CN111067965B - Preparation method of bacopa monnieri saponin - Google Patents
Preparation method of bacopa monnieri saponin Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention provides a preparation method of bacopaside, which comprises the steps of extracting the bacopaside by adopting high-selectivity acetonitrile-n-butyl alcohol according to the polarity of the bacopaside, adsorbing the bacopaside by sodium chloride, chitosan and diatomite, removing a large amount of impurities by ethyl acetate-ethanol hot washing, further enriching the bacopaside, finally recrystallizing by adopting isopropanol, transferring the components into isopropanol solution, and crystallizing, thereby purifying the bacopaside content to be more than 88%. The method has the advantages of simple process, low cost, short period, high recovery rate and stable quality, is suitable for industrial production, avoids complex processes such as resin separation and the like, and ensures that the content of the product of the bacopaside is purified to be more than 88 percent.
Description
Technical Field
The invention belongs to the field of natural medicines, and relates to a preparation method of bacopaside.
Background
The herba Bacopae Monnieri is whole plant of Bacopa monnieri (L.) Wettst of Scrophulariaceae, and is also named as herba Lysimachiae Christinae. Bacopa monnieri has been used in India for 3000 years to nourish the brain, improve memory and intelligence, such as anxiety, cognitive decline, mental confusion, and the like, and is currently used for the treatment of psychosis and epilepsy. The main chemical components of the bacopa monnieri for promoting memory are bacopa monnieri saponin A and bacopa monnieri saponin B, and the most important functions of the bacopa monnieri saponin A and the bacopa monnieri saponin B are to improve the cognitive and memory functions and the biological activities of tranquilizing, resisting anxiety, resisting convulsion, resisting epilepsy and the like. The method is mainly distributed in Fujian, Taiwan, Guangdong, Yunnan and Sichuan provinces in China.
The bacopa monnieri medicinal material with the highest content is bacopa monnieri saponin A, wherein Bacopaside I (Baco I for short, molecular weight 979), Bacopaside II (Baco II for short, molecular weight 929), Bacoside A3 (Baco A3 for short, molecular weight 929), bacopaaponin C (Baco C for short, molecular weight 899) and bacopaaponin C isomer (Baco Ci for short, or Baco X for short, molecular weight 899) are mainly contained, the mother core structures of the Bacopaside I, the Bacopaside II and the Baco C are similar, aglycones are jujubogenin or pseudojujubogenin, and Baco A3, Baco C and Baco Ci are isomers. The content of the bacopaside detected after the separation and purification of the patent refers to the total content of the liquid phase of the 5 components.
Patent CN1303098C discloses a method for preparing pseudo-portulaca oleracea saponins, which comprises the steps of extracting raw materials with ethanol, separating by macroporous resin, purifying by RP-18 low-pressure column or HPLC high performance liquid chromatography to obtain 3 pseudo-portulaca oleracea saponins, namely Baco I, Baco C and BacoD. The purification is carried out by adopting a multi-step column chromatography method, so that the method has the advantages of long period, high cost and unsuitability for industrial production.
Patent CN1833692B discloses a method for preparing bacopaside, which comprises extracting raw materials with 40-80% ethanol, concentrating, purifying with macroporous adsorbent resin, directly eluting with 10-95% ethanol, or eluting with 10-60% ethanol first, then eluting with 60-95% ethanol, and increasing the content of bacopaside to 20-80% (calculated as bacopaside a). The method has the advantages of resin purification, long process period, large reagent loss, low recovery rate of effective components (80% of the product, the recovery rate is only 1.17%), and no contribution to industrial production.
Disclosure of Invention
The invention aims to provide a preparation method of bacopaside, which mainly solves the problems of complex process, low recovery rate and unsuitability for large-scale industrial production in the prior art.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
the preparation method of the bacopaside comprises the following steps:
and 4, adding the insoluble precipitate II obtained in the step 3 into an isopropanol solution, heating, adding activated carbon for decoloring, filtering, concentrating, standing for crystallization, centrifuging, and finally crystallizing and drying to obtain the bacopa saponins.
