CN100443095C - Compound formulation of breviscapine and asarum herb for treating cardiovascular and cerebrovascular diseases and its application - Google Patents

Abstract

The present invention provides a compound formula of a fleabane preparation for treating cardiovascular and cerebrovascular diseases, and a preparation method and the application of the compound formula. The compound formula of the present invention is prepared from 1 to 99% of fleabane, 99 to 1% of red peony root and proper percent of auxiliary materials. Compared with the prior art, the compound formula of the present invention conforms to the principle that 'in order to treat wind, blood needs to be treated firstly; when the blood is unobstructed, the wind can be eliminated'; a method for promoting blood circulation, removing blood stasis and dredging channels is adopted. The fleabane which is bitter and warm is used as a main material, and is used for removing blood stasis and dredging channels; the red peony root is bitter and slightly cold; when the red peony root is mixed with blood, the red peony root has the efficiency of cooling blood heat, expelling blood stasis, dredging channels, and treating blood heat stasis; the red peony root is matched with the main material to achieve the efficiency of promoting blood circulation, removing blood stasis, expelling blood stasis and dredging channels, and is used as an auxiliary material. The fleabane and the red peony root are mixed to be used to realize the functions of promoting blood circulation, removing blood stasis, dredging channels and activating collaterals. The compound formula of the present invention has the advantages of less consumption of medicines, special effect, direct action channels, no disadvantage of overheating or overcooling sensation, and obvious characteristic of medicine use. The safety and the availability of a product are ensured through reasonable feasible preparation technology, and thus, the compound formula of the present invention can be suitable for patients to use for a long time.

Description

Compound recipe fleabane preparation of treatment cardiovascular and cerebrovascular disease and its production and application

Technical field:

The present invention is a kind of compound recipe fleabane preparation for the treatment of cardiovascular and cerebrovascular disease and its production and application, belongs to technical field of Chinese medicine.

Technical background: cerebrovascular disease mainly is divided into hemorrhagic apoplexy and ischemic cerebrovascular clinically, ischemic cerebrovascular (Atherosclerosis and thrombosis and cerebral infarction, cerebral embolism and lacunar infarction) is the most common, is about 3 times of hemorrhagic apoplexy.A large amount of epidemiological studies show that cerebral thrombosis accounts for 40～60% of whole cerebrovascular again, and cerebral hemorrhage accounts for 16～39%, and subarachnoid hemorrhage accounts for about 10%, and cerebral embolism accounts for 3～8%.6391 examples of 1971～1974 years statistics of World Health Organization (WHO) are by the apoplexy of diagnosis classification, and cerebral infarction is the most common as a result, on average accounts for more than 40%; Secondly for intracerebral hemorrhage, account for 16%; Subarachnoid hemorrhage is more than 8%; Cerebral embolism accounts for 3.5～5%.U.S. Framingham18 follows up a case by regular visits to and finds that cerebral thrombosis accounts for 57%, and cerebral embolism accounts for 15%, and cerebral hemorrhage accounts for 5%, and subarachnoid hemorrhage accounts for 12%.Along with the growth of industrialization degree, and people's growth in the living standard, the change of dietary structure, hypertension, hyperglycemia, hyperlipidemia, smoking, crowd such as drink increases, and the sickness rate of cerebrovascular is ascendant trend year by year.Ischemic cerebrovascular is the common disease of middle-aged and elderly people, and it falls ill anxious, and mortality rate, disability rate are all higher, are one of middle-aged and elderly people three big underlying cause of deaths.To reduce thrombosis, reduce the cerebral tissue necrosis for the Therapeutic Principle of cerebral infarction, the protection brain cell is main, and the treatment of western modern medicine mainly contains thrombolytic, anticoagulant, antiplatelet aggregation, brain protection, vasodilation, the treatment of calcium antagonism.These medicines are as most of chemical medicines, and clinical practice also is subjected to the restriction of more and more side effect that display.Antithrombotic therapy commonly used has thrombolytics (urokinase, streptokinase etc.), anticoagulant (heparin, dicoumarol, warfarin, acenocumarol tablet etc.), calcium ion antagonist (flunarizine, nimodipine, nicardipine, cinnarizine, nifedipine, lidoflazine etc.), the characteristics that have instant effect mostly, but be prone to hemorrhage, gastrointestinal reaction and allergy, headache, dizziness, feel sick, untoward reaction such as hypotension, erythra, and usually be subjected to the influence of complication and use limited, so clinical practice is very careful.And the disclosed number of patent application of Chinese patent communique is: 03136116, name is called the application of " a kind of Chinese medicine preparation for the treatment of Blood Ropy disease and preparation method thereof ", many Chinese crude drug compatibilities have been used in this part invention, the preparation more complicated, it is very difficult particularly preparing Chinese medicine, is unfavorable for controlling its quality of production.Given this, very be necessary to research and develop a kind of compatibility advantages of simple, the Chinese prescription that curative effect is feasible.

Summary of the invention:

The objective of the invention is to: a kind of compound recipe Herba Erigerontis Chinese medicine preparation for the treatment of cardiovascular and cerebrovascular disease and its production and application is provided; The present invention is directed to prior art, we flee meridians according to wind-phlegm, the resistance of blood vessels numbness, and channel tunnel is obstructed, the gas unable to walk, blood can not moisten, so limbs come into disuse into the etiology and pathogenesis of hemiplegia, abide by the reason of " control wind and control blood earlier, blood sector-style go out ", take the method for activating blood circulation to dissipate blood stasis and dredge the collateral.Herba Erigerontis with the bitter temperature of nature and flavor is monarch, the disperse blood stasis and dredge collateral of using; The Radix Paeoniae Rubra bitter in the mouth is slightly cold, and goes into blood system, removing heat from blood heat, and dissipating blood stasis blood, the meridian dredging is controlled the heat in blood stasis of blood and is stagnated, and principal drug assistance is to reach the effect of blood circulation promoting and blood stasis dispelling, dissipating blood stasis collateral dredging, and what use is minister.Two medicines share has blood circulation promoting and blood stasis dispelling, the function of dredge the meridian passage.The useful YAOJING letter of the prescription length that power is special, action pathway is direct does not have the cooler fraud of temperature partially, and comparatively significantly administration features is arranged.The present invention not only has curative effect preferably for treating cardiovascular and cerebrovascular disease such as coronary heart disease, angina pectoris, arrhythmia, cerebral thrombosis, alzheimer disease etc., can also be used for the treatment of diseases such as hepatorenal syndrome, heart and lung diseases, diabetes and complication thereof.And the present invention Herba Erigerontis Herba Asari, the Radix Paeoniae Rubra selected for use, mainly containing Herba Erigerontis florigen, peoniflorin, technology is rationally feasible, can not produce the anaphylaxis material in the preparation process, and is more safe and effective, but the patients life-time service.

The present invention constitutes like this: calculate according to percentage by weight, it is to be prepared from by Herba Erigerontis 1～99% and Radix Paeoniae Rubra 99～1% and suitable adjuvant.

Be preferably: calculate according to percentage by weight, it is to be made by Herba Erigerontis 20～80% and Radix Paeoniae Rubra 80～20% and suitable adjuvant.

Say accurately: calculate according to percentage by weight, it is to be made by Radix Paeoniae Rubra 60% and Herba Erigerontis 40%.

Radix Paeoniae Rubra in the described prescription and Herba Erigerontis can be medical material or the extract that prepared by the corresponding proportion medical material, and wherein Herba Erigerontis extract can be the highly finished product of Herba Erigerontis alcohol extract, Herba Erigerontis water extract, Herba Erigerontis water extract-alcohol precipitation extract, Herba Erigerontis semi-bionic extraction thing, Herba Erigerontis supercritical extract or above each extract; Radix Paeoniae Rubra extract can be the highly finished product of Radix Paeoniae Rubra alcohol extract, Radix Paeoniae Rubra water extract, Radix Paeoniae Rubra water extract-alcohol precipitation extract, Radix Paeoniae Rubra semi-bionic extraction thing, Radix Paeoniae Rubra supercritical extract or above each extract.

Described preparation be directly used in the injection with small volume of drug administration by injection, directly for the venous transfusion of intravenous drip, need be used for the concentrated solution for injection of intravenous drip and injectable sterile powder and the aseptic block that makes with freeze-drying or spray drying method, tablet, capsule, granule, drop pill, pill, soft capsule, oral liquid, oral cavity disintegration tablet or dispersible tablet after the dilution.

Contain flavones ingredient and glycoside composition in the preparation, calculate by weight percentage, in the injection flavones ingredient and glycoside component content sum be not less than deduction adjuvant amount and water quantities in the preparation total solid 25%.

The preparation method of the compound recipe fleabane preparation of described treatment cardiovascular and cerebrovascular disease: get the Radix Paeoniae Rubra medical material, adding entry or alcoholic solution after the pulverizing extracts, merge extractive liquid,, filter, concentrate the Radix Paeoniae Rubra crude extract, also can adopt in ethanol precipitation, column chromatography, extraction, the flocculent precipitation one or more to unite on this basis to use carry out suitably refining, the Radix Paeoniae Rubra extract; Get the Herba Erigerontis medical material, adding entry or alcoholic solution after the pulverizing extracts, merge extractive liquid,, filter, concentrate the Herba Erigerontis crude extract, also can adopt in ethanol precipitation, column chromatography, extraction, the flocculent precipitation one or more to unite on this basis to use carry out suitably refining, the Herba Erigerontis extract, with Herba Erigerontis crude extract or extract and Radix Paeoniae Rubra crude extract or extract mix homogeneously, add adjuvant and make different preparations.

Be preferably: Herba Erigerontis adds 5～12 times of water gagings and decocts 1～5 time, each 0.5～2 hour, filters, merging filtrate, filtrate decompression concentrate, and add ethanol and make and contain the alcohol amount and reach 30～80%, constantly stir, left standstill sucking filtration 6～24 hours, decompression filtrate recycling ethanol also concentrates, and uses the hydrochloric acid adjust pH, insulation, the tipping supernatant, sucking filtration, precipitation washes with water to pH value, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 3～10 times of water gagings and decocts 1～5 time, each 0.5～2 hour, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40～80%, stir, left standstill sucking filtration 6～24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1～5 time, merge n-butyl alcohol liquid, wash with water 1～3 time, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, and last polyamide column is used ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, add adjuvant and make different preparations.

Say accurately: Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate, when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and make and contain the alcohol amount and reach 60%, stir, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20 is with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, reclaim under reduced pressure n-butyl alcohol, residue add 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, absorption flow velocity: 0.5BV/h, wet 5 times of volumes of column volume=dried resin weight of BV-are with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, add adjuvant and make different preparations.

Injectable sterile block in the described preparation prepares like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract and 10g mannitol, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, add the injection water to 2000ml, mixing, boiled 30 minutes, be sub-packed in the infusion bottle of having handled well, fill in butyl rubber bung and thin film, roll aluminium lid, flowing steam sterilization 30 minutes, cold preservation 24 hours filters the filtrate packing with 0.45 μ m and 0.22 μ m microporous filter membrane under aseptic condition, every bottle of 2.0ml, lyophilization, the equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, and froze the jail fully, promptly begin evacuation until product, enter drying program, under 45 ℃ of constant temperature-and evacuation, make the air pressure of hothouse reduce to 13.332～266.64Pa, under this pressure, slowly heat up, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, heating up, dry the time is about 12～15 hours to remove residual moisture, keeps more than 35 ℃ dry 3 hours, gland, promptly.

Injection and concentrated solution for injection in the described preparation prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate adds the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, to boil, 4 ℃ of cold preservations are spent the night, coarse filtration, fine straining, add the injection water, divide to install to pacify and cut open bottle, seal sterilization and promptly get injection with small volume or concentrated solution for injection.

Glucose or sodium chloride intravenous infusion in the described preparation prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, add the glucose or the sodium chloride of ormal weight, by volume add 1.0% active carbon behind the mixed dissolution, boil, keep little 30min that boils, cold slightly filtration, filtrate add the injection water to ormal weight, the saturated sodium hydroxide solution of reuse adjust pH to 7.0～7.5, boil, 4 ℃ of cold preservations are spent the night, and coarse filtration, fine straining add the injection water, packing, sterilization promptly.

