CN108299300B - Method for purifying aconitine A and deacetyl aconitine A in Aconitum carmichaeli - Google Patents
Method for purifying aconitine A and deacetyl aconitine A in Aconitum carmichaeli Download PDFInfo
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- CN108299300B CN108299300B CN201810222429.3A CN201810222429A CN108299300B CN 108299300 B CN108299300 B CN 108299300B CN 201810222429 A CN201810222429 A CN 201810222429A CN 108299300 B CN108299300 B CN 108299300B
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Abstract
The invention provides a purification method of aconitine A and deacetyl aconitine A in Aconitum carmichaeli Debx. The purification method of the aconitine A and the deacetyl aconitine A in the Aconitum yunnanense comprises the following steps: step one, extracting medicinal materials; step two, extraction and separation; step three, column chromatography purification; step four, crystallization and purification; step five, hydrolyzing the aconitine A; step six, column chromatography purification of the deacetyl aconitine A. The purification method for the mesaconitine and the deacetyl mesaconitine in the aconitum sinomontanum provided by the invention purifies the mesaconitine and the deacetyl mesaconitine in the aconitum sinomontanum, provides a certain foundation for quality control and pharmacological research of the mesaconitine, and provides a related material foundation for further utilizing the mesaconitine to reduce toxic and side effects.
Description
Technical Field
The invention belongs to the field of medicine production and manufacture, and particularly relates to a purification method of aconitine A and deacetyl aconitine A in Aconitum carmichaeli.
Background
Aconitum yunnanense is dried root of Aconitum yunnanense Bulleyanum Diels of Aconitum of Ranunculaceae, and is mainly distributed in Lijiang and Dali regions of western Yunnan province for treating rheumatic arthritis, numbness, paralysis and various pain diseases. Wherein the component diester alkaloid aconitine A has good analgesic effect and no addiction, but has high toxicity. In the clinical application of the radix aconiti and the radix aconiti lateralis, the diester alkaloids aconitine, hypaconitine and the aconitine thereof are hydrolyzed to obtain the benzoylaconine, the benzoylmesaconine and the benzoylhypaconine, which can effectively reduce the toxic and side effects.
Disclosure of Invention
In order to solve the technical problem that the existing aconitum sinomontanum nakai contains the aconitine A and the deacetylated aconitine A, the invention provides a method for purifying the aconitine A and the deacetylated aconitine A in the aconitum sinomontanum nakai.
The purification method of the aconitine A and the deacetyl aconitine A in the Aconitum yunnanense provided by the invention comprises the following steps:
step one, extraction of medicinal materials: pulverizing dried radix Aconiti Yunnanensis, soaking in appropriate amount of ethanol, percolating with ethanol, mixing filtrates, and concentrating to obtain paste;
step two, extraction and separation: dissolving the obtained paste with hydrochloric acid solution, filtering, adjusting pH of the obtained acid water solution to alkaline, extracting the alkaline solution with petroleum ether for multiple times, extracting with ethyl acetate for multiple times, mixing the petroleum ether or ethyl acetate extracts, and recovering under reduced pressure to obtain total alkali;
step three, column chromatography purification, namely dissolving the total alkali obtained in the extraction separation step by using dichloromethane, adsorbing the dissolved total alkali on silica gel, stirring the mixture, volatilizing the mixture, performing silica gel column chromatography, performing gradient elution by using petroleum ether, ethyl acetate and diethylamine, collecting eluent, performing tracking detection by using thin-layer chromatography T L C by using crude stem aconitine A as a contrast, combining fractions mainly containing the crude stem aconitine A, and recovering the solvent until the fractions are dry to obtain a fraction mainly containing the crude stem aconitine A;
step four, crystallization and purification: dissolving the part containing the aconitine A with ethanol, crystallizing and recrystallizing to obtain aconitine A;
step five, hydrolyzing the crude stem aconitine A, namely adding ethanol into the crude stem aconitine A to dissolve the crude stem aconitine A, adding an equal amount of saturated disodium hydrogen phosphate aqueous solution, heating in a water bath, sampling T L C every half an hour to monitor the reaction condition, stopping the reaction when only a small amount of crude stem aconitine A is detected by a thin layer, concentrating the reaction solution to a small amount, extracting the reaction solution by dichloromethane for multiple times, combining dichloromethane layers, and concentrating the reaction solution to obtain white-like powder;
and sixthly, purifying the deacetylated aconitine A by column chromatography, namely taking the hydrolyzed powder, adding a small amount of dichloromethane for dissolving, adsorbing the dissolved powder on silica gel for stirring, volatilizing, carrying out silica gel column chromatography, eluting with petroleum ether, ethyl acetate and diethylamine, collecting single-point fractions of the deacetylated aconitine A T L C, and recovering the solvent until the solvent is dry to obtain white powder.
