CN102749413A

CN102749413A - Quality detection method of traditional Chinese medicine composition for treating headache

Info

Publication number: CN102749413A
Application number: CN2012102120117A
Authority: CN
Inventors: 付立家; 付建家
Original assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Current assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Priority date: 2008-02-22
Filing date: 2008-02-22
Publication date: 2012-10-24
Anticipated expiration: 2028-02-22
Also published as: CN102749413B

Abstract

The invention provides a traditional Chinese medicine composition for treating headache and its preparation method and quality control method. The traditional Chinese medicine composition comprises cassia seed, szechuan lovage rhizome, dahurian angelica root, prepared aconite root and gambir plant. The traditional Chinese medicine composition can be added with auxiliary materials and be prepared into clinically acceptable tablets, capsules, an oral liquid, dropping pills, soft capsules and granules by conventional processes. The quality control method of the traditional Chinese medicine composition comprises the following steps of carrying out qualitative detection of szechuan lovage rhizome and dahurian angelica root, carrying out qualitative amount-limited detection of aconitine, carrying out quantitative detection of alcohol soluble extract, carrying out detection of nitrogen content, and carrying out quantitative detection of ferulic acid. The traditional Chinese medicine composition has an obvious effect of treating headache.

Description

The quality determining method of the Chinese medicine composition of treatment headache

The present invention is for dividing an application, and the original bill application number is 200810057967.8, and the original bill applying date is on February 22nd, 2008, and the original bill name is called Chinese medicine composition of treatment headache and preparation method thereof and method of quality control.

Technical field

The present invention relates to a kind of pharmaceutical composition, relate in particular to a kind of Chinese medicine composition of treating headache and preparation method thereof and method of quality control, belong to technical field of traditional Chinese medicines.

Background technology

Headache is common clinically illness, is that primary symptom person then is found in fever caused by infection property disease at present with the headache, diseases such as high blood pressure, encephalic illness, neurosis, cerebral concussion and antimigraine.

All diseases caused by external factors six external factors which cause diseases, in viscerotrauma, cause yang-energy to block, saw on the turbid pathogenic factor, liver-yang hyperactivity, the marrow deficiency of vital energy and blood, the not normal person of channels and collaterals running all can have a headache.By cause of disease branch, headache has diseases caused by external factors, internal injury not.Exogenous headache has cold, wind-heat, rheumatism, hinders heat, and fire-evil causes pain and cold headache etc.Beadache with internal injury has the deficiency of vital energy, the deficiency of blood, the deficiency of yang, the deficiency of Yin, liver-yang, gets ill from overeating, hemostasis causes pain etc.From the channels and collaterals branch, three-yang headache (taiyang headache, yangming headache, shaoyang headache), headache of triple yin (taiyin headache, shaoyin headache, jueyin headache) etc. are arranged.By state of an illness weight, course of disease length, outbreak rule and painful area branch, unendurable headache a, wind, antimigraine, thunder headache, brain wind, parietal headache, chronic headache etc. are arranged.

Headache is that excuse neck pain sensation tip receptor is upset and produces unusual nerve impulse and be communicated to due to the brain.Cranium is organized outward except that skull itself, and is all responsive to pain until face, oral cavity from periosteum; Intracranial tissue has only venous sinus and reflux veins, the hard brain of basis cranii and arteria basilaris to the pain sensitivity, and the brain remaining tissue is all insensitive to the pain sensation.To cranial nerve and the 2nd～3 pair of cervical nerve conduction, still can conduct through sympathetic nerve except that above-mentioned nerve by the outer pain sensation of cranium through V, IV, X for the encephalic pain sensation.

Treatment at first is active prevention and the various protopathy of treatment.Symptomatic treatment then can use the analgesic drug product except that the morphine class, like various antipyretic analgesics, can according to the state of an illness pause clothes or 2-3 time/d of short-term take, severe patient can be taken codeine, Rotundine or dihydroetorphine etc. on a small quantity.But above these medicines are taken for a long time and are not only had habituation property, and tolerance is poor, can't fundamentally solve patient's headache problem.The treatment by Chinese herbs headache not only can reach the effect of treating both principal and secondary aspect of disease, and can effectively treat various protopathy, and can not produce dependence, has improved the security that the patient uses.

Summary of the invention

The object of the present invention is to provide a kind of Chinese medicine composition of treating headache; Another object of the present invention is to provide the preparation method of this Chinese medicine composition; The 3rd purpose of the present invention is to provide the method for quality control of this Chinese medicine composition.

The present invention seeks to realize through following technical scheme:

The bulk drug of the Chinese medicine composition of treatment headache of the present invention consists of:

Goat's horn 1300-1500 weight portion, Ligusticum wallichii 200-400 weight portion, root of Dahurain angelica 300-500 weight portion, aconiti preparata,radix 100-300 weight portion.

The above-mentioned raw materials optimum ratio is:

Goat's horn 1350-1450 weight portion, Ligusticum wallichii 200-300 weight portion, root of Dahurain angelica 400-500 weight portion, aconiti preparata,radix 100-200 weight portion.

The Chinese medicine composition of treatment headache of the present invention also can be processed by the bulk drug of following weight ratio:

Goat's horn 1300-1500 weight portion, Ligusticum wallichii 200-400 weight portion, root of Dahurain angelica 300-500 weight portion, aconiti preparata,radix 100-300 weight portion, yncaria stem with hooks 100-300 weight portion.

The above-mentioned raw materials optimum ratio is:

Goat's horn 1350-1450 weight portion, Ligusticum wallichii 200-300 weight portion, root of Dahurain angelica 400-500 weight portion, aconiti preparata,radix 100-200 weight portion, yncaria stem with hooks 150-250 weight portion.

The above-mentioned raw materials optimum ratio is:

Goat's horn 1400 weight portions, Ligusticum wallichii 250 weight portions, the root of Dahurain angelica 450 weight portions, aconiti preparata,radix 150 weight portions, yncaria stem with hooks 200 weight portions.

In the invention described above traditional Chinese medicinal composition raw materials:

Goat's horn can be substituted by mother-of-pearl, raddle, oyster, tribulus terrestris or bright rhizoma Gastrodiae; The root of Dahurain angelica can be substituted by windproof, the notopterygium root or the achene of Siberian cocklebur.

Composition of the present invention can add auxiliary material by common process and process clinical acceptable forms such as tablet, capsule, oral liquid, pill, soft capsule, granule; Said auxiliary material comprises solvent, disintegrant, flavouring, antiseptic, colorant, bonding agent, lubricant, matrix etc.

