CN111329905A - Total alkaloid component extracted from coptis chinensis and refining method thereof - Google Patents
Total alkaloid component extracted from coptis chinensis and refining method thereof Download PDFInfo
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Abstract
The invention mainly provides a total alkaloid component in a coptis extract and a refining method thereof. The preparation method provided by the invention specifically relates to water extraction, clarification and filtration, a high-efficiency preparation liquid phase technology and a concentration and drying process. The method takes coptis as a raw material, adopts water as a solvent to extract to obtain a crude extract containing coptis alkaloid, and combines a membrane filtration technology and high-efficiency reverse phase preparative chromatography for refining, so that the enrichment and refining preparation of coptis total alkaloid can be realized, and the purity of coptis total alkaloid in the obtained final product can reach more than 85 percent. The process method is simple, high in purity and yield, safe and controllable, and suitable for large-scale production; the method provides a new technical scheme for the development of alkaloid component medicaments.
Description
Technical Field
The invention relates to the technical field of Chinese herbal medicine processing, in particular to a component of total alkaloid extracted from coptis and a refining method thereof.
Background
The Coptidis rhizoma is a perennial herb, and the base source is dried rhizome of Coptidis rhizoma, Coptidis rhizoma or Coptis Teeta wall of Ranunculaceae. The main effective component of the coptis is a alkaloid compound, has obvious physiological activity, and can reduce blood fat and blood sugar, and treat diabetes, enteritis, damp-heat fullness, vomiting and acid regurgitation, diarrhea, conjunctival congestion, toothache, diabetes, carbuncle, swelling, furuncle, eczema, ear purulence and the like.
The preparation of the high-purity compound-like component is very important for clarifying the material basis and deep activity evaluation of the traditional Chinese medicine component. The separation and purification of the coptis alkaloid are mostly realized by adopting macroporous adsorption resin, and the research on the process for purifying the coptis total alkaloid by using the macroporous adsorption resin is used for purifying the coptis total alkaloid, so that the final products with the berberine purity of 73.2 percent and the total alkaloid purity of 67.71 percent are respectively obtained.
The macroporous adsorption resin has the advantages of reasonability, simple operation and the like in the separation and refining of the coptis total alkaloids, but the obtained final product has lower purity and has the following defects in practical application: the pretreatment and regeneration are complex, the commercial macroporous resin lacks uniform and strict quality standard, is easy to break, has poor repeatability, poor selectivity, serious organic residue and potential safety hazard. The process conditions of pretreatment, regeneration and purification and the like of the macroporous resin also lack standard evaluation indexes, and no uniform operation rules are provided for ensuring the stability. Therefore, the development of a method for selectively enriching and efficiently preparing the coptis total alkaloids is very important for the research of coptis.
The invention is provided in view of the above.
Disclosure of Invention
The invention mainly provides a total alkaloid component in a coptis extract and a refining method thereof, which are used for solving the problems of low purity of target components, personal harm and environmental pollution caused by residual organic solvents, uncontrollable quality and the like in the current final product.
In order to realize the purpose, the invention provides a method for refining total alkaloid components in a coptis extract, which comprises the following steps:
(1) preparing a crude extract: cleaning, drying and crushing the coptis medicinal material, weighing 1-20Kg, adding water which is 5-50 times of the weight of the coptis medicinal material, extracting for 0.1-5h by using a decocting, refluxing, ultrasonic or microwave-assisted process, filtering the liquid medicine by using gauze or a 100-plus-mesh 300-mesh sieve, adding 5-50 times of water into dregs, extracting in the same way, filtering the liquid medicine, and removing the dregs; mixing the two medicinal liquids, performing rotary evaporation, concentration and drying to obtain Coptidis rhizoma crude extract dry powder;
(2) clarifying and filtering: weighing a proper amount of the coptis chinensis crude extract dry powder obtained in the step (1), dissolving the coptis chinensis crude extract dry powder with water of which the mass is 2-20 times that of the coptis chinensis crude extract dry powder, removing particles and pigments by using a filter membrane, and performing rotary evaporation concentration and drying on the obtained membrane permeate to obtain a dry product, namely clear dry powder;
(3) refining total alkaloids of coptis: dissolving the clear dry powder with water to a mass concentration of 5-300mg/mL, purifying by high performance liquid chromatography, collecting eluate, mixing the target components, concentrating, and drying to obtain refined Coptidis rhizoma powder product.
