CN101396544A

CN101396544A - Traditional Chinese medicine composition capable of ventilating the lung and relieving asthma and preparation and quality control method thereof

Info

Publication number: CN101396544A
Application number: CNA2007101224885A
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: 2007-09-26
Filing date: 2007-09-26
Publication date: 2009-04-01
Anticipated expiration: 2027-09-26
Also published as: CN101396544B

Abstract

The invention discloses a pharmaceutical composition for ventilating lung and relieving asthma, a preparation method and a quality control method. The pharmaceutical composition comprises the following pharmaceutical raw materials: ephedra herb, raw ginger, cassia twig, bitter apricot seed, Chinese magnoliavine fruit (prepared) and prepared liquoric root, the preparation method is as follows: the five herbal medicines except the bitter apricot seed are decocted by adding water through a countercurrent circulatory extraction process and concentrated to obtain a clear paste after reducing pressure, ethanol is added, then the mixture is stirred evenly and placed staticly, supernatant liquid is taken, the ethanol is recovered, and the concentration is carried out to obtain a thick paste; the bitter apricot seed is arranged in a hot pressure sterilization pot for hot pressure steaming and boiling and then is arranged in a small amount of cold water for soaking instantly after being taken out, an apricot processed product can be obtained by removing seed coats and being dried; the quality control method is as follows: the thin layer identification is carried out on the ephedra herb, the raw ginger and the liquoric root, the content measurement is carried out on ephedrine in the ephedra herb and schizandrin in the Chinese magnoliavine fruit. The pharmaceutical composition has good effects on ventilating lung and relieving asthma.

Description

Chinese medicine composition of depressed lung-energy dispersing and preparation method thereof and method of quality control

Technical field

The present invention relates to a kind of lung qi dispersing that is used for, Chinese medicine composition of relievining asthma and preparation method thereof and method of quality control relate in particular to a kind of lung qi dispersing that is used for, and capsule of relievining asthma and preparation method thereof and method of quality control belong to technical field of Chinese medicines.

Background technology

Chronic obstructive pulmonary disease is a kind of commonly encountered diseases of internal department, frequently-occurring disease, is a kind of important chronic respiratory system diseases, and ill patient's number is many, dead rate height, because its state of an illness delay has a strong impact on patient's work capacity and quality of life, and is subjected to the generally attention of countries in the world.

At present, medical circle does not still have the development that way is controlled the chronic obstructive pulmonary disease state of an illness effectively, seeks to prevent that the effective ways of chronic obstructive pulmonary disease from being domestic and international medical circle problem anxious to be solved.The chronic obstructive pulmonary disease pathogenesis is not clear fully as yet, generally believe that at present chronic obstructive pulmonary disease is is feature with air flue, pulmonary parenchyma and pulmonary vascular chronic inflammatory disease, increase at different parts alveolation macrophage, T lymphocyte and the neutrophilic granulocyte of lung.Activated inflammatory cell discharges multiple medium, destroy lung structure and (or) promote the neutrophilic granulocyte inflammatory reaction.Suck deleterious particle or gas and can cause pneumonia; Smoking can be induced inflammation and directly be damaged lungs.Except that inflammation, the protease of pulmonary and protease inhibitor system are unbalance, and oxidation antioxidation unbalance also play a part important.Modern medicine is by Drug therapy for chronic obstructive pulmonary disease at present, and prevention and treatment complication are improved symptom, mainly are dyspnea, improve the quality of living, and remove the inducement of chronic obstructive pulmonary disease acute exacerbation, enjoin patient's smoking cessation.Because this patient's immunity of organisms reduces and to breathe defense mechanism impaired, occurs respiratory tract infection easily, repeated infection causes the bronchial mucosa congestion and edema, the glandular hyperplasia hypertrophy, and secretory function is hyperfunction, and tube wall thickens, narrow, increases the weight of airflow obstruction.The pathogen that infects is common to be antibacterial, virus, mycoplasma etc.Western modern medicine mainly adopts antibacterials, hormone at bacterium, and does not still have active drug for infecting due to the virus, in addition antibacterials, hormone have certain toxic and side effects again, in addition, even acute stage is eased through treatment, still can not effectively control its recurrence.Therefore, for better relieve chronic obstructive pulmonary disease development, control its recurrence, improve life quality, make chronic obstructive pulmonary disease treatment standardization more, the prescription medicine of seeking safe and effective treatment Patients with Chronic Obstructive Pulmonary Disease is very necessary.

Summary of the invention

The object of the invention is to provide a kind of lung qi dispersing that has, the Chinese medicine composition of antiasthmatic effect;

The object of the invention also is to provide a kind of lung qi dispersing that has, the Chinese medicine composition preparation method of antiasthmatic effect;

The object of the invention also is to provide a kind of lung qi dispersing that has, the method for quality control of the Chinese medicine composition of antiasthmatic effect.

The present invention seeks to be achieved through the following technical solutions:

Of the present invention have a lung qi dispersing, and the Chinese medicine composition of antiasthmatic effect is to be made by the crude drug of following weight ratio:

Herba Ephedrae 3000-4000 weight portion, Rhizoma Zingiberis Recens 2000-3000 weight portion, Ramulus Cinnamomi 2000-3000 weight portion, Semen Armeniacae Amarum 1000-1500 weight portion, Fructus Schisandrae Chinensis (system) 250-500 weight portion, Radix Glycyrrhizae Preparata 80-160 weight portion

The above-mentioned raw materials optimum ratio is:

Herba Ephedrae 3000-3200 weight portion, Rhizoma Zingiberis Recens 2800-3000 weight portion, Ramulus Cinnamomi 2800-3000 weight portion, Semen Armeniacae Amarum 1400-1500 weight portion, Fructus Schisandrae Chinensis (system) 300-400 weight portion, Radix Glycyrrhizae Preparata 100-120 weight portion

The above-mentioned raw materials optimum ratio is:

The preparation method of Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Ramulus Cinnamomi 3000g, Semen Armeniacae Amarum 1500g, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g Chinese medicine composition of the present invention may further comprise the steps:

Above medical material is except that Semen Armeniacae Amarum, and other five tastes medical materials adopt the circulated in countercurrent extraction process to decoct with water 1-3 time, each 2-4 hour, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 12-36 hour, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃); Semen Armeniacae Amarum rinsing dedust soil, draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.01～0.03kpa/cm ²(103 ℃), hot pressing steaming and decocting 20-40 minute, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries, and promptly gets the Semen Armeniacae Amarum processed product.With above-mentioned thick paste and Semen Armeniacae Amarum processed product vacuum drying, pulverize, make the preparation of clinical acceptance according to this area routine techniques, as: capsule, tablet, granule, soft capsule, pill.

