CN1507907A - Medicinal injection for treating cardio-cerebrovascular disease and preparing method thereof - Google Patents
Medicinal injection for treating cardio-cerebrovascular disease and preparing method thereof Download PDFInfo
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Abstract
The present invention relates to a Chinese medicine injection preparation for curing cardio-cereb ral vascular diseases. Said injection preparation is made up by using the Chinese medicinal materials of peach kernel, rey peony root and ligusticum root through a certain preparation process including different extraction methods. Said invention also provides its quality control method, also provides fingerprint chromatogram quality control method of its freeze-dried powder injection.
Description
Technical field
The present invention relates to a kind of pharmaceutical preparation and preparation method thereof, particularly relate to pharmaceutical preparation for the treatment of cardiovascular and cerebrovascular disease and preparation method thereof.
Background technology
Cardiovascular disease is to cause human first dead sick the kind, and along with the raising of social life level and the aging of population, the research pay attention to day by day of cardiovascular drugs is used also increasingly extensive.The emergency treatment of ischemia apoplexy (cerebral infarction) in acute attack stage is the crucial opportunity of its treatment, therefore developing the Chinese medicine traditional advantage, and it is significant to develop Chinese medicine quick-acting, efficient, the emergency treatment medication.
Summary of the invention
The object of the invention is to provide a kind of pharmaceutical preparation for the treatment of cardiovascular and cerebrovascular disease and preparation method thereof; The present invention also aims to provide a kind of method of quality control of ejection preparation.
The present invention seeks to be achieved through the following technical solutions:
Semen Persicae 1-4 weight portion Radix Paeoniae Rubra 1-4 weight portion Rhizoma Chuanxiong 1-4 weight portion, optimum ratio is
Semen Persicae 1 weight portion Radix Paeoniae Rubra 1 weight portion Rhizoma Chuanxiong 1 weight portion can also be
Semen Persicae 2 weight portion Radix Paeoniae Rubra 3 weight portion Rhizoma Chuanxiongs 2 weight portions
Press practice of pharmacy, preparation of pharmaceutical compositions of the present invention can be become the various clinical pharmaceutical dosage form, comprise the dosage form of oral formulations or parenterai administration.Said oral formulations is selected from a kind of in the middle of the tablet, capsule, pill, granule, suspensoid, drop pill, oral liquid; Said parenterai administration dosage form is selected from a kind of in the middle of injection, aerosol, suppository or the subcutaneous administration dosage form.
Medicine of the present invention also can add conventional drug excipient, as solvent, disintegrating agent, correctives, antiseptic, coloring agent etc.
The present invention makes the preparation method of injection: above 3 flavors, get Rhizoma Chuanxiong 1 flavor, and be ground into coarse powder, put in the supercritical extraction still CO
2Supercritical extraction 2-3 time is extracted still pressure 20-40Mpar at every turn, temperature 40-60 ℃; Separation reactor I pressure 10-14Mpar, temperature 30-40 ℃; Separation reactor I I pressure 4-8Mpar, temperature 30-40 ℃; Extracted 1-2 hour, and collected, get extractive of volatile oil from separating still.After the last supercritical extraction, continous way pumps into 95% ethanol of 1.2 times of amounts of medical material among the separation reactor I I, extracts altogether 1 hour, collects extract from separating still, gets ethanol extract.The Rhizoma Chuanxiong volatile oil extract adds the double distilled water that medical material 4-10 doubly measures, and vapor distillation is collected the distillate that medical material 2-5 doubly measures, and distillate adds the NaOH of medical material 0.1-0.3% amount, leaves standstill, and filters, and gets medicinal liquid, and is standby.Get Semen Persicae, Radix Paeoniae Rubra 2 flavors add 6-12 times of water gaging, decoct 1-2 hour, filter, medicinal residues add 6-10 times of water gaging again, decoct 1-2 hour, filter, merging filtrate, medicinal liquid are evaporated to relative density 40 ℃ following 1.19, add clear paste 2-6 and doubly measure 85-95% ethanol, make medicinal liquid contain alcohol amount 70-80%, precipitate with ethanol, left standstill 8-18 hour, and filtered decompression filtrate recycling ethanol, be concentrated into relative density and descend 1.24 for 40 ℃, add clear paste 7-10 and doubly measure 85-95% ethanol, make medicinal liquid contain alcohol amount 80-85%, precipitate with ethanol, pure liquid transfers to pH value 6-8 with 15-25%NaOH liquid, leaves standstill 8-18 hour, filter, medicinal liquid adds the active carbon of 1-2% amount, stirs, filter, medicinal liquid and above-mentioned Rhizoma Chuanxiong alcohol extract merge, and reclaim ethanol to there not being the alcohol flavor, get clear paste, clear paste adds above-mentioned Rhizoma Chuanxiong reserve liquid, stirs, and filters, medicinal liquid is adjusted to water for injection and contains crude drug 0.6g/ml, add the sodium sulfite of 0.1-0.3% amount and the active carbon of 0.2-0.4% amount, 100 ℃ are incubated 8-12 minute, filtration or centrifugal, medicinal liquid ultrafiltration sterilization, promptly.
The method of quality control of injection of the present invention is:
Discrimination method in the method for quality control is selected from one or more in the following method:
A. get this product 1ml, as need testing solution.Other gets the amygdaloside reference substance, adds methanol 6ml and makes dissolved solution, in contrast product solution.According to thin layer chromatography (" 2000 editions appendix VIB of Chinese pharmacopoeia) test, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 12-18: 30-50: 15-25: 3-6 dichloromethane-ethyl acetate-methanol-water is developing solvent, launch, take out, spray is heated to clear spot with the phosphomolybdic acid sulfuric acid solution at 100-105 ℃.In the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color.Wherein the phosphomolybdic acid sulfuric acid solution is to add water 20ml by phosphomolybdic acid 2g to make dissolving, slowly adds sulphuric acid 30ml again, and mixing makes.