The preparation method of the tablet of the Chinese medicine composition of treatment headache of the present invention is:

Goat's horn pound sheet, the boiling secondary adds 8 times of water gagings for the first time and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, merging filtrate, relative density is 1.32～1.35 thick paste when being concentrated into 50 ℃; All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, relative density is 1.32～1.35 thick paste when being concentrated into 50 ℃; With goat's horn cream and dextrin 100～150g mixing, granulate drying; Be pressed into 1000, dressing promptly gets.

Goat's horn pound sheet is meant scleroid goat's horn raw medicinal material, becomes superfine medicine materical crude slice according to traditional processing procedure with special pound cutter pound.

Pharmaceutical composition method of quality control of the present invention comprises one or more in following qualitative checking method and/or the quantitative detecting method:

(1) qualitative detection of Ligusticum wallichii

Get pharmaceutical composition content of the present invention, porphyrize, the sonicated that adds diethyl ether filters, the filtrating evaporate to dryness, residue adds ethanol makes dissolving, as need testing solution;

Get the Ligusticum wallichii control medicinal material, add the ethanol sonicated and be prepared into control medicinal material solution;

According to the thin-layered chromatography test, draw need testing solution, control medicinal material solution, put respectively on same silica gel g thin-layer plate, be developping agent with cyclohexane-ethyl acetate, launch, take out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(2) qualitative detection of the root of Dahurain angelica

Get pharmaceutical composition content of the present invention, porphyrize adds absolute ethyl alcohol, and sonicated filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol and disperses, and gets supernatant as need testing solution;

Get root of Dahurain angelica control medicinal material, be prepared into control medicinal material solution according to the need testing solution preparation method;

According to the thin-layered chromatography test, draw above-mentioned two kinds of solution respectively, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-methyl alcohol-formic acid, launch, take out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(3) the qualitative limit examine of aconitine

Get pharmaceutical composition content of the present invention, be ground into fine powder, precision takes by weighing, and adds ammonia solution, places, and adds diethyl ether, and jolting is placed, filter, and the filtrating evaporate to dryness, residue is used anhydrous alcohol solution, as need testing solution;

Get the aconitine reference substance, add absolute ethyl alcohol and process reference substance solution;

According to the thin-layered chromatography test, draw above-mentioned two kinds of solution points on same silica G plate, be developping agent with cyclohexane-ethyl acetate-diethylamine, launch, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot colors that occurs should be shallower than the reference substance spot or spot not occur;

(4) detection by quantitative of forulic acid:

According to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; With acetonitrile-water-glacial acetic acid is moving phase; The detection wavelength is 322nm; Number of theoretical plate calculates by the forulic acid peak should be not less than 1500;

The preparation of reference substance solution: precision takes by weighing the forulic acid reference substance, puts in the measuring bottle, with dissolve with methanol and be diluted to scale, shakes up, and promptly gets;

The preparation of need testing solution: get pharmaceutical composition content of the present invention, porphyrize, the accurate title, decide, and adds methyl alcohol, and reflux is put cold. filters; Residue is washed 2 times with methyl alcohol, filters, and merging filtrate, evaporate to dryness, residue add time dissolving of water-ammonia moisture, put in the separating funnel; Wash 2 times with ether, discard ether solution, water liquid is transferred pH with watery hydrochloric acid, with extracted with diethyl ether 4 times, merges ether solution; With washing, discard water liquid, the ether solution evaporate to dryness, residue adds dissolve with methanol, is transferred in the measuring bottle; Be diluted to scale with methyl alcohol, shake up, filter, get subsequent filtrate, promptly get;

Determination method: accurate respectively reference substance solution and the need testing solution drawn, inject liquid chromatograph, measure, promptly get.

Pharmaceutical composition method of quality control of the present invention comprises one or more in following preferred qualitative checking method and/or the quantitative detecting method:

(1) qualitative detection of Ligusticum wallichii

Pharmaceutical composition content of the present invention is equivalent to crude drug 49g, porphyrize, and the 30ml sonicated that adds diethyl ether 30 minutes filters, the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets Ligusticum wallichii control medicinal material 1g, adds ethanol 30ml sonicated 30 minutes, filter, and the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as control medicinal material solution; According to the thin-layered chromatography test, draw need testing solution 8 μ l, control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate; Cyclohexane-ethyl acetate with the 7:3 ratio is a developping agent, launches, and takes out; Dry, put under the ultraviolet lamp that wavelength is 365nm and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(2) qualitative detection of the root of Dahurain angelica

Pharmaceutical composition content of the present invention is equivalent to crude drug 15g, and porphyrize adds absolute ethyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol 2ml and disperses, and gets supernatant as need testing solution; Other gets root of Dahurain angelica control medicinal material 1g, shines medicinal material solution in pairs with the method preparation; According to thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with the methenyl choloride-methyl alcohol-formic acid of 9:1:0.1 ratio, launch, take out, dry, put under the ultraviolet lamp that wavelength is 365nm and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(3) the qualitative limit examine of aconitine

Pharmaceutical composition content of the present invention is ground into fine powder, and precision takes by weighing 5g, adds ammonia solution 5ml, placed 2 hours, and the 100ml that adds diethyl ether, jolting 1 hour was placed 24 hours, filters, the filtrating evaporate to dryness, residue is with absolute ethyl alcohol 2ml dissolving, as need testing solution; It is an amount of that other gets the aconitine reference substance, adds absolute ethyl alcohol and process the reference substance solution that every 1ml contains 2mg; According to thin-layered chromatography test, draw each 2 μ l point of above-mentioned two kinds of solution on same silica G plate, be developping agent with the cyclohexane-ethyl acetate-diethylamine of 7:2:0.5 ratio, launch, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot colors that occurs should be shallower than the reference substance spot or spot not occur;

(4) detection by quantitative of forulic acid:

According to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; Acetonitrile-water-glacial acetic acid with the 20:80:1 ratio is a moving phase; The detection wavelength is 322nm; Number of theoretical plate calculates by the forulic acid peak should be not less than 1500;

The preparation of reference substance solution: precision takes by weighing forulic acid reference substance 3mg, puts in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up, and processes to contain forulic acid 30 μ g among every 1ml, promptly gets;

The preparation of need testing solution: it is an amount of to get pharmaceutical composition content of the present invention, and porphyrize is got 2.5 g, and accurate the title decides, and adds methyl alcohol 50ml, reflux 1h; Put cold. filter, residue is washed 2 times with the methyl alcohol of 5ml, filters, and merging filtrate, evaporate to dryness, residue add the water-ammonia water 20ml gradation dissolving of 20:2 ratio; Put in the separating funnel, wash 2 times with the ether of 20ml, discard ether solution, water liquid is transferred pH to 2 with watery hydrochloric acid, with extracted with diethyl ether 4 times, and 20ml at every turn; Merge ether solution,, discard water liquid with the washing of 30ml, the ether solution evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10 ml measuring bottles; Be diluted to scale with methyl alcohol, shake up, filter, get subsequent filtrate, promptly get;

Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.