The preparation method of Chinese medicinal composition capsules agent of the present invention is: get crude drug Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Ramulus Cinnamomi 3000g, Semen Armeniacae Amarum 1500g, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g

It is characterized in that this method may further comprise the steps:

Above medical material is except that Semen Armeniacae Amarum, and other five tastes medical materials adopt the circulated in countercurrent extraction process to decoct with water 2 times, each 2 hours, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃); Semen Armeniacae Amarum rinsing dedust soil, draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.01～0.03kpa/cm ²(103 ℃), hot pressing steaming and decocting 30 minutes, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries, and promptly gets the Semen Armeniacae Amarum processed product.With above-mentioned thick paste and Semen Armeniacae Amarum processed product vacuum drying, pulverize, it is an amount of to add dextrin, and mixing incapsulates, and makes 1000, promptly.

Pharmaceutical composition method of quality control of the present invention comprises one or more in following discrimination method and/or the assay:

(1) get medicine of the present invention and be equivalent to crude drug 9-11g, add strong ammonia solution 1-3ml, add chloroform 30-50ml again, reflux 0.5-2 hour, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets ephedrine hydrochloride, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (15-25: 3-10: 0.5-2) be developing solvent, launch, take out, dry, spray is with 5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical aubergine speckle.

(2) get medicine of the present invention and be equivalent to crude drug 200-220g, the 20-40ml that adds diethyl ether, ultrasonic 20-40 minute (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80-100ml and decocts 1 hour, filters, and filtrate is concentrated into 20-30ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw control medicinal material solution 5 μ l, need testing solution 15 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (3-7: 0.5-2) be developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(3) get medicine of the present invention and be equivalent to crude drug 200-220g, add petroleum ether (60～90 ℃) 40-60ml, reflux 0.5-2 hour, filter, residue adds methanol 40-60ml, reflux 0.5-2 hour, filter, filtrate evaporate to dryness, residue add water 30-50ml makes dissolving, extracts 2-4 time with water saturated n-butyl alcohol jolting, each 10-30ml, merge n-butyl alcohol liquid, use the saturated water washing of n-butyl alcohol 2-4 time, each 20-40ml, get n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, (10-15: 5-10: lower floor's solution 1-3) is developing solvent with chloroform-methanol-water, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

[assay]

(1) measures according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With 0.1% phosphoric acid (containing 0.1% triethylamine)-acetonitrile (90-100:0-10) is mobile phase; The detection wavelength is 210nm.Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 2500.

It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, accurate claims surely, makes the solution that every 1ml contains 50 μ g with dissolve with methanol, product solution in contrast, promptly.

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 3-5g, and accurate the title decides, to the 50ml volumetric flask, and the accurate hydrochloric acid solution 1ml that adds 0.5mol/L, add methanol again to scale, jam-pack, supersound process 20-40 minute, put coldly, microporous filter membrane (0.45 μ m) filters, promptly.

Accurate respectively reference substance solution and each the 5 μ l～10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The daily metering of medicine of the present invention contains Herba Ephedrae with ephedrine hydrochloride (C ₁₀H ₁₅NOHCl) the amount meter must not be less than 8.0mg.(2) measure according to high performance liquid chromatography (appendix VID).

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-methanol-0.1% phosphoric acid (35-40:15-20:40-50) is mobile phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schisandrin peak should be not less than 3000.

Schisandrin reference substance 1.5mg is got in the preparation of reference substance solution, accurate claims surely, puts in the 50ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up, and promptly gets (every 1ml contains schisandrin 0.03mg).

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 100-110g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 50ml that adds, claim to decide weight, supersound process (power 250W, frequency 20kHz) 10-30min, take out, put coldly, claim to decide weight again, add methanol and supply the weight of minimizing, filter, the accurate subsequent filtrate 20ml that draws, evaporate to dryness, residue add methanol makes dissolving, and fixed molten to 5ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each the 10 μ l of test solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

Medicine of the present invention contains schisandrin (C ₂₄H ₃₂O ₇) must not be less than 0.40%.

Pharmaceutical composition method of quality control of the present invention is preferably as follows one or more in discrimination method and/or the assay:

(1) get medicine of the present invention and be equivalent to crude drug 10.8g, add strong ammonia solution 2ml, add chloroform 40ml again, reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets ephedrine hydrochloride, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (20: 5: 0.5) is developing solvent, launch, take out, dry, spray is with 5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical aubergine speckle.

(2) get medicine of the present invention and be equivalent to crude drug 216.25g, the 30ml that adds diethyl ether, ultrasonic 30 minutes (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80ml and decocts 1 hour, filters, and filtrate is concentrated into 20ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw control medicinal material solution 5 μ l, need testing solution 15 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (5: 1) is developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(3) get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour, filter, residue adds methanol 50ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, extract 3 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol 3 times, each 30ml gets n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution with chloroform-methanol-water (13: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

[assay]

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With 0.1% phosphoric acid (containing 0.1% triethylamine)-acetonitrile (97:3) is mobile phase; The detection wavelength is 210nm.Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 2500.

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 4.3g, and accurate the title decides, to the 50ml volumetric flask, and the accurate hydrochloric acid solution 1ml that adds 0.5mol/L, add methanol again to scale, jam-pack, supersound process 30 minutes, put coldly, microporous filter membrane (0.45 μ m) filters, promptly.

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-methanol-0.1% phosphoric acid (39:16.7:44.3) is mobile phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schisandrin peak should be not less than 3000.

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 108.1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 50ml that adds, claim to decide weight, supersound process (power 250W, frequency 20kHz) 20min, take out, put coldly, claim to decide weight again, add methanol and supply the weight of minimizing, filter, the accurate subsequent filtrate 20ml that draws, evaporate to dryness, residue add methanol makes dissolving, and fixed molten to 5ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.

The specific embodiment

Following experimental example and embodiment further specify but are not limited to the present invention down

Experimental example 1. Chinese drug-treated group of the present invention and composite capsule preparation technology's optimization experiment

1. water is proposed the screening of amount of water

Dispose every part of 648.8g of 5 parts of medical materials, dividing three groups tests, 6 times of amounts of first group of amount of water, 4 times of amounts, 8 times of amounts of second group of amount of water, 6 times of amounts, 10 times of amounts of the 3rd group of amount of water, 8 times of amounts, the 4th group of circulated in countercurrent extraction process adds 2 times of amounts of water, the 5th group of circulated in countercurrent extraction process adds 4 times of amounts of water, and the 6th group of circulated in countercurrent extraction process adds 6 times of amounts of water, and going out the clear paste amount after carrying with water is index, determine amount of water, the results are shown in Table 1:

Table 1: water is carried amount of water

With the paste volume is index, and the water consumption of circulated in countercurrent extraction process obviously reduces as can be seen, and paste-forming rate is improved, and can reduce concentration time, selects for use the circulated in countercurrent extraction process to add 4 times of amounts of water in the production.