B. get this product 1ml, as need testing solution.Other gets the peoniflorin reference substance, makes the solution that every 1ml contains 2mg with methanol, in contrast product solution.According to thin layer chromatography (" 2000 editions appendix VIB of Chinese pharmacopoeia) test, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 30-50: 3-6: 8-12: 0.1-0.3 dichloromethane-ethyl acetate-methanol-formic acid is developing solvent, launch, take out, dry, spray is heated to clear spot with 1-2% vanillin sulfuric acid solution under 100-105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
C. get this product 1ml, as need testing solution.Other gets the ferulic acid reference substance, makes the solution that every 1ml contains 0.5mg with methanol, in contrast product solution.According to thin layer chromatography (appendix VIB) test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with 25-35: 1-3: 1-2 toluene-methanol-glacial acetic acid is developing solvent, launches, and takes out, dry, spray is with 50% alcoholic solution of 2% ferric chloride-2% potassium ferricyanide.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay in the method for quality control:
Measure according to high performance liquid chromatography (2000 editions one appendix VI D of Chinese Pharmacopoeia), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica, and the methanol-water of 25-29: 71-76 is a mobile phase; The detection wavelength is 218nm, and number of theoretical plate calculates by amygdaloside peak and peoniflorin peak all should be not less than 3000; The preparation of reference substance solution, it is an amount of to be taken at 60 ℃ of amygdaloside, peoniflorin reference substances that are dried to constant weight, and accurate the title, decide, and adds mobile phase respectively and make the solution that every 1ml contains 0.4mg, promptly; This product 5ml is got in the preparation of need testing solution, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Every of this product contains Semen Persicae with amygdaloside (C
20H
27NO
11) meter, must not be less than 22mg, contain Radix Paeoniae Rubra with peoniflorin (C
23H
28O
11) meter, must not be less than 36mg.
Injection method of quality control of the present invention also can adopt the method for finger printing:
Measure according to high performance liquid chromatography (" 2000 editions appendix VID of Chinese pharmacopoeia), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 24-28: 74 methanol-0.1% trifluoroacetic acid aqueous solution is a mobile phase; The detection wavelength is 218nm.Theoretical cam curve is pressed object of reference amygdaloside peak and is calculated, and should be not less than 3000; The preparation of object of reference solution, it is an amount of to be taken at 60 ℃ of amygdaloside objects of reference that are dried to constant weight, and accurate the title, decide, and adds methanol and make the solution that every 1ml contains 0.4mg, promptly; This product 2ml is got in the preparation of test solution, puts in the 25ml measuring bottle, and thin up shakes up to scale, promptly; Algoscopy, accurate object of reference solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid respectively, write down 1 hour chromatogram, be the S peak with corresponding peak, object of reference peak in the test sample finger printing, calculate relative retention time and peak area ratio, should meet the following requirements: this product finger printing should be similar to standard finger-print; Finger printing should have 8 total peaks;
Relative retention time (peak number): 0.350 ± 30% (1), 0.554 ± 30% (2), 1 (S), 1.413 ± 30% (3), 1.552 ± 30% (4), 2.022 ± 30% (5), 3.600 ± 30% (6), 3.985 ± 30% (7);
Peak area ratio (peak number): 1.218 ± 30% (5), 0.451 ± 40% (7); Non-total peak area must not be crossed 5% of total peak area.
Instrument: Agilent 1100 chromatograph of liquid; Chromatographic column: Agillent HypersilODS C
18(4.6 * 250mm, 5 μ m); Integral way: paddy paddy integration.
Amygdaloside object of reference chromatogram
Promoting blood circulation to remove obstruction in the collateral injection standard finger-print
Powder injection formulation of the present invention (promoting blood circulation to remove obstruction in the collateral injection) is Pharmacodynamic test of active extract result show: the promoting blood circulation to remove obstruction in the collateral injection can obviously reduce cerebral index and the brain water content that the ligation bilateral common carotid arteries causes acute experiment imperfection rats with cerebral ischemia; Obviously reduce behavioristics's scoring and the cerebral infarction rate of middle cerebral artery blocking-up (MCAO) rat; Obviously reduce behavioristics's scoring, cerebral infarction rate and the brain water content of middle cerebral artery ischemia-reperfusion rat; Can reduce systolic pressure, diastolic pressure, the mean arterial pressure of anesthetized dog, increase cardiac output, coronary flow, reduce left indoor pressure and left chamber EDP, reduce total oxygen consumption index and total peripheral resistance, the cerebral blood flow increasing amount reduces cerebral vascular resistance; Can obviously reduce rat arteriovenous shut wet weight of thrombus and thrombosis dry weight, rising wet weight of thrombus suppression ratio and thrombosis dry weight suppression ratio; Obviously reduce the rat platelet maximum agglutination rate, raise and assemble suppression ratio; Obviously reduce ADP, AA, the inductive rabbit platelet aggregation rate of Coll, the hematoblastic depolymerization rate of the rising ADP inductive rabbit of institute; The clotting time of significant prolongation rat and mice; The plasma viscosity that can effectively suppress rat; Can significantly reduce the erythrocyte FI of rat; Total number of blood platelet, activated partial prothrombin time, prothrombin time and plasma fibrinogen to rat do not have remarkable change.The promoting blood circulation to remove obstruction in the collateral injection can improve neurobehavioral obstacle behind the cerebral infarction, dwindle infarct size; Reduce the swelling degree of ischemia brain; Cardiovascular system is had blood vessel dilating, reduce Peripheral resistance, bring high blood pressure down, lightening heart load increases coronary flow and cardiac output, reduces myocardial oxygen consumption; Brain circulation system is had expansion of cerebral vascular, reduce cerebral vascular resistance, the cerebral blood flow increasing amount is improved effects such as cerebral circulation.Can obviously suppress thrombosis and anticoagulant; Prolong clotting time, blood viscosity lowering, improve erythrocyte deformability.These effects are highly effective for its clinical treatment ischemia apoplexy.
Following experimental example is used to further specify the present invention.
Experimental example 1:Protective effect to acute experiment imperfection rats with cerebral ischemia
Test objective: observe and be subjected to of the influence of reagent thing, determine whether medicine has protective effect to acute experiment imperfection rats with cerebral ischemia to ligation bilateral common carotid arteries rat brain index and brain water content.
