CN101396486A

CN101396486A - Traditional Chinese medicine composition for treating cough and asthma and preparation and quality control method thereof

Info

Publication number: CN101396486A
Application number: CNA2007101224902A
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: CN101396486B

Abstract

The invention discloses a traditional Chinese medicine composition for treating cough and asthma diseases, a preparation method and a quality control method thereof. The composition is made from the bulk drugs blackberrylily rhizome, platycodon root, epimedium, hawthorn fruit (crude), peppergrass, houttuynia, astragalus root, Chinese ephedra (roasted), earthworm, Chinese yam rhizome, common anemarrhena rhizome, gecko and the like, high performance liquid chromatography is adopted in the quality control method to assay the epimedium, and thin layer chromatography is employed to quanlitatively identify the blackberrylily rhizome, the platycodon root, the epimedium, the astragalus root, the Chinese ephedra (roasted) and the earthworm. The composition has outstanding curative effect and the functions of nourishing qi and kidney, removing heat from the lung and dissolving phlegm, and relieving cough and asthma. The composition has very good curative effect on the cough and asthma caused by the qi deficiency of lung and kidney as well as the phlegm-heat stagnating in lung, acute bronchitis, chronic bronchitis and bronchial asthma with the syndromes.

Description

Chinese medicine composition of treatment cough illness and preparation method thereof and method of quality control

Technical field

The present invention relates to a kind of Chinese drug-treated group for the treatment of cough illness and thing and preparation method thereof and method of quality control, relate in particular to the cough with asthma of a kind of treatment insufficiency of QI of the lung and kidney, expectorant heat stagnation lung, and the Chinese drug-treated group of acute/chronic bronchitis, bronchial asthma and thing and preparation method thereof and method of quality control, belong to technical field of Chinese medicines.

Background technology

Cough illness comprises diseases such as asthma, tracheitis, is one group of worldwide commonly encountered diseases, difficult disease.According to World Health Organization (WHO): " the summation that harm that asthma causes human health and financial burden have surpassed tuberculosis and acquired immune deficiency syndrome (AIDS)." because of the ecological environmental pollution degree constantly increases the weight of, pathogenesis of asthma rate and mortality rate are and increase trend year by year.At present there are asthma patient 1.5 hundred million～200,000,000 people in the whole world, and this numeral also increases continuing, and the people who dies from asthma every year reaches 180,000 more than.Wherein China's asthma prevalence is 1%～5%, and 3,000 ten thousand asthma patients are arranged approximately.In recent years, along with the development with chemical engineering industry of increasing the weight of of the improving constantly of industrialization degree, atmospheric pollution, the pathogenesis of asthma rate has increased trend gradually.Persistent ailment when outbreak clinical manifestation is: cough with asthma, asthma, abundant expectoration, uncomfortable in chest, itching of the throat, dry cough, dyspnea, cardiopalmus hyperhidrosis, seriously fidgety, stridulate sound, heart beating is disorderly, blood pressure drops even death by suffocation.

In the medicine of treatment cough illness, a chemical drugs symptomatic treatment can't effect a radical cure, and most Chinese medicine is long the course of treatment, and dose is big, the curative effect instability.

Summary of the invention

The object of the invention is to provide a kind of Chinese medicine composition for the treatment of cough illness;

The object of the invention also is to provide a kind of preparation method for the treatment of the Chinese medicine composition of cough illness;

The object of the invention also is to provide a kind of method of quality control for the treatment of the Chinese medicine composition of cough illness.