Injectable sterile powder in the described preparation prepares like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, adds the injection water to 2000ml, mixing, boiled 30 minutes, and divided to install in the enamel tray lyophilization, equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, and froze the jail fully, promptly begin evacuation until product, enter drying program, under 45 ℃ of constant temperature-and evacuation, make the air pressure of hothouse reduce to 13.332～266.64Pa, under this pressure, slowly heat up, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, heating up, dry the time is about 12～15 hours to remove residual moisture, keeps more than 35 ℃ dry 3 hours, under aseptic condition, divide and install in the cillin bottle, promptly.

Injectable sterile powder in the described preparation can also prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, mixing adds an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate add the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, boil, 4 ℃ of cold preservations are spent the night, and coarse filtration, fine straining are 150 ℃ in inlet temperature, leaving air temp is 60 ℃, and air velocity is 20ms ^-1Condition under spray drying get powder, packing, promptly.

Among the present invention: Herba Erigerontis extract can be a flavones ingredient content greater than 90% total flavone valid target, and Radix Paeoniae Rubra extract can be the glycoside component content greater than total glycosides effective part of 90%.

Among the present invention: Radix Paeoniae Rubra and Radix Paeoniae Rubra extract can also substitute with the Radix Paeoniae Alba and the Radix Paeoniae Alba extract of equivalent.

Described preparation also can be used to prepare the medicine of diseases such as treatment hepatorenal syndrome, heart and lung diseases, diabetes and complication thereof.

That is to say that the product of the present invention's preparation can be used for the treatment of diseases such as hepatorenal syndrome, heart and lung diseases, diabetes and complication thereof.

Compared with prior art, we flee meridians according to wind-phlegm, the resistance of blood vessels numbness, and channel tunnel is obstructed, the gas unable to walk, blood can not moisten, so limbs come into disuse into the etiology and pathogenesis of hemiplegia, abide by the reason of " control wind and control blood earlier, blood sector-style go out ", take the method for activating blood circulation to dissipate blood stasis and dredge the collateral.Herba Erigerontis with the bitter temperature of nature and flavor is monarch, the disperse blood stasis and dredge collateral of using; The Radix Paeoniae Rubra bitter in the mouth is slightly cold, and goes into blood system, removing heat from blood heat, and dissipating blood stasis blood, the meridian dredging is controlled the heat in blood stasis of blood and is stagnated, and principal drug assistance is to reach the effect of blood circulation promoting and blood stasis dispelling, dissipating blood stasis collateral dredging, and what use is minister.Two medicines share has blood circulation promoting and blood stasis dispelling, the function of dredge the meridian passage.The useful YAOJING letter of the prescription length that power is special, action pathway is direct does not have the cooler fraud of temperature partially, and comparatively significantly administration features is arranged, by reasonable feasible technology, can not produce the anaphylaxis material in the preparation process, guarantee the safe and effective of product, but the patients life-time service.

The applicant has carried out detailed research to each link in development process:

Extraction process route research: by the contained chemical constitution study of the other side's Chinese medicine, analyze the effective ingredient and the pharmacological action of every flavor medical material, in conjunction with the different pharmacodynamics test research of solvent, Different Extraction Method and the rates of transform of index components extracted, determined our basic technology route.

Prescription proportioning and the side's of tearing open research: we are evaluation index with the protective effect of chmice acute cerebral ischemia, and two flavor medical materials are formed proportionings and carried out the best prescription screening among the other side, and the result shows: the ratio of Herba Erigerontis and Radix Paeoniae Rubra consumption is to be this side's optimum formula at 1: 1.5.Simultaneously act as the research of the index side of tearing open with the chmice acute cerebral ischemia, the result shows that the breathing that each administration group all can prolong the acute cerebral ischemia mice holds time, and the action intensity that two medicines share significantly is better than single medication group (P＜0.01).

The selection of extracting method and extraction conditions: two flavor medical materials are through different solvent, Different Extraction Method and independent extraction, the united extraction of extracting in the side, carry out dynamic tracking with indexs such as chemical index composition and pharmacodynamics tests, the unsuitable mixed extraction of result of the test prompting two flavor medical materials, sample physiologically active with the water boiling and extraction preparation is stronger, extraction ratio of effective constituents height in the preparation, the quality of the pharmaceutical preparations are easy to control.In the test solvent load, extraction time, extraction time etc. are carried out orthogonal test, screened optimum extraction process.

Separation, purifying process: in the extract of medical material, except that effective ingredient, still contain many impurity components, be quality that guarantees product and the quality control that helps product, need further refining purification, remove the impurity such as macromole phlegmatic temperament, protein, starch and solid particle in the extract that anhydrates as much as possible, improve content of effective.Through the distinct methods contrast test, the decoction liquor of two flavor medical materials all can adopt Ethanol Treatment.The Herba Erigerontis acidify is refining; (test adopts methods such as absorption with macroporous adsorbent resin separation, n-butanol extraction to carry out physicochemical property research and pharmacodynamics test to Radix Paeoniae Rubra with n-butanol extraction; the result shows that one of the active component in Radix Paeoniae Rubra liposoluble ingredient mainly is present in water lotion and stream is worn in the liquid; and show all that to the influence of focal cerebral ischemia in rats (MCAO) cerebral blood flow and to the protective effect result of the test of chmice acute cerebral ischemia water lotion and stream wears liquid and still have stronger pharmacologically active, illustrating that macroporous adsorbent resin separates can not be with the effective enrichment of effective ingredient).Purified purified sample aggregative indicator is good, and the purity and the yield of main component are higher, and product quality is easy to control, shows that through the investigation of experimental study method with to the results of pharmacodynamic test made from extra care component (semi-finished product) this technology is reasonable, feasible.

Removing the tannin method selects: contain tannin in the Radix Paeoniae Rubra, must remove tannin and handle.Through the various comparative studies that remove the tannin method, adopt the extremely strong polyamide of tannin absorption affinity is removed tannin.The result shows, it is better that this method is removed the tannin effect, loss of effective components is few, the rate of transform height of peoniflorin, tannin in the sample is checked up to specification, the undue toxicity of mice checks and can reach (through commercially available several Chinese medicines are compared inspection, the undue toxicity of mice is mostly about 60 times) more than 120 times, removes the tannin method apparently higher than other.Illustrate that this product is feasible with the method for polyamide removal tannin, the safety of preparation is effectively guaranteed.

The selection of semi-finished product drying means: through the test contrast to different drying conditions, the result shows that vacuum drying is less to the influence of effective ingredient, and rate of drying is fast, and temperature is low, and the sample quality is loose, is easy to dissolving.

Preparations shaping technical study:, determined the basic parameter of preparations shaping technology and the prescription that the preparation semi-finished product feed intake through tests such as preferred, freeze drying process to the influence of the selection of additives and consumption, pH value, sterilising conditions.

In addition, the present invention also provides the detailed processing technology of other extraction processes, several formulations in embodiment, can be directly used in and instruct actual production.

The applicant has carried out a series of experiments, can prove that medicine provided by the invention has effective effect.

Experimental example 1: the pharmacodynamics comparative study of different formulations

(1) to the influence of stasis syndrome rat blood rheological characteristic: 70 of rats, male and female half and half, body weight 270 ± 20g successive administration 12 days, 1h after the last administration, all the other respectively organize equal sc injection epinephrine 0.8mg/kg, totally twice, two minor tick 4h except that the normal control group.(front and back each 2 hours at interval) immerse 5min in the frozen water, fasting with rat between twice.Femoral artery blood sampling in morning next day, each index of hemorheology is measured in the heparin sodium anticoagulant.

Group plasma viscosity packed cell volume reduced viscosity

Normal control 1.52 ± 0.34 0.43 ± 0.11 7.78 ± 0.29

Model contrast 2.15 ± 0.18 0.55 ± 0.04 8.39 ± 3.43

Radix Paeoniae Rubra 2g/kg 1.96 ± 0.24 0.48 ± 0.27 8.20 ± 2.54

Herba Erigerontis 2g/kg 1.81 ± 0.18 0.45 ± 0.08 8.10 ± 1.72

Radix Paeoniae Alba 2g/kg 1.98 ± 0.27 0.49 ± 0.39 8.21 ± 1.33

Oil lamp Radix Paeoniae Alba extracting solution 1.70 ± 0.45 0.44 ± 0.87 7.85 ± 1.33

Oil lamp Radix Paeoniae Rubra extracting solution 1.68 ± 0.19 0.43 ± 0.15 7.81 ± 1.64

The result shows that the Herba Erigerontis main component is the breviscapine total flavones, and chemistry is by name 4,5,6-trihydroxyflavone-7-glucuronide, have suppress that platelet is gathered, anticoagulant function, microcirculation improvement effect, can suppress the transhipment of calcium pump to calcium ion; Radix Paeoniae Rubra can reduce the blood plasma lipide peroxide, reduces calcium and is deposited on arterial wall, influences calcium metabolism, the balance atherosclerosis; Behind Herba Erigerontis and the Radix Paeoniae Rubra compatibility ability of improving the blood stasis model blood status is more by force arranged, blood stasis rat plasma viscosity, packed cell volume, platelet aggregation are significantly reduced, prescription of the present invention can synergy.This experiment also proves: substitute the preparation that Radix Paeoniae Rubra and Radix Paeoniae Rubra extract obtain with the Radix Paeoniae Alba of equivalent and Radix Paeoniae Alba extract and have essentially identical effect.

(2) to the influence of blood stasis model rabbit blood rheological characteristic

64 of rabbit are divided into 8 groups at random, and 8 every group, ♀ ♂ half and half.Each treated animal elder generation auricular vein is injected (iv) administration.Blank and model control group injecting normal saline (NS) 1ml/kg, positive drug group iv puerarin injection 30mg/kg (being made into 30mg/ml) with NS, experimental group is injected the medicinal liquid (being made into NF) of 0.25mg/ml respectively, dosage is 1ml/kg, two weeks of successive administration (14d), in administration the 2nd, 13d, except that the blank group, each rabbit is annotated 10% high molecular dextran 5ml/kg through auricular vein respectively, every day twice, causes blood stasis model.After the last administration, inject 10% high molecular dextran 5ml/kg once more, behind the 15min, heart is taked fasting blood 6ml, carrying out hemorheology index detects, wherein platelet aggregation rate adopts turbidimetry for Determination: the rotary cone-plate viscosity apparatus mensuration of other employing LBY-N6A+ with LBY-NJ2 type platelet aggregation instrument.

To rabbit platelet aggregation and fibrinogenic influence (x ± s, n=6)

Group dosage body weight (kg) platelet aggregation (%) Fibrinogen (g/L)

Blank group-2.41 ± 0.22 16.33 ± 1.73 2.87 ± 3.18

Model group-2.49 ± 0.16 39.06 ± 17.13 2.65 ± 1.29

Positive drug group 30mg/kg 2.38 ± 0.15 12.48 ± 0.36 3.04 ± 2.88

Oil lamp: Radix Paeoniae Rubra (1: 99) 2g/kg 2.40 ± 0.27 13.11 ± 0.23 2.91 ± 1.53

Oil lamp: Radix Paeoniae Rubra (1: 4) 2g/kg 2.39 ± 0.35 13.02 ± 3.05 2.95 ± 1.32

Oil lamp: Radix Paeoniae Rubra (2: 3) 2g/kg 2.37 ± 0.13 12.50 ± 0.40 3.03 ± 0.87

Oil lamp: Radix Paeoniae Rubra (4: 1) 2g/kg 2.40 ± 0.15 13.19 ± 0.30 2.90 ± 0.69

Oil lamp: Radix Paeoniae Rubra (99: 1) 2g/kg 2.41 ± 0.27 13.23 ± 0.42 2.96 ± 0.58

Preliminary experiment shows, oil lamp, Radix Paeoniae Rubra compatibility are effective, and the applicant has also carried out detailed prescription proportioning and the side's of tearing open research in the technical study process.

Experimental example 2: Study on extraction

For understanding effective site and the main effective ingredient of we to the cerebrovascular disease effect, we extract solvent, extracting method (close and fry in shallow oil, singly fry in shallow oil) to we, and prescription proportioning, different way of administration, different extraction separation, the pharmacodynamics of refining purification process and the dynamic change of chemical constituent are studied.Result of the test shows: its pharmacologically active is relevant with the material base in the medicinal liquid, and the compound recipe drug effect is better than any single medicinal material; The intravascular administration effect obviously is better than extravascular administration; Not only kept effective ingredient behind the two flavor medical material purification refines, and the quality of the pharmaceutical preparations effectively improves, reach the prescription of used for intravenous injection preparation.

1 selection of extracting solvent, extracting method is an evaluation index with the protective effect of chmice acute cerebral ischemia, adopts systematic solvent extraction, to two flavor medical material extractions respectively, mixed extraction, extract the back biased sample respectively and test.

