CN114853840A - Preparation method and application of akebia saponin D bulk drug - Google Patents
Preparation method and application of akebia saponin D bulk drug Download PDFInfo
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- CN114853840A CN114853840A CN202210648663.9A CN202210648663A CN114853840A CN 114853840 A CN114853840 A CN 114853840A CN 202210648663 A CN202210648663 A CN 202210648663A CN 114853840 A CN114853840 A CN 114853840A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- C—CHEMISTRY; METALLURGY
- 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
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention discloses a preparation method and application of akebia saponin D bulk drug, wherein the preparation method comprises the following steps: pulverizing raw materials to obtain radix Dipsaci powder, extracting with supercritical fluid to remove impurities with low polarity, recovering the rest phase, concentrating under reduced pressure, performing polyamide resin column chromatography, gradient eluting with eluting solvent, checking with silica gel thin layer chromatography or HPLC, collecting eluate containing akebia saponin D, concentrating under reduced pressure to obtain akebia saponin D concentrate, and recrystallizing. The method adopts the steps of solvent extraction, supercritical fluid extraction, chromatographic separation, recrystallization and the like which are commonly used in industrialization, has simple and practical process, low toxicity and environmental protection, optimizes the process conditions in the steps, for example, removes the impurities with smaller polarity such as volatile oil, aliphatic hydrocarbon and the like by using the supercritical fluid extraction method, removes the impurities such as phenolic acids, iridoids and the like by using the polyamide resin column chromatography, obtains the ASD with the yield of up to 7.51 percent and the purity of up to 99.8 percent, and has wide application prospect.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a preparation method and application of a akebia saponin D bulk drug.
Background
Akebia Saponin D (ASD) is a triterpene Saponin compound contained in the dried root of Dipsacus asperoides (Dipsacus asperWall.) belonging to the family Dipsacaceae, and has the following structural formula:
【CAS】39524-08-8。
[ molecular formula and molecular weight ] C 47 H 76 O 18 ；929.10。
[ physicochemical properties ] white crystals (anhydrous methanol), readily soluble in water, methanol, ethanol, acetone, dimethyl sulfoxide, sparingly soluble in diethyl ether, insoluble in petroleum ether; melting point: 234 ℃ and 235 ℃.
ASD is triterpenoid saponin compound, is one of main components of radix Dipsaci, and has pharmacological activities of resisting tumor, resisting senile dementia, protecting heart, promoting bone healing, and preventing and treating metabolic syndrome.
CN102924545A discloses a method for extracting ASD by using macroporous resin, which comprises the following steps: drying and crushing teasel roots to obtain coarse powder, performing reflux extraction on the coarse powder by using a solvent, treating resin, and filling a column, loading a sample, enriching liquid and the like; the disadvantages of this method include: 1) the ASD obtained according to the patent method has lower maximum purity and yield; 2) the preparation process has large requirements on resin fillers, and a large amount of fillers can only purify a small amount of ASD, so that large-scale and industrial production cannot be realized.
In view of this, the invention is particularly proposed.
Disclosure of Invention
In order to overcome the defects in the technology, the invention provides a preparation method and application of akebia saponin D bulk drug.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a preparation method of akebia saponin D bulk drug comprises the following steps:
s1, crushing the teasel root medicinal decoction pieces to obtain teasel root powder, adding an extraction solvent to perform reflux extraction to obtain an extracting solution, filtering by using an ultrafiltration membrane to remove macromolecular impurities in the extracting solution, and concentrating under reduced pressure to obtain a concentrated solution;
s2, injecting the concentrated solution into a supercritical extraction kettle, sequentially extracting by using entrainers with different polarities respectively at a specific temperature and pressure, after each static extraction for 0.5-1h, dynamically extracting for 1-2h at a speed of 0.1-1L/min, removing impurities with smaller polarity (such as volatile oil, aliphatic hydrocarbon, aromatic compounds and partial monoterpene compounds), and collecting to obtain an extraction residual phase;
s3, concentrating the residual phase under reduced pressure, performing polyamide resin column chromatography, performing gradient elution with eluting solvent (to remove impurities which are easily adsorbed by polyamide resin filler, such as caffeoylquinic acid, ursolic acid, isochlorogenic acid, caffeic acid, loganin, swertisin, and phenomenoside, such as phenolic acid and iridoid), performing silica gel thin layer chromatography or HPLC, collecting eluate containing akebia saponin D, and concentrating under reduced pressure to obtain akebia saponin D concentrated solution;
s4, recrystallizing the akebia saponin D concentrated solution, and drying to obtain the akebia saponin D.