The flat liver of goat's horn is relieving convulsion in the medicine of the present invention, is applicable to liver-yang hyperactivity, have a dizzy spell, and liver-fire flaming, red eye, swell pain, diseases such as convulsion are monarch drug in a prescription; Ligusticum wallichii, the root of Dahurain angelica, aconiti preparata,radix are dispelled rheumatism, and promoting qi circulation and relieving pain is a ministerial drug; The yncaria stem with hooks heat-clearing, flat liver, breath wind is adjutant; Each medicine share plays flat liver altogether, and the effect of analgesia is used for antimigraine, vascular headache, tension headache and nervous headache, and effect is remarkable.

Owing to contain a large amount of volatile ingredients in the prescription, in the preparation process of medicine of the present invention, select the percolation effective component extracting, be the key that curative effect of medication is improved; And the extract powder that contains in the preparation has very strong hydroscopicity, if only process plain sheet, the not only easy moisture absorption influences the dissolving of medicine, and distribution, absorption and stability of formulation are processed coating tablet and then can effectively be avoided above problem.

Through method of quality control of the present invention; Not only qualitative detection Ligusticum wallichii, the root of Dahurain angelica have also carried out the content limit inspection to aconitine, and quantitatively control content of ferulic acid in the medicine; Control the quality and the curative effect of product significantly, guaranteed the security of medicine of the present invention.

In order to make those skilled in the art understand content of the present invention better, carry out detailed explanation in the face of pharmaceutical composition of the present invention and preparation method thereof with method of quality control down.

Beneficial effect

Following Test Example and embodiment are used to further specify the present invention but are not limited to the present invention.

Test Example 1 process test research

1, diacolation speed trial:

Press three parts of medicinal materials of embodiment 1 prescription configuration, every part contains Ligusticum wallichii 250g, root of Dahurain angelica 450g; Aconiti preparata,radix 150g, yncaria stem with hooks 200g add 30% ethanol and make solvent; Cold soaking 24 hours, diacolation is divided into five groups; Diacolation speed is respectively: 100ml/min.kg, 120ml/min.kg, 150ml/min.kg, 180 ml/min.kg, 200 ml/min.kg, confirm diacolation speed according to paste volume.The result sees table 1.

Table 1: diacolation speed trial

According to above data, can find out that diacolation speed is 150ml/min.kg, this is complete to go out cream base, and diacolation speed is decided to be 150ml/min.kg in the actual production.

2, pulverize the smashing fineness test of medicinal material

Press embodiment 1 recipe quantity configuration Ligusticum wallichii, the root of Dahurain angelica, aconiti preparata,radix.Divide four groups to do experiment respectively: smashing fineness is respectively: 10 orders, 24 orders, 50 orders, 65 orders are that index is confirmed smashing fineness to pulverize loss respectively.The result sees table 2:

Table 2: pulverize test findings

Above result shows: when smashing fineness was 24 orders, each item index was better, so select 24 orders for use in producing.

3, amount of water test

Press three parts of medicinal materials of embodiment 1 prescription preparation, every part contains goat's horn pound sheet 1400g and divides into groups to make an experiment, and first group of amount of water is 6 times of amounts of medicinal material, 4 times of amounts; Second group of amount of water is 10 times of amounts of medicinal material, 8 times of amounts; The 3rd group of amount of water is 8 times of amounts of medicinal material, 6 times of amounts.With the paste volume is that leading indicator is confirmed amount of water.See table 3:

Table 3: amount of water test

Above result shows: be that 8 times of amounts of index amount of water, 6 times of amount extractions are complete basically with the paste volume, confirm as 8 times of amounts, 6 times of amounts according to the needs of production amount of water.

4, the selection of auxiliary material:

In view of drug substance contents of the present invention is full medicinal extract, have very strong viscosity and moisture absorption, be unfavorable for preparations shaping, need to add suitable filling agent, respectively dextrin, starch, lactose are examined or check, the examination result sees table 4:

The examination result of table 4 filling agent

?	The Comprehensive Assessment result
		Dextrin	Compressibility is strong, and is cheap.
Starch	Separately as filling agent, poor compressibility.
		Lactose	Price is more expensive, and specification is also inconsistent.

[0083]So select dextrin for use, both can change the viscosity of sheet, be easy to compressing tablet again, select sugar coated tablet, have certain protection against the tide, secluding air effect and can cover up bad smell, improve outward appearance and be easy to and swallow.

5, sugar coated tablet art for coating

5.1 separation layer: place coating pan to roll plain sheet; (35% peach gum: 70% syrup=1:5) makes and evenly adheres to unilateral (the plain sheet: mixed pulp=50 g:1 ml) of going up to add mixed pulp; Blowing hot-air is dry, for preventing that tablet is inter-adhesive or sticking on the coating pan, adds talcum powder; 40～50 ℃ of hot blasts are dry down, twice of repetitive operation.

5.2 sub coat: slice, thin piece continues in coating pan, to roll, and adds 70% syrup, make the sheet sub-surface evenly wetting after; Add talcum powder; Make to adhere to the sheet sub-surface, continue to roll and blowing dry (50～55 ℃) repetitive operation; Till the slice, thin piece faceted pebble disappeared, heavy by plain sheet: 70% syrup: talcum powder=3:1.7:1 fed intake.

5.3 sugarcoating layer: slice, thin piece rolls in coating pan, and it is slowly dry to add 70% syrup (plain sheet: 70% syrup=15:1, the syrup addition is descending successively decreases) sheet sub-surface, forms fine and smooth sugar crystal clothing layer, increases clothing layer fastness and sweet taste.

5.4 coloured sugarcoating layer: be diluted to the mill base of 2mg/ml with 70% syrup, mill base is diluted to not to concentration with 70% syrup, concentration is ascending, adds coating pan, and is dry layer by layer.Plain sheet and pigment amount ratio are 7.5kg:1g.

