic acid block copolymer (mPEG-PLA; mPEG section molecular weight 2000, PLA section molecular weight 2000-.
Experimental example 1.
According to the detection method disclosed in Shanghai Jia Kai Biotechnology Co., Ltd, Standard Q/C-H0026-4, horse chestnut extract, high performance liquid chromatography is used to analyze the purity of the product. The sodium aescinate prepared in example 1 and comparative example 1 was subjected to chromatographic detection.
Accurately weighing 5.0 mg of aescin control reagent with identified content of aescin A, and diluting to constant volume with methanol to obtain aescin standard solution. The chromatographic conditions for sample detection are as follows: c18 column (4.6 mm. times.250 mm, 5 μm); the mobile phase is acetonitrile: 0.3% phosphoric acid in water (33: 67 by volume); a detection wavelength of 220nm (nanometers); the column temperature is 35 ℃; flow rate: l ml/min.
And (3) respectively injecting 1 microliter (microliter), 2 microliter, 4 microliter, 8 microliter and 10 microliter of aescin standard solution for chromatographic analysis, and respectively drawing standard curves of aescin A and aescin B components by taking the concentration of each aescin as a horizontal coordinate and taking a peak area as a vertical coordinate.
Performing chromatographic analysis on 10 mu l of sodium aescinate prepared in example 1 and comparative example 1 by feeding, determining the peak area according to retention time and ultraviolet spectrogram, obtaining the concentration of sodium aescinate A, C in the solution to be detected according to an aescinate A standard curve, and obtaining the concentration of sodium aescinate B, D in the solution to be detected according to an aescinate B standard curve; taking the sum of the peak areas of the aescin A, B, C, D as the aescin response value, and calculating the amount of related substances by a normalization method; taking logarithm of sum of peak areas of aescin A, B, C, D, and performing regression with logarithm of concentration, and calculating total content of aescin (sum of A, B, C, D) by standard curve method; and calculating the content proportion of the beta-aescine sodium according to the sum of peak areas of the aescine A, B. The results are shown in table 1 below.
Table 1:
as can be seen from comparison of experimental data in table 1, in the method for preparing aescine sodium by buckeye extraction in example 1 of the present invention, mPEG-PCL can selectively form micelles with β -aescine to increase the proportion of β -aescine in the product, the weight proportion of β -aescine sodium in total saponin sodium is as high as 71.35%, while the weight proportion of β -aescine sodium in mPEG-PLA total saponin sodium in comparative example 1 is only 52.31%.
Experimental example 2.
According to the detection method disclosed in Chinese pharmacopoeia (2020 edition, one part, 305 pages, buckeye), the content of beta-aescine in buckeye raw materials is measured by adopting high performance liquid chromatography (general rule 0512).
Octadecylsilane chemically bonded silica is used as a filling agent; the mobile phase is acetonitrile: 0.2% phosphoric acid solution (36: 64); the detection wavelength was 220nm (nanometers). Adding methanol into sodium aescinate reference substance (labeled content of aescinate A and aescinate B) to obtain reference solution. Taking about 1 g of buckeye raw material adopted in example 1, accurately weighing, placing in a Soxhlet extractor, adding diethyl ether, heating and refluxing for 1 hour, removing the diethyl ether solution, placing the decoction dregs and a filter paper cylinder into a conical flask with a plug, accurately adding 50 ml of methanol, weighing, carrying out ultrasonic treatment (power 250W, frequency 33kHz) for 30 minutes, cooling, weighing again, complementing the weight loss with methanol, shaking up, filtering, accurately taking 25 ml of subsequent filtrate, placing in an evaporation dish, concentrating on a water bath at 40 ℃ to a proper amount, transferring to a 10 ml measuring flask, adding methanol to dilute to scale, shaking up, filtering, and taking the subsequent filtrate.
Respectively and precisely sucking 10 mu l of reference solution and buckeye seed solution, injecting into a liquid chromatograph, measuring, and calculating according to peak areas of corresponding peaks at the positions of aescin A and aescin B in the reference solution to obtain the content of beta-aescin in the buckeye seed medicinal material. The results are shown in table 2 below.