In a preferred embodiment of the purification method of mesaconitine A and deacetyl mesaconitine A in Aconitum carmichaeli provided by the invention, in the sixth step, the elution mass ratio of petroleum ether, ethyl acetate and diethylamine is 30:5: 0.1.
In a preferred embodiment of the method for purifying aconitine a and deacetylaconitine a in aconitum sinomontanum provided by the present invention, in the third step and the sixth step, the sample adsorption on silica gel is a sample 2 times as much as the amount of silica gel.
In a preferred embodiment of the purification method of mesaconitine A and deacetyl mesaconitine A in Aconitum carmichaeli provided by the invention, in the fifth step, the water bath temperature is 50-100 ℃, and the ratio of the ethanol for dissolving to the mesaconitine A is 100: 1.
In a preferred embodiment of the purification method of mesaconitine A and deacetyl mesaconitine A in Aconitum carmichaeli, the elution ratio of petroleum ether, ethyl acetate and diethylamine in the step III is 100-20:10: 0.1.
In a preferred embodiment of the purification method of aconitine A and deacetyl aconitine A in Aconitum carmichaeli provided by the present invention, the second step comprises dissolving the paste obtained in the extraction step with 2% hydrochloric acid solution, filtering, adjusting pH of the acid water solution to 9 with 25% concentrated ammonia water, extracting the alkali solution with petroleum ether for 2 times, and extracting with ethyl acetate for 3 times.
In a preferred embodiment of the purification method of mesaconitine A and deacetyl mesaconitine A in Aconitum carmichaeli provided by the invention, in the first step, the amount of ethanol is 6-10 times of the amount of the medicinal materials.
The purification method of the aconitine A and the deacetylated aconitine A in Aconitum yunnanense has the following beneficial effects:
the method mainly aims at the separation and purification of the aconitine A and the deacetylated aconitine A serving as a hydrolysate thereof, provides a material basis for subsequent pharmacological research, provides a certain basis for the quality control and the pharmacological research of Aconitum yunnanense, and provides a related material basis for further utilizing the aconitine A to reduce the toxic and side effects of the aconitine A.
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In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without inventive efforts, wherein:
FIG. 1 is a process flow diagram of an embodiment of the purification method of mesaconitine A and deacetyl mesaconitine A in Aconitum carmichaeli of the present invention;
FIG. 2 is a crude aconitine A HNMR spectrum;
FIG. 3 is an MS spectrum of deacetyl aconitine A;
FIG. 4 is a CNMR spectrum of deacetyl mesaconine A;
FIG. 5 is a HNMR spectrum of deacetyl aconitine A.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1 to 5, fig. 1 is a process flow chart of an embodiment of the purification method of aconitine A and deacetyl aconitine A in Aconitum yunnanense of the present invention; FIG. 2 is a crude aconitine A HNMR spectrum; FIG. 3 is an MS spectrum of deacetyl aconitine A; FIG. 4 is a CNMR spectrum of deacetyl mesaconine A; FIG. 5 is a HNMR spectrum of deacetyl aconitine A.