5.5 polishing: for the sugar coated tablet surface-brightening is attractive in appearance, have moisture-proof role concurrently, add insect wax, press plain sheet: insect wax=3kg:5g adds, and rotates coating pan insect wax is wrapped on the slice, thin piece uniformly, takes out coating tablet, after dry 24 hours, promptly gets.

Experimental example 2 quality determining method experimental studies

1, the thin layer of Ligusticum wallichii is differentiated

(1) preparation of need testing solution and reference substance solution

Get according to 20 totally four parts in the preparation of embodiment 1 preparation, remove sugar-coat, porphyrize, the 30ml sonicated that adds diethyl ether respectively filters, the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution.

Get totally four parts of Ligusticum wallichii control medicinal material 1g, add ethanol 30ml sonicated, filter, the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as control medicinal material solution.

Prepare the negative sample that lacks Ligusticum wallichii according to embodiment 1 preparation method, prepare negative control solution according to the need testing solution preparation method again.

According to the thin-layered chromatography test, draw each 8 μ l of need testing solution and negative control solution, control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate; With cyclohexane-ethyl acetate (7:3) is developping agent, launches, and takes out; Dry, put under the ultraviolet lamp (365nm) and inspect.Relatively different extraction times, need testing solution and the color developing effect of control medicinal material solution on thin layer plate, the result sees the following form 5:

The comparison of table 5 ultrasonic Extraction time

(2) selection of developping agent

Prepare need testing solution, reference substance solution and negative sample solution according to above-mentioned preferable methods,, draw each 8 μ l of need testing solution and negative control solution, control medicinal material solution 2 μ l according to the thin-layered chromatography test; Put respectively on same silica gel g thin-layer plate; With developping agent, launch, take out; Dry, put under the ultraviolet lamp (365nm) and inspect.More different developping agents, need testing solution and the control medicinal material solution expansion effect on thin layer plate, the result sees the following form 6:

The selection of table 6 developping agent

Can find out that from last table selecting cyclohexane-ethyl acetate 7:3 is developping agent, launch effectively that and feminine gender is noiseless, the Pass Test requirement.Through revision test repeatedly, prove that this method stability and reappearance are all good, so with the qualitative detection of Ligusticum wallichii as one of method of quality control of pharmaceutical composition of the present invention.

2, the thin layer of the root of Dahurain angelica is differentiated

Get by 6 in the preparation of embodiment 1 preparation, porphyrize adds absolute ethyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol 2ml and disperses, and gets supernatant as need testing solution.

Get root of Dahurain angelica control medicinal material 1g, shine medicinal material solution in pairs with the method preparation.

Prepare the negative sample that lacks the root of Dahurain angelica according to embodiment 1 preparation method, prepare negative control solution according to the need testing solution preparation method again.

(1) comparison of developping agent

Prepare test sample, control medicinal material solution and negative control article solution by above-mentioned method, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With methenyl choloride-methyl alcohol-formic acid is developping agent; Proportioning is respectively 7:3:0.2,8:2:0.1,9:1:0.2,9:1:0.1, launches, and takes out; Dry, put under the ultraviolet light (365nm) and inspect.Compare need testing solution and the expansion effect of reference substance solution on thin layer plate, the result sees the following form 7:

The selection of table 7 developping agent proportioning

Can find out from last table, so that methenyl choloride-methyl alcohol-when formic acid 9:1:0.1 was developping agent, need testing solution and reference substance solution are launched effect on thin layer plate best, and negative noiseless, the Pass Test requirement.

(2) selection of point sample amount

Prepare test sample and control medicinal material solution as stated above, draw need testing solution 2 μ l, 5 μ l, 8 μ l, 10 μ l, reference substance solution 10 μ l; Putting respectively in same and contain on the silica gel g thin-layer plate that sodium carboxymethyl cellulose is a bonding agent, is developping agent with benzene-ethyl acetate (8:2), launches; Take out; Dry, after smoked 3 minutes, put under the ultraviolet light (365nm) and inspect with liquor ammoniae fortis.Compare need testing solution and the color developing effect of reference substance solution on thin layer plate, the result sees the following form 8:

The selection of table 8 test sample point sample amount

The point sample amount	2μl	5μl	8μl	10μl
					Color developing effect	Spot does not develop the color	The spot colour developing is very shallow	The spot colour developing is shallow	The spot colour developing is clear

Can find out that from last table reference substance solution point sample amount is 10 μ l, need testing solution and reference substance solution thin layer plate relevant position, the spot colour developing is clear, the Pass Test requirement, then the point of sample diameter is excessive to continue to strengthen the point sample amount, has influenced the quality of point sample.

3, the qualitative limit examine of aconitine

The inventor studies the content of medicine mesaconitine of the present invention before the qualitative limit examine of confirming aconitine.The result is following:

Assay is measured according to high performance liquid chromatography (appendix VID).

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; Methyl alcohol-0.5% triethylamine (4:1) is a moving phase; The detection wavelength is 235nm; Flow velocity: 0.5ml/min; Column temperature: room temperature; Column type: Agilent, C18,150mm.

It is an amount of that the preparation precision of reference substance solution takes by weighing the aconitine reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.25mg, promptly gets.

The preparation of need testing solution is got according to the pharmaceutical preparation of the present invention of embodiment 1 preparation an amount of, and the desaccharification clothing is ground into fine powder, and precision takes by weighing powder 2g; Add the 25ml that adds methylene chloride behind the wetting 5min of ammoniacal liquor 2ml, 30min is extracted in jolting, and inclining extract, divides several to wash the dregs of a decoction and container with the 25ml methylene chloride again; Combined dichloromethane, water-bath (80 ℃) evaporate to dryness, residue is used dissolve with methanol; And being settled to 10ml, micro porous filtration promptly gets.

The result surveys medicine mesaconitine content of the present invention and is lower than ten thousand/, and inferior separating effect, so detection method does not comprise the content detection of aconitine.

Though the detection by quantitative to aconitine is not listed in the method for quality control; But because aconitine belongs to toxic component; So to carrying out the detection of aconitine in the quality control of medicine of the present invention, then aconitine has been carried out qualitative limit examine test, the result is following:

The preparation of need testing solution is got according to the pharmaceutical preparation of the present invention of embodiment 1 preparation an amount of, and the desaccharification clothing is ground into fine powder, and precision takes by weighing 5g; Add ammonia solution 5ml, placed 2 hours, 100ml adds diethyl ether; Jolting 1 hour was placed 24 hours, filtered; The filtrating evaporate to dryness, residue is with absolute ethyl alcohol 2ml dissolving, as need testing solution.

