CN117582464A - Extraction method and application of active ingredients for treating cerebral infarction in Va drug Nie Liang administration - Google Patents
Extraction method and application of active ingredients for treating cerebral infarction in Va drug Nie Liang administration Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/69—Polygalaceae (Milkwort family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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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
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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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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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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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
- A61K2236/55—Liquid-liquid separation; Phase separation
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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
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Abstract
The invention discloses an extraction method and application of an effective component for treating cerebral infarction in the middle-jiao of a host of a medicine Nie Liang, and belongs to the field of medicine in the general field. The invention discloses a method for extracting an effective component in the administration of Va Nie Liang, which can effectively extract the effective component in the administration of Nie Liang and is convenient to use. The extraction method is simple, industrial production is easy to realize, and when the active ingredients prepared by the invention are applied to clinically treating cerebral infarction, the invention has the advantages of quick response, good curative effect, small side effect, convenient administration and the like.
Description
Technical Field
The invention belongs to the application of the field of Va medicine, and particularly relates to an extraction method of an effective component for treating cerebral infarction in the middle warmer of a Va medicine Nie Liang and application thereof.
Background
The medicine Nie Liang is given by the academic name Japanese polygala, namely Japanese polygala, largetrifoliolious bugle herb, glaucescent fissistigma root, tenuifolia, golden key and herba schizophragmatis integrifolium, is a polygala plant of polygalaceae, has the effects of eliminating phlegm, relieving cough, promoting blood circulation, reducing swelling, detoxifying and relieving pain, and is commonly used for treating symptoms such as excessive cough phlegm, sore throat, external treatment of traumatic injury, furuncle, swelling, snake bite and insect bite; nie Liang has the effects of strengthening tendons and bones and tonifying yang; modern pharmacological researches show that the melon seed hardware has the effects of treating acute upper respiratory tract infection, resisting depression and regulating lipid metabolism, and has an improvement effect on Alzheimer's disease and Parkinson.
Cerebral infarction is also called cerebral infarction and ischemic cerebral apoplexy, and refers to a disease caused by ischemic and anoxic pathological changes and necrosis of brain tissue due to local brain tissue blood supply disorder caused by various reasons, so as to generate clinical corresponding nerve function deficiency. The physician considers that the paralysis of the limbs and the inability to speak caused by cerebral infarction are weak manifestations, the treatment is that the disease is deficient and strong when the disease is normal, and the Japanese polygala has the effects of strengthening tendons and bones and strengthening yang, so that the Japanese polygala possibly has a certain treatment effect on cerebral infarction.
At present, no clinical research and report on medicines of the Japanese polygala in the aspect of cerebral infarction treatment exist in the field, and the effective components in the Japanese polygala and the mechanism for treating cerebral infarction are not clear, so that the extraction method of the effective components for treating cerebral infarction in the administration of the drug Nie Liang is provided, and the research of the disease treatment mechanism of the Japanese polygala on cerebral infarction according to the effective components becomes a problem to be solved urgently in the field.
Disclosure of Invention
In view of the technical defects, the invention discloses an extraction method and application of an effective component for treating cerebral infarction in the administration of a relief drug Nie Liang, and a foundation is laid for exploring the action mechanism of Nie Liang on treating cerebral infarction by extracting the effective component in Nie Liang.
The extraction method of the active ingredients for treating cerebral infarction in the administration of the Va drug Nie Liang comprises the following steps:
s1: drying the harvested polygala japonica to obtain a dried polygala japonica raw material;
s2: crushing Japanese polygala, adding an entrainer for leaching and filtering to obtain filter residues and filtrate;
s3: extracting the filtrate obtained in the step S2 to obtain an extract, and drying the extract to obtain an extract;
s4: carrying out alcohol extraction treatment on the filter residue obtained in the step S2, concentrating, adsorbing and eluting the extracting solution, and collecting the eluent to obtain an extract;
s5: purifying the extractum obtained in S3 and S4, detecting and collecting target components, and recovering reagent to obtain the target product saponin.
Further, the specific operation in S2 is as follows:
the crushing is that the powder is crushed to 80-100 meshes, and the entrainer is added and leached for 2-4 hours at room temperature;
the entrainer includes, but is not limited to, one or more of methanol, ethanol, acetone, and ethyl acetate;
the filtering is filtering by using a ceramic membrane to obtain filter residues and filtrate.