1.1 Herba Erigerontis, Radix Paeoniae Rubra medical material are got in sample preparation, extract successively with ethyl acetate, n-butyl alcohol, second alcohol and water respectively, the segmentation sample preparation gets sample TR1 (A), TR1 (B), TR1 (C), TR1 (D), TR2 (A), FR2 (B), TR2 (C), TR2 (D), TR3 (A), TR3 (B), TR3 (C), TR3 (D), TR4 (A), TR4 (B), TR4 (C), TR4 (D) extract.Above-mentioned each sample is further removed residual solvent in water-bath, add the dissolving of suitable quantity of water slight fever, adjust pH to 6, fully stir, Dropwise 5 % gelatin solution does not produce to there being precipitation, adds ethanol again and makes that to contain the alcohol amount be 75%, leaves standstill 12 hours, filter, filtrate recycling ethanol concentrates near doing, and thin up extremely every 1ml contains the 2.0g crude drug, adjust pH to 7.0 boils, and puts cold, filter, cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, the filtrate packing, sterilization, standby.Above-mentioned sample is carried out pharmacodynamics test relatively, investigate this side's extraction solvent, extracting method.

1.2 experimental technique and result

Get about 180 of Kunming mouse, body weight 18～22g, the male and female dual-purpose is divided into 18 groups at random, 10 every group (male and female half and half).Sample TR-1:2.4g crude drug/kg; Sample TR-2:3.6g crude drug/kg; Sample TR-3:6.0g crude drug/kg; Sample TR-4:6.0g crude drug/kg; Positive control drug: 4.0ml/kg; Model group gives isometric normal saline.Each group is all by the tail intravenously administrable, and the administration volume is 10ml/kg.Behind the intravenously administrable 15min, dorsal position is fixed, with cutting off in the middle of 0 trumpeter's art suture two ends ligation bilateral common carotid arteries (merging vagus nerve) back.The breathing of record mice is held time, and the results are shown in following table.

Result of the test shows, except that TR-1 (A), TR-1 (B), TR-1 (D), TR-2 (A), TR-3 (A), six samples of TR-3 (D), each administration group all can prolong the mouse breathing of acute cerebral ischemia holds time, with model group comparing difference remarkable (P＜0.05, P＜0.01).Wherein ethanol and n-butanol extracting liquid play a role clearly, and with model group significant difference (P＜0.05, P＜0.01) is arranged relatively, and the effect of the more above-mentioned two kinds of solvents of effect of ethyl acetate and water extract are poor.

Result of the test prompting: first, effective ingredient among this side is mainly the bigger material of polarity, simultaneously organic solvent extraction is described after, still have active component in the water extract, use the influence of priority to pharmacological action for getting rid of solvent, two flavor medical materials select water, ethanol etc. to carry out demonstration test respectively.The second, the effect of sample 4 is obviously than sample the last 3, and it is more effective than united extraction to illustrate that two medicines separately extract.

The influence that the acute cerebral ischemia mouse breathing is held time (x ± s, n=10)

Sample	Dosage (the g crude drug/kg)	Number of animals (only)	Breathing hold time (min)
				Model group positive drug group TR-1 (A)	- 4.0ml 2.4	10 10 10	2.41±1.22 4.17±1.90 * 3.18±1.43

(B) (C) (D) TR-2(A) (B) (C) (D) TR-3(A) (B) (C) (D) TR-4(A) (B) (C) (D)

3.6 6.0 6.0

10 10 10 10 10 10 10 10 10 10 10 10 10 10 10

3.71±1.75 3.83±1.71 * 3.05±1.18 3.14±1.14 4.00±2.02 * 4.03±1.87 * 3.98±1.96 * 2.62±1.04 3.98±1.97 * 4.08±2.15 * 3.03±1.34 3.64±1.19 * 4.07±2.03 * 4.87±2.23 ** 4.10±2.07 *

Compare * P＜0.05 * * P＜0.01 with model group

Annotate: sample TR1 is that the Herba Erigerontis medical material extracts separately, and sample TR2 is that the Radix Paeoniae Rubra medical material extracts separately.Sample TR3 is two flavor medical material mixed extraction, after sample TR4 extracts for difference is independent, and extract mixing sample preparation.The A-ethyl acetate extraction, B-n-butanol extraction, C-are ethanol extraction, D is that decocting in water extracts.Positive control drug: FUFANG DANSHEN ZHUSHEYE, Shanghai General Pharmaceutical Co., ltd.'s product, lot number 0302182.

2 extract separately or the united extraction process choice

Being further to determine we's extraction process, is evaluation index with the protective effect of chmice acute cerebral ischemia, and two flavor medical materials extract separately or united extraction technology is tested comparison among the other side.

2.1 sample preparation

Independent water boiling and extraction: get Herba Erigerontis 100g, add 12 times of water gagings and decoct each 0.5 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 24 hours, filter, concentrate, add ethanol again and make that to contain the alcohol amount be 70%, left standstill 24 hours, and filtered filtrate recycling ethanol, concentrate near doing, get Herba Erigerontis extract; Get Radix Paeoniae Rubra medical material 150g again, add 10 times of water gagings and decoct each 1 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 12 hours, filter, reclaim solvent and be concentrated into every 1ml and contain the 2g crude drug, Dropwise 5 % gelatin solution does not produce to there being precipitation, add ethanol again and make that to contain alcohol amount be 75%, left standstill 12 hours, filter, filtrate recycling ethanol concentrates near doing, and gets Radix Paeoniae Rubra extract.Get Radix Paeoniae Rubra and Herba Erigerontis extract and add the dissolving of injection water 120ml slight fever, adjust pH to 7.0 is added water for injection to 125ml, boils, put coldly, filter, cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, the filtrate packing, sterilization promptly gets (every 1ml contains crude drug 2.0g).

Merge water boiling and extraction: get Herba Erigerontis 100g, Radix Paeoniae Rubra medical material 150g adds 10 times of water gagings and decocts each 1 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 24 hours, filter, filtrate recycling ethanol to and be concentrated into every 1ml and contain crude drug 2g, Dropwise 5 % gelatin solution does not produce to there being precipitation, add ethanol again and make that to contain alcohol amount be 75%, left standstill 12 hours, filter, filtrate recycling ethanol, concentrate near doing, get extract, add the dissolving of injection water 120ml slight fever, adjust pH to 7.0, add water for injection to 125ml, boil, put cold, filter, cold preservation 24 hours filters the filtrate packing with 0.45 μ m microporous filter membrane, sterilization promptly gets (every 1ml contains crude drug 2.0g).

Positive control drug: FUFANG DANSHEN ZHUSHEYE, specification: 2ml/ props up.The 3rd company of Shanghai General Pharmaceutical Co., ltd., lot number 0302182.

The sodium chloride injection that the drug dilution method is faced with preceding usefulness 0.9% is diluted to suitable concentration, and is standby.

2.2 pharmacodynamics test method and result

About 40 of Kunming mouse is got in the protective effect of chmice acute cerebral ischemia, body weight 18～22g, the male and female dual-purpose is divided into 4 groups at random, 10 every group (male and female half and half).Sample TF1,2 dosages are: 2.5g crude drug/kg; Positive control drug: 4.0ml/kg; Model group gives isometric normal saline.Each group is all by the tail intravenously administrable, and the administration volume is 10ml/kg.Behind the intravenously administrable 15min, dorsal position is fixed, with cutting off in the middle of 0 trumpeter's art suture two ends ligation bilateral common carotid arteries (merging vagus nerve) back.The breathing of record mice is held time, and result of the test sees the following form.

The influence that the acute cerebral ischemia mouse breathing is held time (x ± s, n=10)

Compare * P＜0.05, * * P＜0.01 with model group

TF-1: united extraction sample, TF-2: extract sample separately, down together

Result of the test shows, each administration group all can prolong the mouse breathing of acute cerebral ischemia and hold time, with model group comparing difference remarkable (P＜0.05, P＜0.01), wherein two flavor medical materials separate extraction effect obviously (P＜0.01), compare there was no significant difference (P＞0.05) between group.

2.3 the rate of transform of index components in the sample

Get above-mentioned sample respectively, measure in the Herba Erigerontis content of paeoniflorin in the scutellarin and Radix Paeoniae Rubra in accordance with the law, calculate the rate of transform, the results are shown in following table.

Different Extraction Method index components rate of transform result of the test

Separately extracting the scutellarin of sample and the peoniflorin rate of transform all apparently higher than the united extraction sample, mainly is that especially the influence to scutellarin is bigger because in decoction process, extracting liquid pH value lower (pH=4～5) influences the extraction of effective ingredient.

To sum up, the second alcohol and water is effective extraction solvent of Herba Erigerontis and Radix Paeoniae Rubra, and the extract of independent extraction and united extraction all can prolong the acute cerebral ischemia mouse breathing and hold time.Although compare there was no significant difference between group, the effect of extracting obviously is better than united extraction separately; Water boiling and extraction scutellarin and the peoniflorin rate of transform in sample is higher, separately extracts the scutellarin and the peoniflorin rate of transform apparently higher than united extraction.In conjunction with the results of pharmacodynamic test analysis, scutellarin and peoniflorin are our active substance in this product, so we should be that solvent decocts extraction with inexpensive water, two flavor medical materials should not merge decoction.

3 prescription proportionings are proved recipe with the side of tearing open research we; clinical practice prescription proportioning is not quite similar; for further inquiring into the reasonability of product of the present invention (freeze-dried powder) prescription proportioning and prescription; with chmice acute cerebral ischemia, anoxybiotic protective effect is evaluation index, and the two flavor medical materials composition proportionings and the side of tearing open carry out pharmacodynamics test research among the other side.

3.1 prescription proportion research

3.1.1 prescription-1 (Herba Erigerontis: Radix Paeoniae Rubra=1: 1): get Herba Erigerontis 120g, add 12 times of water gagings and decoct each 0.5 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 24 hours, filter, concentrate, add ethanol again and make that to contain the alcohol amount be 70%, left standstill 24 hours, and filtered filtrate recycling ethanol, concentrate near doing, get Herba Erigerontis extract.Get Radix Paeoniae Rubra 120g again, add 10 times of water gagings and decoct each 1 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 12 hours, filter, concentrate, Dropwise 5 % gelatin solution does not produce to there being precipitation, add ethanol again and make that to contain alcohol amount be 75%, left standstill 12 hours, filter, filtrate recycling ethanol concentrates near doing, and gets Radix Paeoniae Rubra extract.Get Radix Paeoniae Rubra and Herba Erigerontis extract and add an amount of slight fever dissolving of injection water, adjust pH to 7.0 is added water for injection to 120ml, boils, put coldly, filter, cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, the filtrate packing, sterilization promptly gets (every 1ml contains crude drug 2g).

The prescription-2 (Herba Erigerontis: Radix Paeoniae Rubra=1: 1.5): get Herba Erigerontis 120g, Radix Paeoniae Rubra 180g, respectively by last method operate extract.Get two extracts and add an amount of water for injection slight fever dissolving, adjust pH to 7.0 is added water for injection to 150ml, with the method operation promptly.

(Herba Erigerontis: Radix Paeoniae Rubra=1: 2): get Herba Erigerontis 120g, Radix Paeoniae Rubra 240g is respectively by last method operation for prescription-3.Get two extracts and add an amount of water for injection slight fever dissolving, adjust pH to 7.0 is added water for injection to 180ml, with the method operation promptly.

(Herba Erigerontis: Radix Paeoniae Rubra=2: 1): get Herba Erigerontis 180g, Radix Paeoniae Rubra 90g is respectively by last method operation for prescription-4.Get two extracts and add an amount of water for injection slight fever dissolving, adjust pH to 7.0 is added water for injection to 135ml, with the method operation promptly.

The drug dilution method face with the sodium chloride injection of preceding usefulness 0.9% (Pubei, Guangxi pharmaceutical factory, lot number: A0301153) be diluted to suitable concentration, standby.

3.1.2 test method and result

About 210 of Kunming mouse is got in the protective effect of chmice acute cerebral ischemia, body weight 18～22g, the male and female dual-purpose is divided into 21 groups at random, 10 every group (male and female half and half).Each dosage group according to the form below administration; Model group gives isometric normal saline.Each group is all by the tail intravenously administrable, and the administration volume is 10ml/kg.Behind the intravenously administrable 15min, dorsal position is fixed, with cutting off in the middle of 0 trumpeter's art suture two ends ligation bilateral common carotid arteries (merging vagus nerve) back.The breathing of record mice is held time, and result of the test sees the following form.