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

The Chinese medicine composition of treatment cough illness of the present invention is to be made by the crude drug of following weight ratio:

Rhizoma Belamcandae 20-40 weight portion Radix Platycodonis 10-20 weight portion Herba Epimedii 30-60 weight portion

Radix Astragali 30-50 weight portion Herba Ephedrae (processing) 10-20 weight portion Pheretima 10-20 weight portion

The Chinese medicine composition of treatment cough illness of the present invention can be to be made by the crude drug of following weight ratio:

Rhizoma dioscoreae 20-40 weight portion Semen Lepidii (Semen Descurainiae) 25-40 weight portion

Rhizoma dioscoreae 20-40 weight portion Rhizoma Anemarrhenae 10-30 weight portion

Rhizoma dioscoreae 20-40 weight portion Fructus Crataegi (life) 60-80 weight portion

Fructus Crataegi (life) 60-80 weight portion Semen Lepidii (Semen Descurainiae) 25-40 weight portion

Fructus Crataegi (life) 60-80 weight portion Gecko 5-15 weight portion

The Chinese medicine composition of treatment cough illness of the present invention can be to add Herba Houttuyniae 80-120 weight portion in the above-mentioned raw materials medicine.

Rhizoma Belamcandae 30 weight portion Radix Platycodoniss 15 weight portion Herba Epimedii 50 weight portions

Fructus Crataegi (life) 75 weight portion Semen Lepidii (Semen Descurainiae)s 30 weight portion Herba Houttuyniae 100 weight portions

The Radix Astragali 45 weight portion Herba Ephedraes (processing) 15 weight portion Pheretimas 15 weight portions

The Rhizoma dioscoreae 30 weight portion Rhizoma Anemarrhenaes 15 weight portion Geckos 10 weight portions

Prescription science of the present invention, the Radix Astragali, benefiting QI for strengthening the superficies, Pheretima, the heat clearing away suppressing the hyperactive liver, collateral dredging is relievingd asthma, diuresis, the two medicines strengthening vital QI to eliminate pathogenic factors that is harmonious, specimen is held concurrently solid, is monarch's medicine.Herba Epimedii, Gecko, the kidney invigorating lung benefiting helps the lung improving inspiration by invigorating kidney-QI; Herba Houttuyniae, Rhizoma Belamcandae, heat-clearing and toxic substances removing, the damp eliminating eliminating stagnation, the clear purte the white of the Rhizoma Anemarrhenae, nourishing YIN and moistening the lung helps the principal agent removing heat-phlegm; Herba Ephedrae (processed) matter profit antitussive and antiasthmatic; Six medicines are ministerial drug, help the principal agent main attack primary symptom of concentrating one's efforts altogether.Assistant is with Semen Lepidii (Semen Descurainiae), and the eliminating the pathogens from the lung diuretic helps the monarch and his subjects' medicine resolving phlegm and relieving asthma; Fructus Crataegi, promoting digestion and removing stagnation, activating blood circulation to dissipate blood stasis, Rhizoma dioscoreae, tonifying the spleen and stomach, the lung benefiting kidney helps transporting and transforming function of the spleen and stomach, and reaches building up the spleen to supplement the lung, the disappear effect of painful abdominal mass of the chest stuffiness relieving; Radix Platycodonis, lung qi dispersing eliminates the phlegm, and promoting flow of QI and blood is specially gone into lung meridian, draws the through sick institute of all medicines, is messenger drug in the side.Full side mends void to some extent, dispels to some extent in fact, and strengthening vital QI to eliminate pathogenic factors, giving consideration to both the incidental and fundamental must be answered lung qi, and expectorant heat must be removed, the cough with asthma self-balancing.

The Chinese medicine composition preparation method of treatment cough illness of the present invention is:

Choose described crude drug medical material, decoct with water 1-3 time, each 1-3 hour, collecting decoction filters, and filtrate is concentrated into the extractum that relative density is 1.15～1.20 (50 ℃), add adjuvant, make the various conventional formulations in this area, as: granule, capsule, tablet, soft capsule, pill, promptly.