2, the screening of precipitate with ethanol alcohol adding amount

Dispose every part of 648.8g of 3 parts of medical materials, divide three groups to test, first group adds 0.5 times of amount of amount of alcohol, and second group adds 1 times of amount of amount of alcohol, and the 3rd group adds 2 times of amounts of amount of alcohol, is index to go out the thick paste amount behind the precipitate with ethanol, determines alcohol adding amount, the results are shown in Table 2:

Table 2: precipitate with ethanol alcohol adding amount

With the paste volume is index, and it is better to add 1 times of amount of ethanol paste volume as can be seen, selects the 1 times of amount of amount of alcohol that adds in the production for use.

Table 3: important technological parameters

3, pilot scale

Table 4: batch pilot scale creation data

The optimization experiment of amygdalate processing procedure in experimental example 2. Chinese drug-treated group of the present invention and the thing

The Chinese medicine Semen Armeniacae Amarum contains the antitussive component amygdaloside, under proper condition, as make moist, decoction and water logging etc., the amygdalase that itself contained easily makes amygdaloside decompose, thereby reduces even lose drug effect.Therefore, Semen Armeniacae Amarum needs to reach the purpose that enzyme denaturing is protected glycosides through the process of preparing Chinese medicine.Because present traditional concocting method: decocting cooking method and fry the method enzyme denaturing and protect the glycosides poor effect, and quality is restive, thus introduce the steam heat platen press herein and traditional method compares experiment, to explore better concocting method.

1 instrument and reagent

1.1 instrument: YXQ.GY22.600 type horizontal circular pressurized steam disinfector (Hengyang medical apparatus and instruments factory).

1.2 reagent: silver nitrate is the AR level; Potassium iodide, liquor ammoniae fortis are the CP level; Semen Armeniacae Amarum is given birth to product and goods.

2 methods and result

2.1 the assay of amygdaloside in the Semen Armeniacae Amarum.

Get Semen Armeniacae Amarum (giving birth to product) the about 15g of coarse powder, precision is weighed, and puts in the kjeldahl flask, add water 150ml, close immediately plug is put in 37 ℃ of water-baths and is incubated 2h, connect condensing tube, the water flowing steam distillation, distillate imports in the absorption liquid of water 10ml and ammonia solution 2ml, and receiving flask is put in the ice bath and is cooled off, when reaching 60ml, distillate stops distillation, add potassium iodide test solution (16.5%) 2ml in the distillate, with slowly titration of silver nitrate solution (0.1mol/L), the yellow muddiness of showing to solution does not disappear.1ml silver nitrate solution (0.1mol/L) is equivalent to 91.48mg amygdaloside (C ₂₀H ₂₇NO ₁₁).The results are shown in Table 5.

Amygdaloside assay in table 5 Semen Armeniacae Amarum

* silver nitrate titration liquid is 0.1008mol/L

As shown in Table 1, the average content of amygdaloside is 4.343% in the living Semen Armeniacae Amarum.

2.2 in the traditional process of preparing Chinese medicine method goods amygdaloside and assay respectively with decocting cooking method system with fry the legal system Semen Armeniacae Amarum and grind into coarse powder, respectively get the about 15g of coarse powder.Carry out assay according to said method.The results are shown in Table 6.

Table 6 Semen Armeniacae Amarum tradition process of preparing Chinese medicine method amygdaloside measurement result

As shown in Table 2, the average content of frying amygdaloside in the legal system Semen Armeniacae Amarum is 3.061%; The average content of amygdaloside is 2.989% in the decocting cooking method system Semen Armeniacae Amarum.

2.3 the assay of amygdaloside in the steam heat platen press goods

Semen Armeniacae Amarum 200g cleans post rinse dedust soil, and draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.03kpa/cm ²(103 ℃), hot pressing steaming and decocting 30 minutes, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries.And measure the content of amygdaloside as stated above, the results are shown in Table 7.

Amygdaloside measurement result after the table 7 steam heat platen press process of preparing Chinese medicine Semen Armeniacae Amarum

As shown in Table 3, the average content of amygdaloside is 4.059% in the steam heat platen press process of preparing Chinese medicine Semen Armeniacae Amarum.

3 results

3.1 the concocting method that Semen Armeniacae Amarum is at present commonly used is the decocting cooking method and fries method, these two kinds method is simple, and the suitability is strong, but the enzyme denaturing weak effect, residual enzyme can also continue to decompose glycosides.The decocting cooking method is concocted Semen Armeniacae Amarum and is dropped in the boiling water, causes water temperature to descend, and severe patient can be reduced to 70 ℃, and this provides chance for zymolysis, and water consumption is big, and the part glycosides is water-soluble and lose.The method of frying will be fried to full Huang, just can reach the enzyme denaturing effect, and duration and degree of heating temperature is wayward, and the products appearance yellowing influences drug quality.Because anthropic factor (temperature, time are by artificial judgement) influence is bigger during operation, quality is difficult to homogeneous.

3.2 steam hot pressing enzyme denaturing method is concocted Semen Armeniacae Amarum, temperature is than decocting cooking method height, and is not soaked in water, and enzyme denaturing is effective, and glycosides runs off few.According to experimental result, hot pressing (103 ℃) enzyme denaturing 30min, the enzyme denaturing rate can reach 97.5%.

Experimental example 3. pharmacodynamic experiments

In order to prove curative effect of the present invention, we have carried out following pharmacodynamic experiment.

The used medicine group of the present invention of the following test of pesticide effectiveness is the capsule according to embodiment 4 preparations;

Positive control drug: the granule that following method makes:

Herba Ephedrae 1200g, Rhizoma Zingiberis Recens 1200g, Fructus Schisandrae Chinensis (system) 150g, Radix Glycyrrhizae Preparata 45g

Sucrose 627g, dextrin 260g make 1000g

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, decompression recycling ethanol also is concentrated into the thick paste that relative density is 1.34 ~ 1.40 (60 ~ 70 ℃).With sucrose, dextrin, mixing is made granule, drying, promptly.