Specifically, in step S2, during the static extraction for 0.5-1h, the fluid is ensured to be in full contact with the analyte, and the highly diffusive substance can be used to penetrate through the interaction between the matrix and the analyte, so as to transfer the impurities with small polarity (such as volatile oil, aliphatic hydrocarbon, aromatic compound, etc. of dill, chrysanthenone, trimethylphenol, β -sitosterol, n-tridecanoic acid, n-pentadecanoic acid, etc.) into the fluid as much as possible, and then the dynamic extraction is performed, so as to remove the impurities with small polarity to the maximum extent through the fluid passing through the flow path in one way, and the extraction residual phase is collected.
In the above technical solution, in step S2, the extractant in the supercritical extraction kettle is CO 2 。
In the above technical solution, in step S2, the entrainer is one or more of petroleum ether, ethyl acetate, dichloromethane and chloroform.
Further, in the above technical solution, in step S2, the temperature and pressure in the supercritical extraction kettle are 35-65 ℃ and 10-40MPa, respectively.
Specifically, in the supercritical fluid extraction of step S2, the polarity of ASD and entrainer is fully considered; the step S2 can remove non-polar and low-polar impurities, greatly simplifying the subsequent column chromatography steps.
In the above technical solution, in step S3, the elution solvent is one or more of water, methanol and ethanol.
Further, in the above technical solution, in the step S3, the polyamide resin column is an alcohol-soluble polyamide resin column.
Specifically, in step S3, a polyamide resin column chromatography method is adopted, which facilitates the gradual enrichment and purification of ASD; in consideration of the polarity of ASD, the ethanol water solution is uniformly used as a gradient elution solvent, so that the production cost is reduced, and the environmental pollution is reduced.
Still further, in the above technical solution, in step S1, the addition amount of the extraction solvent is 8-15 times of the mass of the teasel root powder.
Further, in the above technical scheme, in step S1, the number of times of reflux extraction is 2-4, each time of extraction is 2-3 hours, after reflux extraction, the combined extract is sequentially filtered by an ultrafiltration membrane and concentrated under reduced pressure to obtain a concentrated solution with a density of 1.00-1.30 g/ml.
Specifically, in the above technical solution, in step S1, the extraction solvent is one or more of water, methanol and ethanol.
Specifically, in the above technical solution, in step S1, the temperature of the reflux extraction is equal to or higher than 70 ℃.
In detail, in the above technical solution, in step S1, the extraction solvent is an ethanol aqueous solution with a concentration of less than 90 v%.
Specifically, in step S1, during the extraction process, water, methanol, ethanol or a mixture thereof is selected as the extraction solvent for extraction, which has the characteristic of high extraction rate; in particular, a specific extraction solvent (i.e., 35-65 v% ethanol aqueous solution) is preferable for extraction, and since ethanol and water are both solvents that are less toxic and are tolerated by the human body, an extract rich in ASD can be obtained from a 35 v% ethanol aqueous solution, and since the probability of introducing small polar impurities is greatly increased when the ethanol concentration is increased to 65%, in order to enrich to a large amount of pure ASD, considering operability, cost, and efficiency of mass production, a 35-65 v% ethanol aqueous solution is preferable as the extraction solvent.
In a preferred embodiment of the invention, the extraction solvent is 35-90 v% ethanol water, and the addition amount of the extraction solvent is 10 times of the mass of the teasel root powder.
Still further, in the above technical solution, in step S4, the solvent used in the recrystallization process is one or more of dichloromethane, chloroform, ethyl acetate, propyl acetate, acetone, n-butanol, ethanol, and methanol.