Choose described raw medicinal material, decoct with water secondary, the decocting that adds for the first time 10 times of amounts boiled 2 hours, and the decocting that adds 8 times of amounts for the second time boiled collecting decoction 2 hours, filter, filtrate is concentrated into the extractum that relative density is 1.15～1.20 (50 ℃), adds starch, granulates, dry, the magnesium stearate mix homogeneously of adding 0.5%, tabletting is made solvent with 95% ethanol and is prepared 8% stomach dissolution type coating solution, inlet temperature is 40 ℃, hydrojet speed is 25kg/h, selects 4 rev/mins speed when the coating pan rotating speed begins for use, along with the continuous formation of film-coat layer on plain sheet surface, progressively increase rotating speed again, up to 7 rev/mins; After hydrojet finishes, use hot air drying 20 minutes, promptly.

The method of quality control of the Chinese medicine composition of treatment cough illness of the present invention comprises one or more in following discrimination method and/or the assay:

[discriminating]

A. get and be equivalent to crude drug 10-50g drug combination preparation content of the present invention, porphyrize adds methanol 40-80ml, supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add water 20-60ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 1-3ml methanol makes dissolving, as need testing solution.Other gets Rhizoma Belamcandae control medicinal material 0.5-2g, adds methanol 10-20ml, and supersound process 20-60 minute, filter, filtrate is concentrated into 1ml medical material solution in contrast.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw above-mentioned need testing solution 3-10 μ l, control medicinal material solution 1-5 μ l puts in same silica gel G F respectively ₂₅₄On the lamellae, (6-10: 1-5: lower floor's solution 0.5-2) is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp (254nm) and inspects with chloroform-methanol-water.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

B. get and be equivalent to crude drug 40-70g drug combination preparation content of the present invention, porphyrize adds chloroform 10-40ml, and supersound process 10-60 minute, filter, filtrate is concentrated into 0.5-2ml, as need testing solution.Other gets Pheretima control medicinal material 0.5-2g, adds chloroform 10-40ml, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned control medicinal material solution 3-10 μ l, need testing solution 5-20 μ l put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (8-12: 0.5-2) be developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

C. get and be equivalent to crude drug 40-70g drug combination preparation content of the present invention, porphyrize, add 1-3% potassium hydroxide methanol solution 40-60ml, claim to decide weight, reflux, keep little and boiled 0.5-2.0 hour, filter, filtrate adds chloroform-n-butyl alcohol, and (1-3: 0.5-2) mixed solution extracts 1-3 time, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 1-3 time, each 30-50ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 0.5-2.0ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.1-1.0mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution 10 μ l, reference substance solution 1-5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, (10-20: 5-10: 1-5) lower floor's solution of placing below 10 ℃ is developing solvent with chloroform-methanol-water, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

D. get and be equivalent to crude drug 5-20g drug combination preparation content of the present invention, porphyrize adds 30-70% ethanol 30ml, reflux 10-50 minute, take out, put cold, filter, filtrate is steamed near and is done, and adds water 1-5ml and makes dissolving, be added on polyamide column (5g, on the post footpath 10～15mm), water 100ml eluting, discard eluent, reuse ethanol 100ml eluting is collected eluent, evaporate to dryness, residue add ethanol 0.5-2.0ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds ethanol and makes the solution that every 1ml contains 0.5-2.0mg, in contrast product solution.According to thin layer chromatography (an appendix VI of the yellow version in 2005 of middle traditional Chinese medicines B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with dehydrated alcohol-water (55: 44) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical kermesinus speckle.

E. get and be equivalent to crude drug 20-40g drug combination preparation content of the present invention, porphyrize adds strong ammonia solution 2-8ml and makes moistening, add chloroform 20-60ml, reflux 0.5-2.0 hour, filter, filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Other gets the ephedrine hydrochloride reference substance, and chlorination is copied into the solution that every 1ml contains 0.5-2.0mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3-10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with isopropyl alcohol-n-butyl alcohol-strong ammonia solution (10-20: 1-3: 1-5) be developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation.