Further, the specific operation steps of S3 are as follows: the extraction comprises feeding the filtrate into supercritical extraction kettle, extracting under 15-20MPa at 40-60deg.C, and introducing liquid CO 2 And entrainer for critical extraction for 2-3h, and resolving the extract at 50-60deg.C under 5-10MPa
Further, the specific operation in S4 is:
adding 70-90% ethanol solution into the filter residue, extracting with microwave, concentrating the extractive solution, and adding macroporous resin for adsorption;
eluting with 3-7 times of 40-70% ethanol solution, collecting eluate, and concentrating to obtain extract.
Further, the specific operation method in S5 is as follows:
purifying the extractum in S3 and S4 by adopting high-speed countercurrent chromatography, detecting by an evaporative light scattering detector, collecting target components, recovering reagents, and drying under reduced pressure to obtain a target product.
The use of Nie Liang extracted by any of the above methods in medicine.
The beneficial effects of the technical scheme are that: the radix Polygalae contains three mushroom saponins, resin, fatty oil, polygalagenin and tetraacetate; the aerial parts contain the polygalasaponin A, B, C, T and the melon seeds Jin Zaodai I-XIX; she Hanshan Kaempferol-3-O-6/-O- (3-hydroxy-3-methyl-glutaryl) glucoside, astragalin, kaempferol 3- (6 "-acetyl) glucoside, kaempferol 3, 7-diglucoside.
Pharmacological researches show that the melon seed hardware has obvious anti-inflammatory effect; after cerebral ischemia, a strong inflammatory reaction can be generated, and secondary damage to brain tissues is caused, so that early anti-inflammatory treatment can reduce the degree of brain damage.
The invention applies the Japanese polygala to the clinical treatment of cerebral infarction, and has the following beneficial effects
The effect is quick: all cases showed improvement of symptoms after taking herba Polygalae Japonicae granule for 4 weeks;
the curative effect is good: through clinical case observation, the Japanese polygala has curative effect on 98 percent of patients;
the side effect is small: the polygala japonica is applied to the treatment of cerebral infarction, has almost no side effect, and overcomes the defect of large side effect of western medicines or prescription medicines;
therefore, the polygala japonica has a certain treatment effect on cerebral infarction, the invention discloses an extraction method of the effective components in the polygala japonica, lays a foundation for the mechanism exploration of the polygala japonica for treating cerebral infarction, and provides a thought for preparing a medicine for treating cerebral infarction by taking the effective components of the polygala japonica as raw materials.
Detailed Description
The technical solutions of the present invention will be clearly and completely described below in connection with specific embodiments, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1 decoction prepared from Japanese polygala
S1: weighing quantitative herba Polygalae Japonicae, adding purified water according to the weight ratio of 1:1.3, soaking for 1 hr until the medicinal materials become soft, and expanding to restore to its natural state.
S2: adding purified water into the soaked polygala japonica according to the weight ratio of 1:5 for first decoction for 30min; filtering the residue to obtain medicinal liquid;
s3: adding purified water into the medicine residues according to the weight ratio of the medicine residues to the water of 1:3, and performing secondary decoction for 40min; filtering the residue to obtain medicinal liquid;
s4: mixing the decoctions.
Example 2A granule prepared from Japanese polygala
S1: weighing herba Polygalae Japonicae, adjuvants and wetting agent; wherein the auxiliary materials are soluble starch, dextrin, microcrystalline cellulose, mannitol and beta-cyclodextrin, and the proportion of the guar gold to the auxiliary materials is as follows: 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5; the wetting agent is 65% ethanol, 95% ethanol, 98% ethanol, and anhydrous ethanol;
s2: melon seeds Jin Jingao and beta-cyclodextrin are uniformly mixed according to the proportion of 1:3.5, absolute ethyl alcohol is selected as a wetting agent to prepare a soft material, the soft material is sieved by a 20-mesh sieve to prepare particles, the particles are dried to constant weight at 60 ℃, and the particles are sieved by a first sieve and a fifth sieve to prepare particles.
Example 3 extraction method of effective ingredient in Japanese polygala
S1: pulverizing dried Japanese polygala to 80-100 mesh, adding entrainer, leaching at room temperature for 2-4 hr, and ceramic membrane filtering to obtain filter residue and filtrate;
s2: the filtrate enters a supercritical extraction kettle, the extraction pressure is 15-20MPa, the liquid CO2 and entrainer are introduced at the temperature of 40-60 ℃ for critical extraction for 2-3h, and the extract is resolved at the temperature of 50-60 ℃ under the pressure of 5-10 MPa; the extract is dried to obtain extract;
s3: extracting the residue with 70-90% ethanol solution under microwave, concentrating the extractive solution, adsorbing with macroporous resin, eluting with 3-7 times of 40-70% ethanol solution, collecting eluate, and concentrating to obtain extract;
s4: purifying the extract by high-speed countercurrent chromatography, detecting by an evaporative light scattering detector, collecting target components, recovering reagent, and drying under reduced pressure to obtain target product saponin;
preferably, the entrainer includes, but is not limited to, one of methanol, ethanol, acetone, and ethyl acetate.
