阅读说明：本技术 一种从婆罗子中提取分离纯化七叶皂苷的方法 (Method for extracting, separating and purifying aescine from semen Veronicae ) 是由 张大帅 史载锋 陈文豪 陈光英 李晨 于 2021-08-19 设计创作，主要内容包括：本发明公开了一种从婆罗子中提取分离纯化七叶皂苷的方法,该方法采用50％低浓度乙醇,进行闪提--超声提取,过滤分离,得到大极性组分提取液,再用D101大孔吸附树脂反复吸附解吸3次,经水及不同浓度的乙醇洗脱,收集七叶皂苷流分,将第三次纯化的浓缩液放入器皿中,冷冻干燥后,白絮状物黏在器皿上,糖类等黄色物杂质黏在器皿底部,收集白絮状物,得到的七叶皂苷纯度到达96.5％以上。该方法简化了提取纯化步骤,节约了生产成本；操作简单、易于重复,适于实现标准工业化生产。(The invention discloses a method for extracting, separating and purifying aescin from salmons, which comprises the steps of carrying out flash extraction-ultrasonic extraction by adopting 50% low-concentration ethanol, filtering and separating to obtain a large-polarity component extracting solution, repeatedly adsorbing and desorbing for 3 times by using D101 macroporous adsorption resin, eluting by using water and ethanol with different concentrations, collecting aescin fractions, putting a third purified concentrated solution into a vessel, freezing and drying, adhering white floccules on the vessel, adhering yellow impurities such as saccharides and the like at the bottom of the vessel, and collecting white floccules, wherein the purity of the obtained aescin reaches over 96.5%. The method simplifies the extraction and purification steps, and saves the production cost; the method is simple to operate, easy to repeat and suitable for realizing standard industrial production.)

1. A method for extracting, separating and purifying aescine from semen Veronicae is characterized in that: the method comprises the following steps:

1) flash extraction-ultrasonic extraction:

1.1) flash extraction: crushing the salmon to 40-200 meshes, adding an ethanol water solution with the volume concentration of 50% at room temperature, wherein the using amount of the ethanol water solution is 8-12 ml per gram of the salmon, then sequentially carrying out forward extraction and reverse extraction at room temperature, wherein the forward extraction is 5000-15000 r/min, the forward extraction is 8-13 min, the reverse extraction is 5000-15000 r/min, the reverse extraction is 8-13 min, then carrying out static precipitation for 8-12 min, and removing a supernatant to obtain a first precipitate and a first clear liquid;

1.2) ultrasonic extraction: the method comprises a first ultrasonic extraction and a second ultrasonic extraction, wherein the first ultrasonic extraction is as follows: adding an ethanol water solution with the volume concentration of 50% into the first precipitate, wherein the dosage of the ethanol water solution is 5-7 ml per gram of the salmon, carrying out ultrasonic extraction for 25-35 min, carrying out static precipitation for 8-12 min, and removing a supernatant to obtain a second precipitate and a second clear solution; the second ultrasonic extraction is as follows: adding an ethanol water solution with the volume concentration of 50% into the second precipitate, wherein the using amount of the ethanol water solution is 3-5 ml per gram of the salsify, performing ultrasonic extraction for 25-35 min, performing static precipitation for 8-12 min, and removing a supernatant to obtain a third precipitate and a third clear solution; filtering to obtain third precipitate, washing the third precipitate with anhydrous ethanol for 2-3 times, and mixing the first clear solution, the second clear solution, the third clear solution and the washing solution to obtain aescin extractive solution;

2) concentrating an extracting solution: concentrating the aescin extractive solution under reduced pressure to foam to obtain aescin concentrated solution;

3) carrying out first adsorption by macroporous adsorption resin: adding the concentrated solution of aescin into a macroporous adsorption resin column which is 5-7 times of the weight of the salmon seeds for adsorption, eluting with an ethanol water solution with the volume concentration of 25% until the solution is nearly colorless, discarding the solution, washing the column with absolute ethanol, and concentrating the absolute ethanol eluent until the column is foamed to obtain a concentrated solution purified for the first time;

4) and (3) carrying out secondary adsorption by using macroporous adsorption resin: washing the first resin column with distilled water until no alcohol smell exists, adsorbing the first purified concentrated solution on the column, eluting with 25% ethanol water solution until the solution is nearly colorless, discarding, washing the column with anhydrous ethanol, and concentrating the anhydrous ethanol eluate until foaming to obtain second purified concentrated solution;