In some embodiments of the present invention, preferably, the water used in steps (1), (2) and (3) may be purified water, distilled water or water for injection.
Preferably, the drying method in steps (1), (2) and (3) is microwave drying, freeze drying, vacuum drying or spray drying.
Preferably, the extraction process in step (1) is extraction at 80-180 deg.C, reflux extraction at 60-100 deg.C, ultrasonic extraction at 60-100 deg.C, or microwave-assisted extraction at 60-100 deg.C.
Preferably, the type of the filter membrane used in step (2) may be a hollow fiber type, a roll type, a tube type or a plate and frame type, and the area of the filter membrane used is 0.1 to 0.5m2The size of the filter membrane used is 0.1-0.65 μm or 500-1000kD, and the washing and filtering times in the membrane separation are 3-10 times.
Preferably, in the membrane filtration technology used in step (2), when 30-100mL of the trapped fluid remains, 1-30 times of the volume of the trapped fluid is added for washing by 3-10 times, and when 30-100mL of the trapped fluid remains again, 10% -80% of ethanol with 1-30 times of the volume of the trapped fluid is added for washing by 3-10 times.
Preferably, in the high performance liquid chromatography used in step (3), the filler matrix is silica gel or high molecular polymer; the bonding phase is one or more of normal chain alkyl groups with 1-30 carbon atoms; the grain diameter is 8-100 μm; having a pore diameter of
The specific surface area is 100-500m2(ii)/g; the inner diameter of the chromatographic column is 20-1600mm, and the height of the chromatographic column bed is 100-1000 mm.
Preferably, in the high performance liquid chromatography used in step (3), the eluent contains organic solvent and acid with a certain concentration, wherein the organic solvent is methanol, ethanol or acetonitrile with the content of 5% -90%, and the acid is phosphoric acid, formic acid or acetic acid with the concentration of 0.01% -0.5%.
Preferably, in the high performance liquid chromatography used in step (3), the loading amount of the chromatographic column is 0.01-20.0% of the mass of the stationary phase, the loading and elution flow rate is 140-350cm/h, and the elution conditions are isocratic or linear.
The invention has the following beneficial effects:
1) the method for purifying the coptis total alkaloids by adopting the membrane separation and high performance liquid chromatography technology can obviously improve the purity of a target object and the content of effective components, saves the cost of medicinal materials, can provide preparation conditions more favorable for industrial production, ensures the stability and safety of the process, and enables the coptis medicinal materials to exert the effects as much as possible.
2) The membrane separation and high performance liquid preparation technology are combined, so that ineffective components such as lipids, pigments, heavy metals, pesticide residues and the like can be effectively removed, the effective components of the coptis chinensis are retained, and the medicinal value of the coptis chinensis medicinal material can be improved to the maximum extent.
3) The prepared total coptis alkaloid component has the purity higher than 85 percent and the total alkaloid yield higher than 95 percent, which are both higher than the prior art.
Drawings
FIG. 1 is a liquid chromatography contrast chromatogram of a clarified liquid and a stock solution of a clarification filtration experiment;
FIG. 2 is a schematic representation of the collection of fractions purified by the HPLC technique of example 1;
FIG. 3 is a liquid chromatogram of a fraction obtained by purification according to the high performance liquid chromatography technique of example 1;
FIG. 4 is a comparison chromatogram of liquid chromatography analysis of total alkaloid samples from crude extract and purified preparation of Coptidis rhizoma in example 1.
Detailed Description
The method of the invention will now be described in more detail with reference to the schematic drawings, in which preferred embodiments of the invention are shown, it being understood that a person skilled in the art may modify the invention described herein while still achieving the advantageous effects of the invention. Accordingly, the following description should be construed as broadly as possible to those skilled in the art and not as limiting the invention.
Example 1:
weighing 2Kg of pretreated Coptidis rhizoma, adding 20L of water, decocting at 120 deg.C for 1 hr, filtering with gauze, adding 20L of water into the residue, decocting at 120 deg.C for 0.5 hr, filtering with gauze, mixing filtrates, rotary steaming, concentrating, and vacuum drying under reduced pressure to obtain Coptidis rhizoma extract powder.