Step S11, the purification method of the aconitine A and the deacetyl aconitine A in Aconitum yunnanense (Levl.) Kuntze comprises the following steps of S1: pulverizing dried Aconitum carmichaeli root tuber, soaking in appropriate amount of ethanol, percolating with ethanol (6-10 times of the amount of the medicinal materials), mixing percolate, and concentrating to obtain paste;
step S12, extraction and separation: dissolving the obtained paste with 2% hydrochloric acid solution, filtering, adjusting pH of the obtained acid water solution to 9 with 25% concentrated ammonia water, extracting the alkali solution with petroleum ether for 2 times, extracting with ethyl acetate for 3 times, mixing the petroleum ether or ethyl acetate extracts, and recovering under reduced pressure to obtain total alkali;
step S13, column chromatography purification, namely dissolving the total alkali obtained in the extraction separation step by using dichloromethane, adsorbing the total alkali by using 2 times of silica gel, stirring the mixture to be dried, carrying out silica gel column chromatography, carrying out gradient elution by using petroleum ether, ethyl acetate and diethylamine according to the dosage ratio of 100-20:10:0.1, collecting eluent, carrying out tracking detection by using thin-layer chromatography T L C by using aconitine A as a contrast, combining fractions mainly containing the aconitine A, and recovering the solvent to be dry to obtain a fraction mainly containing the aconitine A;
step S14, crystallization and purification: dissolving the part containing the aconitine A with ethanol, crystallizing and recrystallizing to obtain aconitine A;
step S15, hydrolyzing the aconitine A, namely adding ethanol into the aconitine A to dissolve the aconitine A, wherein the ratio of the ethanol to the aconitine A is 100:1, adding an equivalent saturated disodium hydrogen phosphate aqueous solution, heating the mixture in a water bath at the temperature of 50-100 ℃, sampling T L C every half an hour to monitor the reaction condition, stopping the reaction when a thin layer detects that only a small amount of aconitine A exists, concentrating the mixture to a small amount, extracting the dichloromethane for multiple times, combining dichloromethane layers, and concentrating the dichloromethane layers to obtain white-like powder;
and step S16, column chromatography purification of the deacetyl aconitine A, which is to take the hydrolyzed powder, add a small amount of dichloromethane to dissolve the powder, adsorb the dissolved powder to 2 times of silica gel for sample mixing, volatilize the solution to dryness, carry out silica gel column chromatography, elute the sample with petroleum ether, ethyl acetate and diethylamine according to the mass ratio of 30:5:0.1, collect the single-point fraction of the deacetyl aconitine A T L C, and recycle the solvent to dryness to obtain white powder.
The purification method S1 of the aconitine A and the deacetyl aconitine A in Aconitum carmichaeli has the following beneficial effects:
the method mainly aims at the separation and purification of the aconitine A and the deacetylated aconitine A serving as a hydrolysate thereof, provides a material basis for subsequent pharmacological research, provides a certain basis for the quality control and the pharmacological research of Aconitum yunnanense, and provides a related material basis for further utilizing the aconitine A to reduce the toxic and side effects of the aconitine A.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (5)
1. A method for purifying aconitine A and deacetyl aconitine A in Aconitum carmichaeli is characterized by comprising the following steps:
step one, extraction of medicinal materials: pulverizing dried radix Aconiti Yunnanensis, soaking in appropriate amount of ethanol, percolating with ethanol, mixing filtrates, and concentrating to obtain paste;
step two, extraction and separation: dissolving the obtained paste with hydrochloric acid solution, filtering, adjusting pH of the obtained acid water solution to alkaline, extracting the alkaline solution with petroleum ether for multiple times, extracting with ethyl acetate for multiple times, mixing the petroleum ether or ethyl acetate extracts, and recovering under reduced pressure to obtain total alkali;
