CN105311119A - Refined rhizoma corydalis decumbentis total alkaloid product, preparation method thereof, pharmaceutical composition and application of refined rhizoma corydalis decumbentis total alkaloid product and pharmaceutical composition - Google Patents

Abstract

The invention discloses refined summer free total alkaloids, a preparation method, a pharmaceutical composition and applications. The present invention provides a summer-free refined product of total alkaloids, the percentages by weight of each component in the refined product of total alkaloids are as follows: original opioid: 15% to 40%; tetrahydropalmatine: 10% to 40%; Ling: 0-10%; Palmatine Hydrochloride: 0.01%-2%; Corydalis Ermin: 2%-10%; Chardonine: 15%-30%; Other alkaloids and extracts: 1%-5% %. The present invention establishes a more scientific and efficient high-purity summer-free total alkaloid extraction method, which is efficient, simple and easy to implement, and the HPLC purity of the prepared summer-free total alkaloid refined product is greater than 98%, which can play summer more safely and effectively No pharmacological effects of total alkaloids.

Description

Refined summer free total alkaloids, preparation method, pharmaceutical composition and application

technical field

The present invention relates to summer free total alkaloid refined product, preparation method, pharmaceutical composition and application.

Background technique

Summer Wu is the dried tuber or whole plant of Corydalis decumbens (Thunb.) Pers., a plant of the genus Corydalis of the family Papaveraceae. It is mainly used for the treatment of stroke and hemiplegia caused by high blood pressure and cerebrovascular disease, and has a good effect on rheumatoid arthritis, sciatica, and sequelae of polio. Its main active ingredients are total alkaloids of isoquinolines. Literature data show that more than 30 isoquinoline alkaloids have been isolated from Xia Wu, mainly containing protopine, tetrahydropalmatine, bicuculline, and palmatine , jatrorrhizine and other alkaloids.

At present, the commonly used extraction and refining methods of summer non-alkali include water extraction and alcohol precipitation, percolation, alcohol extraction, and acid-base method. With the application of modern preparation technology, methods such as macroporous adsorption resin, ultrasound, ultrafiltration membrane separation, and supercritical _CO2 extraction are gradually applied to the extraction and refining of summer free total alkali. Mahonda et al. studied the process conditions for extracting total alkaloids in summer mushrooms by supercritical CO ₂ extraction. Taking extraction pressure, extraction temperature and extraction time as the investigation factors, taking the comprehensive score of total alkaloids, proto-opioid, and tetrahydropalmatine content in summer and the evaluation index of dry cream rate, the optimal process conditions were optimized by orthogonal experiments. Results: The optimal process conditions were extraction pressure 30MPa, extraction temperature 55℃, extraction time 2.5h. The total alkaloid content in the extract is about 67%, the original opioid content is about 17%, and the tetrahydropalmatine content is about 10%. (Ma Hongda, An Ye, Guo Tao, etc. Research on the process of extracting total alkaloids in summer by supercritical CO ₂ extraction method [J]. ×103～1.0×104 ultrafiltration membrane prepared summer without injection clear, good color, showing stronger efficacy. Zhu Caiqing, Yu Hua, Fan Qikun, et al. Purification of summer free total alkaloids by ultrafiltration membrane. Chinese Herbal Medicine, 2005, 36(9):1319-1325.

CN101058576 invention discloses a summer free total alkaloid extract and its preparation method, its preparation method is to use summer free as raw material, utilize supercritical _CO2 fluid extraction condition to extract, collect extract, dry to obtain summer free total alkaloid extract . The average content of summer free total alkaloids extracted by this method is 90% as determined by UV method, wherein the total alkaloids mainly include protopine, corydalidin, bicorkolin, palmatine hydrochloride, corydalis urmin, summer Without nicotine and other alkaloids, the content of the six main alkaloids accounts for 75% to 90% of the extract.

In CN101623332, the present invention discloses a preparation method of summer free total alkaloid acid salt freeze-dried product, using summer free supercritical _CO2 total alkaloid extract as raw material, adding acid aqueous solution to dissolve it, and obtaining summer free total alkaloid acid aqueous solution; after filtration or centrifugation, the clarified solution is freeze-dried to obtain the freeze-dried product without total alkaloid salts in summer.

