CN101683363B - Composition preparation of ginkgo biloba extract and dipyridamole and preparation method thereof - Google Patents
Composition preparation of ginkgo biloba extract and dipyridamole and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a composition preparation of ginkgo biloba extract and dipyridamole and preparation method thereof, which is a traditional Chinese medicine injection preparation prepared by combining 20 to 100 parts of ginkgo biloba extract and 2 to 10 parts of dipyridamole. Wherein the ginkgo biloba extract is obtained by proceeding respective alcohol extraction and water extraction to ginkgo biloba medicinal material, the extracting solution is then concentrated, micro-filtration by inorganic ceramic film, refining by polyamide resin and then being made by organic film ultrafiltration. Comparing with the prior art, the invention adopts a method of membrane separation technology combined with resin refining treatment to proceed purifying, separating and condensation to the extract, active ingredient is enriched, physical method is used to replace conventional chemical method can maintain characteristics of traditional Chinese medicine compound preparation better, which reduces adverse effects of drugs and improves curative effect, and thereby quality of the product is improved and assured.
Description
Technical field: the present invention relates to composite preparation of a kind of Folium Ginkgo extract and dipyridamole and preparation method thereof, belong to technical field of traditional Chinese medicine pharmacy.
Background technology: traditional medicine Injectio is to be raw material with the Chinese crude drug, the sterilization preparation of making through steps such as extracting, separate, be refining.It has changed the traditional administering mode of Chinese medicine, and it is rapid to possess drug effect, the bioavailability height, advantages such as dosage is more accurate, be applicable to the case of unsuitable oral or emergency and severe disease, and can make some drugs performance orientation or site-specific delivery of drugs, brought into play the effect of treatment by Chinese herbs acute disease serious symptom and difficult disease preferably.But, increasing along with clinical practice, Chinese medicine causes that the incident of adverse effect occurs repeatedly, and has caused vast medical worker's concern.The main cause of traditional medicine Injectio generation untoward reaction is: the chemical analysis of Chinese medicine raw material is very complicated, and materials such as some of them such as tannin, acid very easily cause allergic reaction; In addition, because the production technology of Chinese medicine owes maturation, cause indivedual lot number medicinal liquid purity not good enough, and then cause allergic reaction.
Ginkgo Leaf Extract and Dipyridamole Injection is the herbal species that the applicant develops, make by Folium Ginkgo extract and dipyridamole, can the expansion artery blood vessel, reduce the blood vessel wall permeability, improve edema, reduce ischemia viscosity, suppress platelet and erythrocytic high aggregation, reduce myocardial oxygen consumption, strengthen myocardial cell to the toleration of hypoxic-ischemic etc., it is evident in efficacy to treat cerebral thrombosis, cerebral vasospasm, ischemic heart desease, peripheral blood vessel etc. clinically.Record in " National Drug Administration's national drug standards " first 213 pages.Wherein Folium Ginkgo extract is made through extracting processing by Folium Ginkgo, contains multiple composition in the Folium Ginkgo, comprises total flavones glycoside, terpenoid, alkyl phenols, polyene alcohols and alkaloid, sugar, starch, protein and inorganic salt.To effective ingredient such as the total flavone glucoside in the Folium Ginkgo and total terpene lactoness, present extraction process has a variety of, general normal employing organic solvent extraction, resin or steps such as activated carbon adsorption, precipitate and separate, new methods such as supercritical carbon dioxide extraction method and high-speed countercurrent chromatography extraction method have appearred again recently, but organic solvent kind that back two kinds of methods are introduced and consumption are complicated, simultaneously solvent has residual allergy and the toxicity of causing easily, and this is for big production and to put on market be totally unfavorable.
Application number is that 200410039123.2 patent documentation discloses a kind of ginkgo damo powder injection and preparation method thereof, and the preparation method of its Folium Ginkgo extract is that ethanol extract from water precipitation is just carried, and purification by macroporous resin obtains Ginkgo total flavones, CO then
2Supercritical extraction obtains Ginkgo total lactones, and the effective ingredient that this method obtains increases, but complex operation has all greatly influenced obtaining effective ingredient to the temperature in the purifying process, pressure, flow and choice of Solvent etc.; Owing to the component complexity of Chinese crude drug extracting solution, wherein except effective ingredient, also contain a large amount of impurity simultaneously, the structure difference of each component, molecular size is also different.Therefore study a kind of easy and simple to handle, remove impurity good separating effect, can not destroy active ingredient, the technology that be suitable for big production, saves production cost is the problem that the inventor need solve.