Specifically, in step S4, for further purification and impurity removal, a recrystallization method is adopted, and ASD screened by previous experiments is easy to recrystallize and has a good crystal form in water, ethanol or a mixture thereof, and water and ethanol (or a mixture thereof) are preferred in consideration of the fact that water and ethanol are more used and less toxic in mass production.
The preparation method provided by the application takes dry roots of Dipsacus asperoides (Dipsacus asper Wall.) of the family teasel as a raw material, and obtains ASD through the steps of solvent extraction, supercritical fluid extraction, polyamide resin column chromatography, recrystallization, drying and the like, and identifies impurities such as akebia saponin B, akebia saponin PA, Dipsacus asperoides saponin M and the like.
The invention also provides application of the preparation method of the akebia saponin D bulk drug in extraction of the akebia saponin D bulk drug.
Compared with the prior art, the invention has the following advantages:
the preparation method of the akebia saponin D bulk drug provided by the invention adopts the steps of industrial common solvent extraction, supercritical fluid extraction, chromatographic separation, recrystallization, drying and the like, has simple and practical process flow and low toxicity and is environment-friendly, and specific process conditions are adopted in each step, such as the supercritical fluid extraction method is utilized to remove most of volatile oil, aliphatic hydrocarbon and other impurities with smaller polarity, and the polyamide resin column chromatography is utilized to remove phenolic acid, iridoid and other impurities, so that the ASD with high yield and high purity is obtained, the ASD yield is up to 7.51%, the purity is up to 99.8%, and the ASD can be directly used as the bulk drug.
Drawings
Fig. 1 is a high performance liquid chromatogram of akebia saponin D prepared in example 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments.
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the examples, the means used are conventional in the art unless otherwise specified.
The terms "comprises," "comprising," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
A preparation method of akebia saponin D bulk drug comprises the following steps:
(1) pulverizing radix Dipsaci into powder, adding 10 times of 40 v% ethanol water solution, reflux extracting at 80 deg.C for 4 times (4 hr each time), mixing extractive solutions, filtering with ultrafiltration membrane (13mm × 0.22 μm) to remove macromolecular impurities, and concentrating under reduced pressure to 1/10 of original volume to obtain concentrated solution;
(2) injecting the concentrated solution into a supercritical extraction kettle (extracting agent is CO) 2 ) Adjusting the constant temperature in the kettle to be 45 ℃ and the pressure to be 35MPa, sequentially adopting petroleum ether, ethyl acetate and dichloromethane as entrainers for extraction, after each static extraction is carried out for 0.6h, carrying out dynamic extraction for 1h at the speed of 0.5L/min, and discarding the extraction liquid (containing impurities with lower polarity) to obtain the extraction residual phase;
(3) concentrating the residual phase of the extraction under reduced pressure to 1/10 of the original volume, loading the concentrated solution on 60-100 mesh alcohol-soluble polyamide resin column, loading the column by wet method (the ratio of sample amount to resin amount loaded on the column is 1: 6), eluting with 30 v% ethanol water solution for 5BV, discarding, eluting with 50 v% ethanol water solution for 5BV, collecting the eluate in half or one third as one fraction, checking by silica gel thin layer chromatography or HPLC, collecting and combining the eluates containing ASD, and concentrating under reduced pressure until no alcohol smell exists to obtain ASD concentrated solution;
(4) dissolving the ASD concentrated solution with 90 v% ethanol aqueous solution with 2 times volume, standing for 3h for crystallization, washing off impurities on the crystal surface with ethanol, repeatedly recrystallizing for 4 times, detecting the ASD purity to be 99.8 wt% by using a high performance liquid chromatograph, and drying under reduced pressure to obtain the ASD bulk drug with the yield of 7.51%.