And (3) performing a clarification filtration experiment by using a hollow fiber membrane of 750kD, adding 0.8L of pure water for 8 times when about 50mL of trapped fluid remains, adding 200mL of pure water for circular washing when about 50mL of trapped fluid remains again, and washing by using 200mL of 50% ethanol. And carrying out rotary evaporation concentration and spray drying on the obtained sample liquid respectively to obtain a dried product. The product obtained in the clarifying filtration experiment and the dry powder of the crude extract are respectively dissolved into a mass concentration of 1mg/mL by pure water, the comparative chart of HPLC analysis is shown in figure 1, and the six main alkaloids are hardly lost in the clarifying filtration experiment as can be seen from figure 1.
2g of the clarified product is taken and prepared into a sample solution of 100mg/mL by using pure water. After preparation of the column (C18HCE 50 x 230mm, 10 μm, 300g,
) The refining experiment of the rhizoma coptidis total alkaloids is carried out, and 0.1% acetic acid water solution (A) and ethanol (B) are used as a mobile phase system. The elution condition is 0-60min, 10% B; 60-65min, 10% -95% B; 65-80min, 95% B; the detection wavelength is 345 nm; the flow rate is 60 mL/min; the sample volume was 10mL (1000 mg solids, 0.33% sample loading). Fraction collection schematic as shown in fig. 2, the eluate was collected for a total of 7 fractions; the liquid phase analysis of each fraction is shown in FIG. 3, from which FIG. 3 it can be seen that Fr.1 and Fr.7 are non-alkaloid moieties, and Fr.2-Fr.6 are alkaloid moieties; a comparison graph of liquid phase analysis of the sample solution and the obtained refined alkaloid (with the same prepared mass concentration) is shown in FIG. 4, and it can be seen from FIG. 4 that the purity of six main alkaloids in the obtained refined alkaloid component is greatly improved. And carrying out rotary evaporation concentration and freeze drying on each obtained fraction to obtain a dried product. The purity of the finally obtained coptis total alkaloid is 87.76%.
Example 2:
weighing 2Kg of pretreated Coptidis rhizoma, adding 20L of water, decocting at 120 deg.C for 1 hr, filtering with gauze, adding 20L of water into the residue, decocting at 120 deg.C for 0.5 hr, filtering with gauze, mixing filtrates, rotary steaming, concentrating, and vacuum drying under reduced pressure to obtain Coptidis rhizoma extract powder.
And (3) performing a clarification filtration experiment by using a hollow fiber membrane of 750kD, adding 0.8L of pure water for 8 times when about 50mL of trapped fluid remains, adding 200mL of pure water for circular washing when about 50mL of trapped fluid remains again, and washing by using 200mL of 50% ethanol. And carrying out rotary evaporation concentration and spray drying on the obtained sample liquid respectively to obtain a dried product.
2g of the clarified and filtered product is taken and prepared into a sample solution of 100mg/mL by pure water. After preparation of the column (C18YE-a (300g, 60 μm, 50 x 240mm,
) ) refining Coptidis rhizoma total alkaloid at flow rate of 50mL/min with 0.04% acetic acid water and about 3BV, loading 210mL (loading amount of about 7%), rinsing with 0.04% acetic acid water and 3BV, and eluting ① 30%Collecting eluate with 3BV ethanol (1% formic acid) and ② 70% ethanol (1% formic acid) according to column volume, concentrating by rotary evaporation, and lyophilizing to obtain dried rhizoma Coptidis total alkaloids with purity of 86.05%.
Example 3:
weighing 2Kg of pretreated Coptidis rhizoma, adding 20L of water, decocting at 120 deg.C for 1 hr, filtering with gauze, adding 20L of water into the residue, decocting at 120 deg.C for 0.5 hr, filtering with gauze, mixing filtrates, rotary steaming, concentrating, and vacuum drying under reduced pressure to obtain Coptidis rhizoma extract powder.
A clarifying filtration experiment is carried out by using a 300kD hollow fiber membrane, 0.8L of pure water is added for 8 times when about 50mL of trapped fluid remains, 200mL of pure water is added for circular washing when about 50mL of trapped fluid remains again, and then 200mL of 50% ethanol is used for washing. And carrying out rotary evaporation concentration and spray drying on the obtained sample liquid respectively to obtain a dried product.