1 material

1.1 reagent ammonia, inflexible rafter acid, phenol red, histamine phosphate, acecoline, histamine phosphate.

1.2 instrument 721 spectrophotometers, Shanghai the 3rd analytical tool factory produces.

1.3 the animal Kunming mouse, male and female half and half, body constitution amount 18-22g; Rat, male and female half and half, body constitution amount 200-250g.Cavia porcellus 150 ~ 200g, ♀ ♂ dual-purpose.

The influence that 2 antitussive effects are cough caused to mice ammonia.48 of mices, male and female half and half, be divided into 4 groups at random, according to the form below dosage gastric infusion, the 1st the sky, afternoon each 1 time place mice in the 5000ml glass bell jar during 30min after the administration in the 2nd day, and constant voltage is with ammonia (28%) spray people bell jar, spraying 5s observes the cough latent period of mice and the number of times (3min) of coughing.The results are shown in Table 8.

Table 8 medicine group of the present invention to mice because of the cough caused influence of ammonia (x ± s)

Annotate: * P＜0.05 of comparing with the normal saline group, * * P＜0.01.

The result: medicine group of the present invention can make the cough caused prolongation of latency of mice ammonia, and 3min cough number of times reduces.3 phlegm-dispelling functions are to the influence of the phenol red expectoration amount in mouse breathing road, 48 of mices, male and female half and half are divided into 4 groups at random, press the various dose of table 1 and irritate the stomach medicine, the 1st the sky, afternoon each 1 time, gave medicine on the 2nd day after 0.5h lumbar injection phenol red solution 0.5ml/ only.Behind the 30min, take off vertebra and put to death, insert about 0.3cm in people's trachea, draw NaHCO with the 1ml syringe from thyroid cartilage ₃Solution 0.5ml injects in the trachea, push away continuously repeatedly and take out 3 times, with syringe irrigating solution is extracted out at last and annotated in people's test tube, operate as stated above 3 times, wash 9 times, draw sodium carbonate liquor 1.5ml altogether, merge the about 1.2-1.5ml of eluate, get supernatant after centrifugal, the phenol red secretory volume of 721 spectrophotometer eudiometers the results are shown in Table 9.

Table 9 medicine group of the present invention is to the influence of the phenol red expectoration amount in mouse breathing road (x ± s)

Annotate: compare with the normal saline group: * P＜0.05, ※ P＜0.05 is compared with positive controls in * * P＜0.01.

The result: medicine group of the present invention can make the phenol red expectoration amount in mouse breathing road increase.

4 antiasthmatic effects

4.2 drawing Cavia porcellus, medicine group of the present invention breathes heavily preclinical influence

The guinea pig asthmatic model preparation causes the method for breathing heavily by spraying, selects Cavia porcellus childhood for use, body weight＜200g, insert in the lucite case of 20cm * 20cm * 150cm, ultrasonic atomizatio sprays into 2% acetylcholine and 0.2% histamine phosphate mixed liquor 15s, screens, and draws to breathe heavily to surpass 120s person incubation period and will not select for use.Next day, the Cavia porcellus random packet, medication group per os gives medicine group of the present invention and positive controls (press the crude drug amount and calculate, down together), and negative control group gives isometric normal saline, aminophylline group lumbar injection aminophylline (25mgkg ^-1).The 1h ultrasonic atomizatio sprays into 2% acetylcholine and 0.2% histamine phosphate mixed liquor 15s and measures to draw and breathe heavily incubation period (promptly begin to asthma attack from spraying, dyspnea is until the time that tic falls down to the ground) after the medication, the results are shown in Table 10.

Table 10 medicine group of the present invention to the preclinical influence of Cavia porcellus asthma (x ± s, n=8)

Compare with the normal saline group ^*P＜0.01

The result: medicine group of the present invention can make Cavia porcellus asthma obviously prolong incubation period, and asthma had antagonism due to histamine's acetylcholine mixing was stimulated.

4.3 influence to the contraction of guinea-pig isolated tracheal smooth muscle bar

The shrinkage test of guinea-pig isolated tracheal smooth muscle bar prepares isolated helical strips of guinea, places to fill in oxygenation Ke-Heng Shi liquid isolated organ mensuration bath (37 ℃), and load 2.0g balance, (final concentration is respectively 2.5 * 10 to inject choline or histamine respectively ^-7With 5 * 10 ^-7MmolL ^-1), record maximum collapse height adds medicine group of the present invention and positive controls, aminophylline group on this basis respectively, and record shrinks height after the balance, the results are shown in Table 11.

On this basis, set up 4 dosage groups according to a certain percentage separately, write down it and shrink height.The contraction inhibiting rate corresponding with it with the log10 dose of oral liquid carries out regression Calculation, draws its IC50 and is respectively 0.095gL ^-1(choline, n=4, r=0.989) and 0.345gL ^-1(histamine, n=4, r=0.965).

The influence that table 11 medicine group of the present invention is shunk guinea-pig isolated tracheal smooth muscle (x ± s, n=6)

Acetylcholine final concentration: 2.5 * 10 ^-7MmolL ^-1Histamine final concentration: 5 * 10 ^-7MmolL ^-1Compare * P＜0.05, * * P＜0.01 with the normal saline group

The result: acetylcholine or histamine can cause the tracheal smooth muscle bar and shrink strongly, and medicine group of the present invention has tangible relexation to the tracheal smooth muscle of spasm.

4.4 influence to the contraction of isolated ileum segments in guinea pigs smooth muscle

The shrinkage test of isolated ileum segments in guinea pigs smooth muscle prepares isolated ileum segments in guinea pigs 2cm, place and fill in the oxygenation tyrode's solution organ mensuration bath (37 ℃), trace its shrinkage curve, treat that intestinal tube begins experiment after the spontaneous easypro stable equilibrium that contracts, (final concentration is 5 * 10 to inject choline or histamine respectively ^-7MmolL ^-1), record maximum collapse height adds medicine group of the present invention and positive controls, atropine group on this basis respectively, and record shrinks height after the balance, the results are shown in Table 12.

On this basis, set up 4 dosage groups according to a certain percentage separately, write down it and shrink height, carry out regression Calculation, draw its IC50 and be respectively 0.183gL with log10 dose and its corresponding contraction inhibiting rate of medicine group of the present invention ^-1(choline, n=4, r=0.982) and 0.249gL ^-1(histamine, n=4, r=0.948).