The influence that the acute cerebral ischemia mouse breathing is held time (x ± s, n=10)

Sample	Dosage (the g crude drug/kg)	Breathing hold time (min)	Effective percentage (%)	ED 50(g/kg)
					Model group CF-1 (1: 1)	- 0.60 0.86 1.23 1.75 2.50	2.47±1.11 2.51±0.90 2.58±1.15 3.00±1.36 3.84±1.72 * 3.98±1.32 *	10 20 30 50 70	1.60#

CF-2 (1∶1.5) CF-3 (1∶2) CF-4 (2∶1)

0.60 0.86 1.23 1.75 2.50 0.60 0.86 1.23 1.75 2.50 0.60 0.86 1.23 1.75 2.50

2.64±1.03 3.22±1.35 3.67±1.09 * 3.88±1.06 ** 4.73±1.20 ** 2.31±1.01 2.67±1.03 3.67±1.39 * 3.77±1.49 * 3.91±1.15 * 2.39±0.93 2.49±0.84 3.38±1.23 3.73±1.40 * 3.89±1.26 *

20 30 60 80 90 10 20 40 50 70 10 10 30 50 70

1.09 1.54 1.66#

Compare with model group: * P＜0.05, * * P＜0.01, compare with prescription 2: #P＜0.05, ##P＜0.01

CF-1～CF-4: prescription 1～prescription 4, down together.

The result shows that the extract of 4 kinds of different proportioning prescriptions all can prolong the acute cerebral ischemia mouse breathing in various degree and hold time.By improving the median effective dose (ED that karber's method calculates each proportioning prescription ₅₀) ascending being followed successively by: prescription 2 (the 1.09g crude drug/kg)＜prescription 3 (the 1.54g crude drug/kg)＜prescription 1 (the 1.60g crude drug/kg)＜prescription 4 (the 1.66g crude drug/kg).The ED of prescription 2 ₅₀Significantly less than prescription 1,4; Prescription 2 is less than prescription 3, but difference with insignificance.As from the foregoing, 2 best results of writing out a prescription, promptly Herba Erigerontis is 1: 1.5 with the ratio of Radix Paeoniae Rubra consumption.

3.2 the prescription side of tearing open research

3.2.1 sample preparation

Herba Erigerontis sample: get Herba Erigerontis 300g, add 12 times of water gagings and decoct each 0.5 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 24 hours, filter, concentrate, add ethanol again and make that to contain the alcohol amount be 70%, left standstill 24 hours, and filtered filtrate recycling ethanol, concentrate near doing, get Herba Erigerontis extract.Add the dissolving of an amount of slight fever of injection water, adjust pH to 7.0 is added water for injection to 150ml, boils, and puts coldly, filters, and cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, and packing is sterilized, and promptly gets (every 1ml contains crude drug 2g).

Radix Paeoniae Rubra sample: get Radix Paeoniae Rubra 450g, add 10 times of water gagings and decoct each 1 hour 3 times, filter, merging filtrate is concentrated into the concentration that every 1ml contains crude drug 2g, add ethanol and make that to contain alcohol amount be 60%, stir, left standstill 12 hours, filter, concentrate, Dropwise 5 % gelatin solution does not produce to there being precipitation, add ethanol again and make that to contain alcohol amount be 75%, left standstill 12 hours, filter, filtrate recycling ethanol concentrates near doing, and gets Radix Paeoniae Rubra extract.Add the dissolving of an amount of slight fever of injection water, adjust pH to 7.0 is added water for injection to 225ml, boils, and puts coldly, filters, and cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, and packing is sterilized, and promptly gets (every 1ml contains crude drug 2g).

Two medicines share sample: get Herba Erigerontis 300g, Radix Paeoniae Rubra 450g, extract as stated above respectively, get extract.Two extracts merge, and add an amount of slight fever dissolving of injection water, and adjust pH to 7.0 is added water for injection to 375ml, boils, and put coldly, filter, and cold preservation 24 hours filters with 0.45 μ m microporous filter membrane, and packing is sterilized, and promptly gets (every 1ml contains crude drug 2g).

3.2.2 test method and result

3.2.2.1 protective effect to the chmice acute cerebral ischemia

Get 50 of Kunming mouses, body weight 18～22g, the male and female dual-purpose is divided into 5 groups at random, 10 every group (male and female half and half).Sample CFYJ-1 dosage is 1.0g crude drug/kg, and sample CFYJ-2 dosage is 1.5g crude drug/kg, and sample CFYJ-3 dosage is 2.5g crude drug/kg; Positive controls: 4.0ml/kg; Model group gives isometric normal saline.Each group is all by the tail intravenously administrable, and the administration volume is 10ml/kg.Behind the intravenously administrable 15min, dorsal position is fixed, with cutting off in the middle of 0 trumpeter's art suture two ends ligation bilateral common carotid arteries (merging vagus nerve) back.The breathing of record mice is held time, and result of the test sees the following form.

The influence that the acute cerebral ischemia mouse breathing is held time (x ± s, n=10)

Compare with model group: * P＜0.05, * * P＜0.01

Compare with the CFJY-3 group: #P＜0.05, ##P＜0.01

CFJY-1: Herba Erigerontis CFJY-2: Radix Paeoniae Rubra CFJY-3: Herba Erigerontis+Radix Paeoniae Rubra

Result of the test shows that each administration group all can prolong the breathing of acute cerebral ischemia mice holds time, with model group comparing difference remarkable (P＜0.05, P＜0.01); Herba Erigerontis and Radix Paeoniae Rubra drug combination group, action intensity significantly are better than single medication group (P＜0.01), illustrate that the effect of compound recipe obviously is better than folk prescription.

3.2.2.2 prescription interactive analysis

Reciprocal action for existing after further research two medicines share adopts L4 (2 ³) orthogonal experiment carries out interactive analysis, the results are shown in following table.

L4 (2 ³) orthogonal design table and result (x ± s, n=10)

From the visual result analysis of last table, the order of each factor affecting drug effect is, Herba Erigerontis is slightly larger than Radix Paeoniae Rubra (A＞B) for the contribution of drug effect; The orthogonal experiment ANOVA showed significant, Herba Erigerontis or Radix Paeoniae Rubra all have utmost point appreciable impact (P＜0.01) for drug effect, and there is obvious interaction (P＜0.01) in two medicines.Interactive analysis is as follows:

Interactive analysis	Without Radix Paeoniae Rubra (∑ x)	With Radix Paeoniae Rubra (∑ x)
			Without oil lamp (∑ x)	23.60	32.23
With oil lamp (∑ x)	32.66	58.73

With not medication test group as radix (23.60), Herba Erigerontis drug effect=(32.66-23.60), Radix Paeoniae Rubra drug effect=(32.23-23.60), oil lamp+Radix Paeoniae Rubra drug effect=(58.73-23.60).Because (58.73-23.60)＞(32.66-23.60)+(32.23-23.60),, illustrate that two medicines share and have synergy so there are reciprocal action in Herba Erigerontis and Radix Paeoniae Rubra.

4. system process research

4.1 Herba Erigerontis Study on extraction

4.1.1 the orthogonal test of decocting process

The design of orthogonal test factor level table: according to result of the test, amount of water, decocting time, decoction number of times are the principal elements that influence decocts efficient, by each factor 3 level, use L ₉(3 ⁴) orthogonal table arrangement test.With the scutellarin yield serves as to investigate index, and extraction process is inquired into.

The factor level table

Herba Erigerontis decocting process condition L ₉(3 ⁴) orthogonal test arrangement and result

Analysis of variance table

F _0.05(2，2)＝19.00 F _0.01(2，2)＝99.00

Interpretation of result:

Visual result analysis from last table, the order of each factor affecting extraction efficiency are C＞B＞A, and wherein extraction time and extraction time are bigger to the extraction process influence; The results of analysis of variance shows that extraction time has utmost point significant difference (P＜0.01), and there are significant difference (P＜0.05), both conclusion unanimities extraction time.Optimum process condition is A ₃B ₁C ₃, the extreme difference of optimised process and time good technology is less in the A factor, considers that from saving the energy and the angle of time inferior good technology is A ₂B ₁C ₃So with optimised process A ₃B ₁C ₃With inferior good technology A ₂B ₁C ₃Carry out repeated trials (the results are shown in following table).

The orthogonal test reproducible results

From last table result as seen, the scutellarin yield no significant difference of first and second liang of technologies from saving of work and time, the angle that cuts down the consumption of energy, selects second technology to produce.Promptly add the water (adding 3 times of amounts for the first time) of 10 times of medical material weight, decoct each 0.5 hour 3 times.

4.1.2 the selection of Herba Erigerontis purification process:

The effective ingredient of Herba Erigerontis is a flavone compound, and this compounds is a polyatomic phenol, water soluble, ethanol, under acid condition, separate out precipitation, water insoluble, under neutral and weak basic condition, dissolve, so adopt the method refining and edulcoration of ethanol precipitation, acidification.

4.1.2.1 the orthogonal test of alcohol precipitation process

The design of orthogonal test factor water-glass: the relative density of medicinal liquid, alcohol precipitation concentration are (according to result of the test, scutellarin dissolubility in 50% ethanol is better, so range of alcohol concentration fixes on 45-65% test), be the principal element that influence alcohol precipitation process standing time, presses each factor 3 level, uses L ₉(3 ⁴) orthogonal table arrangement test (following table).With the scutellarin extraction ratio serves as to investigate index, and extraction process is inquired into.

The factor level table

Sample preparation: get 9 parts of decocting liquid that are equivalent to the 100g crude drug, be concentrated into desired concn respectively, test by condition shown in the orthogonal test table.Sampling and measuring scutellarin concentration is calculated the scutellarin amount, is that index is estimated with the scutellarin rate of transform, the results are shown in following table.

Herba Erigerontis alcohol precipitation process condition L ₉(3 ⁴) orthogonal test arrangement and result

Analysis of variance table

F _0.05(2，2)＝19.00 F _0.01(2，2)＝99.00。

Interpretation of result:

Visual result analysis from last table, the order of each factor affecting precipitate with ethanol efficient is A＞B＞C, wherein relative density and concentration of alcohol are bigger to rate of transform influence; The results of analysis of variance shows that relative density has utmost point significance influence (P＜0.01) to the rate of transform, and concentration of alcohol has significance influence (P＜0.05) to the rate of transform, both conclusion unanimities, and optimum process condition is A ₃B ₁C ₃, inferior good technology A ₂B ₂C ₂In A, B factor, the extreme difference of optimised process and time good technology is less, and concentration of alcohol is lower than difficulty of alcohol deposit fluid filtration in 50% o'clock in the B factor, and the C factor is a secondary cause, takes all factors into consideration, with optimised process A ₃B ₁C ₃With inferior good technology A ₂B ₂C ₂Carry out repeated trials (the results are shown in following table).

The orthogonal test reproducible results

From last table result as seen, the scutellarin rate of transform no significant difference of first and second liang of technologies, but first technology is low excessively owing to concentration of alcohol, and solution is difficult for heavy clear, and is difficult during sample filtering, so selection second technology is produced.Be decoction liquor to be concentrated into relative density be 1.10 (50 ℃), add ethanol makes that to contain the alcohol amount be 55%, stir, placed filtration, decompression filtrate recycling ethanol 12 hours.

4.1.2.2 the optimization of acidulated condition

Flavonoid is a polyatomic phenol in the Herba Erigerontis, can separate out precipitation under acid condition, and is water insoluble, dissolves under neutral and weak basic condition, so adopt the method for acidification to prepare Herba Erigerontis extract.

The design of acidification technique orthogonal test factor water-glass: the relative density of medicinal liquid, pH value, holding temperature, standing time are the principal elements that influences acidification technique, by each factor 3 level, use L ₉(3 ⁴) orthogonal table arrangement test (following table).Investigate index with the scutellarin extraction ratio, acidification technique is investigated.

The factor level table

Sample preparation: get 18 parts of medicinal liquids that are equivalent to the 50g crude drug, be concentrated into desired concn, press condition test shown in the orthogonal test table, the collecting precipitation thing, to 500ml, measuring scutellarin content with 70% dissolve with ethanol, is that evaluation index is estimated with the rate of transform of scutellarin.