2g of the clarified product is taken and prepared into a sample solution of 100mg/mL by using pure water. After preparation of the column (C18HCE 50 x 250mm, 10 μm, 300g,
) The refining experiment of the rhizoma coptidis total alkaloids is carried out, and 0.1% formic acid water solution (A) and ethanol (B) are used as a mobile phase system. The elution condition is 0-60min, 10% B; 60-65min, 10% -95% B; 65-80min, 95% B; the detection wavelength is 345 nm; the flow rate was 60mL/min (183 cm/h); the sample volume was 10mL (1000 mg solids, 0.33% sample loading). Collecting the eluent, and respectively carrying out rotary evaporation concentration and freeze drying on the obtained eluent to obtain a dried product. The purity of the coptis total alkaloid is 90.34 percent through weighing calculation analysis.
Example 4:
weighing 2Kg of pretreated Coptidis rhizoma, adding 20L of water, decocting at 120 deg.C for 1 hr, filtering with gauze, adding 20L of water into the residue, decocting at 120 deg.C for 0.5 hr, filtering with gauze, mixing filtrates, rotary steaming, concentrating, and vacuum drying under reduced pressure to obtain Coptidis rhizoma extract powder.
A clarification filtration experiment is carried out by using a 500kD hollow fiber membrane, 0.8L of pure water is added for 8 times when about 50mL of trapped fluid remains, 200mL of pure water is added for circular washing when about 50mL of trapped fluid remains again, and then 200mL of 50% ethanol is used for washing. And carrying out rotary evaporation concentration and spray drying on the obtained sample liquid respectively to obtain a dried product.
2g of the clarified product is taken and prepared into a sample solution of 100mg/mL by using pure water. After preparation of the column (C1850 x 250mm, 60 μm, 300g,
) The refining experiment of the rhizoma coptidis total alkaloids is carried out, and 0.1% acetic acid water solution (A) and ethanol (B) are used as a mobile phase system. The elution condition is 0-60min, 10% B; 60-65min, 10% -95% B; 65-80min, 95% B; the detection wavelength is 345 nm; the flow rate was 60mL/min (183 cm/h); the sample volume was 10mL (1000 mg solids, 0.33% sample loading). Collecting the eluent, and respectively carrying out rotary evaporation concentration and freeze drying on the obtained eluent to obtain a dried product. The purity of the coptis total alkaloid is 88.65 percent by weighing, calculating and analyzing.
Example 5:
weighing 2Kg of pretreated Coptidis rhizoma, adding 20L of water, decocting at 120 deg.C for 1 hr, filtering with gauze, adding 20L of water into the residue, decocting at 120 deg.C for 0.5 hr, filtering with gauze, mixing filtrates, rotary steaming, concentrating, and vacuum drying under reduced pressure to obtain Coptidis rhizoma extract powder.
A clarification filtration experiment is carried out by using a 100kD hollow fiber membrane, 0.8L of pure water is added for 8 times when about 50mL of trapped fluid remains, 200mL of pure water is added for circular washing when about 50mL of trapped fluid remains again, and then 200mL of 50% ethanol is used for washing. And carrying out rotary evaporation concentration and spray drying on the obtained sample liquid respectively to obtain a dried product.
2g of the clarified product is taken and prepared into a sample solution of 100mg/mL by using pure water. After preparation of the column (C1850 x 250mm, 40 μm, 300g,
) The refining experiment of the rhizoma coptidis total alkaloids is carried out, and 0.1% formic acid water solution (A) and methanol (B) are used as a mobile phase system. The elution condition is 12 percentB isocratic elution for 85 min; the detection wavelength is 345 nm; the flow rate was 60mL/min (183 cm/h); the loading volume was 100mL (solid content 10g, loading 3.3%). Collecting the eluent, and respectively carrying out rotary evaporation concentration and freeze drying on the obtained eluent to obtain a dried product. The purity of the coptis total alkaloid is 85.48 percent through weighing calculation analysis.