Summary of the invention:
The applicant finds the thick desilver Folium Pruni of the method that adopts alcohol, moisture to carry under study for action, more can guarantee the extraction fully and the utilization of effective ingredient, adopts the ceramic membrane filter method of not destroying active ingredient simultaneously, then shortens process cycle greatly, enhances productivity.
Therefore, the objective of the invention is to: composite preparation of a kind of Folium Ginkgo extract and dipyridamole and preparation method thereof is provided.Said preparation is an ejection preparation, comprises liquid drugs injection, transfusion, powder pin and freeze-dried powder.The present invention be with the Folium Ginkgo medical material carry out alcohol, moisture is carried, adopting membrane separation techniques such as inorganic ceramic membrane microfiltration, organic membrane ultrafiltration and resin purification to handle the method that combines then carries out purification, separates and concentrates extracting solution, obtain Folium Ginkgo extract, mix with dipyridamole then, mixed active component is prepared into different ejection preparations according to conventional method.
The present invention is achieved in that according to composition by weight and calculates, it is made as follows by Folium Ginkgo extract 20-100 part, dipyridamole 2-10 part: get Folium Ginkgo and pulverize, use alcohol reflux, extracting solution reclaims ethanol, and it is standby to get the Folium Ginkgo alcohol extract; The medicinal residues water extraction, extracting solution mixes with aforementioned alcohol extract, and mixed liquor suitably concentrates, and filters with inorganic ceramic film, the rate of filtration is 10-15L/h, filtered liquid medicine is directly adsorbed by the resin column of having handled well, and effluent is discarded, is that deionized water or the 1%-30% ethanol of 2-6 is washed post with pH value, the reuse pH value is the 60%-95% ethanol elution of 4-6, collect color and divide eluent than the deep, reclaim ethanol, concentrate drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 5%-40% ethanol, and stirring and dissolving is carried out ultrafiltration then, and filtrate concentrates, and drying is mixed with dipyridamole, is dissolved in water, and adds or do not add adjuvant, makes different ejection preparations with conventional method.
Concrete preparation method is: get Folium Ginkgo and pulverize, use 40%-80% soak with ethanol 8-14 hour, reflux, extract, is 2-4 time then, adds ethanol 6-12 at every turn and doubly measures, extracts 1-3 hour, and merge extractive liquid, reclaims ethanol, and it is standby to get the Folium Ginkgo alcohol extract; Medicinal residues add 4-6 times of water gaging and extract 2-4 time, each 0.5-2 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, filtered liquid medicine is directly adsorbed by the resin column of having handled well, effluent is discarded, with the speed of per second 0.5-2ml, is that 1-5 times of column volume deionized water or the 1%-30% ethanol of 2-6 is washed post with pH value, the reuse pH value is the 60%-95% ethanol elution of 4-6, collect color and divide eluent, reclaim ethanol, concentrate with vacuum evaporator than the deep, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 5%-40% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, filtrate concentrates with vacuum evaporator, and drying is mixed with dipyridamole, be dissolved in water, add adjuvant or do not add adjuvant, make different ejection preparations with conventional method.
Preferred manufacturing procedure is: get Folium Ginkgo and pulverize, with 60% soak with ethanol 12 hours, reflux, extract, was 3 times then, and add 9 times of amounts of ethanol at every turn, extracted 2 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 5 times of water gagings and extract 3 times, each 1 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, add the water washing effective ingredient of 2-3 times of medicine liquid volume amount during filtration several times, filtered liquid medicine is directly adsorbed by the resin column of having handled well, and effluent is discarded, speed with per second 1ml, with pH value is that 2 times of column volume deionized waters or 10% ethanol of 2-6 is washed post, and the reuse pH value is 80% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 20% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, filtrate concentrates with vacuum evaporator, and drying is mixed with dipyridamole, be dissolved in water, add adjuvant or do not add adjuvant, make different ejection preparations with diverse ways.
Composite preparation of Folium Ginkgo extract of the present invention and dipyridamole and preparation method thereof calculates according to composition by weight, and it is to be made as stated above for 4 parts by 40 parts of Folium Ginkgo extract, dipyridamole.
The purification resin column is meant CD180 post, DA201 post, D101 post, polyamide resin column among the present invention, preferred DA201 post.
Wherein, hydro-acupuncture preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration is mixed with dipyridamole,, regulate pH to 4.5-6.0 with the water for injection dissolving, and embedding, sterilization, promptly.
Infusion preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds sodium chloride, calcium chloride, glucose or dextran adjusting etc. and ooze, regulate pH to 4.5-6.0 with the water for injection dissolving, and sterilization, promptly.
Powder injection formulation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and drying, promptly.
Freeze-dried powder is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and lyophilization, promptly.