Quadrature engineer testing result

Acidification technique condition L ₉(3 ⁴) orthogonal test arrangement and result

Analysis of variance table

F _0.05(2，9)＝4.26 F _0.01(2，9)＝8.02。

Interpretation of result

From the visual result analysis, the order of each factor affecting acidify efficient is B＞C＞A＞D, and wherein pH value, holding temperature and relative density are bigger to technogenic influence; The results of analysis of variance shows that each factor all has utmost point significant difference (P＜0.01), and optimum process condition is A ₂B ₂C ₃D ₂, the extreme difference of optimised process and inferior good technology is less in C, D factor, takes all factors into consideration, and choosing time good technology is A ₂B ₂C ₂D ₁With optimised process A ₂B ₂C ₃D ₂With inferior good technology A ₂B ₂C ₂D ₁Carry out repeated trials, the results are shown in following table.

The orthogonal test reproducible results

From last result as seen, the scutellarin yield no significant difference of first and second liang of technologies from the angle of saving of work and time, selects second technology to produce.Be that medicinal liquid is concentrated into relative density 1.11 (50 ℃), with hydrochloric acid adjust pH to 2, in 55 ℃ of insulations 6 hours.

4.1.3 the selection of Herba Erigerontis extract drying means

Get Herba Erigerontis 1500g, add 10 times of water gagings and decoct each 0.5 hour 3 times, filter, merging filtrate, filtrate decompression is concentrated into relative density 1.10 (50 ℃), add 2 times of amount 85% ethanol, constantly stir, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.10 (50 ℃), with hydrochloric acid adjust pH to 2,55 ℃ are incubated 6 hours, tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, is divided into 3 parts.According to condition carry out drying, get the loss rate that dry product is measured extract dry back scutellarin under different condition, result of the test sees the following form.

Different drying conditions are to the influence of scutellarin

Result of the test shows, the rate of transform no significant difference of scutellarin under each drying condition, and vacuum drying speed is fast, and drying time is short, and the sample quality is loose, so select 60～80 ℃ of vacuum dryings for use.

4.2 the Study on extraction of Radix Paeoniae Rubra

4.2.1 the orthogonal test of decocting process

The design of orthogonal test factor level table: amount of water, decocting time, decoction number of times are the principal elements that influence decocts efficient, by each factor 3 level, use L ₉(3 ⁴) orthogonal table arrangement test.With peoniflorin extraction ratio and solids yield serves as to investigate index, and extraction process is inquired into.

The factor level table

Radix Paeoniae Rubra decocting process condition L ₉(3 ⁴) orthogonal test arrangement and result

Peoniflorin yield analysis of variance table

F _0.05(2，2)＝19.00 F _0.01(2，2)＝99.00。

Total solid yield analysis of variance table

F _0.05(2，2)＝19.00 F _0.01(2，2)＝99.00。

Interpretation of result:

From the visual result analysis, serve as that the order of each factor affecting extraction efficiency is C＞B＞A when investigating index with the peoniflorin yield, wherein extraction time and extraction time are bigger to the extraction process influence; The results of analysis of variance shows that extraction time has utmost point significant difference (P＜0.01), and there is significant difference (P＜0.05) extraction time, both conclusion unanimities, and optimum process condition is A ₃B ₂C ₃, the extreme difference of optimised process and time good technology is less in the A factor, considers that from saving the energy and the angle of time inferior good technology is A ₂B ₂C ₃From the visual result analysis, serve as that the order of each factor affecting extraction efficiency is C＞B＞A when investigating index with the total solid yield, wherein extraction time and extraction time are bigger to the extraction process influence; The results of analysis of variance shows that there are significant difference (P＜0.05) extraction time and extraction time, both conclusion unanimities, and optimum process condition is A ₂B ₂C ₃, these process conditions are consistent with the inferior good technology that with the peoniflorin yield serves as the investigation index.Above-mentioned two of analysis-by-synthesis is investigated index, with optimised process A-grade in the first class ₃B ₂C ₃With second A ₂B ₂C ₃Carry out repeated trials (the results are shown in following table).

The orthogonal test reproducible results

From the result as seen, the peoniflorin of first and second liang of technologies and total solid yield no significant difference from saving of work and time, the angle that cuts down the consumption of energy, select second technology to produce.Promptly add the water (adding 2 times of amounts for the first time) of 8 times of medical material weight, decoct 3 times, each 1 hour, filter, merging filtrate suitably concentrates, and is standby.

4.2.2 Radix Paeoniae Rubra extracting solution separation and purification

The selection of isolation and purification method: in Radix Paeoniae Rubra water boiling and extraction thing, except that effective ingredient, still contain many impurity components, be quality that guarantees product and the quality control that helps product, need further refining purification, remove the impurity such as macromole phlegmatic temperament, protein, starch and solid particle in the extract that anhydrates as much as possible.Test once adopted ethanol precipitation, placed cold preservation, centrifugal, n-butanol extraction, cross D ₁₀₁Extracting method such as resin are handled, and result of the test shows that the aggregative indicator of the method purification of samples of usefulness ethanol precipitation, n-butanol extraction is good, and the yield of main component is higher, and product quality is easy to control.

4.2.2.1 the orthogonal test of alcohol precipitation process

The design of orthogonal test factor water-glass: the relative density of medicinal liquid, alcohol precipitation concentration, standing time are the principal elements that influences alcohol precipitation process, by each factor 3 level, use L ₉(3 ⁴) orthogonal table arrangement test.With the peoniflorin rate of transform serves as to investigate index, and alcohol precipitation process is investigated.

The factor level table

Sample preparation: get 9 parts of decocting liquid that are equivalent to the 100g crude drug, be concentrated into desired concn respectively.Test by orthogonal test table, medicinal liquid filters, and measures cumulative volume.Sampling and measuring peoniflorin concentration is calculated the peoniflorin amount, is that evaluation index is estimated with the rate of transform of peoniflorin, the results are shown in following table.

Radix Paeoniae Rubra alcohol precipitation process condition L ₉(3 ⁴) orthogonal test arrangement and result

Analysis of variance table

F _0.05(2，2)＝19.00 F _0.01(2，2)＝99.00。

Interpretation of result:

From the visual result analysis, the order of each factor affecting precipitate with ethanol efficient is B＞A＞C, and wherein relative density and concentration of alcohol are bigger to the extraction process influence; The results of analysis of variance shows that relative density and concentration of alcohol have significance meaning (P＜0.05), both conclusion unanimities, and optimum process condition is A ₁B ₃C ₁In remarkable sex B factor was arranged, the extreme difference of optimised process and time good technology was less, and C factor extreme difference is less, considered from the energy-and time-economizing, so time good technology is A ₁B ₂C ₂For further confirming the reasonability of this technology, with optimised process A ₁B ₃C ₁With inferior good technology A ₁B ₂C ₂Carry out repeated trials.

The orthogonal test reproducible results

The result as seen, the peoniflorin rate of transform no significant difference of first and second liang of technologies, and second technology has and significantly saves time, the advantage of low consumption selects second technology to produce.Be decoction liquor to be concentrated into relative density be 1.07 (50 ℃), add ethanol makes that to contain the alcohol amount be 60%, stir, placed 12 hours, filter, decompression filtrate recycling ethanol, concentrated, standby.

4.2.2.2 n-butyl alcohol separation and Extraction condition is investigated

Mainly contain monoterpenes and phenolic acid compound in the Radix Paeoniae Rubra, this compounds polarity is bigger, and refining purification should be selected the bigger organic solvent of polarity.Result of the test shows that n-butyl alcohol has selective dissolution preferably to mentioned component, so select n-butyl alcohol to make with extra care purification.

The research of n-butanol extraction number of times

Get this product 600g, the decocting boiling down ethanol precipitation that contracts is handled, and reclaims ethanol, and (50 ℃, every milliliter contains crude drug 1.5 grams to be concentrated into relative density 1.19; PH4), be divided into 4 parts, 1 part is used to measure total solid matters weight, paeoniflorin content, 3 parts respectively extract 5 times with 1/2 times of saturated n-butyl alcohol of water gaging respectively in addition, divide and get n-butyl alcohol liquid, and it is heavy to measure paeoniflorin content and total solid matters, calculate the peoniflorin and the total solid matters rate of transform, result of the test sees the following form.

Result of the test shows, n-butanol extraction 5 times, and the total solid matters rate of transform is 17.37%, peoniflorin total transfer rate is greater than 97%, illustrates that n-butanol extraction can remove a large amount of impurity.N-butanol extraction 4 times, the rate of transform of peoniflorin total amount have reached more than 95%, and the total solid matters rate of transform is 16.39%, n-butanol extraction 5 times, and the rate of transform of peoniflorin total amount only increases by 1.55%, and the total solid matters rate of transform increases by 0.98%.For reducing production costs, reduce operation, ensure the quality of products, can determine with 1/2 times of saturated n-butanol extraction of water gaging 4 times.For further investigating the efficient of n-butanol extraction, the n-butyl alcohol extraction process is carried out demonstration test.

The n-butanol extraction extraction time is investigated result of the test (n=3)

The washing n-butanol extracting liquid is to the influence of paeoniflorin content

In above-mentioned process treatment process, still residual fraction sugar and the stronger composition of inorganic salt isopolarity in the sample are purification of samples as far as possible, n-butyl alcohol liquid is washed with a small amount of, result of the test shows that washing back total solid matters can reduce about 12%, and the retention rate of peoniflorin total amount reaches more than 95%.

The investigation of n-butanol extracting liquid washing consumption

Get n-butanol extraction demonstration test extracting solution mixing, the micrometric measurement total solid matters is heavy, paeoniflorin content.Get 4 parts of n-butanol extracting liquids, every part of 200ml uses not commensurability washing respectively 1 time, divides and gets n-butyl alcohol liquid, measures peoniflorin and total solid matters, calculates retention rate, and result of the test sees the following form.

Different water gagings are washed n-butyl alcohol liquid result of the test

Result of the test shows that with 1/8 times of amount (25ml) washing n-butyl alcohol liquid, the retention rate of peoniflorin total amount is higher, and total solid matters reduces more relatively.

The investigation of n-butanol extracting liquid washing times

Get 3 parts of n-butanol extracting liquids, every part of 200ml uses the washing 3 times of 1/8 times of amount respectively, divides a water intaking layer liquid, measures peoniflorin and total solid matters loss rate, and result of the test sees the following form.

N-butyl alcohol liquid washing times result of the test (n=3)

Result of the test shows that n-butyl alcohol liquid washes with water 1 time, and it is more obvious that total solid matters reduces, wash 2 times after, the total solid matters variation is less, and the peoniflorin accumulating losses are relatively large, so can determine to wash n-butyl alcohol liquid 1 time with 1/8 times of water gaging.

4.2.2.3 remove the selection of tannin method

Contain tannin in the Radix Paeoniae Rubra, the tannin of n-butanol extraction sample before unprocessed checked defective, must remove tannin and handle.The method of removing tannin mainly contains milk of lime process, polyamide method, gelatin method, improvement gelatin method and the alkaline alcohol sedimentation method, acidic precipitation method etc., characteristics according to this product, once adopted the gelatin method to test in the test, and found that the loss rate of peoniflorin was bigger, it is undesirable to remove the tannin effect.Tannin is a polyphenol compound, easily adsorbed by polyamide, and absorption affinity is extremely strong, be difficult for by pure eluting, therefore, utilize monoterpene glycosides and micromolecule phenolic acid in alcoholic solution, to be difficult for being adsorbed, and the character that tannin still can be adsorbed in alcoholic solution is separated with tannin by polyamide.Result of the test shows that the tannin effect that this method is removed in the Chinese medicine injection is better, and loss of effective components is few, and the tannin in the sample is checked up to specification.In abnormal toxicity test, find that residual tannin is bigger to the toxicity of mice, remove tannin by polyamide, can make undue toxicity's (no dead mouse) of mice be added to 120 times (multiples of lethasl concentration), illustrate that this product is feasible with the method for polyamide removal tannin from traditional 30 multiplications except that the tannin method.Below by test the condition that polyamide removes tannin is investigated:

The Radix Paeoniae Rubra medical material is got in sample preparation, prepares the residue that reclaims behind the n-butyl alcohol by preparation technology, and drying under reduced pressure is pulverized, and is standby.

The investigation sample thief 3.10g of adsorption capacity adds 45% dissolve with ethanol to 120ml, and (about 0.6g crude drug/ml) polyamide 1,2,3,4,5,6g have been handled in adding well respectively to get 20ml respectively, stir frequently, static adsorption 3 hours filters, the filtrate water-bath is waved near and is done, and residue adds water and dissolves in right amount, transfers pH to 7, add water and be settled to 10ml, filter, get filtrate 1ml, add 1% Ovum Gallus domesticus album 5ml of new preparation, shake up, placed 10 minutes, observe solution muddiness or precipitation situation, the results are shown in following table.