Further, the step 1 specifically includes: pulverizing herba Bacopae Monnieri raw material into 80 mesh, adding mixed solvent 5-8 times of the raw material, stirring and extracting for 2 times, mixing extractive solutions, and concentrating the extractive solution to specific gravity of 1.03-1.05; the mixed solvent is a mixed solvent of acetonitrile and n-butanol, and the volume ratio of the acetonitrile to the n-butanol is 1: 1-3.
Further, the mass concentration of the chitosan solution is 2%; wherein the solvent is glacial acetic acid with the mass concentration of 1%.
Further, the mass concentration of the sodium chloride aqueous solution is 10-15%.
Further, the volume ratio of the extracting solution concentrated in the step 1 to the chitosan solution to the sodium chloride aqueous solution is 1:0.1-0.2: 0.05-0.08.
Further, the dissolution in the above step 3 is carried out at 50 ℃.
Further, the mass ratio of the insoluble precipitate I, the organic solvent and the surfactant in the step 3 is 1:15-20: 0.001-0.003;
further, the mass concentration of the ethanol is 90-95%.
Further, the volume ratio of the ethyl acetate to the ethanol in the organic solvent is 10-15: 1.
Furthermore, the mass ratio of the insoluble precipitate II, the isopropanol solution and the activated carbon in the step 4 is 1:8-10: 0.05-0.1.
The invention has the following technical effects:
the method has the advantages of simple process, low cost, short period, high recovery rate and stable quality, is suitable for industrial production, avoids complex processes such as resin separation and the like, and ensures that the content of the product of the bacopaside is purified to be more than 88 percent.
Drawings
FIG. 1 is a flow chart of the preparation of the present invention;
FIG. 2 is a high performance liquid chromatography chromatogram of the Bacopa monnieri raw material of the present invention;
FIG. 3 is a high performance liquid chromatography chromatogram of the product bacopaside of the present invention.
Detailed Description
According to the polarity of the bacopaside, the invention adopts high-selectivity acetonitrile-n-butanol to extract, then sodium chloride, chitosan and diatomite or activated clay are used for adsorption, the bacopaside is adsorbed and settled, thereby removing a large amount of impurities, further according to the physical property of the bacopaside, ethyl acetate-ethanol is selected for hot washing, the impurities with small polarity are removed, finally, the bacopaside is dissolved from the sediment through isopropanol, decolored, concentrated and crystallized, thereby purifying the content of the bacopaside to be more than 88%.
Example 1:
pulverizing 100Kg of Bacopa monnieri, sieving with 80 mesh sieve, adding 5 times of acetonitrile-n-butanol (volume ratio 1:1), stirring at room temperature for 2 times, each for 1.5 hr, mixing extractive solutions, concentrating under reduced pressure to small amount, adding water to remove solvent, and concentrating to specific gravity of 1.05-1.08.
Adding 10% of chitosan solution and 5% of NaCl aqueous solution with mass concentration of 10% into the liquid, heating and stirring for 30min, then adding 3% of diatomite, stirring, filtering, and collecting insoluble precipitate I.
Adding 15 times of ethyl acetate-95% ethanol (volume ratio 15:1) and 0.1% sodium dodecyl sulfate into the precipitate I, dissolving at 50 deg.C for 10min, filtering, and collecting insoluble precipitate II.
Adding the precipitate II into isopropanol solution with the mass of 8 times of that of the precipitate II, heating for dissolving, filtering, adding 5% active carbon for decoloring for 1h, filtering, concentrating to a small amount, standing for crystallizing to obtain 2.58Kg of bacopaside with the content of 89.14%.
Example 2:
pulverizing 100Kg of Bacopa monnieri, sieving with 80 mesh sieve, adding 8 times of acetonitrile-n-butanol (volume ratio 1:3), stirring at room temperature for 2 times, each for 1.5 hr, mixing extractive solutions, concentrating under reduced pressure to small amount, adding water to remove solvent, and concentrating to specific gravity of 1.05-1.08.
Adding 15% of chitosan solution and 8% of NaCl aqueous solution with mass concentration of 15% into the liquid, heating and stirring for 30min, then adding activated clay with liquid volume of 5%, stirring, filtering, and collecting insoluble precipitate I.