5) and (3) adsorbing by macroporous adsorption resin for the third time: washing the resin column with distilled water for the second time until no alcohol smell exists, adsorbing the concentrated solution purified for the second time on the column, eluting with 35% ethanol water solution until the solution is nearly colorless, discarding, washing the column with anhydrous ethanol, and concentrating the anhydrous ethanol eluate until the solution foams to obtain the concentrated solution purified for the third time;

6) and (3) freeze drying and color removal: putting the concentrated solution purified for the third time into a beaker, freeze-drying, sticking white floccules on the wall of the beaker, sticking yellow impurities at the bottom of the beaker, and collecting the white floccules which are the aescin.

2. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1, wherein the method comprises the steps of: in the step 1), the forward rotation extraction comprises the following steps: sequentially extracting at 4500-5500 r/min for 4-6 min, 9500-10500 r/min for 2-4 min, and 14500-15500 r/min for 2-3 min; the inversion extraction is: sequentially extracting at 4500-5500 r/min for 4-6 min, 9500-10500 r/min for 2-4 min, and 14500-15500 r/min for 2-3 min.

3. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 1.2), the ultrasonic frequency of the first ultrasonic extraction and the ultrasonic frequency of the second ultrasonic extraction are both 53KHz, the first ultrasonic extraction temperature is 38-44 ℃, and the second ultrasonic extraction temperature is 41-45 ℃.

4. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 1.2), the dosage of absolute ethyl alcohol for washing the third precipitate every time is 0.8-1.2 ml per gram of salmon.

5. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 2), the aescin extract is decompressed and concentrated to 30-40% of the original volume, and the aescin concentrated solution is obtained.

6. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 2), the pressure of the reduced pressure concentration is 0.05-0.09 MPa, and the temperature is 65-75 ℃.

7. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 3), the macroporous resin is D101 macroporous resin, the flow rate is 1-2mL/min when the aescin concentrated solution is absorbed, the consumption of absolute ethyl alcohol is 3-6 times of the column volume when the aescin concentrated solution is washed, and the flow rate is 8-10 BV/h; in the step 4), the flow rate of the first purified concentrated solution is 1-2mL/min during adsorption, the consumption of absolute ethyl alcohol is 3-6 times of the column volume during washing, and the flow rate is 8-10 BV/h; in the step 5), the flow rate of the concentrated solution purified for the second time is 1-2mL/min during adsorption, the consumption of the absolute ethyl alcohol is 3-6 times of the column volume during washing, and the flow rate is 8-10 BV/h.

8. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the steps 3) to 5), the mixture is concentrated to 30 to 40 percent of the original volume.

9. The method for extracting, separating and purifying aescine from salmon as claimed in claim 1 or 2, wherein the method comprises the steps of: in the step 6), the freeze drying temperature is-35 to-45 ℃ and the pressure is 10-20 Pa.

Technical Field

The invention relates to a method for extracting, separating and purifying aescin from salmons, belonging to the technical field of medicines.

Background

Chinese buckeye (Aesculi) is one of traditional Chinese medicines collected in Chinese pharmacopoeia, and the Chinese traditional medicine Chinese buckeye refers to dry mature seeds of horse chestnut (A esculus chinensis Bge.), Zhejiang horse chestnut (A esculus chinensis var. chekiangensis Fang), Tianshi chestnut (A esculus wilsonii Rehd), and the like, and the 3 medicinal plants are special products in China. Semen aesculi is warm in nature and sweet in taste, enters liver and stomach meridians, has the effects of regulating qi, relieving epigastric distention, harmonizing stomach and relieving pain, and is mainly used for treating chest and abdomen swelling and stuffiness and stomach pain.

Aescin is an important effective component in extract of buckeye seeds, has obvious effects of resisting inflammation, resisting exudation, eliminating swelling and the like, can restore normal permeability of capillary vessels, increase venous tension, improve microcirculation and has high treatment value on brain trauma and peripheral vascular diseases. At present, the technology for extracting aescine from buckeye seeds is still to be improved.