1 group and dosage:
(1) sham operated rats: 0.9% sodium chloride injection.(2) model group: 0.9% sodium chloride injection.(3) positive drug group: 0.02% nimodipine solution (2mg/kg).(4) positive drug group: 80% compound Salviae Miltiorrhizae injection solution (8g/kg).(5) low dose group: 2.5% promoting blood circulation to remove obstruction in the collateral injection (0.25g/kg).(6) dosage group in: 5% promoting blood circulation to remove obstruction in the collateral injection (0.5g/kg).(7) high dose group: dosage group during 10% promoting blood circulation to remove obstruction in the collateral injection (1g/kg) (8) is oral: 5% promoting blood circulation to remove obstruction in the collateral injection (0.5g/kg).(9) oral high dose group: 10% promoting blood circulation to remove obstruction in the collateral injection (1g/kg).The administration volume is 10ml/kg.
2 experimental implementation:
Get 90 of SD male rats, body weight 200～250g, be divided into 9 groups at random, i.e. the oral middle dosage group of sham operated rats, model group, Western medicine positive drug group, Chinese medicine positive drug group, basic, normal, high 3 the dosage groups of promoting blood circulation to remove obstruction in the collateral injection and promoting blood circulation to remove obstruction in the collateral injection, high dose group.1～7 treated animal intravenous administration every day 1 time, for three days on end; 8～9 treated animal oral administration gavage every day administrations 1 time, for three days on end.After the last administration 30 minutes, each Mus is with 10% chloral hydrate anesthesia (400mg/kg, ip), it is fixing to face upward the position, the positive middle part of neck percutaneous incision skin, isolate bilateral carotid, in the middle of behind 0 trumpeter's art suture two ends ligation common carotid artery, cut off (wherein not ligation behind the sham operated rats threading), skin suture respectively.Take off cervical vertebra after 3 hours and put to death, get brain, putting weighs and dried to the weighing botle of constant weight claims its weight in wet base; 106 ℃ even weighing botles dry to constant weight, claim its total constant weight, its total constant weight is deducted weighing botle heavily is dry weight, calculate cerebral index and brain water content by following formula, and the t method of inspection carries out the significance test comparison with model group between the employing group.Heavy (g) * 100/ body weight (g) of cerebral index=cutaneous horn, brain water content (%)=heavy (g) * 100% of (heavy (the g)-brain stem of cutaneous horn heavy (g))/cutaneous horn
3 experimental results:
Influence to cerebral index: the cerebral index of iv promoting blood circulation to remove obstruction in the collateral injection 0.5g/kg group, 1g/kg treated animal is starkly lower than model group, learns by statistics and handles, and is significant difference (P＜0.05, P＜0.05); The cerebral index of ig promoting blood circulation to remove obstruction in the collateral injection 0.5g/kg group, 1g/kg treated animal and model group be not statistically signigicant relatively.
Influence to brain water content: the brain water content of iv promoting blood circulation to remove obstruction in the collateral injection 0.5g/kg group, 1g/kg treated animal is starkly lower than model group, learns by statistics and handles, and be significant difference (P＜0.05, P＜0.01), and drug effect strengthens with dosage; The brain water content of ig promoting blood circulation to remove obstruction in the collateral injection 0.5g/kg group, 1g/kg treated animal and model group be not statistically signigicant relatively.The results are shown in Table 1.
1: to the influence of ligation bilateral common carotid arteries rat brain index and brain water content (X ± S)
Dosed administration number of animals brain water content
The group cerebral index
(g/kg) approach (only) (%)
Sham-operation--iv 10 0.744 ± 0.028 77.91 ± 0.58
Model--iv 10 0.786 ± 0.021
△ △79.83 ± 1.23
△ △ △
Nimodipine 0.002 iv 10 0.722 ± 0.066** 78.41 ± 0.62**
XIANGDAN ZHUSHEYE 8 iv 10 0.754 ± 0.079 78.83 ± 0.69*
Promoting blood circulation to remove obstruction in the collateral liquid 0.25 iv 10 0.771 ± 0.074 79.12 ± 0.93
Promoting blood circulation to remove obstruction in the collateral liquid 0.5 iv 10 0.748 ± 0.053* 78.71 ± 0.88*
Promoting blood circulation to remove obstruction in the collateral liquid 1 iv 10 0.742 ± 0.046* 78.41 ± 0.79**
Promoting blood circulation to remove obstruction in the collateral liquid 0.5 ig 10 0.781 ± 0.034 79.24 ± 0.70
Promoting blood circulation to remove obstruction in the collateral liquid 1 ig 10 0.768 ± 0.020 79.05 ± 1.63
Compare with sham operated rats:
△ △: P＜0.01,
△ △ △: P＜0.001.
Compare with model group: *: P＜0.05, * *: P＜0.01.
Experimental example 2:Protective effect to middle cerebral artery blocking-up (MCAO) rat
Test objective: observe being subjected to of the influence of reagent thing, determine whether medicine has protective effect to rats with cerebral ischemia to scoring of middle cerebral artery blocking-up (MCAO) rat behavior and cerebral infarction rate.
1 group and dosage:
(1) sham operated rats: 0.9% sodium chloride injection.(2) model group: 0.9% sodium chloride injection.(3) positive drug group: 0.02% nimodipine solution (2mg/kg).(4) positive drug group: 80% compound Salviae Miltiorrhizae injection solution (8g/kg).(5) low dose group: 2.5% promoting blood circulation to remove obstruction in the collateral injection (0.25g/kg).(6) dosage group in: 5% promoting blood circulation to remove obstruction in the collateral injection (0.5g/kg).(7) high dose group: 10% promoting blood circulation to remove obstruction in the collateral injection (1g/kg) administration volume is 10ml/kg.