Both of the above-mentioned patents provide a method for preparing summer free total alkaloids, but both have disadvantages. CN101058576 only provides the method of no supercritical extraction in summer without total organisms in summer, and the prepared extract contains oily components, which is difficult to dry, which hinders its subsequent development and application, and the extraction time and CO _The flow rate is limited, so it is difficult to apply to industrial production.

CN101623332B patent is based on CN101058576B the method of preparing the total alkaloid salt lyophilizate by using the summer non-supercritical extraction summer non-total alkaloid crude extract as raw material through acid solution treatment and freeze-drying to prepare the total alkaloid salt freeze-dried product. The content of alkali is more than 80%. This patent does not conduct further research on the enrichment and purification of the total summer alkaloids, and the purity of the prepared summer non-alcoholic extract is not high enough, which limits the application range of the total summer non-alkaloids.

Summer-free total alkaloid preparations are mainly used to treat senile dementia, cerebral infarction, pseudomyopia of primary and middle school students, etc., and have great market prospects, but the current extraction and separation technology of summer-free total alkaloids mostly uses acid extraction through resin columns method, the process is complicated, it takes a long time, and the purity of the total alkaloids obtained is not high. Therefore, it is an urgent technical problem to find an extraction method with simple process, high extraction rate, good purity of the extracted summer free of total alkaloids, and suitable for large-scale industrial production.

Contents of the invention

The technical problem to be solved by the present invention is to overcome the defects of the existing summer-free total alkaloid extraction method, such as cumbersome steps, low extraction rate, low purity of the extracted extract, high impurity content, unsuitable for medicinal use, and unsuitable for industrial production. , and provide a summer free total alkaloid refined product, preparation method, pharmaceutical composition and application. The present invention establishes a more scientific and efficient high-purity extraction method of summer free total alkaloids. Pharmacological effects of alkaloids.

The present invention provides a refined product without total alkaloids in summer, wherein the percentages by weight of each component in the refined product of total alkaloids are: protopine: 15%-40%; tetrahydropalmatine: 10%-40% ; Bikoukouling: 0-10% (excluding 0); Palmatine hydrochloride: 0.01%-2%; Corydalis Ermin: 2%-10%; Chardonine: 15%-30%; other biological Alkali and extract: 1% to 5%.

In the present invention, the refined summer free total alkaloids are measured by HPLC method, and the total alkaloid content is 60% to 100%, wherein the total content of the six main alkaloids accounts for 95% of the summer free total alkaloid refined products above.

The present invention also provides the preparation method of the summer free total alkaloid refined product, which comprises the following steps:

Step 1: use the tuber of summer fruit (Papaveraceae plant Viola corydalis), pulverize and alkalize to obtain alkalized summer fruit; in the alkalization, the quality of the alkali and the described summer fruit The ratio is 0.001-1, preferably 0.001-0.1.

Step 2: Extract and separate the alkalized summer free obtained in step 1 in an extractor coupled with supercritical _CO2 extraction and membrane separation, and dry to obtain refined summer free total alkaloids.

In step 1, the crushing is preferably crushed until the particle size of the summer grass (Papaceae plant Corydalis viticum) is 10 mesh to 60 mesh (that is, 1.74 mm to 0.273 mm). Described alkalization, preferably adopts inorganic base to carry out alkalization, and described inorganic base is preferably one or more in ammonia solution, calcium hydroxide, sodium carbonate and sodium bicarbonate, more preferably ammonia solution, described ammonia The mass concentration of the aqueous solution is preferably 1%-10%, and the mass concentration refers to the percentage of the mass of ammonia gas in the total mass of the ammonia solution. The ammonia solution can be obtained by mixing ammonia reagent with water, the volume ratio of the ammonia reagent to water is preferably 5% to 10%, and the volume ratio refers to the percentage of the volume of the ammonia reagent to the volume of water. The ammonia water reagent can be conventional commercially available ammonia water reagent in the field, and the concentration of the ammonia water reagent is preferably 2%-30%. The inorganic base is preferably used in the form of its aqueous solution, and the concentration of the aqueous solution of the inorganic base is preferably 1% to 10%, and the concentration refers to the percentage of the mass of the inorganic base in the total mass of the aqueous solution of the inorganic base.