F. get and be equivalent to crude drug 20-50g drug combination preparation content of the present invention, add 7% sulphuric acid alcohol-water (0.5-2.0: 1-5) mixed solution 40ml, reflux 1-5 hour, put coldly, add the chloroform jolting and extract 1-3 time, at every turn 10-30ml, combined chloroform liquid adds water 20-40ml washing, discards water liquid, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methanol 1-3ml makes dissolving, add activated carbon and handle in right amount, filter, filtrate is as need testing solution.Other gets Radix Platycodonis control medicinal material 0.5-2.0g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 2-10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (0.5-2.0: 0.5-2.0) be 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 principal spot of same color.

[assay]

The assay of Herba Epimedii

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol-water (40-60:60-40) is mobile phase; The detection wavelength is 200-300nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

It is an amount of that the icariin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 0.05-0.5mg, promptly.

The preparation of need testing solution is got and is equivalent to crude drug 30-50g drug combination preparation content of the present invention, and porphyrize is got about 0.5-2.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 10-20ml that adds, claim to decide weight, supersound process (power 250W, frequency 50kHz) 0.5-2.0 hour, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each the 2-10 μ l of need testing solution of drawing of algoscopy injects chromatograph of liquid, measures, promptly.

This product taking dose every day contains Herba Epimedii with icariin (C ₃₃H ₄₀O ₁₅) meter, must not be less than 1.0-5.0mg.

[discriminating]

A. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, porphyrize adds methanol 50ml, supersound process 30 minutes, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 2ml methanol makes dissolving, as need testing solution.Other gets Rhizoma Belamcandae control medicinal material 1g, adds methanol 15ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml medical material solution in contrast.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) test, draw above-mentioned need testing solution 5 μ l, control medicinal material solution 2 μ l put in same silica gel G F respectively ₂₅₄On the lamellae, be developing solvent, launch, take out, dry, put under the ultra-violet lamp (254nm) and inspect with lower floor's solution of chloroform-methanol-water (8: 2.5: 1).In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

B. get and be equivalent to crude drug 56g drug combination preparation content of the present invention, porphyrize adds chloroform 30ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml, as need testing solution.Other gets Pheretima control medicinal material 1g, adds chloroform 30ml, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned control medicinal material solution 5 μ l, need testing solution 10 μ l put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (9: 1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

C. get and be equivalent to crude drug 56g drug combination preparation content of the present invention, porphyrize adds 2% potassium hydroxide methanol solution 50ml, claim to decide weight, reflux keeps little and boiled 1 hour, filter, filtrate adds chloroform-n-butyl alcohol (2: 1) mixed solution extracts 2 times, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 2 times, each 40ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution 10 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution of placing below 10 ℃ with chloroform-methanol-water (15: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

D. get and be equivalent to crude drug 11g drug combination preparation content of the present invention, porphyrize adds 50% ethanol 30ml, reflux 30 minutes is taken out, and puts cold, filter, filtrate is steamed near and is done, and adds water 3ml and makes dissolving, be added on polyamide column (5g, on the post footpath 10～15mm), water 100ml eluting, discard eluent, reuse ethanol 100ml eluting is collected eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of the yellow version in 2000 of middle traditional Chinese medicines B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with dehydrated alcohol-water (55: 44) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical kermesinus speckle.

E. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, porphyrize adds strong ammonia solution 4ml and makes moisteningly, adds chloroform 40ml, and reflux 1 hour filters, and filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Other gets the ephedrine hydrochloride reference substance, and chlorination is copied into 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 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with isopropyl alcohol-n-butyl alcohol-strong ammonia solution (15: 2: 2.5) is developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation.

F. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, add 7% sulphuric acid alcohol-water (1: 3) mixed solution 40ml, reflux 3 hours, put coldly, add the chloroform jolting and extract 2 times, each 20ml, combined chloroform liquid adds water 30ml washing, discards water liquid, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methanol 2ml makes dissolving, add activated carbon and handle in right amount, filter, filtrate is as need testing solution.Other gets Radix Platycodonis control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (1: 1) 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 principal spot of same color.