2 experimental implementation:
Get 70 of SD male rats, body weight 250～350g, be divided into 7 groups at random, be sham operated rats, model group, Western medicine positive drug group, Chinese medicine positive drug group, basic, normal, high 3 the dosage groups of promoting blood circulation to remove obstruction in the collateral injection, each treated animal intravenous administration every day 1 time, for three days on end, after the last administration 30 minutes, each Mus with 10% chloral hydrate anesthesia (300mg/kg, ip), with the mid point of lateral position along right external auditory canal and right eye outer canthus line, cut the about 2cm of skin perpendicular to line, cut off fascia, the passivity separating muscle exposes zygomatic arch.Bite zygomatic arch broken with mosquito forceps, zygomatic arch and mandibular bone are strutted, be fixed on the Mus plate with little drag hook pulling, expose the major part of squamosal bone, about 2～the 4mm in the front lower place of uniting before cheekbone and squamosal bone holes with desk-top electric car at the place then, open the microcephalia window of the about 2mm of a diameter, see through cerebral dura mater this moment, be middle cerebral artery (MCA) with regard to a visible straight and few ramose little blood vessel.It is almost vertically passed by tractus olfactorius and upwards goes; puncture cerebral dura mater with the fine needle acupuncture needle; expose medium-sized artery; after the affirmation electric knife is put the bipolar coagulation position, select the maximum electricity to coagulate switch, except that sham operated rats is not done the electric coagulation; all the other each treated animal electricity coagulate the interior 1mm of tractus olfactorius to one section middle cerebral artery between the inferior cerebral vein; electricity coagulated 4～6 seconds, avoided hemorrhage when electricity coagulates and the injury cerebral tissue, available wet cotton balls protection.Dab on the cranium window with fritter muscular tissue behind the blocking-up middle cerebral artery, then the layer-by-layer suture wound.Postoperative steams again raises.Above process is all carried out under room temperature constant (24～25 ℃) situation, is beneficial to estimate the cerebral ischemia situation.Observation is to the influence of focal cerebral ischemia in rats behavior scoring and cerebral infarct size.
1. focal cerebral ischemia in rats behavior scoring method
After treating that the MCAO rat anesthesia is clear-headed, carry out behavioristics and detect.Method is behind MCAO 4 hours and 24 hours, marks by following standard:
(1) carries overhead about 1 chi of Mus tail, observe the forelimb situation.Normal rat two forelimbs stretch to ground symmetrically.Left side shoulder inward turning is arranged, and receipts person in the left fore is chosen as 4 fens.Otherwise 0 minute;
(2), push away a left side (or right) shoulder respectively and check the resistance that opposing promotes to side shifting with on the sliding floor of animal horizontalization.Normal rat bilateral resistance is symmetry obviously.When right shoulder is mobile to the left, find the resistance descender,, be chosen as 1～3 fen according to the difference of decline degree;
(3) animal two forelimbs are put on the wire netting, observed the muscular tension of two forelimbs, normal rat two muscle of anterior limb tension force are symmetry obviously.The obvious descender of left fore muscular tension is taken place, and the weight according to descending is chosen as 1～3 fen.According to above standard scoring, full marks 10 minutes, mark is high more, illustrates that the behavior disorder of animal is serious more.Single blind method is adopted in scoring, and the t method of inspection carries out the significance test comparison with model group between the employing group.
2. cerebral infarct size is measured in TTC dyeing
24 hours broken ends are got brain behind the MCAO, and naked eyes are further proved conclusively the right side middle cerebral artery and blown between tractus olfactorius and inferior cerebral vein, and the harmless person of brain essence, brain is put (2～3) 10min in the ice-cold normal saline, after removing olfactory bulb, cerebellum and low brain stem, the crown four blade of cutting is cut into five.First cutter is before brain in the middle of the utmost point and the optic chiasma line; Second cutter is at the optic chiasma position; The 3rd cutter is at the infundibular stalk position; Four blade is between the infundibular stalk and the posterior lobe tail utmost point.Rapidly the brain sheet is put then in the phosphate buffer solution that 5ml contains 1%TTC, the lucifuge temperature was incubated 30 minutes, wherein stirred once every 7～8 minutes, dyed after, normal cerebral tissue is rose, and blocking tissue is white in color, and boundary is clearly demarcated.After temperature is incubated and finished the brain sheet is taken a picture, cut the calculating of weighing of red white colour district.Then according to the weight area method, totally 10 planar gross areas and infraction are removed the area in zone to calculate five brain sheets respectively, obtain the percentage ratio that the infarct area accounts for the hemisphere gross area, i.e. t method of inspection and model group are carried out significance test relatively between infraction rate, and employing group.
3 experimental results:
The result shows that model group is compared with sham operated rats, and infarct size is 21.81%, shows rat MCAO model modeling success.
Behavioristics scoring: after 4 hours, the scoring of the behavioristics of each treated animal of promoting blood circulation to remove obstruction in the collateral injection relatively decreases with model group, but zero difference statistically; After 24 hours, the behavioristics of each treated animal of promoting blood circulation to remove obstruction in the collateral injection scoring compare 4 hours on a declining curve, model group then rises to some extent; Behavioristics's scoring of each treated animal of promoting blood circulation to remove obstruction in the collateral injection all is starkly lower than model group, learns by statistics and handles, and all is significant difference (P＜0.05, P＜0.05, P＜0.01).
Influence to the cerebral infarction rate: the cerebral infarction rate of each treated animal of promoting blood circulation to remove obstruction in the collateral injection all is starkly lower than model group, learns by statistics and handles, and all be significant difference (P＜0.01, P＜0.001, P＜0.001), and drug effect strengthens with dosage.The results are shown in Table 2.
2: the influence (X ± S) that the MCAO rat behavior is learned scoring and cerebral infarction rate
Dosage number of animals behavioristics scoring cerebral infarction rate
Group
(g/kg) (only) 4h 24h (%)
Sham-operation--10 2.0 ± 0.0 2.0 ± 0.0 0.00 ± 0.00
Model--10 6.6 ± 2.7
△ △ △7.1 ± 2.3
△ △ △21.81 ± 7.30
△ △ △
Nimodipine 0.002 10 4.7 ± 2.3 3.2 ± 1.8*** 5.07 ± 3.93***
XIANGDAN ZHUSHEYE 8 10 5.7 ± 2.3 5.2 ± 1.8 11.10 ± 11.70*
Promoting blood circulation to remove obstruction in the collateral liquid 0.25 10 5.6 ± 2.0 4.9 ± 2.0* 12.47 ± 5.51**
Promoting blood circulation to remove obstruction in the collateral liquid 0.5 10 5.4 ± 2.3 4.6 ± 2.1*, 8.54 ± 7.64***
Promoting blood circulation to remove obstruction in the collateral liquid 1 10 4.9 ± 2.3 4.2 ± 1.8**, 2.91 ± 2.35***
Compare with sham operated rats:
△ △ △: P＜0.001.