The ceramic membrane microfiltration is to serve as a kind of efficient, energy-conservation, the free of contamination new separation technology of filtration medium with the novel inorganic ceramic membrane, has the separation accuracy height, chemical property is stable, the enriched product or the efficient height that filters out impurities, need not that heating concentrates, active ingredient is not destroyed, shorten the production cycle, advantage such as enhance productivity; Resin purification is the treatment technology of new development both at home and abroad in recent years, and field of medicaments particularly natural drug refining in widely utilization, be a kind of effective ways that extract aqueous soluble active constituent in the refining Chinese herbal medicine.Therefore, the method purification refine traditional medicine Injectio that the present invention adopts inorganic ceramic film, resin absorption and organic membrane to combine, its characteristics are: replaced traditional chemical method with physical method, can effectively remove the macromolecular substances in the Chinese medicine extraction liquid, do not introduce simultaneously other inorganic ions, the characteristics that kept compound Chinese medicinal preparation preferably can effectively reduce the untoward reaction of medicine.
Technical solutions according to the invention are that the applicant obtains through a large amount of experimental studies, and main content of the test is as follows:
Experimental example 1, Folium Ginkgo extraction process are analyzed
1.1 extracting method is selected
Get the Folium Ginkgo medical material, adopt respectively decoct with water, medicinal residues decoct with water three kinds of methods and extract behind the alcohol reflux, alcohol reflux, with Ginkgo total flavones alcohol glycosides content, terpene lactone contents, extracting solution color and luster, clarity is index, investigates the extraction ratio of ginkgo flavone and lactone.
Folium Ginkgo extraction process table
Method | Evaluation index | Total flavonoids (%) | Terpene lactone (%) | Color and luster | Clarity |
1 | Water is carried | 6.4 | 2.17 | More shallow | Clear and bright |
2 | Alcohol extraction | 11.3 | 2.62 | Shallow | Clear and bright |
3 | Alcohol, water combination | 15.2 | 2.88 | Color and luster is between between the two | Clear and bright |
Experimental result shows, all can measure ginkgetin and bilobalide in water extract and the alcohol extract, and employing is pure, moisture is put forward the method that combines and extracted the Folium Ginkgo medical material, and extracting solution is all obtained good result at aspects such as color and luster, content, clarity.Consider the refining pure metallization processes of next stage, preferred alcohols, the bonded extracting method of water.
1.2 alcohol extraction process research
1.2.1 concentration of alcohol is selected
Get Folium Ginkgo 50g, pulverize, add 20%, 40%, 60%, 80%, 90% ethanol respectively and test, reflux 1 hour, backflow filters, and detects.
Concentration of alcohol-extraction counting rate meter
Extract solvent | 20% ethanol | 40% ethanol | 60% ethanol | 80% ethanol | 90% ethanol |
Extraction ratio | 45.7% | 77.5% | 86.2% | 78.7% | 76.1% |
Relative extraction ratio | 53.4% | 92.3% | 100% | 94.1% | 94.6% |
The result shows, selects for use 40%-80% ethanol as extracting solvent, and extraction ratio is all better, and preferred 60% ethanol is advisable.
1.2.2 extraction time is investigated
Sample preparation: get Folium Ginkgo 50g, pulverize, add 460% alcohol heating reflux 5 times, each 1 hour, collect each backflow respectively, filter.
Extraction time-extraction counting rate meter
Extraction time | 1 | 2 | 3 | 4 | 5 |
Extraction ratio (%) | 45.8 | 81.3 | 83.7 | 84.2 | 84.5 |
Data show in the table when extracting 4 times, has almost been extracted fully, determines that therefore extraction time is 2-4 time, wherein extracts 3 times and is optimised process, and less expensive is reasonable.
1.2.3 return time is selected
Sample preparation: get Folium Ginkgo 50g, pulverize, add 60% ethanol, the according to the form below time is carried out reflux, and backflow filters, and collects and detects.The result is as follows:
Return time-extraction counting rate meter
Return time | 30min | 1hr | 1.5hr | 2hr | 2.5hr | 3hr | 3.5hr |
Extraction ratio (%) | 46.9 | 70.6 | 78.5 | 82.7 | 83.4 | 84.1 | 84.3 |
The result shows that it is little to extract 1-3 hour changes of contents, extracts to reach balance substantially in two hours, thus selective extraction scope 1-3 hour, preferred 2 hours.