Adding 20 times of ethyl acetate-90% ethanol (volume ratio 15:1) and 0.3% sodium dodecyl benzene sulfonate into the precipitate I, dissolving at 50 deg.C for 10min, filtering, and collecting insoluble precipitate II.
Adding the precipitate II into 10 times of isopropanol solution, heating for dissolving, filtering, adding 10% active carbon for decolorizing for 1 hr, filtering, concentrating to a small amount, standing, and crystallizing to obtain 2.57Kg of bacopaside with 90.27% content.
Example 3:
pulverizing 100Kg of Bacopa monnieri, sieving with 80 mesh sieve, adding 6 times of acetonitrile-n-butanol (volume ratio of 1:2), stirring at room temperature for 2 times, each for 1.5 hr, mixing extractive solutions, concentrating under reduced pressure to small amount, adding water to remove solvent, and concentrating to specific gravity of 1.05-1.08.
Adding 20% by volume of chitosan solution and 7% by volume of NaCl aqueous solution with mass concentration of 12% into the above liquid, heating and stirring for 30min, then adding activated clay 3% by volume of the liquid, stirring, filtering, and collecting insoluble precipitate I.
Adding 18 times of ethyl acetate-95% ethanol (volume ratio 12:1) and 0.3% sodium dodecyl benzene sulfonate into the precipitate I, dissolving at 50 deg.C for 10min, filtering, and collecting insoluble precipitate II.
Adding the precipitate II into 10 times of isopropanol solution, heating for dissolving, filtering, adding 6% active carbon for decolorizing for 1 hr, concentrating to a small amount, standing, and crystallizing to obtain 2.63Kg of bacopaside with 88.93% content.
Example 4:
pulverizing 100Kg of Bacopa monnieri, sieving with 80 mesh sieve, adding 6 times of acetonitrile-n-butanol (volume ratio of 1:2), stirring at room temperature for 2 times, each for 1.5 hr, mixing extractive solutions, concentrating under reduced pressure to small amount, adding water to remove solvent, and concentrating to specific gravity of 1.05-1.08.
Adding 15 volume percent of chitosan solution and 8 volume percent of NaCl aqueous solution with the mass concentration of 15 percent into the liquid, heating and stirring for 30min, then adding diatomite with the volume of 5 percent of the liquid, stirring, filtering, and collecting insoluble precipitate I.
Adding 20 times of ethyl acetate-90% ethanol (volume ratio of 10:1) and 0.3% sodium dodecylbenzenesulfonate into the precipitate I, dissolving at 50 deg.C for 10min, filtering, and collecting insoluble precipitate II.
Adding the precipitate II into isopropanol solution with the mass of 8 times of that of the precipitate II, heating for dissolving, filtering, adding 8% active carbon for decoloring for 1h, filtering, concentrating to a small amount, standing for crystallizing to obtain 2.70Kg of bacopaside with the content of 90.54%.
Example 5:
pulverizing 100Kg of Bacopa monnieri, sieving with 80 mesh sieve, adding 7 times of acetonitrile-n-butanol (volume ratio 1:1), stirring at room temperature for 2 times, each for 1.5 hr, mixing extractive solutions, concentrating under reduced pressure to small amount, adding water to remove solvent, and concentrating to specific gravity of 1.05-1.08.
Adding 18 volume percent of chitosan solution and 6 volume percent of NaCl aqueous solution with the mass concentration of 13 percent into the liquid, heating and stirring for 30min, then adding diatomite with the volume of 4 percent of the liquid, stirring, filtering, and collecting insoluble precipitate I.
Adding 16 times of ethyl acetate-90% ethanol (volume ratio 12:1) and 0.2% sodium dodecyl sulfate into the precipitate I, dissolving at 50 deg.C for 10min, filtering, and collecting insoluble precipitate II.
Adding the precipitate II into isopropanol solution with the mass of 9 times of that of the precipitate II, heating for dissolving, filtering, adding 9% active carbon for decoloring for 1h, filtering, concentrating to a small amount, standing for crystallizing to obtain 2.59Kg of bacopaside with the content of 89.79%.