Compare with model group: *: P＜0.05, * *: P＜0.01, * * *: P＜0.001.
Embodiment 1:
Semen Persicae 2000g Radix Paeoniae Rubra 2000g Rhizoma Chuanxiong 2000g
More than 3 flavors, get Rhizoma Chuanxiong 1 flavor, be ground into coarse powder, put in the supercritical extraction still CO
2Supercritical extraction twice is extracted still pressure 35Mpar, 50 ℃ of temperature for the first time; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Extracted 1 hour, and collected, get extractive of volatile oil from separating still.Proceed for the second time CO
2Supercritical extraction is extracted still pressure 35Mpar, 50 ℃ of temperature; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Continous way pumps into 95% ethanol of 1.2 times of amounts of medical material, extracts altogether 1 hour, collects extract from separating still, gets ethanol extract.The Rhizoma Chuanxiong volatile oil extract adds the double distilled water of 6 times of amounts of medical material, and vapor distillation is collected the distillate of 3 times of amounts of medical material, and distillate adds the NaOH of medical material 0.1% amount, leaves standstill, and filters, and gets medicinal liquid, and is standby.
Get Semen Persicae (boiling down), Radix Paeoniae Rubra 2 flavors add 10 times of water gagings, decoct 2 hours, filter, medicinal residues add 8 times of water gagings again, decoct 2 hours, filter, merging filtrate, medicinal liquid are evaporated to relative density 1.19, (40 ℃) add 4 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 75%, precipitate with ethanol left standstill 12 hours, filtered, decompression filtrate recycling ethanol, be concentrated into relative density 1.24 (40 ℃), add 9 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 85%, precipitate with ethanol, alcohol liquid transfers to pH value 7.5 with 20%NaOH liquid, leaves standstill 12 hours, filters, medicinal liquid adds the active carbon of 1% amount, stir, filter, medicinal liquid and above-mentioned Rhizoma Chuanxiong alcohol extract merge, reclaim ethanol to there not being the alcohol flavor, get clear paste, clear paste adds above-mentioned Rhizoma Chuanxiong reserve liquid, stirs, filter, medicinal liquid is adjusted to water for injection and contains crude drug 0.6g/ml, adds the sodium sulfite of 0.2% amount and the active carbon of 0.3% amount, and 100 ℃ are incubated 10 minutes, filtration or centrifugal, medicinal liquid ultrafiltration (membrane molecule amount 10,000), packing (every 10ml) contains crude drug 6g, sterilization, promptly get injection, intravenous drip, each 1, once-a-day, or follow the doctor's advice.Before facing usefulness, with sodium chloride injection 100 or 250ml dilution.
Embodiment 2
Semen Persicae 1714g Radix Paeoniae Rubra 2000g Rhizoma Chuanxiong 2572g
More than 3 flavors, get Rhizoma Chuanxiong 1 flavor, be ground into coarse powder, put in the supercritical extraction still CO
2Supercritical extraction twice is extracted still pressure 35Mpar, 50 ℃ of temperature for the first time; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Extracted 1 hour, and collected, get extractive of volatile oil from separating still.Proceed for the second time CO
2Supercritical extraction is extracted still pressure 35Mpar, 50 ℃ of temperature; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Continous way pumps into 95% ethanol of 1.2 times of amounts of medical material, extracts altogether 1 hour, collects extract from separating still, gets ethanol extract.The Rhizoma Chuanxiong volatile oil extract adds the double distilled water of 6 times of amounts of medical material, and vapor distillation is collected the distillate of 3 times of amounts of medical material, and distillate adds the NaOH of medical material 0.1% amount, leaves standstill, and filters, and gets medicinal liquid, and is standby.
Get Semen Persicae (boiling down), Radix Paeoniae Rubra 2 flavors add 10 times of water gagings, decoct 2 hours, filter, medicinal residues add 8 times of water gagings again, decoct 2 hours, filter, merging filtrate, medicinal liquid are evaporated to relative density 1.19, (40 ℃) add 4 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 75%, precipitate with ethanol left standstill 12 hours, filtered, decompression filtrate recycling ethanol is concentrated into relative density 1.24 (40 ℃), adds 9 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 85%, precipitate with ethanol, pure liquid transfers to pH value 7.5 with 20%NaOH liquid, left standstill 12 hours, and filtered, medicinal liquid adds the active carbon of 1% amount, stir, filter, medicinal liquid and above-mentioned Rhizoma Chuanxiong alcohol extract merge, recovery ethanol gets clear paste to there not being the alcohol flavor.Clear paste adds above-mentioned Rhizoma Chuanxiong reserve liquid, stirs, and filters, and with sucrose 50g, stevioside 1g flavoring transfers total amount to 10000ml, filters, fill, and sterilizing promptly gets oral liquid; Every bottle of 10ml, every bottle contains crude drug 6g, each 1-2 bottle, 1-3 time on the one, or follow the doctor's advice.
Embodiment 3:
The method of quality control of injection of the present invention
Differentiate: get this product 1ml, as need testing solution.Other gets the amygdaloside reference substance, adds methanol and makes the solution that every 1ml contains 3mg, in contrast product solution.According to thin layer chromatography (" 2000 editions appendix VIB of Chinese pharmacopoeia) test, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 15: 40: 20: 5 dichloromethane-ethyl acetate-methanol-water is developing solvent, launches, take out, spray is phosphomolybdic acid 2g with the phosphomolybdic acid sulfuric acid solution, adds water 20ml and makes dissolving, slowly adds sulphuric acid 30ml again, mixing makes, and is heated to clear spot at 105 ℃.In the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color.