The high performance liquid chromatography qualitative and quantitative method of the coptis alkaloid components obtained by the method comprises the following steps: watersalliance high performance liquid chromatography system on an XCharge C18 column, run at A: acetonitrile (containing 0.1% of acetic acid) and water (containing 0.1% of acetic acid) are used as mobile phases for gradient elution, and the elution conditions are as follows: 0-16min, 10% -11%; 16-30min, 11% -60% A; 30-32min, 60% -90% A; 32-35min, 90% A. The flow rate is 1mL/min, the PDA detector, the column temperature is 35 ℃, and the detection wavelength is 345 nm. Taking berberine hydrochloride as a standard substance to make an external standard working curve, taking the mass concentration (X, mg/mL) of the berberine hydrochloride as a horizontal coordinate, and taking a peak area (Y/1000, uV sec) as a vertical coordinate to establish a standard curve. The result shows that the berberine has good linear relation within the range of 0.01-1.2 mg/mL, and the linear correlation coefficient is R2And the linear regression equation is Y18,363.46X-366.46 as 1.00.
In conclusion, compared with the crude extract, the clarified product prepared by the preparation method of the invention has bright color and luster and fine and uniform texture because a plurality of dark black substances are trapped in the trapped liquid, and toxic and harmful substances such as pesticide residue, heavy metal, endotoxin and the like can be removed by an efficient preparation technology. The quality conservation is satisfied in the whole experimental process, the purity of the prepared coptis total alkaloid component is more than 85%, the total alkaloid yield is more than 95%, and the total alkaloid yield is higher than that of the prior art and is shown in table 1. The method has simple whole operation process, is environment-friendly, can be recycled and reused due to the small dosage of the used eluent and the regenerant, low-concentration ethanol and the like, saves the cost, and can be used for large-scale production. Provides a good technical scheme for the basic research of the alkaloid component substances and the development of the traditional Chinese medicine preparation, which are safe, effective and controllable.
TABLE 1 comparison of Total alkaloid content and yield obtained in the examples of the invention and the prior art preparation methods
| Preparation method | Content of total alkaloids of Coptidis rhizoma (%) | Total base yield (%) |
| Example 1 | 87.76 | 95.39 |
| Example 2 | 86.05 | 97.65 |
| Example 3 | 90.34 | 98.35 |
| Example 4 | 88.65 | 97.78 |
| Example 5 | 85.48 | 95.66 |
| Prior Art | 60-75 | 50-85 |
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for refining total alkaloid components in a coptis extract is characterized by comprising the following steps:
(1) preparing a crude extract: cleaning, drying and crushing the coptis medicinal material, weighing 1-20Kg, adding water which is 5-50 times of the weight of the coptis medicinal material, extracting for 0.1-5h by using a decocting, refluxing, ultrasonic or microwave-assisted process, filtering the liquid medicine by using gauze or a 100-plus-mesh 300-mesh sieve, adding 5-50 times of water into dregs, extracting in the same way, filtering the liquid medicine, and removing the dregs; mixing the two medicinal liquids, performing rotary evaporation, concentration and drying to obtain Coptidis rhizoma crude extract dry powder;
(2) clarifying and filtering: weighing a proper amount of the coptis chinensis crude extract dry powder obtained in the step (1), dissolving the coptis chinensis crude extract dry powder with water of which the mass is 2-20 times that of the coptis chinensis crude extract dry powder, removing particles and pigments by using a filter membrane, and performing rotary evaporation concentration and drying on the obtained membrane permeate to obtain a dry product, namely clear dry powder;
(3) refining total alkaloids of coptis: dissolving the clear dry powder with water to a mass concentration of 5-300mg/mL, purifying by high performance liquid chromatography, collecting eluate, mixing the target components, concentrating, and drying to obtain refined Coptidis rhizoma powder product.
2. The method for refining total alkaloid component from Coptidis rhizoma extract as claimed in claim 1, wherein the water used in steps (1), (2) and (3) can be purified water, distilled water or water for injection.
3. The method for refining the total alkaloid component of coptis chinensis extract according to claim 1, wherein the drying manner in steps (1), (2) and (3) is microwave drying, freeze drying, vacuum drying or spray drying.
4. The method for refining the total alkaloid components in the coptis extract according to claim 1, wherein the extraction process in the step (1) is extraction at 80-180 ℃, reflux extraction at 60-100 ℃, ultrasonic extraction at 60-100 ℃ or microwave-assisted extraction at 60-100 ℃ by using an extraction tank or a decoction machine.