In addition, the applicant finds under study for action, and Folium Ginkgo was soaked 8-14 hour before extracting, and the extraction of its total flavonoids and terpene lactone is more complete.So comprehensive The above results, the extraction process of decision Folium Ginkgo is: use 40%-80% soak with ethanol 8-14 hour, reflux, extract, is 2-4 time then, adds ethanol 6-12 at every turn and doubly measures, extracts 1-3 hour.Be preferably:, add 9 times of amounts of ethanol at every turn, extracted 2 hours with 60% soak with ethanol 12 hours, reflux, extract, 3 times.
Experimental example 2, inorganic ceramic film separation purifying technique
Choose inorganic ceramic film the medicinal liquid after extracting carried out microfiltration, to before filtering and filtered liquid medicine done contrast at aspects such as clarity, precipitation, color and luster, changes of contents, and the technological parameter of inorganic membrane filtration is investigated.
2.1 test method
Equipment: 1#, 2#, 3# ceramic membrane, operating parameter is: (P:0.35~0.4MP; T:35~42 ℃; V:10~15L/h), the average flux scope is 70~90L.M-2.H-1.1#, 2#, 3# film carry out microfiltration excessively respectively with the medicinal liquid after extracting.
2.2 result of the test:
In experiment, find to use the inorganic ceramic membrane filtration, effective ingredient distributed density basically identical in permeate and the mother solution, mother solution is the dense medicinal liquid that does not see through remaining in the equipment, and can add pure water aborning and wash out effective ingredient, and the reuse nanofiltration concentrates can improve recovery rate.And the amount that at every turn adds entry is more little, and eluting rate is high more, and amount of water is that 2~3 times of eluting resultant effects are best, and loss of effective components generally is no more than 5%.This is considered as 0 with concentration difference before and after the membrane filtration.
Experimental result shows: obviously improve with the medicinal liquid clarity behind the inorganic membrane filtration, more clear and bright and color and luster obviously shoals, and sterilization is placed and can be produced small amount of precipitate.With gel chromatography check feedback result be: just filter molecular weight through inoranic membrane and can not detect greater than 10000 macromolecular substances.When microfiltration, can add a spot of water washing effective ingredient several times,, extracting solution suitably can be concentrated with vacuum decompression in addition, can reduce filtration yield during microfiltration again, to save time, to raise the efficiency for improving the production efficiency of back workshop section.
Experimental example 3, Choice of Resin
3.1 the primary election of resin kind
Resin: D101, D201, D390, CD180, CAD45, AB-8 macroporous adsorbent resin, LSA-14 resin, ADS series plastics, polyamide, active carbon etc.With total flavonoids content is the adsorption effect that index is investigated resin, as following table:
Resin kind option table
The resin title | Total flavonoids content (g/100g) | The resin title | Total flavonoids content (g/100g) |
D201 | 16.3 | D390 | 6.9 |
D101 | 15.7 | CD180 | 17.4 |
AB-8 | 6.5 | LSA-14 | 9.5 |
ADS series | 11.2 | Activated carbon | 7.6 |
Polyamide | 15.6 | CAD45 | 10.4 |
The result shows, adopts CD180, DA201, D101, polyamide more suitable.
3.2 select the purification resin
Four kinds of resins just electing are done comprehensively test, the results are shown in following table.
Different resins purification result synopsis
The mensuration project | CD180 | DA201 | D101 | Polyamide |
Total flavonoids content (%) | 40.9 | 36.3 | 30.1 | 33.7 |
Terpene lactone contents (%) | 12.3 | 12.1 | 11.9 | 12.2 |
The extract color | More shallow | Deeply | Shallow | Shallow |
Resin proportion | Greatly | Little | Little | Little |
Resin absorption amount (mg/mL) | 6.0 | 9.5 | 44.0 | 46.5 |
Resin size (order) | 80-100 | 20-50 | 20-50 | 20-50 |
Above result shows, no matter all better at aspects such as active constituent content, extract color and adsorbances above-mentioned four kinds of resins are, the purification resin that all can be used for Folium Ginkgo extract, consider from extract flavone glycosides content, should select CD180, but granularity is too little, adopts this resin may cause the phenomenon that can't cross post in suitability for industrialized production, so preferred DA201 is as the purification resin.
Experimental example 4, organic membrane separation purifying technique
4.1 technology: organic membrane is handled and is divided into membrance concentration and two operating procedures of membrane ultrafiltration:
Membrance concentration condition: P:0.5~1MPa; T:30~40 ℃, V: 〉=2250L/h.
The organic membrane ultrafiltration: ultrafilter membrane used in the experiment is the rolling ultrafilter membrane, is provided by Nanjing University of Technology, and molecular cut off is 10000.Operating condition is P:0.5~0.9MPa; T:30~40 ℃; V: 〉=2250L/h.Can add suitable quantity of water washing active ingredient in the ultra-filtration process, effective ingredient is held back, loss rate is at 15%~20%.