In step 2, the time for the extraction and separation of the supercritical _CO2 is preferably 0.5 to 5h; the entrainer used during the extraction and separation of the supercritical _CO2 is preferably methanol and/or ethanol, more preferably ethanol, and the Ethanol is preferably an aqueous ethanol solution with a mass percentage of 95%, and the mass percentage refers to the percentage of the mass of ethanol in the total mass of the aqueous ethanol solution. The membrane in the "supercritical _CO extraction and membrane separation coupled extractor" is preferably an ultrafiltration membrane, a nanofiltration membrane, a reverse osmosis membrane or a hollow fiber membrane, more preferably a nanofiltration membrane; the nanofiltration membrane The pore size is preferably 1 nm to 3 nm. The pore size of the ultrafiltration membrane is preferably 10 nm to 40 nm. The pore size of the reverse osmosis membrane is preferably 1 nm to 5 nm. The pore diameter of the hollow fiber membrane is preferably 1 nm to 40 nm.

In step ₂ , described supercritical CO Extraction and the extractor of membrane separation phase coupling are according to the schematic diagram recorded in the book "New Technology for Extraction and Separation of Chinese Medicine" (April 2010, published by Chemical Industry Press, page P353) Assembled, the extraction apparatus coupled with supercritical CO ₂ extraction and membrane separation includes the following components: extraction device, separation device, high-pressure pump, membrane device, cooler and gas storage tank.

In the present invention, the method for preparing the refined summer free total alkaloids preferably further includes step 3, more preferably further includes step 3 and step 4, and still more preferably further includes step 3, step 4 and step 5.

Step 3: adjust the pH of the refined summer free total alkaloids obtained in step 2 to 2-5, extract by ultrasonic wave, and then filter by ultrafiltration or microfiltration membrane to obtain the purified summer free total alkaloid refined solution.

Step 4: adjusting the pH of the refined summer free total alkaloid solution obtained in step 3 to 8-12, filtering, washing the solid phase with water until neutral, and obtaining further purified summer free total alkaloid refined product;

Step 5: Recrystallize the further purified refined product of summer without total alkaloids obtained in step 4 to obtain the refined product of summer without total alkaloids after recrystallization.

In step 3, the frequency of the ultrasonic extraction is preferably 30 Hz to 80 Hz, and the time of the ultrasonic extraction is preferably 0.5 hours to 3 hours. The pore size of the ultrafiltration or microfiltration membrane is preferably 0.3 micron to 0.8 micron; the pH adjustment is preferably adjusted with an acid, and the acid is preferably an inorganic acid and/or an organic acid, and the inorganic acid is preferably hydrochloric acid. Described organic acid is preferably tartaric acid. The acid is preferably used in the form of its aqueous solution. When the acid is used in the form of its aqueous solution, the mass percent concentration of the acid aqueous solution is preferably 1% to 10%, more preferably 1% to 5%. , the mass percentage refers to the percentage of the mass of the acid in the total mass of the aqueous acid solution. The volume ratio of the aqueous acid solution to the total alkaloid-free refined product obtained in step 2 is preferably 5-10, more preferably 5-8. Step 3 preferably includes a drying step, drying the solution of the purified summer summer without total alkaloids refined product to obtain the purified summer summer without total alkaloids refined product; the drying can be performed according to this type of operation in the art Conventional Methods and Conditions.

In step 4, the pH is preferably 9-11. The pH adjustment is preferably adjusted with an inorganic base, preferably sodium hydroxide or potassium hydroxide, the inorganic base can be used in the form of its aqueous solution, when the inorganic base is in the form of its aqueous solution When used, the mass concentration of the aqueous solution of the inorganic base is preferably 3% to 15%, more preferably 4% to 10%. The mass concentration refers to the percentage of the mass of the inorganic base to the total mass of the aqueous solution of the inorganic base. Step 4 preferably includes a drying step, and the drying can be performed according to conventional methods and conditions of this type of operation in the art.