step three, column chromatography purification, namely dissolving the total alkali obtained in the extraction separation step by using dichloromethane, adsorbing the dissolved total alkali on silica gel, stirring the mixture, volatilizing the mixture, performing silica gel column chromatography, performing gradient elution by using petroleum ether, ethyl acetate and diethylamine, collecting eluent, performing tracking detection by using thin-layer chromatography T L C by using crude stem aconitine A as a contrast, combining fractions mainly containing the crude stem aconitine A, and recovering the solvent until the fractions are dry to obtain a fraction mainly containing the crude stem aconitine A;
step four, crystallization and purification: dissolving the part containing the aconitine A with ethanol, crystallizing and recrystallizing to obtain aconitine A;
step five, hydrolyzing the aconitine A, namely adding ethanol into the aconitine A to dissolve, wherein the ratio of the ethanol to the aconitine A used for dissolving is 100:1, adding an equivalent amount of saturated disodium hydrogen phosphate aqueous solution, heating in a water bath, the temperature of the water bath is 50-100 ℃, sampling T L C to monitor the reaction condition every half hour, stopping the reaction when only a small amount of aconitine A is detected by a thin layer, concentrating to a small amount, extracting with dichloromethane for multiple times, combining dichloromethane layers, concentrating and drying to obtain white-like powder;
and sixthly, purifying the deacetylated aconitine A by column chromatography, namely taking the hydrolyzed powder, adding a small amount of dichloromethane for dissolving, adsorbing the powder on silica gel for stirring, volatilizing, carrying out silica gel column chromatography, eluting with petroleum ether, ethyl acetate and diethylamine, wherein the mass ratio of the petroleum ether to the ethyl acetate to the diethylamine is 30:5:0.1, collecting fractions of single points of the deacetylated aconitine A T L C, and recovering the solvent until the fractions are dried to obtain white powder.
2. The method for purifying aconitine A and deacetyl aconitine A in Aconitum yunnanense according to claim 1, wherein the sample adsorption to silica gel in the third and sixth steps is carried out by mixing a sample with 2 times the amount of silica gel.
3. The method for purifying mesaconitine A and deacetyl mesaconitine A in Aconitum yunnanense according to claim 1, wherein the elution amount ratio of petroleum ether, ethyl acetate and diethylamine in the step three is 100-20:10: 0.1.
4. The method for purifying aconitine A and deacetyl aconitine A in Aconitum yunnanense of claim 1, wherein the second step comprises dissolving the paste obtained in the extraction step with 2% hydrochloric acid solution, filtering, adjusting pH of the acid water solution to 9 with 25% concentrated ammonia water, extracting the alkali solution with petroleum ether for 2 times, and extracting with ethyl acetate for 3 times.
5. The method for purifying mesaconitine A and deacetyl mesaconitine A in Aconitum yunnanense according to claim 1, wherein in the first step, the amount of ethanol is 6-10 times of the amount of the medicinal materials.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555227A (en) * | 2009-05-19 | 2009-10-14 | 昆明制药集团股份有限公司 | Preparation method of high purity bulleyaconitine A |
| CN102775349A (en) * | 2012-07-02 | 2012-11-14 | 云南农业大学 | Preparation method for bulleyaconitine A |
| CN103819404A (en) * | 2014-03-15 | 2014-05-28 | 湖北博瑞生物科技有限公司 | Energy-efficient process for extracting lappa-conitine |
| CN106008344A (en) * | 2016-06-03 | 2016-10-12 | 云南中医学院 | Bulleyaconitine A preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH0827116A (en) * | 1994-07-11 | 1996-01-30 | Tsumura & Co | Yunaconitine derivative and analgesic and antiinflammatory containing said compound as active ingredient |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555227A (en) * | 2009-05-19 | 2009-10-14 | 昆明制药集团股份有限公司 | Preparation method of high purity bulleyaconitine A |
| CN102775349A (en) * | 2012-07-02 | 2012-11-14 | 云南农业大学 | Preparation method for bulleyaconitine A |
| CN103819404A (en) * | 2014-03-15 | 2014-05-28 | 湖北博瑞生物科技有限公司 | Energy-efficient process for extracting lappa-conitine |
| CN106008344A (en) * | 2016-06-03 | 2016-10-12 | 云南中医学院 | Bulleyaconitine A preparation method |
Non-Patent Citations (1)
| Title |
|---|
| 康定乌头根的化学成分研究;吕光华等;《中草药》;19991231;第30卷(第3期);第164-167页 * |
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