Get this product 1ml, as need testing solution.Other gets the peoniflorin reference substance, makes the solution that every 1ml contains 2mg with methanol, in contrast product solution.According to thin layer chromatography (" 2000 editions appendix VIB of Chinese pharmacopoeia) test, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 40: 5: 10: 0.2 dichloromethane-ethyl acetate-methanol-formic acid was developing solvent, launch, take out, dry, spray is heated to clear spot with 2% vanillin sulfuric acid solution under 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Get this product 1ml, as need testing solution.Other gets the ferulic acid reference substance, makes the solution that every 1ml contains 0.5mg with methanol, in contrast product solution.According to thin layer chromatography (appendix VIB) test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with 30: 2.5: 1 toluene-methanol-glacial acetic acid was developing solvent, launched, and took out, dry, spray is with 50% alcoholic solution of 2% ferric chloride-2% potassium ferricyanide.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay: chromatographic condition and system suitability test are filler with the octadecylsilane bonded silica, and 27: 73 methanol-water is a mobile phase; The detection wavelength is 218nm; Number of theoretical plate calculates by amygdaloside peak and peoniflorin peak all should be not less than 3000; The preparation of reference substance solution, it is an amount of to be taken at 60 ℃ of amygdaloside, peoniflorin reference substances that are dried to constant weight, and accurate the title, decide, and adds mobile phase respectively and make the solution that every 1ml contains 0.4mg, promptly; This product 5ml is got in the preparation of need testing solution, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Every of this product contains Semen Persicae with amygdaloside (C
20H
27NO
11) meter, must not be less than 22mg, contain Radix Paeoniae Rubra with peoniflorin (C
23H
28O
11) meter, must not be less than 36mg.
Embodiment 4:
Injection of the present invention adopts fingerprint pattern quality control method
Measure according to high performance liquid chromatography (" 2000 editions appendix VID of Chinese pharmacopoeia), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-0.1% trifluoroacetic acid aqueous solution was a mobile phase in 26: 74; The detection wavelength is 218nm.Theoretical cam curve is pressed object of reference amygdaloside peak and is calculated, and should be not less than 3000; The preparation of object of reference solution, it is an amount of to be taken at 60 ℃ of amygdaloside objects of reference that are dried to constant weight, and accurate the title, decide, and adds methanol and make the solution that every 1ml contains 0.4mg, promptly; This product 2ml is got in the preparation of test solution, puts in the 25ml measuring bottle, and thin up shakes up to scale, promptly; Algoscopy, accurate object of reference solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid respectively, write down 1 hour chromatogram, be the S peak with corresponding peak, object of reference peak in the test sample finger printing, calculate relative retention time and peak area ratio, should meet the following requirements: this product finger printing should be similar to standard finger-print; Finger printing should have 8 total peaks;
Relative retention time (peak number): 0.350 ± 30% (1), 0.554 ± 30% (2), 1 (S), 1.413 ± 30% (3), 1.552 ± 30% (4), 2.022 ± 30% (5), 3.600 ± 30% (6), 3.985 ± 30% (7);
Peak area ratio (peak number): 1.218 ± 30% (5), 0.451 ± 40% (7); Non-total peak area must not be crossed 5% of total peak area.
Instrument: Agilent 1100 chromatograph of liquid; Chromatographic column: Agillent HypersilODS C
18(4.6 * 250mm, 5 μ m); Integral way: paddy paddy integration.
Claims (8)
1, a kind of Chinese medicine injection for the treatment of cardiovascular and cerebrovascular disease is characterized in that this injection made by following method:
Semen Persicae 1-4 weight portion Radix Paeoniae Rubra 1-4 weight portion
Rhizoma Chuanxiong 1-4 weight portion is got Rhizoma Chuanxiong 1 flavor, is ground into coarse powder, puts in the supercritical extraction still CO
2Supercritical extraction 2-3 time is extracted still pressure 20-40Mpar at every turn, temperature 40-60 ℃; Separation reactor I pressure 10-14Mpar, temperature 30-40 ℃; Separation reactor I I pressure 4-8Mpar, temperature 30-40 ℃; Extracted 1-2 hour, and collected, get extractive of volatile oil from separating still; After the last supercritical extraction, continous way pumps into 95% ethanol of 1.2 times of amounts of medical material among the separation reactor I I, extracts altogether 1 hour, collects extract from separating still, gets ethanol extract; The Rhizoma Chuanxiong volatile oil extract adds the double distilled water that medical material 4-10 doubly measures, and vapor distillation is collected the distillate that medical material 2-5 doubly measures, and distillate adds the NaOH of medical material 0.1-0.3% amount, leaves standstill, and filters, and gets medicinal liquid, and is standby; Get Semen Persicae, Radix Paeoniae Rubra 2 flavors add 6-12 times of water gaging, decoct 1-2 hour, filter, medicinal residues add 6-10 times of water gaging again, decoct 1-2 hour, filter, merging filtrate, medicinal liquid are evaporated to relative density 40 ℃ following 1.19, add clear paste 2-6 and doubly measure 85-95% ethanol, make medicinal liquid contain alcohol amount 70-80%, precipitate with ethanol, left standstill 8-18 hour, filter, decompression filtrate recycling ethanol is concentrated into 40 ℃ following 1.24 of relative densities, add clear paste 7-10 and doubly measure 85-95% ethanol, make medicinal liquid contain alcohol amount 80-85%, precipitate with ethanol, pure liquid transfers to pH value 6-8 with 15-25%NaOH liquid, left standstill 8-18 hour, filter, medicinal liquid adds the active carbon of 1-2% amount, stirs, filter, medicinal liquid and above-mentioned Rhizoma Chuanxiong alcohol extract merge, and recovery ethanol gets clear paste to there not being the alcohol flavor; Clear paste adds above-mentioned Rhizoma Chuanxiong reserve liquid, stirs, and filters, medicinal liquid is adjusted to water for injection and contains crude drug 0.6g/ml, adds the sodium sulfite of 0.1-0.3% amount and the active carbon of 0.2-0.4% amount, and 100 ℃ are incubated 8-12 minute, filtration or centrifugal, medicinal liquid ultrafiltration sterilization, promptly.