At present, the existing purification method of the seven-leaf soap shake at home and abroad mainly comprises the following steps:

(1) liudongfeng et al, a method for extracting aescin by ultrasonic-microwave synergistic extraction, which obtains the aescin by the steps of degreasing, ultrasonic-microwave simultaneous extraction, silica gel column chromatography, acetone washing, drying and the like. The method is complicated, the extract has low purity of about 88%, and toxic solvents such as petroleum ether, n-butanol, and acetone are used.

(2) Liudongfeng et al, a method for extracting and purifying aescin from semen Veronicae, which obtains the aescin by the steps of degreasing, ultrasonic extraction, membrane separation, macroporous resin adsorption, recrystallization, drying and the like. The purity of the aescin obtained by the method is about 95%, but the method is complex, a microfiltration membrane is required, the cost is high, and toxic solvents such as diethyl ether, acetone and the like are required.

(3) Luqiang, etc. a method for extracting and purifying aescin from veronica extract, which dissolves the veronica extract by low molecular alcohol, and precipitates and degreases by acetone; after pure water is dissolved, macroporous resin is used for adsorption, and ethanol is used for elution; washing with acetone, decolorizing, and vacuum drying to obtain aescin. The method is complicated, the extract has low purity of only about 85%, and toxic solvents such as acetone are required.

(4) Shihua and the like, a method for extracting aescine from salmon under ultrahigh pressure based on freeze drying, which comprises the steps of crushing and soaking the medicinal material of salmon, freeze drying and then extracting under ultrahigh pressure. The invention changes the internal structure of the medicinal material tissue by freeze-drying the semen juglandis, so that the cell wall is easier to break, and the components of the semen juglandis are easier to exude. Although the aescin in the salmon can be fully extracted, the extract components are too complex, an ultrahigh pressure extraction device is needed, pressure is repeatedly increased and released, the extraction cost of the aescin is increased, and the difficulty in purifying the aescin is increased.

Disclosure of Invention

The invention provides a method for extracting, separating and purifying aescin from salmon, which only uses ethanol and ethanol water solution without using toxic solvents such as acetone, ether and the like in the whole process, improves the operation environment and avoids environmental pollution; the method has mild conditions, does not damage the structure of the saponin, and obviously improves the yield and the purity of the aescin by combining flash extraction and ultrasound and adsorbing by macroporous adsorption resin for three times.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

a method for extracting, separating and purifying aescine from semen Veronicae includes the following steps:

1) flash extraction-ultrasonic extraction:

1.1) flash extraction: crushing the salmon to 40-200 meshes, adding an ethanol water solution with the volume concentration of 50% at room temperature, wherein the using amount of the ethanol water solution is 8-12 ml per gram of the salmon, then sequentially carrying out forward extraction and reverse extraction at room temperature, wherein the forward extraction is 5000-15000 r/min, the forward extraction is 8-13 min, the reverse extraction is 5000-15000 r/min, the reverse extraction is 8-13 min, then carrying out static precipitation for 8-12 min, and removing a supernatant to obtain a first precipitate and a first clear liquid;

1.2) ultrasonic extraction: the method comprises a first ultrasonic extraction and a second ultrasonic extraction, wherein the first ultrasonic extraction is as follows: adding an ethanol water solution with the volume concentration of 50% into the first precipitate, wherein the dosage of the ethanol water solution is 5-7 ml per gram of the salmon, carrying out ultrasonic extraction for 25-35 min, carrying out static precipitation for 8-12 min, and removing a supernatant to obtain a second precipitate and a second clear solution; the second ultrasonic extraction is as follows: adding an ethanol water solution with the volume concentration of 50% into the second precipitate, wherein the using amount of the ethanol water solution is 3-5 ml per gram of the salsify, performing ultrasonic extraction for 25-35 min, performing static precipitation for 8-12 min, and removing a supernatant to obtain a third precipitate and a third clear solution; filtering to obtain third precipitate, washing the third precipitate with anhydrous ethanol for 2-3 times, and mixing the first clear solution, the second clear solution, the third clear solution and the washing solution to obtain aescin extractive solution;

2) concentrating an extracting solution: concentrating the aescin extractive solution under reduced pressure to foam to obtain aescin concentrated solution;