Table 12 medicine group of the present invention to the influence of isolated ileum segments in guinea pigs smooth muscle contraction (x ± s, n=6)

Acetylcholine final concentration: 5 * 10 ^-7MmolL ^-1Histamine final concentration 5 * 10 ^-7MmolL ^-1Compare * * P＜0.01 with matched group

The result: medicine group of the present invention can have tangible diastole effect to the contraction of stripped ileum smooth muscle due to anti-acetylcholine or the histamine.

4.5 acetylcholine is caused the volumetrical influence of guinea-pig isolated trachea

The imitative complete trachea capillary tube method that exsomatizes of guinea-pig isolated trachea volume test, the stunning Cavia porcellus prepares the complete trachea specimen that exsomatizes, and places the bath of the oxygenation Ke-Heng Shi liquid of constant temperature (37 ℃), observes to connect liquid level in the 0.1cm capillary tube.After injecting choline, trachea shrinks, and tracheal volume reduces, and liquid level is raised, reinject medicine group of the present invention or normal saline, and the liquid level of 5min is poor after the record administration, the results are shown in Table 13.

Table 13 medicine group of the present invention to acetylcholine cause the volumetrical influence of guinea-pig isolated trachea (x ± s, n=6)

Acetylcholine final concentration: 2.5 * 10 ^-7MmolL ^-1Compare * P＜0.05, * * P＜0.01 with matched group

The result: acetylcholine shrinks tracheal smooth muscle, and volume reduces, and medicine group of the present invention can make the liquid level of raising descend, and the remarkable effect that the guinea-pig isolated trachea volume dwindles due to pair anti-acetylcholine is arranged.

4.6 influence to the Cavia porcellus lung airway perfusion that exsomatizes

The Guinea pig lung bronchus perfusion experiment Cavia porcellus of exsomatizing hits dizzy, and the carotid artery blood-letting separates trachea and takes out in the lump together with cardiopulmonary, with 37 ℃, gives and contains the oxygen locke solution and carry out perfusion, by perfusion bottle altitude mixture control perfusion flow (10mlmin ^-1About).After treating that perfusion flow is stable, inject a certain amount of acetylcholine solution (3 * 10 ^-7MmolL ^-1), perfusion flow obviously reduces, and injects a certain amount of medicine group (0.16gL of the present invention more respectively ^-1) and aminophylline (0.025gL ^-1), observe the variation of perfusion flow, the results are shown in Table 14.

Table 14 medicine group of the present invention to the influence of guinea pig in vitro lung airway perfusion (x ± s, n=6)

Choline concentration: 3 * 10 ^-7MmolL ^-1Medicine group concentration of the present invention: 0.16gml ^-1Aminophylline concentration: 0.025gml ^-1Compare * * P＜0.01 with matched group+choline group.

The result: after adding choline, because of the stripped lung airway perfusion amount of trachea and bronchus smooth muscle contraction obviously reduces, medicine group of the present invention has the effect of obviously anti-acetylcholine being shunk and increasing the lung airway perfusion amount.

The screening of experimental example 4 discrimination tests

One, Rhizoma Zingiberis discrimination test screening

The preparation of need testing solution:

Need testing solution one: get medicine of the present invention and be equivalent to crude drug 216.25g, the 30ml that adds diethyl ether, ultrasonic 30 minutes (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution;

Need testing solution two: get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution;

Need testing solution three: get medicine of the present invention and be equivalent to crude drug 216.25g, the 50ml that adds diethyl ether, reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution;

Need testing solution four: get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 30ml, ultrasonic 30 minutes (power 300W, frequency 50kHz) filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.

Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80ml and decocts 1 hour, filters, and filtrate is concentrated into 20ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw control medicinal material solution 5 μ l, need testing solution 15 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (5: 1) is developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, the results are shown in Table 15.

The The selection result of table 15 need testing solution

Need testing solution	1	2	3	4
Need testing solution	1	2	3	4	Color developing effect	Test sample is good at corresponding reference substance position speckle color developing effect	Test sample is very shallow in corresponding reference substance position spot colors	Test sample is shallow in corresponding reference substance position spot colors	Test sample is at corresponding reference substance position immaculate

The preparation of control medicinal material solution:

Control medicinal material solution one: Rhizoma Zingiberis control medicinal material 1g, add water 80ml and decocted 1 hour, filter, filtrate is concentrated into 20ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution;

Control medicinal material solution two: Rhizoma Zingiberis control medicinal material 1g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.

Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw control medicinal material solution one, two each 5 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (5: 1) is developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, the results are shown in Table 16.

The The selection result of table 16 control medicinal material solution

Control medicinal material solution	1	2
Control medicinal material solution	1	2	Color developing effect	Color developing effect is good	It is unintelligible to develop the color, and interference is arranged.

The selection of developing solvent

Developing solvent one: petroleum ether (60～90 ℃)-ethyl acetate (5: 1)

Developing solvent two: petroleum ether (60～90 ℃)-ethyl acetate (5: 1)

Developing solvent three: cyclohexane extraction-ether (1: 1)

Developing solvent four: cyclohexane extraction-ether (1: 1)

Get medicine of the present invention and be equivalent to crude drug 216.25g, the 30ml that adds diethyl ether, ultrasonic 30 minutes (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80ml and decocts 1 hour, filters, and filtrate is concentrated into 20ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), on four silica gel g thin-layer plates, point control medicinal material solution 5 μ l, need testing solution 15 μ l, respectively with developing solvent one, two, three, four, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, the results are shown in Table 17.

The The selection result of table 17 developing solvent

Developing solvent	1	2	3	4
Developing solvent	1	2	3	4	Launch effect	Relatively poor	Difference	Good	Difference

Two, Radix Glycyrrhizae discrimination test screening,

The selection of developing solvent

Developing solvent one: lower floor's solution of chloroform-methanol-water (13: 7: 2);

Developing solvent two: n-butyl alcohol-3mol/L ammonia solution-ethanol (5: 2: 1);

Developing solvent three: the upper solution of chloroform-methanol-water (13: 7: 2).

Get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour, filter, residue adds methanol 50ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, extract 3 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol 3 times, each 30ml gets n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), getting with the sodium carboxymethyl cellulose is two of the silica gel g thin-layer plates of adhesive, draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively on lamellae,, launch respectively with developing solvent one, two, three, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, the results are shown in Table 18.