2, injection as claimed in claim 1 is characterized in that the preparation method of this injection is: get Rhizoma Chuanxiong 1 flavor, be ground into coarse powder, put in the supercritical extraction still CO
2Supercritical extraction twice is extracted still pressure 35Mpar, 50 ℃ of temperature for the first time; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Extracted 1 hour, and collected, get extractive of volatile oil from separating still; Proceed for the second time CO
2Supercritical extraction is extracted still pressure 35Mpar, 50 ℃ of temperature; Separation reactor I pressure 12Mpar, 35 ℃ of temperature; Separation reactor I I pressure 6Mpar, 35 ℃ of temperature; Continous way pumps into 95% ethanol of 1.2 times of amounts of medical material, extracts altogether 1 hour, collects extract from separating still, gets ethanol extract; The Rhizoma Chuanxiong volatile oil extract adds the double distilled water of 6 times of amounts of medical material, and vapor distillation is collected the distillate of 3 times of amounts of medical material, and distillate adds the NaOH of medical material 0.1% amount, leaves standstill, and filters, and gets medicinal liquid, and is standby; Get the Semen Persicae down that boils, Radix Paeoniae Rubra 2 flavors add 10 times of water gagings, decoct 2 hours, filter, medicinal residues add 8 times of water gagings again, decoct 2 hours, filter, merging filtrate, medicinal liquid are evaporated to 40 ℃ of following relative densities 1.19, add 4 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 75%, precipitate with ethanol, left standstill 12 hours, and filtered decompression filtrate recycling ethanol, be concentrated into 40 ℃ of following relative densities 1.24, add 9 times of amounts of clear paste, 95% ethanol, make medicinal liquid contain alcohol amount 85%, precipitate with ethanol, pure liquid transfers to pH value 7.5 with 20%NaOH liquid, leaves standstill 12 hours, filter, medicinal liquid adds the active carbon of 1% amount, stirs, filter, medicinal liquid and above-mentioned Rhizoma Chuanxiong alcohol extract merge, and recovery ethanol gets clear paste to there not being the alcohol flavor; Clear paste adds above-mentioned Rhizoma Chuanxiong reserve liquid, stirs, and filters, and medicinal liquid is adjusted to water for injection and contains crude drug 0.6g/ml, add the sodium sulfite of 0.2% amount and the active carbon of 0.3% amount, 100 ℃ are incubated 10 minutes, filtration or centrifugal, the medicinal liquid ultrafiltration, membrane molecule amount 10,000 promptly gets injection.
3, require the method for quality control of 1 or 2 described injection as profit, it is characterized in that discrimination method in this method comprises one or more in the following discriminating:
A. get this product 1ml, as need testing solution; Other gets the amygdaloside reference substance, adds methanol 6ml and makes dissolved solution, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 12-18: 30-50: 15-25: 3-6 dichloromethane-ethyl acetate-methanol-water is developing solvent, launch, take out, spray is heated to clear spot with the phosphomolybdic acid sulfuric acid solution at 100-105 ℃; In the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color; Wherein the phosphomolybdic acid sulfuric acid solution is to add water 20ml by phosphomolybdic acid 2g to make dissolving, slowly adds sulphuric acid 30ml again, and mixing makes;
B. get this product 1ml, as need testing solution; Other gets the peoniflorin reference substance, makes the solution that every 1ml contains 2mg with methanol, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 30-50: 3-6: 8-12: 0.1-0.3 dichloromethane-ethyl acetate-methanol-formic acid is developing solvent, launch, take out, dry, spray is heated to clear spot with 1-2% vanillin sulfuric acid solution under 100-105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
C. get this product 1ml, as need testing solution; Other gets the ferulic acid reference substance, makes the solution that every 1ml contains 0.5mg with methanol, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with 25-35: 1-3: 1-2 toluene-methanol-glacial acetic acid is developing solvent, launches, and takes out, dry, spray is with 50% alcoholic solution of 2% ferric chloride-2% potassium ferricyanide; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
4, require 3 described method of quality control as profit, it is characterized in that discrimination method in this method comprises one or more in the following discriminating:
A. get this product 1ml, as need testing solution; Other gets the amygdaloside reference substance, adds methanol and makes the solution that every 1ml contains 3mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 15: 40: 20: 5 dichloromethane-ethyl acetate-methanol-water is developing solvent, launches, take out, spray is phosphomolybdic acid 2g with the phosphomolybdic acid sulfuric acid solution, adds water 20ml and makes dissolving, slowly adds sulphuric acid 30ml again, mixing makes, and is heated to clear spot at 105 ℃; In the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color;
B. get this product 1ml, as need testing solution; Other gets the peoniflorin reference substance, makes the solution that every 1ml contains 2mg with methanol, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 40: 5: 10: 0.2 dichloromethane-ethyl acetate-methanol-formic acid was developing solvent, launched, and took out, dry, spray is heated to clear spot with 2% vanillin sulfuric acid solution under 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
C. get this product 1ml, as need testing solution; Other gets the ferulic acid reference substance, makes the solution that every 1ml contains 0.5mg with methanol, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with 30: 2.5: 1 toluene-methanol-glacial acetic acid was developing solvent, launched, and took out, dry, spray is with 50% alcoholic solution of 2% ferric chloride-2% potassium ferricyanide; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
5, require the method for quality control of 1 or 2 described injection as profit, it is characterized in that the assay in this method is: according to high effective liquid chromatography for measuring, chromatographic condition and system suitability test, with octadecylsilane chemically bonded silica is filler, and the methanol-water of 25-29: 71-76 is a mobile phase; The detection wavelength is 218nm, and number of theoretical plate calculates by amygdaloside peak and peoniflorin peak all should be not less than 3000; The preparation of reference substance solution, it is an amount of to be taken at 60 ℃ of amygdaloside, peoniflorin reference substances that are dried to constant weight, and accurate the title, decide, and adds mobile phase respectively and make the solution that every 1ml contains 0.4mg, promptly; This product 5ml is got in the preparation of need testing solution, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Every of this product contains Semen Persicae in amygdaloside, must not be less than 22mg, contains Radix Paeoniae Rubra in peoniflorin, must not be less than 36mg.