The preparation of reference substance solution is according to the regulation of in 2005 editions pharmacopeia the aconiti preparata,radix mesaconitine being limited the quantity of, and it is an amount of to get the aconitine reference substance, adds absolute ethyl alcohol and processes the reference substance solution that every 1ml contains 2mg.

According to thin-layered chromatography test, draw each 2 μ l point of above-mentioned two kinds of solution on same silica G plate, be developping agent with the different proportionings of cyclohexane-ethyl acetate-diethylamine, launch, airing, spray is with rare bismuth potassium iodide test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot colors that occurs should be shallower than the reference substance spot or spot not occur.The result sees the following form 9:

The selection of table 9 developping agent proportioning

Can find out from last table, be developping agent with cyclohexane-ethyl acetate-diethylamine 7:2:0.5, launch effective, and good stability, Pass Test requirement.

4, ferulaic acid content is measured in the Ligusticum wallichii

Method one:

Detecting instrument: Tianjin, the island SPD-10ATvp of company type high performance liquid chromatograph

Chromatographic column: Di Ma company (Zorbax C18 4.6 * 150 mm, 5 μ m)

Moving phase: methyl alcohol-2% glacial acetic acid (1:4) flow velocity: 0.800ml/min

Detect wavelength: 323nm column temperature: room temperature

The forulic acid reference substance is purchased lot number: the 0773-9910 in Nat'l Pharmaceutical & Biological Products Control Institute

It is an amount of that the preparation precision of reference substance solution takes by weighing the forulic acid reference substance, adds moving phase and process the solution that every 1ml contains 25 μ g, as reference substance solution.

The preparation of need testing solution is got according to the pharmaceutical preparation of the present invention of embodiment 1 preparation an amount of, and the desaccharification clothing is pulverized, and gets the about 1g of powder; The accurate title, decide, and puts in the tool plug conical flask, the accurate moving phase 25ml that adds, and precision is weighed; Sonicated 30 minutes cools, and weighs, and supplies the weight that subtracts mistake with moving phase; Shake up, filtering with microporous membrane promptly gets

Negative control solution is according to the blank sample of the scarce Ligusticum wallichii of method preparation of embodiment 1, and the preparation method by need testing solution prepares negative controls again.

With miillpore filter (0.45 μ m), accurate respectively each 5～10 μ l of negative controls, reference substance solution and need testing solution that draw inject liquid chromatograph, measure.

The result: forulic acid reference substance appearance time is 6.3min, and the peak is also arranged in test sample chromatogram and negative control chromatogram here, and changing moving phase subsequently is methyl alcohol-2% glacial acetic acid (32:68), feminine gender still have disturb and content very low.

Method two:

Instrument island Tianjin LC one 10A high performance liquid chromatograph, detecting device: SPD-IOA UV-detector.Workstation: Yi Lite EC2003 chromatographic work station.

(Nat'l Pharmaceutical & Biological Products Control Institute provides reagent forulic acid reference substance, lot number: 0773-9910); Medicine of the present invention (according to embodiment 1 self-control); Methyl alcohol, second eyeball are chromatographically pure, and water is redistilled water, and all the other reagent are pure for analyzing.

The chromatographic condition chromatographic column: Hypesil C18 (250 mm x4.6 mm, 5 μ m) is with preposition guard column; Acetonitrile-water-glacial acetic acid (20:80:1) is a moving phase; Flow velocity is 1.0 ml/min; Detecting wavelength is 322 nm.Number of theoretical plate is pressed the forulic acid peak and is calculated, and should be not less than 1500.

The preparation precision of reference substance solution takes by weighing forulic acid reference substance 3mg, puts in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up, and promptly gets (containing forulic acid 30 μ g among every 1ml).

It is an amount of that these article are got in the preparation of need testing solution, and porphyrize is got 2.5 g, and accurate the title decides, and adds methyl alcohol 50ml, reflux 1h; Put cold. filter, residue is washed 2 times with the methyl alcohol of 5ml, filters, and merging filtrate, evaporate to dryness, residue add water-ammonia water (20:2) 20ml gradation dissolving; Put in the separating funnel, wash 2 times with the ether of 20ml, discard ether solution, water liquid is transferred pH to 2 with watery hydrochloric acid, with extracted with diethyl ether 4 times, and 20ml at every turn; Merge ether solution,, discard water liquid with the washing of 30ml, the ether solution evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10 ml measuring bottles; Be diluted to scale with methyl alcohol, shake up, filter, get subsequent filtrate, promptly get.

The preparation of negative control solution is removed Ligusticum wallichii by embodiment 1 recipe quantity, makes the negative control preparation, and the preparation method makes negative control solution by need testing solution.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and promptly get.

The result: the chromatographic peak of forulic acid separates good, does not have the interference of other compositions in the preparation, and retention time is 11min.Investigate through methodology, detection method linearity of the present invention, precision, stability, reappearance and the recovery are all good, can accurately measure content of ferulic acid in the medicine.

According to the investigation result of above two kinds of methods, confirm with the method two to be content of ferulic acid detection method in the medicine of the present invention.

5, the assay of Imperatorin in the root of Dahurain angelica

Chromatographic column: Di Ma company (Zorbax C18 4.6 * 150 mm, 5 μ m)

Moving phase: methanol-water (55:45) flow velocity: 1.000ml/min

Detect wavelength: 300nm column temperature: room temperature

The Imperatorin reference substance is purchased lot number: the 11826-200410 in Nat'l Pharmaceutical & Biological Products Control Institute

It is an amount of that the preparation precision of reference substance solution takes by weighing the Imperatorin reference substance, adds moving phase and process the solution that every 1ml contains 10 μ g, as reference substance solution.

The preparation of need testing solution is got according to the pharmaceutical preparation of the present invention of embodiment 1 preparation an amount of, and the desaccharification clothing is pulverized, and gets the about 1g of powder; The accurate title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 25ml that adds, and precision is weighed; Sonicated 30 minutes cools, and weighs, and supplies the weight that subtracts mistake with methyl alcohol; Shake up, filtering with microporous membrane promptly gets.

Negative control solution is according to the blank sample of the scarce root of Dahurain angelica of method preparation of embodiment 1, and the preparation method by need testing solution prepares negative controls again.

The result: Imperatorin reference substance appearance time is 8.5min; In test sample chromatogram and negative control chromatogram, there is not the peak to occur here; Get part angelica root powder subsequently and prepare sample according to the preparation method of patent medicine; Get single angelica root need testing solution by test sample preparation method place again, the result fails to detect Imperatorin equally, and explaining in 30% ethanol extract of the root of Dahurain angelica does not have Imperatorin.