4.2 analyze: for reducing loss of effective components, should reduce the ultrafiltration number of times, and pressure should be the smaller the better during ultrafiltration, as far as possible so selection ultrafiltration 1-2 time.
4.3 conclusion: the present invention adopts the method for ultrafiltration that Folium Ginkgo extract is further purified, the purpose that reached minimizing technology, shorten the production cycle, economize in raw materials (especially ethanol) reduces cost, keep the composition of original formulation simultaneously, and improved content of effective; Ultrafiltration technology degerming depyrogenation is good in addition, has avoided the heating of medicinal liquid and the precipitation that produces, and also the part that can be removed pigment prepares ejection preparation to the later stage and established good basis.
Experimental example 5, pharmacodynamic experiment
5.1 experiment material:
Medicine: commercially available Ginkgo Leaf Extract and Dipyridamole Injection, beneficial one hundred Pharmacy stock Co., Ltd produces, and 10ml/ props up, positive contrast medicine; Injection by preparation method of the present invention is made is medicine of the present invention.
Animal: Wistar rat, body weight 200~300g; Kunming kind white mice, body weight 18~22g, the male and female dual-purpose, animal housing provides by the Jilin Prov. Inst. of Chinese Medicine and Chinese Medical Science.
Instrument: LBY-NJ2 platelet aggregation instrument
5.2 experimental technique and result
5.2.1 30 of the rats that influence to platelet aggregation are divided 3 groups at random, intravenous administration, every day 1 time, continuous 15 days, 1h after the last administration uses 10% chloral hydrate anesthesia, the ventral aorta blood sampling, adopt the turbidimetry for Determination platelet aggregation rate, derivant is ADP, the results are shown in following table.
5.2.2 influence 20 of rats and be divided into 2 groups at random, intravenous administration, every day 1 time, continuous 15 days to rats in vitro is thrombotic.1h after the last administration, rats in vitro common carotid artery-external jugular vein thrombosis method is adopted in the anesthesia of 3% pentobarbital sodium, the weighing wet weight of thrombus, this product can significantly suppress the formation of rats in vitro thrombosis as a result, the results are shown in following table.
The influence that the inductive platelet aggregation of ADP and external thrombus are formed (x ± s)
Group | Dosage (the mg Furost saponine/kg) | Mus number (only) | Platelet aggregation rate (5min) (%) | Wet weight of thrombus (mg) |
Contrast | 10 | 64.95±16.26 | 26.62±12.32 | |
Commercially available group | 39.38 | 10 | 50.28±10.03* | 15.96±2.30** |
Of the present invention group | 39.38 | 10 | 50.43±10.05* | 15.89±2.40** |
Compare * P<0.05 with matched group, * * P<0.01 (down together)
5.2.3 influence to clotting time of mice
30 of mices are divided 3 groups at random, intravenous administration, every day 1 time, continuous 15 days.1h after the last administration presses slide method and measures clotting time, and preparation of the present invention as a result can obviously prolong clotting time of mice, sees the following form.
To the influence of clotting time of mice (x ± s)
Group | Dosage (the mg Furost saponine/kg) | Number of animals (only) | Clotting time (%) |
Contrast | 10 | 0.83±0.12 | |
Commercially available group | 56.25 | 10 | 1.05±0.18** |
Of the present invention group | 56.25 | 10 | 1.04±0.10** |
5.2.4 influence to acute imperfection cerebral ischemic model
40 of male rats are divided into 4 groups at random, and each intravenous administration of 1h is 1 time before preceding 1 the sky of experiment, afternoon and experiment art on the same day.Broken end is opened cranium and is got brain behind the bilateral ligation 3h, inhales the moisture on decerebrate surface with filter paper, and the weighing weight in wet base places drying baker to dry to constant weight the weighing dry weight for interior 110 ℃ then.Brain water content (%)=(weight in wet base-dry weight)/weight in wet base * 100% the results are shown in following table.
To the influence of acute imperfection cerebral ischemic model rat brain water content and cerebral index (x ± s)
Group | Dosage (the mg Furost saponine/kg) | Mus number (only) | Brain water content (%) | Cerebral index |
The sham-operation contrast | 10 | 68.36±1.96** | 0.65±0.04** | |
The model contrast | 10 | 74.49±1.76 | 0.79±0.05 | |
Commercially available group | 39.38 | 10 | 72.48±1.92** | 0.73±0.03** |
Of the present invention group | 39.38 | 10 | 71.31±2.07** | 0.71±0.04** |
Compare * P<0.05, * * P<0.01 with model group
The result shows, compares with commercially available Ginkgo Leaf Extract and Dipyridamole Injection group, and the ginkgo bilobate extract liquid preparation for preparing by the present invention more can obviously reduce brain water content and the cerebral index of cerebral ischemic model rat.