3) carrying out first adsorption by macroporous adsorption resin: adding the concentrated solution of aescin into a macroporous adsorption resin column which is 5-7 times of the weight of the salmon seeds for adsorption, eluting with an ethanol water solution with the volume concentration of 25% until the solution is nearly colorless, discarding the solution, washing the column with absolute ethanol, and concentrating the absolute ethanol eluent until the column is foamed to obtain a concentrated solution purified for the first time;

4) and (3) carrying out secondary adsorption by using macroporous adsorption resin: washing the first resin column with distilled water until no alcohol smell exists, adsorbing the first purified concentrated solution on the column, eluting with 25% ethanol water solution until the solution is nearly colorless, discarding, washing the column with anhydrous ethanol, and concentrating the anhydrous ethanol eluate until foaming to obtain second purified concentrated solution;

5) and (3) adsorbing by macroporous adsorption resin for the third time: washing the resin column with distilled water for the second time until no alcohol smell exists, adsorbing the concentrated solution purified for the second time on the column, eluting with 35% ethanol water solution until the solution is nearly colorless, discarding, washing the column with anhydrous ethanol, and concentrating the anhydrous ethanol eluate until the solution foams to obtain the concentrated solution purified for the third time;

6) and (3) freeze drying and color removal: putting the concentrated solution purified for the third time into a beaker, freeze-drying, sticking white floccules on the wall of the beaker, sticking yellow impurities at the bottom of the beaker, and collecting the white floccules which are the aescin.

The method comprises the steps of carrying out flash extraction-ultrasonic extraction on 50% low-concentration ethanol water solution, filtering and separating to obtain a large-polarity component extracting solution, repeatedly adsorbing and desorbing for 3 times by using D101 macroporous adsorption resin, eluting by using water and ethanol water solutions with different concentrations, collecting aescin fractions, putting a third purified concentrated solution into a vessel, freezing and drying, sticking white floccules on the vessel, sticking yellow impurities such as saccharides and the like on the bottom of the vessel, collecting white floccules, wherein the purity of the obtained aescin reaches over 96%, and the extraction recovery rate is about 88.00%. The method simplifies the extraction and purification steps, and saves the production cost; the method is simple to operate, easy to repeat and suitable for realizing standard industrial production.

The concentration of the ethanol aqueous solution referred to in the present application is the volume concentration of ethanol.

In the whole process, only ethanol is used, and solvents such as petroleum ether, acetone, ether and the like are not needed, so that the operation environment is improved, and the health and safety of operators are ensured; the method has mild conditions, does not damage the structure of the saponin, and obviously improves the yield and the purity of the aescin and the extraction efficiency by combining flash extraction and ultrasound and adsorbing by macroporous adsorption resin for three times.

In order to further improve the yield of the aescin, in the step 1), the forward extraction is as follows: sequentially extracting at 4500-5500 r/min for 4-6 min, 9500-10500 r/min for 2-4 min, and 14500-15500 r/min for 2-3 min; the inversion extraction is: sequentially extracting at 4500-5500 r/min for 4-6 min, 9500-10500 r/min for 2-4 min, and 14500-15500 r/min for 2-3 min.

In order to further avoid the damage to the aescin and improve the yield of the aescin, in the step 1.2), the ultrasonic frequency of the first ultrasonic extraction and the ultrasonic frequency of the second ultrasonic extraction are both 53KHz, the first ultrasonic extraction temperature is 38-44 ℃, and the second ultrasonic extraction temperature is 41-45 ℃.

In order to improve the yield and reduce the material waste, in the step 1.2), the dosage of absolute ethyl alcohol for washing the third precipitate every time is 0.8-1.2 ml per gram of salmon.

In order to improve the operation efficiency and simultaneously give consideration to the product purity, in the step 2), the aescin extracting solution is decompressed and concentrated to 30-40% of the original volume to obtain the aescin concentrated solution.

In order to further avoid damage to the aescin, in the step 2), the pressure for decompression concentration is 0.05-0.09 MPa, and the temperature is 65-75 ℃.

In order to give consideration to both the yield and the purity of the product, in the step 3), the macroporous resin is D101 macroporous resin, the flow rate is 1-2mL/min when the aescin concentrated solution is adsorbed, the consumption of the absolute ethyl alcohol is 3-6 times of the column volume when the aescin concentrated solution is washed, and the flow rate is 8-10 BV/h; in the step 4), the flow rate of the first purified concentrated solution is 1-2mL/min during adsorption, the consumption of absolute ethyl alcohol is 3-6 times of the column volume during washing, and the flow rate is 8-10 BV/h; in the step 5), the flow rate of the concentrated solution purified for the second time is 1-2mL/min during adsorption, the consumption of the absolute ethyl alcohol is 3-6 times of the column volume during washing, and the flow rate is 8-10 BV/h.