6, require 5 described method of quality control as profit, it is characterized in that the assay in this method is: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica, and 27: 73 methanol-water is a mobile phase; The detection wavelength is 218nm; Number of theoretical plate calculates by amygdaloside peak and peoniflorin peak all should be not less than 3000; The preparation of reference substance solution, it is an amount of to be taken at 60 ℃ of amygdaloside, peoniflorin reference substances that are dried to constant weight, and accurate the title, decide, and adds mobile phase respectively and make the solution that every 1ml contains 0.4mg, promptly; This product 5ml is got in the preparation of need testing solution, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Every of this product contains Semen Persicae in amygdaloside, must not be less than 22mg, contains Radix Paeoniae Rubra in peoniflorin, must not be less than 36mg.
7, the method for quality control of injection as claimed in claim 1 or 2 is characterized in that this method comprises the steps:
Differentiate: a. gets this product, adds methanol and makes every 1ml and contain the 3mg1 bottle, as need testing solution; Other gets the amygdaloside reference substance, adds methanol 6ml and makes dissolved solution, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 12-18: 30-50: 15-25: 3-6 dichloromethane-ethyl acetate-methanol-water is developing solvent, launch, take out, spray is heated to clear spot with the phosphomolybdic acid sulfuric acid solution at 100-105 ℃; In the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color; Wherein the phosphomolybdic acid sulfuric acid solution is to add water 20ml by phosphomolybdic acid 2g to make dissolving, slowly adds sulphuric acid 30ml again, and mixing makes;
B. get this product, add methanol and make every 1ml and contain the 3mg1 bottle, as need testing solution; Other gets the peoniflorin reference substance, makes the solution that every 1ml contains 2mg with methanol, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 2ul of solution, put respectively on same silica gel g thin-layer plate, with 30-50: 3-6: 8-12: 0.1-0.3 dichloromethane-ethyl acetate-methanol-formic acid is developing solvent, launch, take out, dry, spray is heated to clear spot with 1-2% vanillin sulfuric acid solution under 100-105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
C. get this product, add methanol and make every 1ml and contain the 3mg1 bottle, as need testing solution; Other gets the ferulic acid reference substance, makes the solution that every 1ml contains 0.5mg with methanol, in contrast product solution; According to thin layer chromatography (appendix VIB) test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with 25-35: 1-3: 1-2 toluene-methanol-glacial acetic acid is developing solvent, launches, and takes out, dry, spray is with 50% alcoholic solution of 2% ferric chloride-2% potassium ferricyanide; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Assay: according to high effective liquid chromatography for measuring, chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica, and the methanol-water of 25-29: 71-76 is a mobile phase; The detection wavelength is 218nm, and number of theoretical plate calculates by amygdaloside peak and peoniflorin peak all should be not less than 3000; The preparation of reference substance solution, it is an amount of to be taken at 60 ℃ of amygdaloside, peoniflorin reference substances that are dried to constant weight, and accurate the title, decide, and adds mobile phase respectively and make the solution that every 1ml contains 0.4mg, promptly; This product 5ml is got in the preparation of need testing solution, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly; Algoscopy, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Every of this product contains Semen Persicae in amygdaloside, must not be less than 22mg, contains Radix Paeoniae Rubra in peoniflorin, must not be less than 36mg.
8, require the method for quality control of 1 or 2 described injection as profit, it is characterized in that this method has adopted the method for finger printing: according to high effective liquid chromatography for measuring, chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 24-28: 74 methanol-0.1% trifluoroacetic acid aqueous solution is a mobile phase; The detection wavelength is 218nm.Theoretical cam curve is pressed object of reference amygdaloside peak and is calculated, and should be not less than 3000; The preparation of object of reference solution, it is an amount of to be taken at 60 ℃ of amygdaloside objects of reference that are dried to constant weight, and accurate the title, decide, and adds methanol and make the solution that every 1ml contains 0.4mg, promptly; This product 2ml is got in the preparation of test solution, puts in the 25ml measuring bottle, and thin up shakes up to scale, promptly; Algoscopy, accurate object of reference solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid respectively, write down 1 hour chromatogram, be the S peak with corresponding peak, object of reference peak in the test sample finger printing, calculate relative retention time and peak area ratio, should meet the following requirements: this product finger printing should be similar to standard finger-print; Finger printing should have 8 total peaks, relative retention time, i.e. peak number: 0.350 ± 30% (1), 0.554 ± 30% (2), 1 (S), 1.413 ± 30% (3), 1.552 ± 30% (4), 2.022 ± 30% (5), 3.600 ± 30% (6), 3.985 ± 30% (7); Peak area ratio (peak number): 1.218 ± 30% (5), 0.451 ± 40% (7); Non-total peak area must not be crossed 5% of total peak area.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02156776 CN1272036C (en) | 2002-12-18 | 2002-12-18 | Medicinal injection for treating cardio-cerebrovascular disease and preparing method thereof |
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|---|---|---|---|
| CN 02156776 CN1272036C (en) | 2002-12-18 | 2002-12-18 | Medicinal injection for treating cardio-cerebrovascular disease and preparing method thereof |
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| Publication Number | Publication Date |
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| CN1507907A true CN1507907A (en) | 2004-06-30 |
| CN1272036C CN1272036C (en) | 2006-08-30 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526279A (en) * | 2012-02-12 | 2012-07-04 | 贾奎 | Chinese medicinal composition for treating central high fever caused by acute cerebrovascular disease and preparation method for Chinese medicinal composition |
| CN102940702A (en) * | 2012-10-31 | 2013-02-27 | 成都医路康医学技术服务有限公司 | Medicine composition for treating cardia-cerebrovascular diseases |
-
2002
- 2002-12-18 CN CN 02156776 patent/CN1272036C/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526279A (en) * | 2012-02-12 | 2012-07-04 | 贾奎 | Chinese medicinal composition for treating central high fever caused by acute cerebrovascular disease and preparation method for Chinese medicinal composition |
| CN102526279B (en) * | 2012-02-12 | 2013-08-28 | 贾奎 | Chinese medicinal composition for treating central high fever caused by acute cerebrovascular disease and preparation method for Chinese medicinal composition |
| CN102940702A (en) * | 2012-10-31 | 2013-02-27 | 成都医路康医学技术服务有限公司 | Medicine composition for treating cardia-cerebrovascular diseases |
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| CN1272036C (en) | 2006-08-30 |
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