[assay]

The assay of Herba Epimedii

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol-water (53: 47) is mobile phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

It is an amount of that the icariin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 0.1mg, promptly.

The preparation of need testing solution is got and is equivalent to crude drug 37.5g drug combination preparation content of the present invention, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 20ml that adds, claim to decide weight, supersound process (power 250W, frequency 50kHz) 1 hour, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Algoscopy

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

This product taking dose every day contains Herba Epimedii with icariin (C ₃₃H ₄₀O ₁₅) meter, must not be less than 2.25mg.

Prescription science of the present invention, the Radix Astragali, benefiting QI for strengthening the superficies, Pheretima, the heat clearing away suppressing the hyperactive liver, collateral dredging is relievingd asthma, diuresis, the two medicines strengthening vital QI to eliminate pathogenic factors that is harmonious, specimen is held concurrently solid, is monarch's medicine.Herba Epimedii, the kidney invigorating and essence nourishing helps the lung improving inspiration by invigorating kidney-QI, Herba Houttuyniae, Rhizoma Belamcandae, heat-clearing and toxic substances removing, the damp eliminating eliminating stagnation helps the principal agent removing heat-phlegm, Herba Ephedrae (processed) matter profit antitussive and antiasthmatic.Four medicines are ministerial drug, help the principal agent main attack primary symptom of concentrating one's efforts altogether, and assistant is with Semen Lepidii (Semen Descurainiae), and the eliminating the pathogens from the lung diuretic helps the monarch and his subjects' medicine resolving phlegm and relieving asthma.Fructus Crataegi, promoting digestion and removing stagnation, activating blood circulation to dissipate blood stasis helps transporting and transforming function of the spleen and stomach, and reaches building up the spleen to supplement the lung, the disappear effect of painful abdominal mass of the chest stuffiness relieving, Radix Platycodonis, lung qi dispersing eliminate the phlegm, promoting flow of QI and blood.Specially go into lung meridian, draw the through sick institute of all medicines, dispel to some extent for the full side of messenger drug in the side mends in fact void to some extent, strengthening vital QI to eliminate pathogenic factors, giving consideration to both the incidental and fundamental must be answered lung qi, and expectorant heat must be removed, the cough with asthma self-balancing.

Present composition determined curative effect has vital energy benefiting and the kidney invigorating, removing heat from the lung and dissipating phlegm, and the effect of relieving cough and asthma, to the cough with asthma of insufficiency of QI of the lung and kidney, expectorant heat stagnation lung, and acute/chronic bronchitis, bronchial asthma see that above-mentioned patient has better curative effect.Find that by contrast experiment pharmaceutical composition of the present invention has outstanding drug effect with existing preparation.

The method of quality control of Chinese medicine composition provided by the present invention, be by obtaining behind the creative experiment sieving of big measuring, pass through screening in the discrimination method to sample treatment, the selection of developing solvent, make and differentiate that specificity is fine, and method is economic and practical, the result is quick, and can both use different lamellaes.Pass through screening in the content assaying method to sample, test sample processing method, the selection of developing solvent, make content assaying method effectivelyly to carry out quality control, and will compare more stable that product that additive method measures shows on pharmacological effect with the product that this method is measured to product.

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 thing preparation technology's optimization experiment

1, Study on extraction

1.1 water is proposed amount of water research: in former extraction process, stipulate the solvent volume of adding, but in actual production process, the extraction solvent volume that adds is an important parameter, quality to the extractum that extracts has a significant impact, so in research process, former extraction process is carried out perfect, to guarantee the science and the controllability of extraction process.