Table 2:
combining the experimental data of table 1 and table 2, it can be seen that: the content of beta-aescine in the total saponin of the raw material buckeye seeds is 45.48 percent, and the weight proportion of the beta-aescine in the total saponin sodium obtained by the method for extracting and preparing the aescine sodium reaches 71.35 percent; therefore, the method for extracting and preparing sodium aescinate can obviously improve the content of beta-sodium aescinate in the sodium aescinate.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (11)
1. A method for extracting and preparing aescine sodium by adopting buckeye seeds is characterized by comprising the following steps: crushing and breaking the wall of buckeye seed raw materials; leaching the crushed and wall-broken buckeye raw material by using an acidic ethanol aqueous solution to obtain a primary extract; after the primary extract is concentrated, a solvent is adopted to elute the grease to obtain a water phase solution; adding a high molecular polymer surfactant into the aqueous phase solution, and performing rotary evaporation to obtain an aescin concentrated solution; decolorizing the concentrated solution of aescin; carrying out sodium cation exchange resin chromatography on the decolorized aescin concentrated solution, and collecting eluate; filtering, crystallizing and recrystallizing the eluent, and drying to obtain sodium aescinate; the high molecular polymer surfactant is a methoxy polyethylene glycol-polycaprolactone block copolymer, and the molecular weight of the high molecular polymer surfactant is 6000-10000 g/mol; wherein the molecular weight of the methoxy polyethylene glycol section is 2000g/mol, and the molecular weight of the polycaprolactone section is 4000-8000 g/mol.
2. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 1, wherein the method comprises the following steps: the crushed particle size of the buckeye seed raw material is 8-20 meshes; the method for wall breaking comprises the steps of adding acidic water with the pH value of 5.5-6.9 into the crushed buckeye raw materials, freezing, and then performing microwave thawing to break the wall.
3. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 1, wherein the method comprises the following steps: the method for leaching by using the acidic ethanol aqueous solution comprises the following steps: and adding an acidic ethanol aqueous solution into the raw material of the buckeye after wall breaking, and leaching for 3-5 hours at the temperature of 40-45 ℃ to obtain a primary extract.
4. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 3, wherein the method comprises the following steps: the pH value of the acidic ethanol water solution is 5.0-6.9, and the concentration of ethanol is 30-50%.
5. The method for preparing aescine sodium by using buckeye seed extraction as claimed in claim 1, wherein the method for eluting oil and fat by using solvent after the primary extract is concentrated comprises the following steps: and adding an organic solvent into the concentrated solution obtained after the primary extraction liquid is concentrated to elute the grease, separating to obtain an aqueous phase solution and an oil phase solvent, and carrying out subsequent rotary evaporation treatment on the aqueous phase solution.
6. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 5, wherein the method comprises the following steps: the volume ratio of the concentrated solution after the primary extraction solution is concentrated to the organic solvent is 1 (0.2-1); the number of times of grease elution is 1-3; the organic solvent comprises dichloromethane and/or trichloromethane.
7. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 1, wherein the method for performing rotary evaporation by adding a high molecular polymer surfactant into the aqueous phase solution comprises: adding the high molecular polymer surfactant into the aqueous phase solution, performing rotary evaporation at the temperature of 40-60 ℃, and concentrating a reaction system to 1/3-1/4 of the original volume to obtain an aescin concentrated solution; the weight ratio of the aqueous phase solution to the high molecular polymer surfactant is 100: (5-20).
8. The method for preparing aescine sodium by extracting buckeye seeds according to claim 1, wherein the method for decoloring the concentrated solution of aescine comprises the following steps: adding activated carbon into the aescin concentrated solution for decolorization, and then filtering and removing impurities by adopting an ultrafiltration membrane to obtain the decolorized aescin concentrated solution.
9. The method for preparing aescine sodium by extracting with buckeye seeds as claimed in claim 8, wherein the method comprises the following steps: the weight ratio of the aescin concentrated solution to the active carbon is 1000: (1-5); the pore diameter of the ultrafiltration membrane is 50 nanometers.
10. The method for preparing aescine sodium by using buckeye seed extraction as claimed in claim 1, wherein the method for performing sodium cation exchange resin chromatography on the decolorized concentrated solution of aescine comprises: adding equal volume of absolute ethyl alcohol into the concentrated solution of aescin after decolorization treatment; then, the mixture is loaded on a column sodium type cation exchange resin to be exchanged into sodium salt; then eluting by adopting 2 times of 40% ethanol solution, and cleaning the residual high molecular polymer surfactant in the column bed; then using 70% -90% ethanol solution with 2 times of column volume to elute, and collecting eluent.
11. The method for preparing sodium aescinate by extracting semen aesculi according to claim 1, wherein the method for obtaining sodium aescinate comprises the following steps: filtering the eluent by a filter membrane; the aperture of the filter membrane is 0.22 micron, the eluent is concentrated to 1/10-1/8 of the original volume after being filtered, then absolute ethyl alcohol with the volume being 3-5 times of the original volume is added, the mixture is placed at the temperature of minus 10 ℃ to minus 20 ℃ after being dissolved, and the mixture is filtered again after being crystallized to obtain a crystal; repeating the crystallization step for 2 times to obtain recrystallization; and (4) drying the recrystallization in vacuum to obtain sodium aescinate.
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