The polyamide adsorption capacity is investigated the result

"+" expression has muddiness or precipitation, and " ± " expression has not obvious muddiness,

"-" expression does not have muddy or precipitation.

Result of the test shows, when the polyamide adsorption capacity is adsorbed the 4.0g crude drug less than every 1g, can remove the tannin in the sample, crude drug amount/polyamide dry weight in adsorption capacity=upper prop liquid.For guaranteeing the quality of product, the regulation adsorption capacity is that every 3g crude drug uses the 1g polyamide.

Some parts of the selection sample thief of eluting solvent concentration, use 120ml15%, 30%, 45%, 60%, 75% dissolve with ethanol respectively for every part, on handled polyamide column (24g well, blade diameter length ratio: 1: 6, Φ 3cm), use each 360ml eluting of 15%, 30%, 45%, 60%, 75% ethanol respectively, elution speed: 2ml/min.When end opening flows out coloured liquid, collect eluent, eluent is settled to 600ml with the ethanol of respective concentration, and sampling and measuring peoniflorin concentration and total solid matters are heavy.Resampling product solution 100ml, water-bath is waved near and is done, and residue adds water and dissolves in right amount, transfers pH to 7, adds water and is settled to 10ml, filters, and gets filtrate respectively and carries out the tannin inspection in accordance with the law.

Result of the test and elution curve show, the heavy increase with concentration of alcohol of peoniflorin amount and total solid matters increases, and when concentration of alcohol increased to 45%, paeoniflorin content tended to balance, the peoniflorin rate of transform reaches more than 95%, illustrates that 45% ethanol can make peoniflorin effectively resolve; Total solid matters is heavy with this understanding also tends to balance, and the total solid matters rate of transform is 77.27%, has reduced by 22.73%, reaches the purpose of purification refine; And it is all negative to be lower than 60% ethanol elution tannin inspection, illustrates to select less than 60% ethanol elution, and tannin is not resolved gets off.Take all factors into consideration, the eluting solvent concentration is decided to be 45% ethanol.

The eluting solvent concentration is selected result of the test

Some parts of absorption flow velocity sample thief, every part of 3.10g (symphysis medicine 72g, solid content 4.3%), add 45% dissolve with ethanol to 120ml, on polyamide column (24g, the blade diameter length ratio: 1: 6 handled well, Φ 3cm), respectively with 0.5,1,1.5,2,2.5, the flow velocity of 3ml/min carries out dynamic adsorption, adds 45% ethanol liquid eluting, collects eluent 120ml when end opening flows out coloured liquid.Get eluent 20ml, water-bath is waved near and is done, and residue adds water and dissolves in right amount, transfers pH to 7, adds water and is settled to 10ml, filters, and gets filtrate respectively and carries out the tannin inspection in accordance with the law, the results are shown in following table.

Polyamide absorption flow velocity is investigated the result

Annotate: wet column volume (BV) is 5 times of dried resin weight.

240,120,80,60,48,40min adsorption time:

Result of the test shows, when adsorbing flow velocity less than 1.5ml/min (0.75BV/h), tannin is by the polyamide active adsorption, when adsorbing flow velocity greater than 2ml/min (1BV/h), stream is worn in the liquid tannin and is checked defective, be to guarantee product quality, the absorption flow velocity should be controlled at 1ml/min (promptly per hour 0.5BV), begins to flow out until composition.

Eluting solvent consumption sample thief is some part, uses 45% dissolve with ethanol to 120ml respectively for every part, on handled polyamide column (24g well, blade diameter length ratio: 1: 6, Φ 3cm), carry out dynamic adsorption with 1ml/min absorption flow velocity, use 120,240,360,480 respectively, 600ml, 45% ethanol is eluant, carries out dynamic desorption with the flow velocity of 2ml/min, when end opening flows out coloured liquid, collect eluent respectively, sampling and measuring peoniflorin amount and total solid matters are heavy, calculate the rate of transform, the results are shown in following table.

Polyamide eluant consumption is investigated the result

Result of the test shows, the heavy increase with 45% ethanol consumption of peoniflorin amount and total solid matters increases, and when the ethanol consumption increased to 3BV, paeoniflorin content tended to balance, the peoniflorin rate of transform reaches more than 95%, and it is complete substantially to illustrate that 3 times of amount 45% ethanol can make peoniflorin resolve; Total solid matters is heavy with this understanding also tends to balance, and the total solid matters rate of transform is 76.51%, and from increasing work efficiency and the consideration of business efficiency angle, eluting solvent consumption is decided to be 3 times of column volume 45% ethanol.

Some parts of elution flow rate sample thief, use 45% dissolve with ethanol to 120ml respectively for every part, on handled polyamide column (24g well, blade diameter length ratio: 1: 6, Φ 3cm), carrying out dynamic adsorption with 1ml/min absorption flow velocity, is eluant with 360ml (3BV) 45% ethanol, respectively with 1,2,3,4, the flow velocity of 5ml/min carries out dynamic desorption.Collect eluent and be settled to 600ml with 45% ethanol, sampling and measuring peoniflorin concentration and total solid matters are heavy.Extracting sample solution 100ml, water-bath is waved near and is done, and residue adds water and dissolves in right amount, transfers pH to 7, adds water and is settled to 10ml, filters, and gets filtrate respectively and carries out the tannin inspection in accordance with the law, the results are shown in following table.

Polyamide elution flow rate result of the test

Result of the test shows that peoniflorin eluting rate and total solid matters rate of transform best result did not reach 96.78%, 77.51% when elution flow rate was 2ml/min (being 1BV/h), accelerates to reduce with elution flow rate.Elution flow rate is greater than 3ml/min (being 1.5BV/h), the peoniflorin eluting rate and the total solid matters rate of transform descend obviously, and the parsing amount no significant difference of 1ml/min, 2ml/min elution flow rate, each eluting stream tannin inspection is all negative, for shortening the production time, enhance productivity, select the resolution speed of 2ml/min (being 1BV/h) for use.

4.2.3 the selection of Radix Paeoniae Rubra semi-finished product drying means

Get the residual stream of polyamide upper prop process certification test and wear liquid and eluent, merge, mixing is measured paeoniflorin content.Get the stream that is equivalent to the 200g crude drug and wear 3 parts of liquid and eluents, decompression recycling ethanol respectively, residue according to condition carries out drying, and getting dry product, to measure residue dry back paeoniflorin content and total solid matters under different condition heavy, calculate the peoniflorin loss rate, result of the test sees the following form.

Different drying conditions are to the influence of peoniflorin and total solid matters

Result of the test shows that 60～80 ℃ of vacuum dryings are less to the influence of peoniflorin, and loss rate is about 1.5%, and vacuum drying speed is fast, and temperature is low, and the sample quality is loose, is easy to pulverize, so select 60～80 ℃ of vacuum dryings for use.

4.3 preparations shaping technical study

4.3.1 additives are selected and consumption is determined

Through with mannitol, glucose, dextran to the appearance character of product lyophilized preparation, color and luster, porousness, villous shape material formation etc. is observed relatively, the result is when adding a certain amount of mannitol, product appearance shape, good moldability, dissolubility is good, steady quality, result of the test sees the following form.

The relation of filler addition and dried frozen aquatic products molding

Result of the test shows, the amount that adds mannitol in the 100ml medicinal liquid reaches 0.5g when above, can keep sample lyophilizing preceding volume and shape, no duricrust, and color and luster is good, not atrophy, sample is loose porous shape structure, and rehydration is good.Take all factors into consideration from finished product color and luster, dissolubility, formability, economic dispatch aspect, so determine that every recipe quantity can be with mannitol 10g.

4.3.2 the dosing pH value is determined

Two flavor medical materials mainly contain flavonoid, monoterpenes, phenolic acid compound in this product prescription, dissolubility is better under neutrality or alkali condition, so when dosing, regulate suitable pH value, help the dissolving of medicine, increase stability of formulation, but pH value was above 7.5 o'clock, the medicinal liquid mutability is shallow blackish green, color and luster is deepened, and on the trial test basis, the adjusting of pH value is tested during to dosing.

Get 5 parts of extracts, every part of 10.0g (Radix Paeoniae Rubra extract 7.20g, Herba Erigerontis extract 2.30g, mannitol 0.50g), add injection water 60ml respectively, stir and transfer different pH with saturated sodium hydroxide solution, make dissolving, be settled to 100ml, add 0.5% activated carbon, mixing boiling sterilization 30 minutes is put cold.Filter with 0.45 μ m and 0.22 μ m microporous filter membrane, check the variation of clarity, color and luster and the pH value of sterilization back injection, the results are shown in following table.

Different pH value are to the influence of injection clarity and color and luster

Annotate: "+" expression is defective, and "-" is qualified

Result of the test shows, when the dosing pH value is higher than 7.0 when above, the injection clarity is all qualified, and the dosing pH value is higher than 7.5 when above, and medicinal liquid sterilization back color and luster is deepened, and mutability is shallow blackish green.Sterilization back pH value descends about 0.6～1.0, so pH value should be controlled at 6.50～7.50 during this product dosing, the finished product pH value is controlled at 5.50～7.0.

4.3.3 freeze drying process

4.3.3.1 lowest total of the melting point be determined as the quality that guarantees product, the pre-freeze temperature should be controlled at below institute's medicinal liquid eutectic point of surveying about 10～20 ℃, makes solvent, solute curing.Lowest total of the melting point adopts electric-resistivity method to measure.

The measurement result of lowest total of the melting point

From measurement result as can be known, the lowest total of the melting point of this preparation is at-16 ℃.Promptly in refrigerating process, must could begin to carry out sublimation drying with material pre-freeze to below-16 ℃.4.3.3.2 the operation of pre-freeze operation pre-freeze is divided into three phases: the temperature that phase I pre-freeze just forms to ice, i.e. balance solidification point; Second stage pre-freeze freezes product to the minimum eutectic temperature of product; Phase III continues to be cooled to below the lowest total of the melting point temperature, makes product freeze the jail fully.Pre-freeze speed has certain influence to the quality of product, therefore, should measure the parameter of pre-freeze three phases according to concrete sample, and summing up experience is produced to instruct.

Shelf temperature, product temperature and time relation in the freezing process

Equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, needs 1.5 hours approximately, keeps this temperature 1.5 hours, freezes the jail fully until product, promptly begins evacuation, enters drying program.

4.3.3.3 drying program is (45 ℃) evacuation under constant temperature, make the air pressure of hothouse reduce to 13.332～266.64Pa, under this pressure, slowly heat up (2～4 ℃/h) to the lowest total of the melting point temperature, time is about 12 hours, and after sublimation drying was finished, continuation (13.332Pa) under the low pressure condition heated up dry to remove residual moisture, time is about 12～15 hours, kept more than 35 ℃ dry 3 hours, record shelf temperature and product temperature are drawn the time dependent curve of shelf temperature.

Experimental example 3: preparation pharmacodynamic study

(1) to the influence of blood stasis model rabbit blood rheological characteristic

40 of rabbit are divided into 4 groups at random, and 8 every group, ♀ ♂ half and half is respectively blank group, model control group, positive drug control group and injection group of the present invention.Each treated animal elder generation auricular vein is injected (iv) administration.Blank and model control group injecting normal saline (NS) 1ml/kg, positive drug group iv puerarin injection 30mg/kg (being made into 30mg/ml) with NS, experimental group is injected the medicinal liquid (being made into NF) of 0.25mg/ml respectively, dosage is 1ml/kg, two weeks of successive administration (14d), in administration the 2nd, 13d, except that the blank group, each rabbit is annotated 10% high molecular dextran 5ml/kg through auricular vein respectively, every day twice, causes blood stasis model.After the last administration, inject 10% high molecular dextran 5ml/kg once more, behind the 15min, heart is taked fasting blood 6ml, carrying out hemorheology index detects, wherein platelet aggregation rate adopts turbidimetry for Determination: the rotary cone-plate viscosity apparatus mensuration of other employing LBY-N6A+ with LBY-NJ2 type platelet aggregation instrument.