According to above experimental result, therefore the assay of Imperatorin is not listed in the method for quality control of medicine of the present invention.

6, the detection by quantitative of ethanol soluble extractives:

Through research to the detection method of content of medicine mesaconitine of the present invention, forulic acid, Imperatorin; But because above three kinds of component contents are low excessively, so fail in the method for quality control of medicine of the present invention, to set up the detection method of content of aconitine, forulic acid, Imperatorin.The inventor studies extract content again, drafts as follows: use methyl alcohol to be solvent, measure according to the hot dipping under the ethanol soluble extractives determination method item.Three batches of extract testing results are seen table 10:

Table 10: extract testing result

Lot number	The extract result
		1701	27.5％
1802	26.8％
		1903	28.4％

[0170]Can find out from last table, use this method can stably control total active constituent content in the medicine, guarantee curative effect of medication.

7, the mensuration of nitrogen content:

Goat's horn contains a large amount of keratin in the medicine of the present invention; It after hydrolysis several amino acids class material; Therefore can more effectively control the quality of medicine of the present invention to the mensuration of nitrogen content, guarantee security, validity that medicine uses, improve the science of drug standard.

Get according to 3 batches of the medicinal tablets of the present invention of embodiment 1 preparation, remove sugar-coat, be ground into fine powder, precision takes by weighing 0.4g, adopts appendix IXL first method of Chinese Pharmacopoeia version in 2005 to measure, and the result is following:

Table 11: nitrogen content testing result

Lot number	Nitrogen content
		1701	5.9％
1802	6.1％
		1903	6.2％

Can find out from last table, use nitrogen content stability in this method control medicine, reappearance all better.

The research of Test Example 3 pharmacodynamics tests

1, test material

1.1 medicine medicine I of the present invention, II are respectively the tablet according to embodiment 1,2 preparations;

The positive control medicine is the compound cavel sheet, and precious pharmaceutical factory produces by the river, Heilungkiang;

The above medicine time spent grinds and is made into 20% suspension oral gavage.

1.2 animal Kunming mouse and Wistar kind rat are provided by experimental animal chamber, the court.

2, method and result

2.1 analgesic activity

2.1.1 hot plate method: choose 40 of 20 ± 2g female mices, be divided into 4 groups at random, the administration group is irritated stomach and is given and medicine I of the present invention, II, III 5.3g/kg (contained crude drug amount), positive control medicine (compound cavel sheet) 3.9g/kg (contained crude drug amount); Negative control group is given same water by volume, every day 1 time, continuous 7 days; 1h after the last administration places animal on 55 ± 1 ℃ of hot plates, and the record animal is from putting into to the time that begins to lick metapedes (being pain threshold); Every separated 1h measures 1 time, surveys altogether 3 times.The result sees table 12.

The influence of table 12 pair thermostimulation pain threshold (X ± SD)

Compare ##P＜0.01, # P＜0.05 with negative control group; Compare * P＜0.05. with the positive drug control group

Can find out that from above result medicine of the present invention and positive control medicine all can significantly improve the pain threshold of mouse to thermostimulation, wherein the effect of medicine I of the present invention significantly is better than the positive control medicine.

2.1.2 acetic acid twisting method: Kunming mouse male and female half and half, divide into groups and the same 2.1.1 of administration, 30min after the last administration, mouse peritoneal inject 1% acetum 0.1ml/10g, and the body number of times result that turns round of animal sees table 13. in the record 10min

The influence of table 13 Dichlorodiphenyl Acetate writhing response (X ± SD)

Group	Dosage (g/kg*d)	Turn round the body number of times
			Negative control group (10)	0*7	39.8±4.8
Positive control medicine group (10)	3.9*7	33.6±6.9#
			Medicine I of the present invention (10)	5.3*7	27.8±7.3##*
Medicine II of the present invention (10)	5.3*7	29.7±6.8##

Can find out that from above result medicine of the present invention and positive control medicine all can significantly reduce the number of times of mouse acetic acid twisting reaction, the effect of medicine I of the present invention significantly is better than the positive control medicine.

2.2 influence to external thrombus formation and blood viscosity:

2.2.1 to the thrombotic influence of rats in vitro: 40 of adult rats, male and female half and half are divided into groups and the same 2.1.1 of administration, and the 30min extracting vein blood is surveyed thrombus length, weight in wet base and dry weight after the last administration.The result sees table 14:

The thrombotic influence of table 14 pair rats in vitro (X ± SD)

Medicine of the present invention and positive control medicine all can significantly suppress the rats in vitro thrombosis, and thrombus length, weight in wet base, dry weight all have obvious reduction, and the inhibiting effect of medicine of the present invention is superior to the positive control medicine, especially thrombus length are had remarkable inhibiting effect.

2.2.2 the influence to the mouse WBV: the same 2.1.1 of mice group and administration, last administration 30min extracts eyeball and gets blood and place the bottle of heparinize to shake up, and surveys its WBV (ratio) then.The result sees table 15.

The influence of table 15 pair mouse WBV (X ± SD)

Medicine of the present invention and positive control medicine all can reduce the mouse WBV, and the effect of medicine of the present invention is superior to the positive control medicine.

Following examples are used to further specify the present invention.

Embodiment 1

Goat's horn 1400g Ligusticum wallichii 250g root of Dahurain angelica 450g aconiti preparata,radix 150g yncaria stem with hooks 200g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 100g dextrin mixing, granulate drying; Be pressed into 1000, sugar coating promptly gets.

Embodiment 2

Goat's horn 1400g Ligusticum wallichii 250g root of Dahurain angelica 450g aconiti preparata,radix 150g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 116g dextrin mixing, granulate drying; Be pressed into 1000, sugar coating promptly gets.

Embodiment 3

Goat's horn 1300g Ligusticum wallichii 400g root of Dahurain angelica 300g aconiti preparata,radix 300g yncaria stem with hooks 100g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.32 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.32 thick paste (50 ℃); With goat's horn cream and 105g dextrin mixing, granulate drying; Be pressed into 1000, sugar coating promptly gets.

Embodiment 4

Goat's horn 1500g Ligusticum wallichii 200g root of Dahurain angelica 500g aconiti preparata,radix 100g yncaria stem with hooks 300g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 135g dextrin mixing, granulate drying; Be pressed into 1000, sugar coating promptly gets.

Embodiment 5

Goat's horn 1350g Ligusticum wallichii 200g root of Dahurain angelica 400g aconiti preparata,radix 100g yncaria stem with hooks 150g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 118g dextrin mixing, granulate drying; Be pressed into 1000, the bag film-coating promptly gets.