In step 5, the recrystallization can adopt conventional methods and conditions of this type of operation in the art. The solvent used for the recrystallization is preferably an alcohol solvent and/or a halogenated hydrocarbon solvent, the preferred methanol and/or ethanol of the alcohol solvent, and the preferred chlorinated hydrocarbon solvent of the halogenated hydrocarbon solvent, so The above-mentioned chlorinated hydrocarbon solvent is preferably chloroform. Step 5 preferably includes a drying step, and the drying can be performed according to conventional methods and conditions for this type of operation in the art.

The present invention also provides the application of the refined product of summer free of total alkaloids in the preparation of medicines for treating and/or preventing neuron cell damage caused by cerebral ischemia or reperfusion.

The present invention also provides a pharmaceutical composition, which contains a therapeutically effective amount of the refined summer summer free of total alkaloids and a pharmaceutically acceptable carrier. In the present invention, the therapeutically effective amount is preferably 0.1%-99.9% by mass, and the mass percentage refers to the percentage of the total alkaloid-free refined product of summer in the total mass of the pharmaceutical composition. The pharmaceutically acceptable carrier is a conventional pharmaceutically acceptable carrier in the art.

The present invention also provides the application of the pharmaceutical composition in the preparation of medicines for treating and/or preventing neuronal cell damage caused by cerebral ischemia or reperfusion.

In the present invention, the diseases of neuron cell injury caused by cerebral ischemia or reperfusion include cerebral thrombosis and cerebral apoplexy.

On the basis of not violating common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive progress effect of the present invention is: the present invention has established a more scientific and efficient high-purity extraction method of summer free total alkaloids, the method is efficient, simple and easy to implement, and the HPLC purity of the prepared summer free total alkaloids is greater than 98 %, it can more safely and effectively exert the pharmacological effects of no total alkaloids in summer.

Description of drawings

Figure 1 is a schematic diagram of the _CO2 extraction process coupled with supercritical extraction and membrane separation.

detailed description

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions.

The supercritical _CO2 extraction and membrane-coupled extraction apparatus used in the following examples were assembled according to the schematic diagram shown in Figure 1.

Embodiment 1: Preparation of total alkaloid-free refined product in summer

Get the dry tuber that does not have in summer, pulverize, and obtain particle size and be 10 order ~ 40 order medicinal material powder; Get medicinal material powder 300g, add volume ratio and be 6% ammoniacal liquor as alkalizing agent (described volume ratio refers to that the volume of ammoniacal liquor reagent occupies ammonia The percentage of the total volume of the aqueous solution, the mass concentration of the ammonia reagent is 25% to 28%, the mass concentration refers to the percentage of the quality of ammonia gas in the total mass of the ammonia reagent), and the dosage is 50% of the weight of the summer non-medicinal material, soaked for 2h put medicinal material into supercritical _{CO2extraction} and membrane coupling (aperture is the ultrafiltration membrane of 0.8 micron) in the extractor, take methanol as entrainer, after extraction 30 minutes, stop extraction, obtain total alkaloid extract, dry, Instantly. The total alkaloid content determined by high performance liquid chromatography was 98.47%. As determined by HPLC, its composition in weight percent includes: proto-opiate: 34.89%, tetrahydropalmatine: 24.56%, bikkoling: 3.51%, palmatine hydrochloride: 1.84%, chardonine: 19.77%, Corydalis Ming: 9.58%, other alkali content 4.32%.

Embodiment 2: Preparation of total alkaloid-free refined product in summer

Get the dry tuber that does not have in summer, pulverize, and obtain the medicinal material powder that particle size is 20 order～40 order; Get medicinal material powder 300g, add volume ratio and be 10% ammoniacal liquor as alkalizing agent (described volume ratio refers to that the volume of ammoniacal liquor reagent occupies ammonia The percentage of the total volume of the aqueous solution, the mass concentration of the ammonia water reagent is 25% to 28%, and the mass concentration refers to the percentage of the quality of ammonia gas in the total mass of the ammonia water reagent), and the dosage is 60% of the weight of the summer non-medicinal material, soaked for 1h Medical material is dropped into supercritical _CO Extraction and membrane coupling (adopting aperture is the nanofiltration membrane of 2 nanometers) in the extractor, be 95% ethanol as entrainer with mass percent (described mass percent refers to the mass percent of ethanol accounts for The percentage of the total mass of ethanol aqueous solution), the extraction time is 1.5h, and the total alkaloid content determined by high performance liquid chromatography is 98.95%. As determined by HPLC, its composition by weight percentage includes: proto-opiate: 30.52%, tetrahydropalmatine: 28.93%, bikkoling: 0.01%, palmatine hydrochloride: 0.09%, chardonine: 25.64%, and Corydalis Ming: 9.73%, other alkali content 4.03%.