The ASD prepared in example 1 was detected by high performance liquid chromatography under the following specific chromatographic conditions:
the detection system comprises: agilent technologies 1260Infinity type II HPLC apparatus;
a chromatographic column: sonoma C18 (250X 4.6mm, 5 μm);
mobile phase: acetonitrile/water (30: 70, V/V, 40 min);
sample introduction amount: 20 mu L of the solution;
flow rate: 1.0 mL/min;
detection wavelength: 212 nm;
column temperature: at 25 ℃.
The results of the detection are shown in FIG. 1.
Example 2
A preparation method of akebia saponin D bulk drug comprises the following steps:
(1) pulverizing radix Dipsaci into powder, adding 9 times of 70 v% ethanol water solution, reflux extracting at 80 deg.C for 3 times (4 hr each time), mixing extractive solutions, filtering with ultrafiltration membrane (13mm × 0.22 μm) to remove macromolecular impurities, and concentrating under reduced pressure to 1/9 of original volume to obtain concentrated solution;
(2) injecting the concentrated solution into a supercritical extraction kettle (extracting agent is CO) 2 ) In the method, the constant temperature in the kettle is adjusted to be 45 ℃ and the pressure is adjusted to be 35MPa, and petroleum ether, ethyl acetate and dimethyl acetate are adopted in sequenceExtracting with chloromethane as entrainer for 0.6 hr each time, dynamically extracting at 0.5L/min for 1 hr, and discarding extractive solution (containing less polar impurities) to obtain residual phase;
(3) concentrating the residual phase of the extraction under reduced pressure to 1/10 of the original volume, loading the concentrated solution on 60-100 mesh alcohol-soluble polyamide resin column, loading the column by wet method (the ratio of sample amount to resin amount loaded on the column is 1: 6), eluting with 30 v% ethanol water solution for 5BV, discarding, eluting with 50 v% ethanol water solution for 5BV, collecting the eluate in half or one third as one fraction, checking by silica gel thin layer chromatography or HPLC, collecting and combining the eluates containing ASD, and concentrating under reduced pressure until no alcohol smell exists to obtain ASD concentrated solution;
(4) dissolving the ASD concentrated solution by using 90 v% ethanol aqueous solution with 2 times volume, standing for 3h for crystallization, washing impurities on the surface of the crystal by using ethanol, repeatedly recrystallizing for 4 times, detecting the purity of the ASD to be 99.7 wt% by using a high performance liquid chromatograph (the chromatographic condition is as in example one), and drying under reduced pressure to obtain the ASD bulk drug with the yield of 7.42%.
Example 3
A preparation method of akebia saponin D bulk drug comprises the following steps:
(1) pulverizing radix Dipsaci into powder, adding 12 times of 40 v% ethanol water solution, reflux extracting at 80 deg.C for 3 hr for 4 times, mixing extractive solutions, filtering with ultrafiltration membrane (13mm × 0.22 μm) to remove macromolecular impurities, and concentrating under reduced pressure to 1/12 of original volume to obtain concentrated solution;
(2) injecting the concentrated solution into a supercritical extraction kettle (extracting agent is CO) 2 ) Adjusting the constant temperature in the kettle to 45 ℃ and the pressure to 35MPa, sequentially adopting petroleum ether, ethyl acetate and dichloromethane as entrainers for extraction, after 0.6h of static extraction each time, dynamically extracting for 1h at 0.5L/min, and removing the extract (containing impurities with smaller polarity) to obtain the extraction residual phase;
(3) concentrating the residual phase under reduced pressure to 1/10 of the original volume, loading the concentrated solution on 60-100 mesh alcohol-soluble polyamide resin column, loading the column by wet method (the ratio of sample amount to column loading resin amount is 1: 6), eluting with 20 v% ethanol water solution for 5BV, discarding; eluting with 30 v% ethanol water solution for 5BV, and discarding; eluting with 70 v% ethanol water solution for 5BV, collecting the eluate in half or one third as one fraction, detecting with silica gel thin layer chromatography or HPLC, collecting and mixing ASD-containing eluates, and concentrating under reduced pressure until no alcohol smell exists to obtain ASD concentrated solution;
(4) dissolving the ASD concentrated solution by using 90 v% ethanol aqueous solution with 2 times volume, standing for 3h for crystallization, washing impurities on the surface of the crystal by using ethanol, repeatedly recrystallizing for 4 times, detecting the purity of the ASD by using a high performance liquid chromatograph to be 99.5 wt% (the chromatographic condition is as in example one), and drying under reduced pressure to obtain the ASD bulk drug with the yield of 7.23%.