Embodiment 6

Goat's horn 1450g Ligusticum wallichii 300g root of Dahurain angelica 500g aconiti preparata,radix 200g yncaria stem with hooks 250g

Goat's horn pound sheet, the boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 100g dextrin mixing, granulate drying; Be pressed into 1000, the bag film-coating promptly gets.

Embodiment 7 capsules

Mother-of-pearl 1400g Ligusticum wallichii 250g root of Dahurain angelica 450g aconiti preparata,radix 150g yncaria stem with hooks 200g

Mother-of-pearl boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 100g dextrin mixing, granulate drying; Whole grain, 1000 capsules of packing into promptly get.

Embodiment 8 granules

Bright rhizoma Gastrodiae 1400g Ligusticum wallichii 250g notopterygium root 450g aconiti preparata,radix 150g yncaria stem with hooks 200g

Bright rhizoma Gastrodiae boiling secondary adds 8 times of water gagings for the first time and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With goat's horn cream and 200g dextrin, 400g sucrose mixing, granulate drying; Whole grain is processed particle 800g, promptly gets.

Embodiment 9 oral liquids

The windproof 450g aconiti preparata,radix of oyster 1400g Ligusticum wallichii 250g 150g yncaria stem with hooks 200g

Oyster boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.15 ~ 1.20 clear cream (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.15 ~ 1.20 clear cream (50 ℃); With the clear cream mixing of goat's horn, other gets sucrose 650g and adds water boil, after the dissolving, filters; Concentrate and process syrup,, boil, put cold with above-mentioned concentrated clear cream mixing; Add antiseptic and essence, and be diluted to 1000ml, promptly get with cold boiling water.

Embodiment 10 dripping pills

Raddle 1400g Ligusticum wallichii 250g achene of Siberian cocklebur 450g aconiti preparata,radix 150g yncaria stem with hooks 200g

Raddle boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With the clear cream mixing of goat's horn, vacuum drying, dried cream powder is broken to 150 orders; Even with the Macrogol 4000 of fusion according to the mixed of 3:7, process dripping pill, promptly get.

Embodiment 11 soft capsules

Speech puncture vine 1400g Ligusticum wallichii 250g root of Dahurain angelica 450g aconiti preparata,radix 150g yncaria stem with hooks 200g

Tribulus terrestris boiling secondary adds for the first time 8 times of water gagings and decocted 3 hours, adds 6 times of water gagings for the second time and decocts 3 hours, and gradation filters, and merging filtrate is condensed into 1.35 thick paste (50 ℃); All the other Ligusticum wallichiis etc. are crushed to 24 orders, according to the percolation under liquid extract and the extract item, are solvent with 30% ethanol, carry out diacolation with the speed of 150ml/min*kg; Till the colourless or inanimate object alkali reaction, collect the liquid of filtering to the liquid of filtering, reclaim ethanol, be condensed into 1.35 thick paste (50 ℃); With the clear cream mixing of goat's horn, vacuum drying, dried cream powder is broken to 150 orders; Even with soybean oil according to the mixed of 1:2, be pressed into soft capsule, promptly get.

The method of quality control of the medicinal tablet of the present invention of embodiment 12 embodiment 1-6 preparation

Differentiate:

(1) get 20 of these article, porphyrize, the 30ml sonicated that adds diethyl ether 30 minutes filters, the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution.Other gets Ligusticum wallichii control medicinal material 1g, adds ethanol 30ml sonicated 30 minutes, filter, and the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as control medicinal material solution.According to thin-layered chromatography (" Chinese pharmacopoeia version appendix in 2005 VI B) test; Drawing need testing solution 8 μ l, control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7:3); Launch; Take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get 6 of these article, porphyrize adds absolute ethyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol 2ml and disperses, and gets supernatant as need testing solution.Other gets root of Dahurain angelica control medicinal material 1g, shines medicinal material solution in pairs with the method preparation.According to thin-layered chromatography (" Chinese pharmacopoeia version appendix in 2005 VI B) test; Drawing each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with methenyl choloride-methyl alcohol-formic acid (9:1:0.1); Launch; Take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

The aconitine limit inspection:

These article of getting desaccharification clothing is ground into fine powder, and precision takes by weighing 5g, adds ammonia solution 5ml; Placed 2 hours, the 100ml that adds diethyl ether, jolting 1 hour was placed 24 hours; Filter, the filtrating evaporate to dryness, residue dissolves with absolute ethyl alcohol 2ml; As need testing solution, it is an amount of that other gets the aconitine reference substance, adds absolute ethyl alcohol and process the reference substance solution that every 1ml contains 2mg.According to thin-layered chromatography test (" Chinese pharmacopoeia version appendix in 2005 VIB); Drawing each 2 μ l point of above-mentioned two kinds of solution on same silica G plate, is developping agent with cyclohexane-ethyl acetate-diethylamine (7:2:0.5), launches; Airing, spray is with rare bismuth potassium iodide test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should or spot appear less than the reference substance spot.

Extract content is measured:

Measure according to the hot dipping under the ethanol soluble extractives determination method item, make solvent, must not be less than 18.0% with methyl alcohol.

Nitrogen analysis:

These article of getting are an amount of, remove sugar-coat, are ground into fine powder, and precision takes by weighing 0.4g, measure (an appendix IX of Chinese Pharmacopoeia version in 2005 L first method) according to n2 method, and these article nitrogen content must not be less than 4.0%.

Ferulaic acid content is measured

Measure according to high performance liquid chromatography (" Chinese pharmacopoeia version appendix in 2005 VI D).

Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; Acetonitrile-water-glacial acetic acid (20:80:1) is a moving phase; The detection wavelength is 322nm.Number of theoretical plate calculates by the forulic acid peak should be not less than 1500.

It is an amount of that these article are got in the preparation of need testing solution, removes sugar-coat, and porphyrize is got 2.5 g, and accurate the title decides, and adds methyl alcohol 50ml; Reflux 1h is put cold. and filter, residue is washed 2 times with the methyl alcohol of 5ml, filters merging filtrate, evaporate to dryness; Residue adds water-ammonia water (20:2) 20ml gradation dissolving, puts in the separating funnel, washes 2 times with the ether of 20ml, discards ether solution, and water liquid is with watery hydrochloric acid accent pH to 2, with extracted with diethyl ether 4 times; Each 20ml merges ether solution, with the washing of 30ml, discards water liquid, the ether solution evaporate to dryness, and residue adds dissolve with methanol; Be transferred in the 10 ml measuring bottles, be diluted to scale, shake up, filter, get subsequent filtrate, promptly get with methyl alcohol.