In the steps 3) to 5), the concentration is carried out until the volume is 30 to 40 percent of the original volume. Volume before concentration in original volume.

Preferably, in the step 6), the freeze drying temperature is-35 to-45 ℃ and the pressure is 10-20 Pa.

The prior art is referred to in the art for techniques not mentioned in the present invention.

The method for extracting, separating and purifying the aescin from the salmon seeds only uses ethanol without using toxic solvents such as acetone, diethyl ether and the like in the whole process, improves the operation environment and avoids environmental pollution; the condition is mild, and the structure of the saponin cannot be damaged; macroporous resin is adopted for adsorption elution for three times, low alcohol is loaded on a column, and ethanol with different concentrations is eluted, so that the impurity removal effect is obviously improved, the resin is renewable, and the production cost is reduced; by combining flash extraction and ultrasonic and adsorbing by macroporous adsorption resin and freeze drying and purifying for three times, the yield (more than 4%) and the purity (more than 96.5%) of the aescin are obviously improved; simple operation, low cost and contribution to industrial production.

Drawings

FIG. 1 is a standard curve diagram of aescin HPLC;

FIG. 2 is a HPLC chart of the extraction of escin at different ethanol concentrations;

FIG. 3 is a chromatogram of aescin obtained in the present invention;

Detailed Description

In order to better understand the present invention, the following examples are further provided to illustrate the present invention, but the present invention is not limited to the following examples.

Example 1

Weighing 200g of buckeye (Aesculus hippocastanum L.) and crushing into 40-200 meshes, adding into a flash extraction-ultrasonic instrument, adding 2L of ethanol water solution with volume concentration of 50%, and performing flash extraction (at room temperature of 25 ℃): forward rotation extraction: extracting at 5000r/min for 5min, 10000r/min for 3min, and 15000r/min for 2 min; reverse extraction: extracting at 5000r/min for 5min, 10000r/min for 3min, and 15000r/min for 2 min; standing for 10min, and removing supernatant to obtain first precipitate and first clear liquid;

adding 1.2L ethanol water solution with volume concentration of 50% into the first precipitate, ultrasonic extracting for 30min (38-44 deg.C, 53KHz), standing for 10min, and removing supernatant to obtain second precipitate and second clear liquid; adding 50 vol% ethanol water solution 0.8L into the second precipitate, ultrasonic extracting for 30min (41-45 deg.C), standing for 10min, and removing supernatant to obtain third precipitate and third clear solution; filtering to obtain third precipitate, washing the third precipitate with anhydrous ethanol for 2 times (200 ml for each time), and mixing the first clear solution, the second clear solution, the third clear solution and the washing solution to obtain aescin extractive solution; as can be seen from fig. 2, the 50% ethanol concentration is optimal.

Concentrating the aescin extractive solution under reduced pressure (negative pressure 0.07Mpa, temperature 70 deg.C) to 34% of the original volume, and foaming to obtain 24.759% extraction rate (HPLC analysis);

adsorption by macroporous adsorption resin: first adsorption: adding the concentrated solution into 2400g D101 macroporous adsorption resin column for adsorption for 0.75h at flow rate of 1.5mL/min, eluting with 25% ethanol water solution (volume concentration) until the solution is nearly colorless, discarding, washing the column with anhydrous ethanol at flow rate of 9BV/h, and concentrating the anhydrous ethanol eluate to obtain the first purified concentrated solution; and (3) second adsorption: washing the resin column used for the first time with distilled water until no alcohol smell exists, adding the concentrated solution purified for the first time into the resin column for adsorption for 0.75h, wherein the flow rate during adsorption is 1.5mL/min, eluting with 25% ethanol water solution (volume concentration) until the concentrated solution is nearly colorless, washing the column with absolute ethyl alcohol, wherein the use amount of the absolute ethyl alcohol is 4 times of the column volume, and the flow rate is 9BV/h, and concentrating the absolute ethyl alcohol eluent to obtain the concentrated solution purified for the second time; and (3) third adsorption: washing the resin column used for the second time with distilled water until no alcohol smell exists, adding the concentrated solution purified for the second time into the resin column for adsorption for 0.75h, wherein the flow rate during adsorption is 1.5mL/min, eluting with 35% ethanol water solution (volume concentration) until the concentrated solution is nearly colorless, discarding, washing the column with absolute ethyl alcohol, wherein the usage amount of the absolute ethyl alcohol is 4 times of the column volume, and the flow rate is 9BV/h, and concentrating the absolute ethyl alcohol eluent to obtain the concentrated solution purified for the third time; and (3) freeze drying and color removal: placing the third purified concentrated solution into a beaker, freeze-drying at-40 deg.C under 15Pa, sticking white floccule on the wall of the beaker, sticking yellow impurities on the bottom of the beaker, collecting total 8.2612g of white floccule, and performing chromatogram shown in FIG. 3. Purity by HPLC analysis was 97.5%.