To rabbit platelet aggregation and fibrinogenic influence (x ± s, n=6)

Group dosage (mg/kg) body weight (kg) platelet aggregation (%) Fibrinogen (g/L)

Blank group-2.31 ± 0.15 15.54 ± 2.46 2.53 ± 1.16

Model group-2.45 ± 0.08 38.27 ± 15.05 2.73 ± 1.41

Positive drug group 30 2.37 ± 0.11 12.74 ± 3.47 3.12 ± 1.83

Experimental group 0.25 2.32 ± 0.24 14.80 ± 1.36 2.69 ± 2.32 of the present invention

The result shows: preparation of the present invention is good to the blood stasis type curative effect of disease.

(2) to the effect of rat chronic hepatic injury

Healthy Wistar rat, body weight 120～160g is divided into 4 groups: normal control group, CCl at random ₄Model group, injectable powder group of the present invention.Adopt CCl ₄Preparation rat liver fibrosis model, colchicine group, compatibility group are gastric infusion, injectable powder 3.40g/kg of the present invention in beginning administration in the 1st day of modeling; Normal control group, CCl ₄Model group gives the 0.9%NaCl solution of equivalent.Each treated animal is sacrificed by decapitation behind modeling 10w.Get blood system from the determination of serum liver function.

Each organizes rats'liver changes of function (x ± s)

Group n ALT (IU/L) Glb (ρ B/g/L) Alb (ρ B/g/L)

Normal group 10 35.50 ± 2.42 25.74 ± 2.13 34.20 ± 1.57

Model group 9 115.27 ± 11.28 32.58 ± 3.19 26.95 ± 2.14

Injectable powder group 11 59.72 of the present invention ± 13.39 26.23 ± 2.80 33.01 ± 3.63

The result shows that preparation of the present invention is the hepatic injury effect of being significantly improved to rat chronic.

Concrete embodiment:

Embodiments of the invention 1: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Embodiments of the invention 2: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Get Herba Erigerontis extract, Radix Paeoniae Rubra extract and 10g mannitol, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, add the injection water to 2000ml, mixing, boiled 30 minutes, be sub-packed in the infusion bottle of having handled well, fill in butyl rubber bung and thin film, roll aluminium lid, flowing steam sterilization 30 minutes, cold preservation 24 hours filters the filtrate packing with 0.45 μ m and 0.22 μ m microporous filter membrane under aseptic condition, every bottle of 2.0ml, lyophilization, the equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, freeze the jail fully until product, promptly begin evacuation, enter drying program, under 45 ℃ of constant temperature-evacuation, make the air pressure of hothouse reduce to 13.332～266.64Pa, under this pressure, slowly heat up, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, and it is dry to remove residual moisture to heat up, and the time is about 12～15 hours, kept more than 35 ℃ dry 3 hours, gland promptly gets the Injectable sterile block, intravenous drip, one time 2,1 time on the one, with using every dress 200mg behind the 250ml0.9% physiological saline solution; Calculate by weight percentage, in the injection flavones ingredient and glycoside component content sum be not less than deduction adjuvant amount and water quantities in the preparation total solid 25%.

Embodiments of the invention 3: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate adds the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, to boil, 4 ℃ of cold preservations are spent the night, coarse filtration, fine straining, add the injection water, divide to install to pacify and cut open bottle, seal sterilization and promptly get injection with small volume or concentrated solution for injection.

Embodiments of the invention 4: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, add the glucose or the sodium chloride of ormal weight, by volume add 1.0% active carbon behind the mixed dissolution, boil, keep little 30min that boils, cold slightly filtration, filtrate add the injection water to ormal weight, the saturated sodium hydroxide solution of reuse adjust pH to 7.0～7.5, boil, 4 ℃ of cold preservations are spent the night, and coarse filtration, fine straining add the injection water, packing, sterilization promptly gets glucose or sodium chloride intravenous infusion.

Embodiments of the invention 5: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, adds the injection water to 2000ml, mixing, boiled 30 minutes, and divided to install in the enamel tray lyophilization, equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, and froze the jail fully, promptly begin evacuation until product, enter drying program, under 45 ℃ of constant temperature-and evacuation, make the air pressure of hothouse reduce to 13.332～266.64Pa, under this pressure, slowly heat up, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, heating up, dry the time is about 12～15 hours to remove residual moisture, keeps more than 35 ℃ dry 3 hours, under aseptic condition, divide to install in the cillin bottle, promptly get injectable sterile powder.

Embodiments of the invention 6: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract;

Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-, with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h collects stream and wears liquid and eluent, reclaims ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract.

Get Herba Erigerontis extract, Radix Paeoniae Rubra extract, mixing adds an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate add the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, boil, 4 ℃ of cold preservations are spent the night, and coarse filtration, fine straining are 150 ℃ in inlet temperature, leaving air temp is 60 ℃, and air velocity is 20ms ^-1Condition under spray drying get powder, packing promptly gets injectable sterile powder.

Embodiments of the invention 7: Herba Erigerontis 100g Radix Paeoniae Rubra 9900g

Herba Erigerontis adds 5 times of water gagings and decocted 0.5 hour, filters merging filtrate, filtrate decompression concentrates, and adds ethanol and makes and contain the alcohol amount and reach 30%, constantly stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, use the hydrochloric acid adjust pH, insulation, tipping supernatant, sucking filtration, precipitation washes with water to pH value, and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 3 times of water gagings and decocted 0.5 hour, filters merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40%, stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1 time, merge n-butyl alcohol liquid, wash with water 1 time, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, and last polyamide column is used ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, add the soybean oil mixing, the pressing pill promptly gets soft capsule.

Embodiments of the invention 8: Herba Erigerontis 9900g Radix Paeoniae Rubra 100g

Herba Erigerontis adds 12 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, filtrate decompression concentrate, and add ethanol and make and contain the alcohol amount and reach 80%, constantly stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, and uses the hydrochloric acid adjust pH, insulation, the tipping supernatant, sucking filtration, precipitation washes with water to pH value, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 10 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 80%, stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 5 times, and merging n-butyl alcohol liquid, wash with water 3 times, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, last polyamide column, use ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying, get Radix Paeoniae Rubra extract,, add the PEG400 mixing Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, the dropping preparation method pill promptly gets drop pill.

Embodiments of the invention 9: Herba Erigerontis 2000g Radix Paeoniae Rubra 8000g

Herba Erigerontis adds 12 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, filtrate decompression concentrate, and add ethanol and make and contain the alcohol amount and reach 80%, constantly stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, and uses the hydrochloric acid adjust pH, insulation, the tipping supernatant, sucking filtration, precipitation washes with water to pH value, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 10 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 80%, stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 5 times, and merging n-butyl alcohol liquid, wash with water 3 times, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, last polyamide column, use ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying, get Radix Paeoniae Rubra extract,, add 2% sodium carboxymethyl cellulose Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, extrude-the spheronization pill, promptly get pellet.

Embodiments of the invention 10: Herba Erigerontis 8000g Radix Paeoniae Rubra 2000g

Herba Erigerontis adds 5 times of water gagings and decocted 0.5 hour, filters merging filtrate, filtrate decompression concentrates, and adds ethanol and makes and contain the alcohol amount and reach 30%, constantly stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, use the hydrochloric acid adjust pH, insulation, tipping supernatant, sucking filtration, precipitation washes with water to pH value, and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 3 times of water gagings and decocted 0.5 hour, filters merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40%, stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1 time, merge n-butyl alcohol liquid, wash with water 1 time, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, last polyamide column, use ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously,, add 3% methylcellulose, it is moistening to add water, make granule, drying adds 1% carboxymethyl starch sodium, tabletting promptly gets dispersible tablet.

Embodiments of the invention 11: Herba Erigerontis 100g Radix Paeoniae Alba 9900g

Herba Erigerontis adds 5 times of water gagings and decocted 0.5 hour, filters merging filtrate, filtrate decompression concentrates, and adds ethanol and makes and contain the alcohol amount and reach 30%, constantly stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, use the hydrochloric acid adjust pH, insulation, tipping supernatant, sucking filtration, precipitation washes with water to pH value, and vacuum drying gets Herba Erigerontis extract; The Radix Paeoniae Alba adds 3 times of water gagings and decocted 0.5 hour, filters merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40%, stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1 time, merge n-butyl alcohol liquid, wash the reclaim under reduced pressure n-butyl alcohol with water 1 time, residue adds dissolve with ethanol, last polyamide column is used ethanol elution, collects stream and wears liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Alba extract, with Herba Erigerontis extract and Radix Paeoniae Alba extract mix homogeneously, pill promptly gets pill.

Embodiments of the invention 12: Herba Erigerontis 9900g Radix Paeoniae Alba 100g

Herba Erigerontis adds 12 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, filtrate decompression concentrate, and add ethanol and make and contain the alcohol amount and reach 80%, constantly stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, and uses the hydrochloric acid adjust pH, insulation, the tipping supernatant, sucking filtration, precipitation washes with water to pH value, vacuum drying gets Herba Erigerontis extract; The Radix Paeoniae Alba adds 10 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 80%, stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 5 times, merge n-butyl alcohol liquid, wash with water 3 times, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, and last polyamide column is used ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Alba extract, add distilled water, promptly get oral liquid.

Embodiments of the invention 13: Herba Erigerontis 2000g Radix Paeoniae Alba 8000g

Herba Erigerontis adds 12 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, filtrate decompression concentrate, and add ethanol and make and contain the alcohol amount and reach 80%, constantly stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, and uses the hydrochloric acid adjust pH, insulation, the tipping supernatant, sucking filtration, precipitation washes with water to pH value, vacuum drying gets Herba Erigerontis extract; The Radix Paeoniae Alba adds 10 times of water gagings and decocts 5 times, each 2 hours, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 80%, stir, left standstill sucking filtration 24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 5 times, and merging n-butyl alcohol liquid, wash with water 3 times, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, last polyamide column, use ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying, get Radix Paeoniae Alba extract, with Herba Erigerontis extract and Radix Paeoniae Alba extract mix homogeneously, granulate, promptly get granule.

Embodiments of the invention 14: Herba Erigerontis 8000g Radix Paeoniae Alba 2000g

Herba Erigerontis adds 5 times of water gagings and decocted 0.5 hour, filters merging filtrate, filtrate decompression concentrates, and adds ethanol and makes and contain the alcohol amount and reach 30%, constantly stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, use the hydrochloric acid adjust pH, insulation, tipping supernatant, sucking filtration, precipitation washes with water to pH value, and vacuum drying gets Herba Erigerontis extract; The Radix Paeoniae Alba adds 3 times of water gagings and decocted 0.5 hour, filters merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40%, stirs, left standstill 6 hours, sucking filtration, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1 time, merge n-butyl alcohol liquid, wash with water 1 time, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, last polyamide column, use ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying, get Radix Paeoniae Alba extract, with Herba Erigerontis extract and Radix Paeoniae Alba extract mix homogeneously, it is moistening to add water, makes granule, encapsulated, promptly get capsule.

Embodiments of the invention 15: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filters, when merging filtrate, filtrate decompression are concentrated into 50 ℃ of relative densities 1.09～1.11, add ethanol and make and contain the alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ of insulations 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and makes and contain the alcohol amount and reach 60%, stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20 with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merges n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, reclaim under reduced pressure n-butyl alcohol, residue add 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, wet 5 times of volumes of column volume=dried resin weight of absorption flow velocity: 0.5BV/h, BV-are with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying, get Radix Paeoniae Rubra extract, mixing, alcohol granulation promptly gets granule.

Embodiments of the invention 16: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, and each 0.5 hour, filter, merging filtrate, filtrate decompression is concentrated into relative density 1.09～1.11 (50 ℃), and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, and each 1 hour, filter, merging filtrate is evaporated to relative density 1.06～1.08 (50 ℃), and vacuum drying gets Radix Paeoniae Rubra extract; Herba Erigerontis extract and Radix Paeoniae Rubra extract are mixed, granulate, tabletting promptly gets tablet.

Embodiments of the invention 17: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filter, merging filtrate, filtrate decompression is concentrated into relative density 1.09～1.11 (50 ℃), add twice of ethanol precipitate with ethanol, make for the first time to contain the alcohol amount and reach 55%, make for the second time to contain the alcohol amount and reach 85%, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.10～1.12 (50 ℃), and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filter, merging filtrate is evaporated to relative density 1.06～1.08 (50 ℃), add twice of ethanol precipitate with ethanol, make for the first time to contain the alcohol amount and reach 50%, make for the second time to contain the alcohol amount and reach 80%, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.10～1.12 (50 ℃), and vacuum drying gets Radix Paeoniae Rubra extract; Herba Erigerontis extract and Radix Paeoniae Rubra extract are mixed, add syrup, promptly get syrup.