Claims (3)
1. A preparation method of bacopa saponins is characterized by comprising the following steps:
step 1, crushing a bacopa monnieri raw material, adding a mixed solvent, stirring and extracting, and concentrating an extracting solution; the mixed solvent is a mixed solvent of acetonitrile and n-butanol, wherein the volume ratio of the acetonitrile to the n-butanol is 1:1-3, and the addition amount of the mixed solvent is 5-8 times of the mass of the raw materials;
step 2, adding a chitosan solution and a sodium chloride aqueous solution into the extracting solution concentrated in the step 1, wherein the volume ratio of the concentrated extracting solution to the chitosan solution to the sodium chloride aqueous solution is 1:0.1-0.2:0.05-0.08, heating and stirring, then adding diatomite or activated clay for stirring, filtering, and collecting insoluble precipitate I, wherein the mass concentration of the chitosan solution is 2%, and the mass concentration of the sodium chloride aqueous solution is 10% -15%;
step 3, adding an organic solvent and a surfactant into the insoluble precipitate I obtained in the step 2, dissolving at 50 ℃, wherein the mass ratio of the insoluble precipitate I to the organic solvent to the surfactant is 1:15-20:0.001-0.003, filtering, and collecting an insoluble precipitate II; the organic solvent is a mixed solvent of ethyl acetate and ethanol, and the volume ratio of the ethyl acetate to the ethanol in the organic solvent is 10-15: 1; the surfactant is sodium dodecyl sulfate or sodium dodecyl benzene sulfonate;
and 4, adding the insoluble precipitate II obtained in the step 3 into an isopropanol solution, heating the insoluble precipitate II, the isopropanol solution and the activated carbon in a mass ratio of 1:8-10:0.05-0.1, adding the activated carbon for decoloring, filtering, concentrating, standing for crystallization, centrifuging, and finally crystallizing and drying to obtain the bacopaside.
2. The method for preparing bacopaside according to claim 1, wherein the step 1 is specifically: pulverizing herba Bacopae Monnieri raw material, sieving with 80 mesh sieve, adding mixed solvent, stirring and extracting for 2 times, mixing extractive solutions, and concentrating the extractive solution to specific gravity of 1.03-1.05.
3. The method of producing bacopaside according to claim 1, wherein the step of extracting comprises: the mass concentration of the ethanol is 90-95%.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101172137A (en) * | 2007-07-03 | 2008-05-07 | 巫军 | Method for high effectively extracting pseudo-purslane saponin by using cavitated suspension extraction and stripping apparatus |
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| AU2003201756A1 (en) * | 2003-01-03 | 2004-07-29 | Ganga Raju Gokaraju | A process for producing enriched fractions containing up to 100% of bacoside a and bacoside b |
| WO2006117795A1 (en) * | 2005-05-03 | 2006-11-09 | Raju Gokaraju Ganga | A process for producing enriched fractions containing upto 100% of bacopasaponins from the plant materials of bacopa species |
| CN101045107A (en) * | 2006-12-26 | 2007-10-03 | 巫军 | Method for extracting pseudo-purslane saponin, and its application in medical field |
| CN106831932A (en) * | 2016-12-29 | 2017-06-13 | 陕西嘉禾生物科技股份有限公司 | It is a kind of that the method for separating bacopa monnieri saponin is extracted from bacopa monnieri |
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| CN101172137A (en) * | 2007-07-03 | 2008-05-07 | 巫军 | Method for high effectively extracting pseudo-purslane saponin by using cavitated suspension extraction and stripping apparatus |
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| Title |
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| 健脑功能的假马齿苋提取物的制备;无;《国外医药.植物药分册》;20051231;第20卷(第2期);第87页 * |
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Application publication date: 20200428 Assignee: SHAANXI JIAHE PHYTOCHEM Co.,Ltd. Assignor: Sanyuan Runhe Biological Technology Co.,Ltd. Contract record no.: X2021980015778 Denomination of invention: A preparation method of pseudopurslane saponin Granted publication date: 20211001 License type: Common License Record date: 20211227 |
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Application publication date: 20200428 Assignee: SHAANXI JIAHE PHARMACEUTICAL CO.,LTD. Assignor: Sanyuan Runhe Biological Technology Co.,Ltd. Contract record no.: X2021980017334 Denomination of invention: A preparation method of pseudopurslane saponin Granted publication date: 20211001 License type: Common License Record date: 20220112 |
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