The The selection result of table 18 developing solvent

Developing solvent	1	2	3
Developing solvent	1	2	3	Launch effect	Good	Difference	Relatively poor

Experimental example 4 Herba Ephedrae content assaying methods are learned and are investigated

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

Chromatographic column: Di Ma company (ZorbaxC184.6 * 250mm, 5 μ m)

Mobile phase: 0.1% phosphoric acid (containing 0.1% triethylamine)-acetonitrile (97:3)

Detect wavelength: 210nm column temperature: room temperature flow rate: 1.000ml/min

The reference substance source: the ephedrine hydrochloride reference substance, purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

Assay method: get the preparation sample liquid; And preparation lacks the blank preparation of Herba Ephedrae, the preparation negative controls.Filter with microporous filter membrane (0.45 μ m), accurate respectively each 5～10 μ l of negative controls, reference substance solution and need testing solution that draw inject chromatograph of liquid, measure, promptly.

1. content assaying method is investigated:

(1) preparation of blank assay blank solution is to get the group's medicine that lacks Herba Ephedrae, make blank preparation by technology, press the preparation of need testing solution preparation method again, above-mentioned chromatographic condition is measured, blank solution does not have chromatographic peak at the place of identical retention time with the ephedrine hydrochloride reference substance solution as a result, so think noiseless.

(2) stability test is got need testing solution, respectively at preparing the back 0,2,4,6,12,24 hour, measures in accordance with the law, and the injection equal volume the results are shown in Table 19.

Table 19 stability test result

The result shows that ephedrine hydrochloride is basicly stable in 24 hours.

(3) linear relationship is investigated and to be got reference substance solution (51.28 μ g/ml) and shake up, accurate respectively 2,4,6,8, the 10 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in Table 20, and drawing standard curve, the result shows that ephedrine hydrochloride is good in 0.103 μ g～0.513 μ g scope internal linear relation, and its regression equation is: Y=1942305X+23805 (r=0.9995).

Table 20 linear relationship is investigated the result

(4) the accurate ephedrine hydrochloride reference substance solution of drawing of precision test repeats sample introduction 5 times, and each 5 μ l try to achieve relative standard deviation＜2%, the results are shown in Table 21:

Table 21 Precision test result

(5) the text method is pressed in repeatability test, gets 5 parts of the same batch samples of lab scale, and every part is measured, and tries to achieve relative standard deviation＜2%, the results are shown in Table 22:

Table 22 reproducible test results

(6) recovery test takes by weighing the lab scale sample 0.2g of known content, and accurate the title decides, accurate ephedrine hydrochloride reference substance solution (0.4024mg/ml) 5ml that adds, press the preparation method operation of text need testing solution, measure its content, and calculate its response rate, measurement result sees Table 23:

Table 23 recovery test result

The result shows: recovery test is qualified, and this test method can be used for detecting Determination of Ephedrine Hydrochloride in this product.

2, sample size is measured

Measure 3 prepared batch samples of embodiment 5 according to the text content assaying method.The results are shown in Table 24:

Table 24 ephedrine hydrochloride assay is table as a result

Conclusion: measure by Determination of Ephedrine Hydrochloride among 3 crowdes of embodiment 5, in every preparation, Herba Ephedrae is counted 2.70mg～2.85mg with the ephedrine hydrochloride amount, contains Herba Ephedrae in the ephedrine hydrochloride amount in the daily metering of medicine group of the present invention, must not be less than 8.0mg.

Experimental example 5 Fructus Schisandrae Chinensis content assaying methods are learned and are investigated

1. instrument and reagent

Day island proper Tianjin LC-10AT high performance liquid chromatograph; The SPD-10A UV-detector; KQ-250 type processor for ultrasonic wave; The schisandrin reference substance; Methanol is chromatographically pure, and other reagent are analytical pure.

2. method and result

2.1 chromatographic condition

Chromatographic column: Shim-pack-C18 post (5 μ m, 4.6mm X 150mm), mobile phase: acetonitrile-methanol-0.1% phosphoric acid (39:16.7:44.3); Flow velocity 1ml/min; The detection wavelength is 250nm; Number of theoretical plate calculates by the schisandrin peak should be not less than 3000; Column temperature is 30 ℃.

2.2 the preparation of reference substance solution

It is an amount of that precision takes by weighing the schisandrin reference substance, adds methanol and be mixed with every 1ml and contain 0.03mg solution, promptly.

2.3 the preparation of need testing solution

Get medicine of the present invention and be equivalent to crude drug 108.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, supersound process 20min, take out, put coldly, claim to decide weight again, add methanol and supply the weight of minimizing, filter, the accurate subsequent filtrate 20ml that draws, evaporate to dryness, residue add methanol makes dissolving, and fixed molten to 5ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.

2.4 the preparation of negative control solution

Do not contain the negative sample of schisandra chinensis medicinal material in prescription ratio preparation, prepare negative control solution by the preparation method of need testing solution.Accurate respectively each the 10 μ l of negative control solution, need testing solution and reference substance solution that draw inject chromatograph of liquid, press liquid phase chromatogram condition and measure, and the result shows that the place of identical retention time with reference substance is noiseless in the negative control solution chromatogram.

2.5 linear relationship is investigated

Accurate schisandrin reference substance solution (0.03mg/ml) 4,8,12,16, the 20 μ l that draw, inject high performance liquid chromatograph analysis respectively, (μ g) is abscissa with the reference substance sample size, with the long-pending integrated value in the sharp side of reference substance is the vertical coordinate mapping, the drawing standard curve, its regression equation is: Y=1327476.55X-9683.109, r=0.9998, the result shows that schisandrin is good linear relationship between 0.11792-0.5896 μ g.

Table 25 linear relationship is investigated the result

2.6 precision test

The accurate need testing solution of drawing repeats sample introduction continuous 6 times, and each 10 μ l try to achieve relative standard deviation＜2%, the results are shown in Table 26:

Table 26 Precision test result

2.7 stability test

Get need testing solution, in placement 0,2,4 and 6h, sample introduction 10 μ l write down chromatogram respectively, and the RSD=0.49% of schisandrin peak area the results are shown in Table 27 as a result.

Table 27 stability test result

The result shows that schisandrin is basicly stable in 6 hours.

2.8 repeatability test

Get same batch sample, press this paper method and independently measure five times, sample introduction is measured successively, schisandrin content in the sample,, try to achieve relative standard deviation＜2%, see Table 28.

Table 28 reproducible test results

The result shows that its repeatability is good.

2.9 recovery test

It is an amount of to get the schisandrin reference substance, joins in the medicine group sample of the present invention of surveying schisandrin content, and press the preparation method preparation of need testing solution, and record schisandrin content, calculate recovery rate, measurement result sees Table 29.

Table 29 average recovery measurement result

Average recovery rate=98.22%, RSD=0.60%

2.10 sample determination

Press this paper 2.2 and 2.3 following reference substance solution and need testing solutions of preparing,, measure in accordance with the law, measure three batch samples altogether, the results are shown in Table 30 by above-mentioned chromatographic condition.