Comparative example 1
The same procedure as in the first example is repeated, except that supercritical fluid extraction is not performed before the polyamide resin column chromatography (i.e., step S2 is not included).
The comparative example shows that the purity of the finally obtained ASD is only 81.6% without adopting supercritical fluid extraction before macroporous adsorption resin column chromatography, and the effect is inferior to that of the application.
Comparative example 2
The same procedure as in example one was repeated except that the polyamide resin column was not used for the chromatography (i.e., step S3 was not included).
This comparative example shows that the purity of the finally obtained ASD was only 82.2% without using a polyamide resin column chromatography, and the effect was inferior to that of the present application.
Comparative example 3
The same procedure as in example one was repeated except that the polyamide resin column was not eluted with ethanol-water.
This comparative example shows that elution of a polyamide resin column without ethanol-water gives a purity of the finally obtained ASD of only 88.7%, and the effect is inferior to that of the present application.
Comparative example 4
The process is as in example one except that recrystallization is not employed.
This comparative example shows that the purity of the ASD finally obtained without recrystallization was only 89.6%, and the effect was inferior to that of the present application.
Comparative example 5
The same as in the first embodiment except that the solvent for extraction in step S1 is water.
This comparative example shows that the yield of ASD finally obtained without extraction with the preferred extraction solvent was only 1.23%, and the effect was inferior to that of the present application.
Comparative example 6
The same procedure as in example one was repeated except that the extraction solvent of step S1 was 20 v% aqueous ethanol.
This comparative example shows that the yield of ASD finally obtained by extraction with a solvent lower than the range of the preferred extraction solvent ratio is only 1.99%, the effect is inferior to that of the present application;
comparative example 7
The same as example 1, except that only petroleum ether was used as the supercritical fluid extraction entrainer in step S2.
This comparative example shows that the yield of ASD finally obtained without gradient supercritical extraction is only 2.72%, the effect is inferior to that of the present application.
Comparative example 8
Example 1 was followed except that the supercritical fluid extraction entrainer of step S2 used ethyl acetate only.
The comparative example shows that the yield of the ASD finally obtained without the gradient supercritical extraction is only 3.29%, the effect is inferior to that of the present application, and the effect is inferior to that of the present application.
Comparative example 9
Example 1 was followed except that only dichloromethane was used as the supercritical fluid extraction entrainer in step S2.
This comparative example shows that the yield of ASD finally obtained without gradient supercritical extraction is only 3.76%, the effect being inferior to that of the present application.
Comparative example 10
The ASD was enriched and purified as described in example 3 of the process described in patent CN 102924545A.
The purity of the ASD finally obtained by this comparative example was 99.2%, and the effect was inferior to that of the present application.
Comparative example 11
The ASD was enriched and purified as described in example 2 of the process described in patent CN 103265601A.
The purity of the ASD finally obtained by this comparative example was 97.1%, and the effect was inferior to that of the present application.
Comparative example 12
The ASD was enriched and purified as described in example 2 of the process described in patent CN 103265604A.
The purity of the ASD finally obtained by this comparative example was 95.6%, and the effect was inferior to that of the present application.
Comparative example 13
The ASD was enriched and purified as described in example 1 of the process described in patent CN 102462688A.
The purity of the ASD finally obtained by this comparative example was 98.0%, the yield was only 5.24%, and the effect was inferior to that of the present application.
Comparative example 14
The ASD was enriched and purified as described in examples 1 and 2 of the process described in patent CN 102180937A.
The purity of the ASD finally obtained by this comparative example was 98.0%, the yield was only 5.49%, and the effect was inferior to that of the present application.