Example 2

Weighing 0.5Kg of buckeye (Aesculus chinensis Baill.) and crushing to 40-200 meshes, adding into a flash extraction-ultrasonic instrument, then adding 5L of 50% ethanol, and performing flash extraction (room temperature 25 ℃): forward rotation extraction: extracting at 5000r/min for 5min, 10000r/min for 3min, and 15000r/min for 2 min; reverse extraction: extracting at 5000r/min for 5min, 10000r/min for 3min, and 15000r/min for 2 min; standing for 10min, and removing supernatant to obtain first precipitate and first clear liquid;

adding 50% ethanol water solution 3L into the first precipitate, ultrasonic extracting for 30min (38-43 deg.C, 53KHz), standing for 10min, and removing supernatant to obtain second precipitate and second clear liquid; adding 50 vol% ethanol water solution into the second precipitate, ultrasonic extracting for 30min (40-43 deg.C, 53KHz) for 2L, standing for 10min, and removing supernatant to obtain third precipitate and third clear solution; filtering to obtain third precipitate, washing the third precipitate with anhydrous ethanol for 2 times (the amount of anhydrous ethanol used is 500ml each time), and mixing the first clear solution, the second clear solution, the third clear solution and the washing solution to obtain aescin extractive solution;

concentrating the aescin extractive solution under reduced pressure (negative pressure 0.07Mpa, temperature 70 deg.C) to 36% of the original volume, and foaming to obtain 24.454% extraction rate (HPLC analysis);

adsorption by macroporous adsorption resin: first adsorption: adding the concentrated solution into a 5000g D101 macroporous adsorption resin column for adsorption for 0.75h, wherein the flow rate during adsorption is 1.5mL/min, eluting with 25% ethanol water solution (volume concentration) until the solution is nearly colorless, discarding the solution, washing the column with absolute ethanol, wherein the absolute ethanol is 4 times the column volume, and the flow rate is 9BV/h, and concentrating the absolute ethanol eluate to obtain the first purified concentrated solution; and (3) second adsorption: washing the resin column used for the first time with distilled water until no alcohol smell exists, adding the concentrated solution purified for the first time into the resin column for adsorption for 0.75h, wherein the flow rate during adsorption is 1.5mL/min, eluting with 25% ethanol water solution (volume concentration) until the concentrated solution is nearly colorless, washing the column with absolute ethyl alcohol, wherein the use amount of the absolute ethyl alcohol is 4 times of the column volume, and the flow rate is 9BV/h, and concentrating the absolute ethyl alcohol eluent to obtain the concentrated solution purified for the second time; and (3) third adsorption: washing the resin column used for the second time with distilled water until no alcohol smell exists, adding the concentrated solution purified for the second time into the resin column for adsorption for 0.75h, wherein the flow rate during adsorption is 1.5mL/min, eluting with 35% ethanol water solution (volume concentration) until the concentrated solution is nearly colorless, discarding, washing the column with absolute ethyl alcohol, wherein the usage amount of the absolute ethyl alcohol is 4 times of the column volume, and the flow rate is 9BV/h, and concentrating the absolute ethyl alcohol eluent to obtain the concentrated solution purified for the third time; concentrating the third eluate to 2080mL, placing into a beaker, freeze drying at-40 deg.C and 15Pa, collecting 20.4917g white floccule, and taking out a small portion of sample for HPLC analysis to obtain 96.5% purity.