Embodiment 3: the preparation method of total alkaloid-free refined product in summer

Get the dry tuber that does not have in summer, make powder, and obtain particle size and be 10 order ~ 20 order medicinal material powder; Get medicinal material powder 200g, add volume ratio and be 7% ammoniacal solution as alkalizing agent (the volume ratio refers to the volume of ammonia water reagent accounts for The percentage of total volume of ammonia solution, the mass concentration of ammonia reagent is 25%～28%, described mass concentration refers to the percentage of the quality of ammonia gas accounting for the total mass of ammonia reagent), and the consumption is 50% of the summer non-medicinal weight, infiltrating 3h; the medicinal material is put into supercritical _CO2 extraction and membrane coupling (using a hollow fiber membrane with a pore size of 0.1 micron) in the extractor, with a mass percentage of 95% ethanol as an entrainer (the mass percentage refers to the quality of ethanol Accounting for the percentage of the total mass of ethanol aqueous solution), the extraction time was 40 minutes, and the mass percentage of the total alkaloid obtained was 1% hydrochloric acid aqueous solution 300mL (the described mass percentage refers to the quality of hydrogen chloride accounts for the percentage of the total mass of hydrochloric acid aqueous solution), ultrasonic ( Ultrasonic frequency is 80H _Z ) extraction 2h, 1.0 μm filter membrane filtration, add mass percentage and be 8% potassium hydroxide solution and adjust pH10～11 (the described mass percentage refers to the quality of potassium hydroxide accounts for the total mass of potassium hydroxide percentage), filtered to obtain a precipitate, and the high-performance liquid chromatography determined that the total alkaloid content was 98.88%. As determined by HPLC, its composition by weight percentage includes: proto-opiate: 35.68%, tetrahydropalmatine: 19.36%, bicorkolin: 4.64%, palmatine hydrochloride: 0.61%, chardonine: 28.71%, Corydalis Ming: 8.75%, other alkali content 1.13%.

Embodiment 4: the preparation method of total alkaloid-free refined product in summer

Get the dry tuber of summer without, powder, and obtain particle size and be 20 mesh～40 mesh without summer medicinal material powder; Get summer without medicinal material powder 300g, add volume ratio 10% ammoniacal liquor as alkalizing agent (described volume ratio refers to the ammoniacal liquor reagent Volume accounts for the percentage of the total volume of ammonia solution, the mass concentration of ammonia water reagent is 25%～28%, described mass concentration refers to the percentage of the quality of ammonia gas accounting for the total mass of ammonia water reagent), and consumption is 50% of the summer non-medicinal material weight , infiltrate 2.5h; Medical material is dropped into supercritical _CO Extraction and membrane coupling (adopting aperture is the composite nanofiltration membrane of 1 nanometer) in the extractor, be 95% ethanol as entrainer with mass percent (the described mass percent is Refers to the mass of ethanol as a percentage of the total mass of the aqueous ethanol solution), which is obtained after 1 hour of extraction. 500ml of 1% hydrochloric acid aqueous solution by mass percentage, ultrasonic (ultrasonic frequency 60H _Z ) extraction for 1h, 0.45 μm filter membrane filtration, adding mass percentage of 10% sodium hydroxide solution to adjust pH9～10 (the mass percentage refers to hydrogen The mass of potassium oxide accounts for the percentage of the total mass of potassium hydroxide), and the precipitate is obtained by filtration, and the total alkaloid content determined by high performance liquid chromatography is 98.68%. As determined by HPLC, its composition by weight percentage includes: proto-opiate: 20.68%, tetrahydropalmatine: 32.16%, bicorkolin: 8.84%, palmatine hydrochloride: 0.50%, chardonine: 28.71%, and Corydalis Ming: 6.75%, other bases 1.04%.