After above-mentioned ischemic tissue of brain is fixing, the crown brain that cuts, conventional dehydration, paraffin embedding film-making, AE dyeing, light microscopic check (* 400).Mirror is observed down, and lacuna is little around Sham-operated control group cerebral tissue neurocyte and the blood vessel, and lacuna obviously becomes big around the cerebral ischemic model group blood vessel, and lacuna changes not fairly obvious around the neurocyte.And two medicine groups can suppress obviously due to the modeling type that lacuna enlarges around the blood vessel, illustrate that the Folium Ginkgo extract of preparation method gained of the present invention and dipyridamole combination preparation have certain protective role to the pathological change of rat ischemia cerebral tissue.
Compared with prior art, the present invention with the Folium Ginkgo medical material carry out alcohol, moisture is carried, and adopt membrane separation technique binding resin refinement treatment technology such as inorganic ceramic membrane microfiltration, organic membrane ultrafiltration, substitute traditional chemical method with physical method, improved the production technology of Folium Ginkgo extract and dipyridamole combination preparation, can effectively remove the macromolecular substances in the Chinese medicine extraction liquid, do not introduce simultaneously other inorganic ions, fully enrichment effective ingredient, effectively reduce the untoward reaction of medicine, improve curative effect, thereby improved and guaranteed the quality of product; And the present invention has reduced processing step, has shortened flow process, can obtain good cost benefit.
The specific embodiment:
Embodiments of the invention 1: get Folium Ginkgo extract 20g, dipyridamole 2g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 40% soak with ethanol 8 hours, reflux, extract, was 2 times then, and add 6 times of amounts of ethanol at every turn, extracted 1 hour, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 4 times of water gagings and extract 2 times, each 0.5 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, filtered liquid medicine is directly adsorbed by the CD180 resin column of having handled well, effluent is discarded, with the speed of per second 0.5ml, is that 3 times of column volume deionized waters or 1% ethanol of 2-6 is washed post with pH value, the reuse pH value is 60% ethanol elution of 4-6, collect color and divide eluent, reclaim ethanol, concentrate with vacuum evaporator than the deep, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 5% ethanol, and stirring and dissolving is carried out ultrafiltration with rolled film then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole, with the water for injection dissolving, add mannitol or lactose or sorbitol, regulate pH to 4.5-6.0, lyophilization promptly gets freeze-dried powder.
Embodiments of the invention 2: get Folium Ginkgo extract 100g, dipyridamole 10g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 80% soak with ethanol 14 hours, reflux, extract, was 4 times then, and add 12 times of amounts of ethanol at every turn, extracted 3 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 6 times of water gagings and extract 4 times, each 2 hours, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, filtered liquid medicine is directly adsorbed by the D101 resin column of having handled well, effluent is discarded, with the speed of per second 2ml, is that 5 times of column volume deionized waters or 30% ethanol of 2-6 is washed post with pH value, the reuse pH value is 95% ethanol elution of 4-6, collect color and divide eluent, reclaim ethanol, concentrate with vacuum evaporator than the deep, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 40% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby
Folium Ginkgo extract is mixed with dipyridamole, with the water for injection dissolving, add mannitol or lactose or sorbitol, regulate pH to 4.5-6.0, drying promptly gets powder injection formulation.
Embodiments of the invention 3: get Folium Ginkgo extract 40g, dipyridamole 4g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 60% soak with ethanol 12 hours, reflux, extract, was 3 times then, and add 9 times of amounts of ethanol at every turn, extracted 2 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 5 times of water gagings and extract 3 times, each 1 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, the water washing effective ingredient that adds 2-3 times of medicine liquid volume amount during filtration several times, filtered liquid medicine directly adsorb by the DA201 resin column of having handled well, and effluent is discarded, speed with per second 1ml, with pH value is that 2 times of column volume deionized waters or 10% ethanol of 2-6 is washed post, and the reuse pH value is 80% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 20% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole,, regulate pH to 4.5-6.0 with the water for injection dissolving, embedding, sterilization promptly gets hydro-acupuncture preparation.