Table 30 sample determination result

Conclusion

(1) Fructus Schisandrae Chinensis is the main flavour of a drug in the medicine group of the present invention, and its contained schisandrin is one of main effective ingredient, therefore with this product schisandrin content as medicine group quality control index of the present invention.Adopt the content of schisandrin in the high effective liquid chromatography for measuring preparation in this article, this method is simple to operate, favorable reproducibility, response rate height.

(2) in the relevant report of schisandrin HPLC content assaying method, mobile phase adopts systems such as methanol, water more.We grope by experiment, determine with methanol, acetonitrile, 0.1% tricresyl phosphate kind system to be mobile phase, and in sample chromatogram, the schisandrin separating effect is comparatively satisfied, the separating degree height, and the peak shape symmetry is better.

Following embodiment all can realize above invention effect (embodiment lacks method of quality control)

Embodiment 1

Herba Ephedrae 3000g Rhizoma Zingiberis Recens 3000g Fructus Schisandrae Chinensis (system) 375g Radix Glycyrrhizae Preparata 112.5g

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).Vacuum drying is pulverized, and makes the preparation of clinical acceptance according to this area routine techniques, as: capsule, tablet, granule, soft capsule, pill.

Embodiment 2

Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Ramulus Cinnamomi 2800 weight portions, Semen Armeniacae Amarum 1000 weight portions, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g

Above medical material is except that Semen Armeniacae Amarum, and other five tastes medical materials decoct with water 3 times, each 2 hours, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃); Semen Armeniacae Amarum rinsing dedust soil, draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.03kpa/cm2 (103 ℃), hot pressing steaming and decocting 30 minutes, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries, and promptly gets the Semen Armeniacae Amarum processed product.With above-mentioned thick paste and Semen Armeniacae Amarum processed product vacuum drying, pulverize, make the preparation of clinical acceptance according to this area routine techniques, as: capsule, tablet, granule, soft capsule, pill.

Embodiment 3

Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).Vacuum drying is pulverized, and adds dextrin 300g, cane sugar powder 600g, and mixing gets medicinal mixture, makes granule, and drying is made 1000 bags, promptly.

Embodiment 4 tablets

Above four Chinese medicine material adopts the circulated in countercurrent extractive technique to decoct with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).Vacuum drying is pulverized, and adds Sodium Tvlose 20g, and mixing gets medicinal mixture, makes granule, drying, and granulate, tabletting, coating is made 1000, promptly.

Embodiment 5 capsules

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).Vacuum drying is pulverized, and adds dextrin 150g, and mixing gets medicinal mixture, and granulation, encapsulated is made 1000, promptly.

Embodiment 6 soft capsules

Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g,

Above four Chinese medicine material decocts with water secondary, and 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), puts and is chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃), drying under reduced pressure, pulverize,, add vegetable oil 200g with the fine powder mixing, stir evenly, make 660 of soft capsules, promptly.

Embodiment 7 granules

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).Vacuum drying is pulverized, and adds cane sugar powder 600g, dextrin 300g, and mixing gets medicinal mixture, makes granule, and drying is made 1000g, promptly.

Embodiment 8 pills

Above four Chinese medicine material decocts with water secondary, 3 hours for the first time, 2 hours for the second time, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃).60 ℃ of drying under reduced pressure are pulverized, with above-mentioned fine powder mixing be that binding agent is made 1000 balls with the alcoholic solution of 3%PVP, promptly.

Claims

1, a kind of have a lung qi dispersing, and the pharmaceutical composition of antiasthmatic effect is characterized in that the crude drug of this pharmaceutical composition consists of:

Herba Ephedrae 3000-4000 weight portion, Rhizoma Zingiberis Recens 2000-3000 weight portion, Ramulus Cinnamomi 2000-3000 weight portion, Semen Armeniacae Amarum 1000-1500 weight portion, Fructus Schisandrae Chinensis (system) 250-500 weight portion, Radix Glycyrrhizae Preparata 80-160 weight portion.

2, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug in this pharmaceutical composition consists of:

Herba Ephedrae 3000-3200 weight portion, Rhizoma Zingiberis Recens 2800-3000 weight portion, Ramulus Cinnamomi 2800-3000 weight portion, Semen Armeniacae Amarum 1400-1500 weight portion, Fructus Schisandrae Chinensis (system) 300-400 weight portion, Radix Glycyrrhizae Preparata 100-120 weight portion.

3, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug of this pharmaceutical composition consists of:

Herba Ephedrae 3000g, Rhizoma Zingiberis Recens 3000g, Ramulus Cinnamomi 3000g, Semen Armeniacae Amarum 1500g, Fructus Schisandrae Chinensis (system) 375g, Radix Glycyrrhizae Preparata 112.5g.

4, as the preparation method of Chinese medicine composition as described in any one of the claim 1-3, it is characterized in that this method is:

Medical material is except that Semen Armeniacae Amarum, and other five tastes medical materials adopt the circulated in countercurrent extraction process to decoct with water 1-3 time, each 2-4 hour, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 12-36 hour, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃); Semen Armeniacae Amarum rinsing dedust soil, draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.01～0.03kpa/cm ²(103 ℃), hot pressing steaming and decocting 20-40 minute, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries, and promptly gets the Semen Armeniacae Amarum processed product; With above-mentioned thick paste and Semen Armeniacae Amarum processed product vacuum drying, pulverize, make the preparation of clinical acceptance according to this area routine techniques, as: capsule, tablet, granule, soft capsule, pill.

5, as the preparation method of Chinese medicinal composition capsules agent as described in the claim 4, it is characterized in that this method is:

Medical material is except that Semen Armeniacae Amarum, and other five tastes medical materials adopt the circulated in countercurrent extraction process to decoct with water 2 times, each 2 hours, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.08 (60 ℃), put and be chilled to room temperature, add equivalent ethanol, stir evenly, left standstill 24 hours, get supernatant, reclaim ethanol and be concentrated into the thick paste that relative density is 1.34～1.40 (60～70 ℃); Semen Armeniacae Amarum rinsing dedust soil, draining the water splits on the dish, puts into the pressure sterilizing pot, and the control steam pressure is 0.01～0.03kpa/cm ²(103 ℃), hot pressing steaming and decocting 30 minutes, taking-up is promptly put into a small amount of cold water and is soaked, and removes kind of a skin, dries, and promptly gets the Semen Armeniacae Amarum processed product; With above-mentioned thick paste and Semen Armeniacae Amarum processed product vacuum drying, pulverize, it is an amount of to add dextrin, and mixing incapsulates, and makes 1000, promptly.