5. The method for refining the total alkaloid component of Coptidis rhizoma extract as claimed in claim 1, wherein the type of filter membrane used in step (2) is hollow fiber type, roll type, tube type or plate and frame type, and the area of the filter membrane used is 0.1-0.5m2The size of the filter membrane used is 0.1-0.65 μm or 500-1000kD, and the washing and filtering times in the membrane separation are 3-10 times.
6. The method for refining the total alkaloid component of coptis chinensis extract according to claim 1, wherein the membrane filtration technology used in step (2) is used to wash the remaining 30-100mL of the retentate by adding 1-30 times of 10% ethanol by adding 1-30 times of the volume of the retentate 3-10 times of water, and wash the remaining 30-100mL of the retentate by adding 10-80% ethanol by adding 1-30 times of the volume of the retentate 3-10 times of water.
7. The method as claimed in claim 1, wherein the high performance liquid chromatography used in step (3) comprises silica gel or high molecular polymer as the filler, one or more of n-alkane groups with 1-30 carbon atoms as the bonding phase, 8-100 μm in particle size, 60-300 Å in pore size, and 100-500m in specific surface area2(ii)/g; the inner diameter of the chromatographic column is 20-1600mm, and the height of the chromatographic column bed is 100-1000 mm.
8. The method for refining the total alkaloid component of coptis chinensis extract according to claim 1, wherein the high performance liquid chromatography used in step (3) is characterized in that an eluent contains 5 to 90 percent of organic solvent and acid, wherein the organic solvent is methanol, ethanol or acetonitrile, and the acid is phosphoric acid, formic acid or acetic acid, and the concentration of the acid is 0.01 to 0.5 percent.
9. The method as claimed in claim 1, wherein the HPLC technique used in step (3) is used, the loading amount of the chromatographic column is 0.01-20.0% of the mass of the stationary phase, the loading and elution flow rates are 140-350cm/h, and the elution conditions are isocratic or linear.
10. The component of total alkaloids extracted from coptis chinensis prepared by the refining method according to claims 1-9.
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| CN113082094A (en) * | 2021-04-26 | 2021-07-09 | 上海中医药大学 | Rhizoma coptidis aqueous extract and application of components thereof in preparation of medicine for preventing and treating diabetes |
| CN114573446A (en) * | 2020-12-02 | 2022-06-03 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus igniarius medicinal material |
| CN114588203A (en) * | 2020-12-02 | 2022-06-07 | 中国科学院大连化学物理研究所 | A kind of chromatographic purification and purification process method for purifying Guanxinquanfang |
| CN115575555A (en) * | 2022-09-16 | 2023-01-06 | 中国烟草总公司郑州烟草研究院 | A multi-stage membrane separation coupling chromatographic online sampling device |
| CN115813984A (en) * | 2022-12-30 | 2023-03-21 | 中国中医科学院中药研究所 | Extraction method of alkaloid in Xuanhuanglian, alkaloid extract and application and pharmaceutical composition thereof |
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| CN114573446A (en) * | 2020-12-02 | 2022-06-03 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus igniarius medicinal material |
| CN114588203A (en) * | 2020-12-02 | 2022-06-07 | 中国科学院大连化学物理研究所 | A kind of chromatographic purification and purification process method for purifying Guanxinquanfang |
| CN114573446B (en) * | 2020-12-02 | 2023-05-30 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus linteus medicinal material |
| CN113082094A (en) * | 2021-04-26 | 2021-07-09 | 上海中医药大学 | Rhizoma coptidis aqueous extract and application of components thereof in preparation of medicine for preventing and treating diabetes |
| CN115575555A (en) * | 2022-09-16 | 2023-01-06 | 中国烟草总公司郑州烟草研究院 | A multi-stage membrane separation coupling chromatographic online sampling device |
| CN115813984A (en) * | 2022-12-30 | 2023-03-21 | 中国中医科学院中药研究所 | Extraction method of alkaloid in Xuanhuanglian, alkaloid extract and application and pharmaceutical composition thereof |
| CN115813984B (en) * | 2022-12-30 | 2023-11-24 | 中国中医科学院中药研究所 | Extraction method of alkaloid in rhizoma coptidis, alkaloid extract, application of alkaloid extract and pharmaceutical composition |
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