Embodiments of the invention 4: get Folium Ginkgo extract 80g, dipyridamole 8g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 70% soak with ethanol 14 hours, reflux, extract, was 2 times then, and add 10 times of amounts of ethanol at every turn, extracted 2 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 4 times of water gagings and extract 4 times, each 1 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 12-15L/h, add the water washing effective ingredient of 2 times of medicine liquid volume amounts during filtration several times, filtered liquid medicine is directly adsorbed by the polyamide resin column of having handled well, and effluent is discarded, speed with per second 1.5ml, with pH value is that 3 times of column volume deionized waters or 25% ethanol of 2-6 is washed post, and the reuse pH value is 75% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 25% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole,, regulate pH to 4.5-6.0 with the water for injection dissolving, embedding, sterilization promptly gets hydro-acupuncture preparation.
Embodiments of the invention 5: get Folium Ginkgo extract 60g, dipyridamole 6g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 50% soak with ethanol 10 hours, reflux, extract, was 3 times then, and add 8 times of amounts of ethanol at every turn, extracted 3 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 4 times of water gagings and extract 4 times, each 1.5 hours, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-12L/h, the water washing effective ingredient that adds 3 times of medicine liquid volume amounts during filtration several times, filtered liquid medicine directly adsorb by the DA201 resin column of having handled well, and effluent is discarded, speed with per second 1ml, with pH value is that 4 times of column volume deionized waters or 15% ethanol of 2-6 is washed post, and the reuse pH value is 70% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 15% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole, with the water for injection dissolving, add sodium chloride, calcium chloride, glucose or dextran adjusting etc. and ooze, regulate pH to 4.5-6.0, sterilization promptly gets infusion preparation.
Embodiments of the invention 6: get Folium Ginkgo extract 90g, dipyridamole 9g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 65% soak with ethanol 9 hours, reflux, extract, was 4 times then, and add 12 times of amounts of ethanol at every turn, extracted 1 hour, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 5 times of water gagings and extract 3 times, each 2 hours, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 12-14L/h, filtered liquid medicine is directly adsorbed by the polyamide resin column of having handled well, effluent is discarded, with the speed of per second 2ml, is that 4 times of column volume deionized waters or 20% ethanol of 2-6 is washed post with pH value, the reuse pH value is 90% ethanol elution of 4-6, collect color and divide eluent, reclaim ethanol, concentrate with vacuum evaporator than the deep, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 30% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole, with the water for injection dissolving, add sodium chloride, calcium chloride, glucose or dextran adjusting etc. and ooze, regulate pH to 4.5-6.0, sterilization promptly gets infusion preparation.
Embodiments of the invention 7: get Folium Ginkgo extract 50g, dipyridamole 5g, according to the following steps preparation:
Get Folium Ginkgo and pulverize, with 55% soak with ethanol 12 hours, reflux, extract, was 3 times then, and add 9 times of amounts of ethanol at every turn, extracted 1.5 hours, merge extractive liquid,, recovery ethanol, must the Folium Ginkgo alcohol extract standby; Medicinal residues add 6 times of water gagings and extract 2 times, each 2 hours, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 12-15L/h, the water washing effective ingredient that adds 2-3 times of medicine liquid volume amount during filtration several times, filtered liquid medicine directly adsorb by the D101 resin column of having handled well, and effluent is discarded, speed with per second 1.5ml, with pH value is that 2 times of column volume deionized waters or 8% ethanol of 2-6 is washed post, and the reuse pH value is 85% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 10% ethanol, and stirring and dissolving is carried out ultrafiltration with rolled film then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Folium Ginkgo extract is mixed with dipyridamole, with the water for injection dissolving, add mannitol or lactose or sorbitol, regulate pH to 4.5-6.0, lyophilization promptly gets freeze-dried powder.
Claims (8)
1. the composite preparation of Folium Ginkgo extract and dipyridamole, according to listed as parts by weight, it is to be made by Folium Ginkgo extract 20-100 part, dipyridamole 2-10 part, it is characterized in that: preparation method is: get Folium Ginkgo and pulverize, use 40%-80% soak with ethanol 8-14 hour, reflux, extract, is 2-4 time then, each ethanol 6-12 that adds doubly measured, extracts 1-3 hour, merge extractive liquid, reclaims ethanol, and it is standby to get the Folium Ginkgo alcohol extract; Medicinal residues add 4-6 times of water gaging and extract 2-4 time, each 0.5-2 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, filtered liquid medicine is directly adsorbed by the resin column of having handled well, effluent is discarded, with the speed of per second 0.5-2ml, is that 1-5 times of column volume deionized water or the 1%-30% ethanol of 2-6 is washed post with pH value, the reuse pH value is the 60%-95% ethanol elution of 4-6, collect color and divide eluent, reclaim ethanol, concentrate with vacuum evaporator than the deep, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 5%-40% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, filtrate concentrates with vacuum evaporator, and drying is mixed with dipyridamole, be dissolved in water, add adjuvant or do not add adjuvant, make different ejection preparations with conventional method.