Every of these article contain Ligusticum wallichii with forulic acid (C ₁₀H ₁₀O ₄) meter, must not be less than 0.08mg.

Embodiment 13

[proterties] these article are sugar coated tablet, remove to show sepia behind the sugar-coat; It is little puckery to distinguish the flavor of.

20 of these article are got in [discriminating] (1), remove sugar-coat, porphyrize, and the 30ml sonicated that adds diethyl ether 30 minutes filters, the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution.Other gets Ligusticum wallichii control medicinal material 1g, adds ethanol 30ml sonicated 30 minutes, filter, and the filtrating evaporate to dryness, residue adds ethanol 1ml makes dissolving, as control medicinal material solution.According to thin-layered chromatography (" Chinese pharmacopoeia version appendix in 2005 VI B) test; Drawing need testing solution 8 μ l, control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7:3); Launch; Take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get 6 of these article, remove sugar-coat, porphyrize adds absolute ethyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol 2ml and disperses, and gets supernatant as need testing solution.Other gets root of Dahurain angelica control medicinal material 1g, shines medicinal material solution in pairs with the method preparation.According to thin-layered chromatography (" Chinese pharmacopoeia version appendix in 2005 VI B) test; Drawing each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with methenyl choloride-methyl alcohol-formic acid (9:1:0.1); Launch; Take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

[inspection] should meet each item relevant under tablet item regulation (" Chinese pharmacopoeia version appendix in 2005 I D page or leaf).

These article desaccharification clothing is got in the aconitine limit inspection, is ground into fine powder, and precision takes by weighing 5g, adds ammonia solution 5ml; Placed 2 hours, the 100ml that adds diethyl ether, jolting 1 hour was placed 24 hours; Filter, the filtrating evaporate to dryness, residue dissolves with absolute ethyl alcohol 2ml; As need testing solution, it is an amount of that other gets the aconitine reference substance, adds absolute ethyl alcohol and process the reference substance solution that every 1ml contains 2mg.According to thin-layered chromatography test (" Chinese pharmacopoeia version appendix in 2005 VIB); Drawing each 2 μ l point of above-mentioned two kinds of solution on same silica G plate, is developping agent with cyclohexane-ethyl acetate-diethylamine (7:2:0.5), launches; Airing, spray is with rare bismuth potassium iodide test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should or spot appear less than the reference substance spot.

[extract] measured according to the hot dipping (Chinese Pharmacopoeia version appendix in 2005 X A) under the ethanol soluble extractives determination method item, makes solvent with methyl alcohol, must not be less than 18.0%.

[assay] photograph high performance liquid chromatography (" Chinese pharmacopoeia version appendix in 2005 VI D) measure.

It is an amount of that these article are got in the preparation of need testing solution, removes sugar-coat, and porphyrize is got 2.5g, and accurate the title decides, and adds methyl alcohol 50ml; Reflux 1h is put cold. and filter, residue is washed 2 times with the methyl alcohol of 5ml, filters merging filtrate, evaporate to dryness; Residue adds water-ammonia water (20:2) 20ml gradation dissolving, puts in the separating funnel, washes 2 times with the ether of 20ml, discards ether solution, and water liquid is with watery hydrochloric acid accent pH to 2, with extracted with diethyl ether 4 times; Each 20ml merges ether solution, with the washing of 30ml, discards water liquid, the ether solution evaporate to dryness, and residue adds dissolve with methanol; Be transferred in the 10 ml measuring bottles, be diluted to scale, shake up, filter, get subsequent filtrate, promptly get with methyl alcohol.

[function with cure mainly] flat liver, analgesia.Be used for antimigraine, vascular headache, tension headache and nervous headache.

Claims

1. quality determining method of treating the Chinese medicine composition of headache is characterized in that this method comprises one or more in the following qualitative checking method:

(1) qualitative detection of Ligusticum wallichii

(2) qualitative detection of the root of Dahurain angelica

According to the thin-layered chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-methyl alcohol-formic acid, launch, take out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(3) limit examine of aconitine

Wherein, said traditional Chinese medicinal composition raw materials consists of:

2. quality determining method as claimed in claim 1 is characterized in that said traditional Chinese medicinal composition raw materials consists of:

3. quality determining method as claimed in claim 1 is characterized in that said traditional Chinese medicinal composition raw materials consists of:

4. quality determining method as claimed in claim 1 is characterized in that said traditional Chinese medicinal composition raw materials consists of:

5. quality determining method as claimed in claim 1 is characterized in that said traditional Chinese medicinal composition raw materials consists of:

6. like each described detection method of claim 1-5, it is characterized in that in the said traditional Chinese medicinal composition raw materials:

7. like each described detection method of claim 1-5, it is characterized in that the preparation method of said Chinese medicine composition tablet is:

8. like the arbitrary described quality determining method of claim 1-5, it is characterized in that this method comprises one or more in the following qualitative checking method:

(1) qualitative detection of Ligusticum wallichii

(2) qualitative detection of the root of Dahurain angelica

Pharmaceutical composition content of the present invention is equivalent to crude drug 15g, and porphyrize adds absolute ethyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue adds absolute ethyl alcohol 2ml and disperses, and gets supernatant as need testing solution; Other gets root of Dahurain angelica control medicinal material 1g, shines medicinal material solution in pairs with the method preparation; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Methenyl choloride-methyl alcohol-formic acid with the 9:1:0.1 ratio is developping agent, launches, and takes out; Dry, put under the ultraviolet lamp that wavelength is 365nm and inspect, in the test sample chromatogram; With the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

(3) the qualitative limit examine of aconitine

Pharmaceutical composition content of the present invention is ground into fine powder, and precision takes by weighing 5g, adds ammonia solution 5ml, placed 2 hours, and the 100ml that adds diethyl ether, jolting 1 hour was placed 24 hours, filters, the filtrating evaporate to dryness, residue is with absolute ethyl alcohol 2ml dissolving, as need testing solution; It is an amount of that other gets the aconitine reference substance, adds absolute ethyl alcohol and process the reference substance solution that every 1ml contains 2mg; According to thin-layered chromatography test, draw each 2 μ l point of above-mentioned two kinds of solution on same silica gel g thin-layer plate, be developping agent with the cyclohexane-ethyl acetate-diethylamine of 7:2:0.5 ratio, launch, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot colors that occurs should be shallower than the reference substance spot or spot not occur.