Embodiment 5: the preparation method of total alkaloid-free refined product in summer

Get the dry tuber of summer without, powder, and obtain particle size and be that 30 order～40 order summer without medicinal material powder; Get summer without medicinal material powder 8000g, add volume ratio and be 10% ammoniacal liquor as alkalizing agent (described volume ratio refers to ammoniacal liquor reagent The volume accounts for the percentage of total volume of ammonia solution, and the mass concentration of ammonia water reagent is 25%～28%, and described mass concentration refers to the percentage that the quality of ammonia gas accounts for the total mass of ammonia water reagent), and consumption is 70% of the summer non-medicinal material weight %, infiltrate 24h; Medical material is dropped into supercritical _CO Extraction and membrane coupling (adopting aperture is 15 nanometer reverse osmosis membrane) in the extractor, be 95% ethanol with mass percentage as entrainer (described mass percentage refers to ethanol The mass of the ethanol solution accounts for the percentage of the total mass of the ethanol aqueous solution), the extraction time is 1 h, and it is dried to obtain. Then use mass percent as 2% hydrochloric acid aqueous solution 300mL (the described mass percent refers to the percentage that the quality of hydrogen chloride accounts for the total mass of hydrochloric acid aqueous solution), ultrasonic (ultrasonic frequency 80H _Z ) extraction 0.5h, aperture is the filter membrane filtration of 0.1 micron , add mass percentage and be 10% sodium hydroxide solution and adjust pH9～10 (the described mass percentage refers to the percentage that the quality of sodium hydroxide accounts for the total mass of sodium hydroxide), filters to obtain precipitation, recrystallizes with methanol, high-efficiency liquid phase The total alkaloid content determined by chromatography is 99.88%. As determined by HPLC, its composition by weight percentage includes: proto-opiate: 29.28%, tetrahydropalmatine: 23.83%, bicorkolin: 8.52%, palmatine hydrochloride: 1.90%, chardonine: 29.71%, and Corydalis Ming: 4.62%, other bases 2.02%.

Claims (17)