Experimental example 1
This example prepared Japanese polygala granule according to example 2 and applied to the clinical treatment of cerebral infarction.
The patients with cerebral infarction take the guarseed gold granules every day, and the curative effect observation and statistics of clinical cases are carried out;
1) Case object: clinical observation shows that NIHSS integral is 5-20 minutes for patients with cerebral infarction recovery blood stasis, 47 cases are present, 25 cases are men and women are 15 cases, the age is 18-80 years, the onset time is 14-180 days, 2 cases fall off, and 45 cases are observed to end;
2) Symptoms and efficacy criteria: evaluating clinical efficacy of the patient by using stroke disease diagnosis scores, NIHSS scale and Barthel index;
3) The medicine taking mode is as follows: the Japanese polygala granule is taken after warm water for half an hour after breakfast, midday and supper, the dosage is 4-8 g/day according to individual and disease difference, and 8 weeks is a treatment course;
4) Curative effect results: 45 cases after the treatment course is finished, the effective rate is 98%, the clinical remission rate is 7%, and the obvious efficiency is 58%;
and (3) calculating curative effect index: efficacy evaluation criteria of Chinese medicine symptoms (efficacy evaluation criteria referring to the guidelines of clinical research on New Chinese medicine): efficacy index = (pre-treatment integral-post-treatment integral)/(pre-treatment integral x 100%;
(1) clinical alleviation: before and after administration, symptoms and signs are obviously improved (the curative effect index is more than or equal to 95 percent);
(2) the effect is shown: after taking the medicine, the symptoms and the signs are obviously improved (the curative effect index is less than or equal to 70 percent and less than 95 percent);
(3) the method is effective: after taking the medicine, symptoms and signs are improved (the curative effect index is less than or equal to 30 percent and less than 70 percent);
(4) invalidation: after administration, symptoms and signs were not significantly reduced or aggravated (efficacy index < 30%);
5) Neurological deficit score:
scoring criteria:
basic cure: the score for neurological deficit decreases by 91% -100% and the disabling level is 0;
the method has the advantages that: the score for neurological deficit is reduced by 46% -90% and the disability level is 1-3;
improvement: the score for neurological deficit decreases by 18% -45%;
no change: a decrease or increase in the neurological deficit score of <17%;
deterioration: a >18% increase in neurological deficit score;
statistical results: of the 45 cases, 7 cases were basically healed, 31 cases were significantly improved, 6 cases were improved, and 1 case was unchanged.
Conclusion: the Japanese polygala can effectively improve the symptom of nerve function defects such as limb hemiplegia, language disorder and the like of patients with the blood stasis and collateral obstruction syndrome in the recovery period of cerebral infarction, has a certain treatment effect on cerebral infarction, and contains effective components for treating cerebral infarction.
Experimental example 2
Acute toxicity test
1) The experimental method comprises the following steps: taking 40 healthy SD rats with the maximum dosage method, wherein the healthy SD rats are male and female; the male and female parts are randomly divided into 2 groups according to sex and weight, and each group comprises 20 male and female halves; a first group, a normal control group; the second group, japanese polygala group, has a drug administration concentration of 0.167 g.ml -1 The maximum feasible administration volume is 20 ml.kg -1 The administration was carried out at a total dose of 6.68 g.kg -1 ·d -1 . Dosing was performed once each of the morning and afternoon on the same day, 6h between doses, and 14d was observed continuously after the last dose.
2) Results
a. General symptomatic observation: animals did not die during the trial period, and animals showed only reduced voluntary activity after dosing and recovery of symptoms 30min after dosing;
b. the mass of the body: before administration, there was no statistical difference (P > 0.05) in the animal mass of the polygala group compared to the vehicle control group. After administration, the animal mass of melon seeds Jin Zu (female) 1 and 3d is obviously reduced (P is more than 0.05) compared with that of a solvent control group, and the animal mass is recovered to be normal after 5d after administration; the quality of the animals in the Japanese polygala group (female) showed no statistical difference (P > 0.05) compared with the vehicle control group;
c. general section: the surviving rats at the end of the test were generally dissected and examined for no obvious abnormalities with the naked eye and no obvious toxic target organs were found.