2. according to the composite preparation of described Folium Ginkgo extract of claim 1 and dipyridamole, it is characterized in that: according to listed as parts by weight, it is made for 4 parts with 40 parts of Folium Ginkgo extract, dipyridamole.
3. according to the composite preparation of claim 1 or 2 described Folium Ginkgo extract and dipyridamole, it is characterized in that: preparation method is: get Folium Ginkgo and pulverize, with 60% soak with ethanol 12 hours, reflux, extract, is 3 times then, 9 times of amounts of each adding ethanol, extraction 2 hours, merge extractive liquid, reclaims ethanol, and it is standby to get the Folium Ginkgo alcohol extract; Medicinal residues add 5 times of water gagings and extract 3 times, each 1 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, add the water washing effective ingredient of 2-3 times of medicine liquid volume amount during filtration several times, filtered liquid medicine is directly adsorbed by the resin column of having handled well, and effluent is discarded, speed with per second 1ml, with pH value is that 2 times of column volume deionized waters or 10% ethanol of 2-6 is washed post, and the reuse pH value is 80% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 20% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, filtrate concentrates with vacuum evaporator, and drying is mixed with dipyridamole, be dissolved in water, add adjuvant or do not add adjuvant, make different ejection preparations with diverse ways.
4. according to the described composite preparation of claim 3, it is characterized in that the purification resin column described in the preparation method is meant CD180 post, DA201 post, D101 post, polyamide resin column.
5. according to the described composite preparation of claim 4, it is characterized in that: the purification resin column described in the preparation method is meant the DA201 post.
6. according to the composite preparation of claim 1 or 2 described Folium Ginkgo extract and dipyridamole, it is characterized in that: described preparation is an ejection preparation, comprises hydro-acupuncture preparation, infusion preparation, powder injection formulation and freeze-dried powder.
7. according to the composite preparation of described Folium Ginkgo extract of claim 6 and dipyridamole, it is characterized in that preparation method is further comprising the steps of:
Hydro-acupuncture preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, regulates pH to 4.5-6.0 with the water for injection dissolving, and embedding, sterilization, promptly;
Infusion preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds sodium chloride, calcium chloride, glucose or dextran adjusting etc. and ooze, regulate pH to 4.5-6.0 with the water for injection dissolving, and sterilization, promptly;
Powder injection formulation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and drying, promptly;
Freeze-dried powder is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and lyophilization, promptly.
8. according to the composite preparation of described Folium Ginkgo extract of claim 1 and dipyridamole, it is characterized in that: according to listed as parts by weight, it is made as follows by Folium Ginkgo extract 20-100 part, dipyridamole 2-10 part: get Folium Ginkgo and pulverize, with 60% soak with ethanol 12 hours, reflux, extract, is 3 times then, adds 9 times of amounts of ethanol at every turn, extracts merge extractive liquid, 2 hours, reclaim ethanol, it is standby to get the Folium Ginkgo alcohol extract; Medicinal residues add 5 times of water gagings and extract 3 times, each 1 hour, merge extractive liquid,, mix with the Folium Ginkgo alcohol extract, suitably concentrate, filter with inorganic ceramic film, the rate of filtration is 10-15L/h, add the water washing effective ingredient of 2-3 times of medicine liquid volume amount during filtration several times, filtered liquid medicine is directly adsorbed by the resin column of having handled well, and effluent is discarded, speed with per second 1ml, with pH value is that 2 times of column volume deionized waters or 10% ethanol of 2-6 is washed post, and the reuse pH value is 80% ethanol elution of 4-6, collects color and divides eluent than the deep, reclaim ethanol, concentrate with vacuum evaporator, drying promptly gets the Folium Ginkgo crude extract; The Folium Ginkgo crude extract is added 20% ethanol, and stirring and dissolving is carried out ultrafiltration with the rolling ultrafilter membrane then, and filtrate concentrates with vacuum evaporator, and drying is standby;
Wherein, hydro-acupuncture preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration is mixed with dipyridamole,, regulate pH to 4.5-6.0 with the water for injection dissolving, and embedding, sterilization, promptly;
Infusion preparation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds sodium chloride, calcium chloride, glucose or dextran adjusting etc. and ooze, regulate pH to 4.5-6.0 with the water for injection dissolving, and sterilization, promptly;
Powder injection formulation is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and drying, promptly;
Freeze-dried powder is preparation like this: the Folium Ginkgo extract behind the ultra filtration mixed with dipyridamole,, adds mannitol or lactose or sorbitol, regulate pH to 4.5-6.0 with the water for injection dissolving, and lyophilization, promptly.
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