1. A refined product without total alkaloids in summer, characterized in that: the percentages by weight of each component in the refined product of total alkaloids are: former opioid: 15% to 40%; tetrahydropalmatine: 10% to 40% ; Bikoukouling: 0-10% (excluding 0); Palmatine hydrochloride: 0.01%-2%; Corydalis Ermin: 2%-10%; Chardonine: 15%-30%; other biological Alkali and extract: 1% to 5%. 2. the preparation method of the total alkaloid refined product of summer tea as claimed in claim 1, is characterized in that: step 1: with the tuber of summer tea, crushing, alkalization, obtains summer tea after alkalization, described alkali The mass ratio with the described summer substance is 0.001～1; Step 2: Extract and separate the alkalized summer free obtained in step 1 in an extractor coupled with supercritical _CO2 extraction and membrane separation, and dry to obtain the refined summer free total alkaloids. 3. The method for preparing the refined total alkaloids of summer powder as claimed in claim 2, characterized in that: in step 1, the pulverization is to crush until the particle size of summer powder is 10 mesh to 60 mesh; and / or, In step 1, the alkalization is carried out with an inorganic base; and / or, In the alkalization described in step 1, the mass ratio of the alkali to the summer moss is 0.001-0.1. 4. the preparation method of summer as claimed in claim 3 has no total alkaloid refinement, it is characterized in that: in step 1, described inorganic alkali is one in aqueous ammonia solution, calcium hydroxide, sodium carbonate and sodium bicarbonate One or more kinds; when ammonia solution is used for alkalization, the mass concentration of the ammonia solution is 1% to 10%, and the mass concentration refers to the percentage of the mass of ammonia gas in the total mass of the ammonia solution. 5. The method for preparing refined summer free total alkaloids as claimed in claim 2, characterized in that: in step 2, the extraction and separation time is 0.5 to 5 hours; and / or, In step 2, the entrainer used during the extraction and separation is methanol and/or ethanol; and / or, In step 2, the membrane in the "extractor coupled with supercritical _CO2 extraction and membrane separation" is an ultrafiltration membrane, nanofiltration membrane, reverse osmosis membrane or hollow fiber membrane. 6. The preparation method of the refined product without total alkaloids in summer as claimed in claim 2, characterized in that: the preparation method of the refined product without total alkaloids in summer also includes step 3, step 3: the obtained in step 2 The refined summer without total alkaloids is adjusted to a pH of 2-5, ultrasonically extracted, and then ultrafiltered or microfiltered to obtain the purified summer without total alkaloids refined solution. 7. The method for preparing refined summer free total alkaloids as claimed in claim 6, characterized in that: in step 3, the frequency of the ultrasonic extraction is 30 Hz to 80 Hz; and / or, In step 3, the time of the ultrasonic extraction is 0.5 hours to 3 hours; and / or, In step 3, the pore size of the ultrafiltration or microfiltration membrane is 0.3 micron to 0.8 micron; and / or, In step 3, the pH adjustment is adjusted with an acid, and the acid is an inorganic acid and/or an organic acid. 8. the preparation method of summer free total alkaloid refined product as claimed in claim 7, is characterized in that: in step 3, described inorganic acid is hydrochloric acid; and / or, In step 3, described organic acid is tartaric acid; and / or, In step 3, the acid is used in the form of its aqueous solution, and when the acid is used in the form of its aqueous solution, the mass percent concentration of the acid aqueous solution is 1% to 10%; the mass The percentage refers to the percentage of the quality of the acid in the total mass of the aqueous acid solution; the volume ratio of the aqueous acid solution to the refined product obtained in step 2 without total alkaloids in summer is 5 to 10; and / or, Step 3 includes a drying step, drying the solution of the purified summer free total alkaloids to obtain the purified summer free total alkaloids. 9. The preparation method of the refined product without total alkaloids in summer as claimed in claim 6, characterized in that: the preparation method of the refined product without total alkaloids in summer also includes step 4, step 4: the obtained in step 3 The pH of the refined summer free of total alkaloids solution is adjusted to 8-12, filtered, and the solid phase is washed with water until neutral to obtain further purified refined summer free of total alkaloids. 10. The method for preparing refined products without total alkaloids in summer as claimed in claim 9, characterized in that: in step 4, the pH is 9-11; and/or, in step 4, the pH adjustment Use inorganic base to adjust. 11. the preparation method of total alkaloid-free refined product in summer as claimed in claim 10, is characterized in that: in step 4, described inorganic base is sodium hydroxide or potassium hydroxide; and / or, In step 4, the inorganic base is used in the form of its aqueous solution, and when the inorganic base is used in the form of its aqueous solution, the mass concentration of the aqueous solution of the inorganic base is 3% to 15%; the Mass concentration refers to the percentage that the quality of inorganic base accounts for the total mass of inorganic base aqueous solution; and / or, Step 4 includes a drying step. 12. The method for preparing the refined product without total alkaloids in summer as claimed in claim 9, characterized in that: the method for preparing the refined product without total alkaloids in summer also includes step 5, step 5: the obtained in step 4 After further purification, the refined summer without total alkaloids was recrystallized to obtain the refined summer without total alkaloids after recrystallization. 13. The method for preparing refined summer free total alkaloids as claimed in claim 12, characterized in that: in step 5, the solvent used in the recrystallization is an alcohol solvent and/or a halogenated hydrocarbon solvent. 14. the preparation method of summer free total alkaloid refined product as claimed in claim 13, is characterized in that: in step 5, described alcoholic solvent is methanol and/or ethanol; and / or, In step 5, the halogenated hydrocarbon solvent is a chlorinated hydrocarbon solvent, preferably chloroform. 15. A pharmaceutical composition, characterized in that: it contains a therapeutically effective amount of the summer free total alkaloid refined product as claimed in claim 1 and a pharmaceutically acceptable carrier; the preferred mass percentage of the therapeutically effective amount is 0.1 % to 99.9%, the mass percentage refers to the percentage of the total alkaloid-free refined product of summer in the total mass of the pharmaceutical composition. 16. summer as claimed in claim 1 without total alkaloid refined product or pharmaceutical composition as claimed in claim 15, in the preparation treatment and/or the disease of the neuron cell injury that prevention cerebral ischemia or reperfusion cause application in medicine. 17. The application according to claim 16, characterized in that the diseases of neuron cell damage caused by cerebral ischemia or reperfusion include cerebral thrombosis and cerebral apoplexy.