Table 1 comparative experiment results table
As can be seen from the table 1, the process flow applied by the invention is simple, the sample loss is less, the environmental pollution is less, the solvent and the time are saved, the yield and the purity of the ASD bulk drug are high, and the method is more suitable for industrial production; in addition, the specific selection of the separation steps of solvent extraction-supercritical fluid extraction-polyamide resin-recrystallization, etc., and the specific selection of the process conditions during the preparation, etc., according to the present invention, has an important influence on the yield and purity of the final product, as compared with comparative examples 1 to 14. The first embodiment has the best effect.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention.
It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A method for preparing akebia saponin D raw material medicine is characterized in that,
the method comprises the following steps:
s1, crushing the teasel root medicinal decoction pieces to obtain teasel root powder, adding an extraction solvent to perform reflux extraction to obtain an extracting solution, filtering by using an ultrafiltration membrane to remove macromolecular impurities in the extracting solution, and concentrating under reduced pressure to obtain a concentrated solution;
s2, injecting the concentrated solution into a supercritical extraction kettle, sequentially extracting by using entrainers with different polarities at a specific temperature and pressure, after each time of static extraction for 0.5-1h, dynamically extracting for 1-2h at a speed of 0.1-1L/min, removing impurities with smaller polarity, and collecting to obtain an extraction residual phase;
s3, concentrating the residual phase under reduced pressure, performing polyamide resin column chromatography, performing gradient elution by using an elution solvent, performing silica gel thin layer chromatography or HPLC (high performance liquid chromatography) inspection, collecting and combining the eluates containing the akebia saponin D, and concentrating under reduced pressure to obtain a akebia saponin D concentrated solution;
s4, recrystallizing the akebia saponin D concentrated solution, and drying to obtain the akebia saponin D.
2. The method for preparing akebia saponin D bulk drug according to claim 1, which is characterized in that,
in the step S2, in the step S,
the extractant in the supercritical extraction kettle is CO 2 ；
The entrainer is one or more of petroleum ether, ethyl acetate, dichloromethane and trichloromethane.
3. The method for preparing akebia saponin D bulk drug according to claim 1 or 2, characterized in that,
in the step S2, in the step S,
the temperature and the pressure in the supercritical extraction kettle are respectively 35-65 ℃ and 10-40 MPa.
4. The method for preparing akebia saponin D bulk drug according to claim 1, which is characterized in that,
in the step S3, in the step S,
the elution solvent is one or more of water, methanol and ethanol.
5. The method for preparing akebia saponin D bulk drug according to claim 1 or 4, characterized in that,
in the step S3, in the step S,
the polyamide resin column is an alcohol-soluble polyamide resin column.
6. The method for preparing a akebia saponin D drug substance according to any one of claims 1-5, wherein,
in the step S1, in the step S,
the adding amount of the extraction solvent is 8-15 times of the mass of the teasel root powder;
the reflux extraction is carried out for 2-4 times, the extraction time of each time is 2-3h, after reflux extraction, the extracting solutions are combined and then filtered by an ultrafiltration membrane and concentrated under reduced pressure in sequence to obtain concentrated solution with the density of 1.00-1.30 g/ml.
7. The method for preparing a akebia saponin D drug substance according to any one of claims 1-6, wherein,
in the step S1, in the step S,
the extraction solvent is one or more of water, methanol and ethanol;
the temperature of reflux extraction is more than or equal to 70 ℃.
8. The method for preparing akebia saponin D bulk drug according to claim 7, which is characterized in that,
in the step S1, in the step S,
the extraction solvent is ethanol water solution with the concentration of less than 90 v%;
preferably, the extraction solvent is 35-90 v% ethanol water solution, and the addition amount of the extraction solvent is 10 times of the mass of the teasel root powder.
9. The method for preparing a akebia saponin D drug substance according to any one of claims 1-8, wherein,
in the step S4, in the step S,
the solvent used in the recrystallization treatment is one or more of dichloromethane, chloroform, ethyl acetate, propyl acetate, acetone, n-butanol, ethanol and methanol.
10. Use of the method of preparation of a akebia saponin D drug substance of any one of claims 1-9 in the extraction of a akebia saponin D drug substance.
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