3) Conclusion(s)
Under the test condition, the maximum dosage method is adopted, after the gastric lavage administration, the autonomous activity of the experimental animal and the quality of the male rats can be slightly influenced, and all symptoms can be gradually recovered to be normal without other obvious toxicity; the maximum dose (MFD) of guar gold to SD rats was 6.68 g.kg -1 Therefore, the herba Polygalae Japonicae has low toxicity, and can be applied to clinical treatment of cerebral infarction.
Experimental example 3 pharmacodynamics
1) Experimental method
Taking 75 healthy male SD rats, randomly dividing the healthy male SD rats into 5 groups of 15 rats; a first group, a Sham (Sham) group; a second group, a focal cerebral ischemia Model (MCAO) group; a third group of guarseed low dose groups; a fourth group of medium dose groups of guar gum; and a fifth group of high dose group of Japanese polygala. The Sham group and the MCAO group are respectively administered according to 1mL/100g gastric lavage physiological saline, and the Japanese polygala group is respectively administered according to 400mg crude drug/kg, 800mg crude drug/kg and 1600mg crude drug/kg gastric lavage. The medicine is taken once a day, and is continuously taken for 7 days, and the cerebral middle artery thrombosis method after Zea Longa improvement is adopted for modeling, ischemia is carried out for 2 hours, and reperfusion is carried out for 24 hours for detection.
2) Results
After 24h of cerebral ischemia reperfusion, neurological deficit scoring was performed, sham rats had no neurological dysfunction; MCAO group neurological deficit score was significantly elevated compared to Sham group; compared with the MCAO group, the nerve function score of the Japanese polygala high-dose group is obviously reduced;
the brain tissue water content result shows that compared with the Sham group, the brain tissue water content of the MCAO group rat is obviously increased; compared with the MCAO group, the water content of the brain tissue of the high-dose Japanese polygala group is obviously reduced;
TTC staining results show that the Sham rats have no infarct zone; compared with the Sham group, the cerebral infarction area of the MCAO group rat is obviously increased; compared with the MCAO group, the cerebral infarction areas of the low, medium and high dose groups of Japanese polygala are obviously reduced.
In conclusion, the cerebral infarction area of the rats taking the Japanese polygala is obviously reduced no matter the dosage is high, medium and low, so that the Japanese polygala contains the effective components for treating cerebral infarction.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.
Claims (6)
1. The extraction method of the effective components for treating cerebral infarction in the administration of the Va drug Nie Liang is characterized by comprising the following steps:
s1: drying the harvested polygala japonica to obtain a dried polygala japonica raw material;
s2: crushing Japanese polygala, adding an entrainer for leaching and filtering to obtain filter residues and filtrate;
s3: extracting the filtrate obtained in the step S2 to obtain an extract, and drying the extract to obtain an extract;
s4: carrying out alcohol extraction treatment on the filter residue obtained in the step S2, concentrating, adsorbing and eluting the extracting solution, and collecting the eluent to obtain an extract;
s5: purifying the extractum obtained in S3 and S4, detecting and collecting target components, and recovering reagent to obtain the target product saponin.
2. The extraction method according to claim 1, wherein the specific operations in S2 are:
the crushing is that the powder is crushed to 80-100 meshes, and the entrainer is added and leached for 2-4 hours at room temperature;
the entrainer includes, but is not limited to, one or more of methanol, ethanol, acetone, and ethyl acetate;
the filtering is filtering by using a ceramic membrane to obtain filter residues and filtrate.
3. The extraction method according to claim 1, wherein,
the specific operation steps of S3 are as follows: the extraction comprises feeding the filtrate into supercritical extraction kettle, extracting under 15-20MPa, and heatingIntroducing liquid CO at 40-60deg.C 2 And carrying out critical extraction for 2-3h by using entrainer, and resolving the extract at the temperature of 50-60 ℃ under the pressure of 5-10 MPa.
4. The extraction method according to claim 1, wherein the specific operations in S4 are:
adding 70-90% ethanol solution into the filter residue, extracting with microwave, concentrating the extractive solution, and adding macroporous resin for adsorption;
eluting with 3-7 times of 40-70% ethanol solution, collecting eluate, and concentrating to obtain extract.
5. The extraction method according to claim 1, wherein the specific operation method in S5 is as follows:
purifying the extractum in S3 and S4 by adopting high-speed countercurrent chromatography, detecting by an evaporative light scattering detector, collecting target components, recovering reagents, and drying under reduced pressure to obtain a target product.
6. Use of Nie Liang as an active ingredient extracted by the method of any one of claims 1 to 5 in a medicament for treating cerebral infarction.
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