By executing 3 parts of medical materials of example 3 prescription proportional arrangement, every part of 187.5g divides four groups to test, 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, 12 times of amounts of the 4th group of amount of water, 10 times of amounts, collecting decoction, filter, be condensed into relative density respectively and be 1.25～1.30 clear paste (75 ℃), vacuum drying, with dried cream yield is index, determines amount of water.The results are shown in Table 1:

Table 1: water is carried amount of water

Above-mentioned experimental result shows, the 3rd group (10 times of amounts of amount of water, 8 times of amounts) and the 4th group of (12 times of amounts of amount of water, 10 times of amounts) dried cream yield result differ less, be energy savings and shortening production time, the 3rd group of parameter adopted in decision in big the production, promptly decoct twice, 2 hours for the first time, amount of water was 10 times of amounts, 2 hours for the second time, amount of water was 8 times of amounts.

1.2, demonstration test: carry out confirmatory experiment by above-mentioned preferred technological parameter, the results are shown in Table 2:

Table 2 water is put forward the demonstration test data

The result shows that demonstration test result is more approaching, and dried cream yield meansigma methods is 10.60%, determines that thus process conditions are more reasonable, is: decoct with water secondary, measured 2 hours for 10 times for the first time, measured 2 hours for 8 times for the second time.

2, the investigation of concentration technology

With reference to former capsular concentration technology parameter, according to the extraction process of determining the water extract is concentrated, being concentrated into relative density is the clear paste of 1.15～1.20 (50 ℃), is that index is carried out confirmatory experiment to the clear paste relative density of determining in the original capsule with the wet granulation complexity.

Press each three parts of nine flavor medical materials such as the embodiment 3 prescription configuration Radixs Astragali; divide three groups by testing under the method for making item; extract by fixed extraction process; being concentrated into relative density is the clear paste of 1.15～1.20 (50 ℃); add starch; mixing is granulated with oscillating granulator, observes the situation of wet granulation.The results are shown in Table 3:

The investigation result of table 3 clear paste density

Determine that according to above demonstration test result it is 1.15～1.20 (50 ℃) that traditional Chinese medicine composition extract liquid of the present invention is concentrated into relative density.

3. tablet formulation technical study

3.1. the selection of supplementary product kind: in view of this tablet content is full extractum, have to a certain degree viscosity and moisture absorption, be unfavorable for preparations shaping, simultaneously for adjusting the amount of making, need to add suitable filler and lubricant, intend selecting moderate and widely used starch and magnesium stearate for use, select for use these two kinds of adjuvants to help granule and granulate, also can change the viscosity of sheet, make it be easy to tabletting.Select the bag film-coat can make product have certain moisture-proof role.

3.2, the selection of granulation mode: select wet granulation (relative density of used clear paste is investigated at the concentration technology item) technology for use.Wet granulation can save vacuum drying and the broken process of dried cream powder simultaneously, makes technology simple and easy to do, can save the energy simultaneously.3.3, the selection of magnesium stearate consumption: the dry gained granule of will granulating adds 0.3%, 0.5%, 1.0% magnesium stearate respectively, is that index is investigated the magnesium stearate consumption with the tabletting difficulty, the results are shown in Table 4:

The investigation of table 4 magnesium stearate consumption

According to above result of the test, when the magnesium stearate consumption was 0.5%, mobility of particle was good, and is unilateral smooth.So select magnesium stearate consumption 0.5% to be used for producing.

3.4 art for coating research:

For protecting product not to be subjected to the influence of factors such as humidity, oxygen, the stability of raising medicine, this product intend adopting the bag film-coat.The bag film-coat is with short production cycle, easy and simple to handle, and rate of drying is fast, and medicine is subjected to damp and hotly to influence for a short time, helps improving the quality of products.

3.4.1 EXPERIMENTAL DESIGN:

With reference to relevant documents and materials, the concentration of determining coating powder is 8%, and the key process parameter (inlet temperature, the hydrojet speed of spray gun and coating pan rotating speed) that influences coating is carried out preferably.Because above-mentioned three factors can influence each other,, carry out the orthogonal test of three factors, three levels so serve as the investigation index with outward appearance, the tablet weight variation of slice, thin piece in the coating process.Factor that test is investigated and level design are as follows:

Table 5 art for coating experimental factor and level design table

3.4.2

Process of the test:

(1) test material: stomach dissolution type coating powder, 95% ethanol, purified water.