Embodiments of the invention 18: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filter, merging filtrate, filtrate decompression is concentrated into relative density 1.09～1.11 (50 ℃), with hydrochloric acid adjust pH to 4, add the equal-volume ethyl acetate, extract combined ethyl acetate 4 times, decompression and solvent recovery, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 10 times of water gagings and decocts 3 times, each 1 hour, filters merging filtrate, be evaporated to relative density 1.06～1.08 (50 ℃),, merge n-butyl alcohol liquid with water saturated n-butanol extraction 4 times, wash 1 time with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, vacuum drying gets Radix Paeoniae Rubra extract; Herba Erigerontis extract and Radix Paeoniae Rubra extract are mixed, add refined honey, pill promptly gets pill.

Embodiments of the invention 18: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filter, merging filtrate is crossed the AB-8 macroporous resin column, earlier with 6 times of resinite hydrops and 4 times of resin volume 10% alcohol flushings, 5 times of resin volume 50% alcohol desorptions of reuse, collect stripping liquid, decompression recycling ethanol, vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 10 times of water gagings and decocts 3 times, each 1 hour, filter, merging filtrate is crossed ZTC-1 type macroporous resin column, earlier with 4 times of resinite hydrops and 3 times of resin volume 10% alcohol flushings, 5 times of resin volume 60% alcohol desorptions of reuse, collect stripping liquid, decompression recycling ethanol, vacuum drying gets Radix Paeoniae Rubra extract; Herba Erigerontis extract and Radix Paeoniae Rubra extract are mixed, add distilled water, promptly get oral liquid.

Embodiments of the invention 19: Herba Erigerontis 3000g Radix Paeoniae Rubra 4500g

Herba Erigerontis adds 10 times of amount 80% alcohol reflux 3 times, and each 1 hour, filter, merging filtrate, decompression recycling ethanol are concentrated into relative density 1.09～1.11 (50 ℃), and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 10 times of amount 70% alcohol reflux 3 times, and each 1 hour, filter, merging filtrate, decompression recycling ethanol are concentrated into relative density 1.09～1.11 (50 ℃), and vacuum drying gets Radix Paeoniae Rubra extract; Herba Erigerontis extract and Radix Paeoniae Rubra extract are mixed, granulate, promptly get granule.

Herba Erigerontis extract can be a flavones ingredient content greater than 90% total flavone valid target or by preparation method system of the present invention or commercially available, and Radix Paeoniae Rubra extract can be the glycoside component content greater than total glycosides effective part of 90% or by preparation method system of the present invention or commercially available.

Claims (17)

1, a kind of compound recipe fleabane preparation for the treatment of cardiovascular and cerebrovascular disease is characterized in that: made by raw material of Chinese medicine medicine and suitable adjuvant, it is 1-99% Herba Erigerontis and 99-1% Radix Paeoniae Rubra that described raw material of Chinese medicine medicine is counted by weight percentage.

2. according to the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 1, it is characterized in that: calculate according to percentage by weight, it is with Herba Erigerontis 20～80% and Radix Paeoniae Rubra 80～20% and adds suitable adjuvant and make.

3. according to the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 2, it is characterized in that: calculate according to percentage by weight, it is with Radix Paeoniae Rubra 60% and Herba Erigerontis 40% and adds suitable adjuvant and make.

4. according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease of claim 1～3, it is characterized in that: Radix Paeoniae Rubra in the described prescription and Herba Erigerontis can be medical material or the extract that prepared by the corresponding proportion medical material, and wherein: Herba Erigerontis extract can be the highly finished product of Herba Erigerontis alcohol extract, Herba Erigerontis water extract, Herba Erigerontis water extract-alcohol precipitation extract, Herba Erigerontis semi-bionic extraction thing, Herba Erigerontis supercritical extract or above each extract; Radix Paeoniae Rubra extract can be the highly finished product of Radix Paeoniae Rubra alcohol extract, Radix Paeoniae Rubra water extract, Radix Paeoniae Rubra water extract-alcohol precipitation extract, Radix Paeoniae Rubra semi-bionic extraction thing, Radix Paeoniae Rubra supercritical extract or above each extract.

5. according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease of claim 1～3, it is characterized in that: described preparation be directly used in the injection of drug administration by injection, directly for the venous transfusion of intravenous drip, the injectable sterile powder, aseptic block, tablet, capsule, granule, pill or the oral liquid that need to be used for after the dilution concentrated solution for injection of intravenous drip, make with freeze-drying or spray drying method.

6. according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 1～3, it is characterized in that: contain flavones ingredient and glycoside composition in the preparation, calculate by weight percentage, in the injection flavones ingredient and glycoside component content sum be not less than deduction adjuvant amount and water quantities in the preparation total solid 25%.

7. according to the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 4, it is characterized in that: Herba Erigerontis extract is a flavones ingredient content greater than 90% total flavone valid target, and Radix Paeoniae Rubra extract is the glycoside component content greater than total glycosides effective part of 90%.

8. according to the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 4, it is characterized in that: the Radix Paeoniae Alba and Radix Paeoniae Alba extract with equivalent substitute Radix Paeoniae Rubra and Radix Paeoniae Rubra extract.

9. as the preparation method of the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 1～5, it is characterized in that: get the Radix Paeoniae Rubra medical material, adding entry or alcoholic solution after the pulverizing extracts, merge extractive liquid,, filter, concentrate the Radix Paeoniae Rubra crude extract, or adopt in ethanol precipitation, column chromatography, extraction, the flocculent precipitation one or more to unite on this basis to use carry out suitably refining, the Radix Paeoniae Rubra extract; Get the Herba Erigerontis medical material, adding entry or alcoholic solution after the pulverizing extracts, merge extractive liquid,, filter, concentrate the Herba Erigerontis crude extract, or adopt in ethanol precipitation, column chromatography, extraction, the flocculent precipitation one or more to unite on this basis to use carry out suitably refining, the Herba Erigerontis extract, with Herba Erigerontis crude extract or extract and Radix Paeoniae Rubra crude extract or extract mix homogeneously, add adjuvant and make different preparations.

10. according to the preparation method of the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 9, it is characterized in that: Herba Erigerontis adds 5～12 times of water gagings and decocts 1～5 time, each 0.5～2 hour, filter, merging filtrate, filtrate decompression concentrates, adding ethanol makes and contains alcohol amount and reach 30～80%, constantly stir, left standstill sucking filtration 6～24 hours, decompression filtrate recycling ethanol also concentrates, use the hydrochloric acid adjust pH, insulation, tipping supernatant, sucking filtration, precipitation washes with water to pH value, and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 3～10 times of water gagings and decocts 1～5 time, each 0.5～2 hour, filters, merging filtrate, concentrating under reduced pressure adds ethanol and makes and contain alcohol amount and reach 40～80%, stir, left standstill sucking filtration 6～24 hours, decompression filtrate recycling ethanol also concentrates, with water saturated n-butanol extraction 1～5 time, merge n-butyl alcohol liquid, wash with water 1～3 time, the reclaim under reduced pressure n-butyl alcohol, residue adds dissolve with ethanol, and last polyamide column is used ethanol elution, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, add adjuvant and make different preparations.

11. preparation method according to the compound recipe fleabane preparation of the described treatment cardiovascular and cerebrovascular disease of claim 10, it is characterized in that: Herba Erigerontis adds 10 times of water gagings and decocts 3 times, each 0.5 hour, filter, merging filtrate is when filtrate decompression is concentrated into 50 ℃ of relative densities 1.09～1.11, adding ethanol makes and contains alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.10～1.12, with hydrochloric acid adjust pH to 2,55 ℃ are incubated 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to pH value 3～4, and vacuum drying gets Herba Erigerontis extract; Radix Paeoniae Rubra adds 8 times of water gagings and decocts 3 times, each 1 hour, filters, merging filtrate, when being evaporated to 50 ℃ of relative densities 1.06～1.08, adding ethanol and make and contain the alcohol amount and reach 60%, stir, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol and when being concentrated into 50 ℃ of relative densities 1.18～1.20 is with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 1 time with 1/8 times of water gaging, reclaim under reduced pressure n-butyl alcohol, residue add 45% ethanol 7500ml dissolving, last polyamide column, 1500g, Φ 12cm, blade diameter length ratio: 1: 6, absorption flow velocity: 0.5BV/h, wet 5 times of volumes of column volume=dried resin weight of BV-are with 45% ethanol 22500ml eluting, elution flow rate: 1BV/h, collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets Radix Paeoniae Rubra extract, with Herba Erigerontis extract and Radix Paeoniae Rubra extract mix homogeneously, add adjuvant and make different preparations.

12. preparation method according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 9～11, it is characterized in that: the Injectable sterile block in the described preparation prepares like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract and 10g mannitol, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, add the injection water to 2000ml, mixing, boiled 30 minutes, be sub-packed in the infusion bottle of having handled well, fill in butyl rubber bung and thin film, roll aluminium lid, flowing steam sterilization 30 minutes, cold preservation 24 hours, under aseptic condition, filter with 0.45 μ m and 0.22 μ m microporous filter membrane, the filtrate packing, every bottle of 2.0ml, lyophilization, equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, and froze the jail fully, promptly begin evacuation until product, enter drying program, evacuation under-45 ℃ of constant temperature makes the air pressure of hothouse reduce to 13.332～266.64Pa, slowly heats up under this pressure, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, heating up, dry the time is about 12～15 hours to remove residual moisture, keeps more than 35 ℃ dry 3 hours, gland, promptly.

13. preparation method according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 9～11, it is characterized in that: injection and concentrated solution for injection in the described preparation prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate adds the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, boil, coarse filtration is spent the night in 4 ℃ of cold preservations, fine straining, add the injection water, divide to install to pacify and cut open bottle, seal sterilization, promptly.

14. preparation method according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 9～11, it is characterized in that: glucose or sodium chloride intravenous infusion in the described preparation prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add an amount of water for injection dissolving, the glucose or the sodium chloride that add ormal weight, by volume add 1.0% active carbon behind the mixed dissolution, boil, keep little 30min that boils, cold slightly filtration, filtrate adds the injection water to ormal weight, boil the saturated sodium hydroxide solution of reuse adjust pH to 7.0～7.5, and 4 ℃ of cold preservations are spent the night, coarse filtration, fine straining, add the injection water, packing, sterilization is promptly.

15. preparation method according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 9～11, it is characterized in that: the injectable sterile powder in the described preparation prepares like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, add 1800ml water for injection, stirring makes dissolving, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, add the injection water to 2000ml, mixing, boiled 30 minutes, divide and install in the enamel tray, lyophilization, the equilibration time when the balance solidification point of phase I is 0 ℃ is 1.5 hours, i.e. the time of shelf temperature and product temperature basically identical; The second stage solidification point is from 0 ℃ during to minimum eutectic temperature-16 ℃, and shelf temperature and product temperature equilibration time are 1.5 hours; Phase III continues to be cooled to-45 ℃, need 1.5 hours approximately, kept this temperature 1.5 hours, and froze the jail fully, promptly begin evacuation until product, enter drying program, evacuation under-45 ℃ of constant temperature makes the air pressure of hothouse reduce to 13.332～266.64Pa, slowly heats up under this pressure, 2～4 ℃/h, to the lowest total of the melting point temperature, the time is about 12 hours, after sublimation drying is finished, continuation is under 13.332Pa low pressure condition, heating up, dry the time is about 12～15 hours to remove residual moisture, keeps more than 35 ℃ dry 3 hours, under aseptic condition, divide and install in the cillin bottle, promptly.

16. preparation method according to the compound recipe fleabane preparation of any described treatment cardiovascular and cerebrovascular disease in the claim 9～11, it is characterized in that: the injectable sterile powder in the described preparation can also prepare like this: get Herba Erigerontis extract, Radix Paeoniae Rubra extract, mixing, add an amount of water for injection dissolving, by volume add 1.0% active carbon, boil, keep little 30min that boils, cold slightly filtration, filtrate adds the injection water to ormal weight, with saturated sodium hydroxide solution adjust pH to 7.0～7.5, to boil, 4 ℃ of cold preservations are spent the night, coarse filtration, fine straining, in inlet temperature is 150 ℃, and leaving air temp is 60 ℃, and air velocity is 20ms ^-1Condition under spray drying get powder, packing, promptly.

17. the application of compound recipe fleabane preparation in the medicine of preparation treatment chronic hepatic injury disease as any described treatment cardiovascular and cerebrovascular disease in the claim 1～4.