6, as the method for quality control of claim 1-3 Chinese medicine composition as described in each, it is characterized in that this method of quality control comprises one or more in the following discrimination method:

Differentiate:

(1) get medicine of the present invention and be equivalent to crude drug 9-11g, add strong ammonia solution 1-3m1, add chloroform 30-50ml again, reflux 0.5-2 hour, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets ephedrine hydrochloride, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (15-25: 3-10: 0.5-2) be developing solvent, launch, take out, dry, spray is with 5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical aubergine speckle;

(2) get medicine of the present invention and be equivalent to crude drug 200-220g, the 20-40ml that adds diethyl ether, ultrasonic 20-40 minute (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80-100ml and decocts 1 hour, filters, and filtrate is concentrated into 20-30ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw control medicinal material solution 5 μ l, need testing solution 15 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (3-7: 0.5-2) be developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(3) get medicine of the present invention and be equivalent to crude drug 200-220g, add petroleum ether (60～90 ℃) 40-60ml, reflux 0.5-2 hour, filter, residue adds methanol 40-60ml, reflux 0.5-2 hour, filter, filtrate evaporate to dryness, residue add water 30-50ml makes dissolving, extracts 2-4 time with water saturated n-butyl alcohol jolting, each 10-30ml, merge n-butyl alcohol liquid, use the saturated water washing of n-butyl alcohol 2-4 time, each 20-40ml, get n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, (10-15: 5-10: lower floor's solution 1-3) is developing solvent with chloroform-methanol-water, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

7, the method for quality control of pharmaceutical composition as claimed in claim 6 is characterized in that this method of quality control comprises one or more in the following discrimination method:

Differentiate: (1) is got medicine of the present invention and is equivalent to crude drug 10.8g, adds strong ammonia solution 2ml, adds chloroform 40ml again, and reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets ephedrine hydrochloride, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (20: 5: 0.5) is developing solvent, launch, take out, dry, spray is with 5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical aubergine speckle;

(2) get medicine of the present invention and be equivalent to crude drug 216.25g, the 30ml that adds diethyl ether, ultrasonic 30 minutes (power 300W, frequency 50kHz) filters, and filtrate volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Zingiberis control medicinal material 1g, adds water 80ml and decocts 1 hour, filters, and filtrate is concentrated into 20ml, adds equal-volume petroleum ether (60～90 ℃) extraction, gets upper strata liquid, and evaporate to dryness, residue add 1ml methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw control medicinal material solution 5 μ l, need testing solution 15 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (5: 1) is developing solvent, secondary launches, and takes out, and dries, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(3) get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour, filter, residue adds methanol 50ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, extract 3 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol 3 times, each 30ml gets n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution with chloroform-methanol-water (13: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

8,, it is characterized in that this method of quality control comprises one or more in the following assay as the method for quality control of the described pharmaceutical composition of claim 1-3:

(1) measures according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D);

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With 0.1% phosphoric acid (containing 0.1% triethylamine)-acetonitrile (90-100:0-10) is mobile phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 2500;

It is an amount of that the ephedrine hydrochloride reference substance is got in the preparation of reference substance solution, accurate claims surely, makes the solution that every 1ml contains 50 μ g with dissolve with methanol, product solution in contrast, promptly;

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 3-5g, and accurate the title decides, to the 50ml volumetric flask, and the accurate hydrochloric acid solution 1ml that adds 0.5mol/L, add methanol again to scale, jam-pack, supersound process 20-40 minute, put coldly, microporous filter membrane (0.45 μ m) filters, promptly;

Accurate respectively reference substance solution and each the 5 μ l～10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

The daily metering of medicine of the present invention contains Herba Ephedrae with ephedrine hydrochloride (C ₁₀H ₁₅NOHCl) the amount meter must not be less than 8.0mg;

(2) measure according to high performance liquid chromatography (appendix VI D);

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-methanol-0.1% phosphoric acid (35-40:15-20:40-50) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates by the schisandrin peak should be not less than 3000;

Schisandrin reference substance 1.5mg is got in the preparation of reference substance solution, accurate claims surely, puts in the 50ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up, and promptly gets (every 1ml contains schisandrin 0.03mg);

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 100-110g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 50ml that adds, claim to decide weight, supersound process (power 250W, frequency 20kHz) 10-30min, take out, put coldly, claim to decide weight again, add methanol and supply the weight of minimizing, filter, the accurate subsequent filtrate 20ml that draws, evaporate to dryness, residue add methanol makes dissolving, and fixed molten to 5ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 10 μ l of test solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

9, the method for quality control of pharmaceutical composition as claimed in claim 8 is characterized in that this method of quality control comprises one or more in the following content assaying method:

(1) measures according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005);

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With 0.1% phosphoric acid (containing 0.1% triethylamine)-acetonitrile (97:3) is mobile phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 2500;

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 4.3g, and accurate the title decides, to the 50ml volumetric flask, and the accurate hydrochloric acid solution 1ml that adds 0.5mol/L, add methanol again to scale, jam-pack, supersound process 30 minutes, put coldly, microporous filter membrane (0.45 μ m) filters, promptly;

(2) measure according to high performance liquid chromatography (appendix VID);

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-methanol-0.1% phosphoric acid (39:16.7:44.3) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates by the schisandrin peak should be not less than 3000;

The preparation of need testing solution is got medicine of the present invention and is equivalent to crude drug 108.1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 50ml that adds, claim to decide weight, supersound process (power 250W, frequency 20kHz) 20min, take out, put coldly, claim to decide weight again, add methanol and supply the weight of minimizing, filter, the accurate subsequent filtrate 20ml that draws, evaporate to dryness, residue add methanol makes dissolving, and fixed molten to 5ml, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly;

Medicine of the present invention contains schisandrin (C ₂₄H ₃₂O ₇₎Must not be less than 0.40%.

10,, it is characterized in that this method of quality control comprises following method as the method for quality control of claim 3 or 5 described pharmaceutical compositions:

(3) get medicine of the present invention and be equivalent to crude drug 216.25g, add petroleum ether (60～90 ℃) 50ml, reflux 1 hour, filter, residue adds methanol 50ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, extract 3 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol 3 times, each 30ml gets n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g shines medical material solution in pairs with legal system in addition; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution with chloroform-methanol-water (13: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

Assay

(2) measure according to high performance liquid chromatography (appendix VID of 2005 editions pharmacopeia);