(2) capital equipment and instrument: Corm Eleocharitis formula coating machine, spray gun, air compressor machine, disintegration time mensuration instrument, analytical balance

(3) coating is write out a prescription substantially: the stomach dissolution type coating powder

95% ethanol

Purified water

The plain sheet of pharmaceutical composition of the present invention

(4) preparation of coating solution:

Get the stomach dissolution type coating powder, the back of weighing adds an amount of 95% ethanol, stirs half an hour, and adding purified water, to be made into concentration be 80% coating solution, continues to stir, and treats that coating powder dissolves fully, promptly.

(5) coating operation:

Get the plain sheet of Chinese medicine composition of the present invention and put in the coating pan, test, keep a close eye on the situation of change of the plain sheet of each group test, its outward appearance and tablet weight variation are checked respectively by designed orthogonal test scheme.The sheet outward appearance is divided into Three Estate: 1. there is honeycomb on a surface; 2. sheet smooth surface; 3. stick together during coating.The check result of tablet weight variation is divided into Three Estate: 1. slice lay particular stress on; 2. meet the requirements; 3. sheet is light partially.

(6) Orthogonal experiment results:

Table 6 orthogonal experiments table

Table 7 an outward appearance analysis of variance table

Result: A〉B〉D〉C

Optimised process: A2 B3 C2 D1

Table 8 tablet weight variation analysis of variance table

Result: B〉A〉D〉C

Optimised process: A2 B3 C3 D1

(7) test result analysis:

Orthogonal experiment results proves, from the sheet outward appearance, A factor inlet temperature has significant difference, optimised process is A2B3C2D1, from tablet weight variation, the hydrojet speed of B factor spray gun has significant difference, optimised process is A2B3C3D1, and C factor coating pan rotating speed there are no significant difference, so take all factors into consideration in conjunction with producing, determine that optimised process is A2B3C2D1, be that inlet temperature is 40 ℃, hydrojet speed is 25kg/h, and the coating pan rotating speed is 7 rev/mins, but consider when the coating process has just begun to carry out, may cause the wearing and tearing of plain sheet, so 4 rev/mins speed the time is selected in beginning for use, along with of the continuous formation of film-coat layer on plain sheet surface, progressively increase rotating speed again, until 7 rev/mins.

Art for coating is carried out confirmatory experiment, the art for coating of determining with above-mentioned orthogonal experiment experimentizes, inlet temperature is 40 ℃, hydrojet speed is 25kg/h, when beginning, selects the coating pan rotating speed 4 rev/mins speed for use, along with of the continuous formation of film-coat layer, progressively increase rotating speed again, up to 7 rev/mins on plain sheet surface.After hydrojet finishes, use hot air drying 20 minutes.The results are shown in following table:

Table 9 art for coating confirmatory experiment is table as a result

Confirmatory experiment is the result show: three confirmatory experiments, every index is all better, parameter stability, can determine that art for coating is: 40 ℃ of inlet temperature, hydrojet speed 25kg/h selects 4 rev/mins speed for use when the coating pan rotating speed begins, along with the continuous formation of film-coat layer on plain sheet surface, progressively increasing rotating speed again, is 7 rev/mins up to rotating speed.After hydrojet finishes, use hot air drying 20 minutes.

The screening of each flavour of a drug discrimination method in experimental example 2. Chinese drug-treated group of the present invention and the thing

Rhizoma Belamcandae

Method one: get this drug combination preparation content and be equivalent to 40g crude drug amount, porphyrize adds methanol 50ml, supersound process 30 minutes, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 2ml methanol makes dissolving, as need testing solution.The negative sample solution that lacks Rhizoma Belamcandae according to test sample preparation and the preparation of sample solution preparation method; Other gets Rhizoma Belamcandae control medicinal material 1g, adds methanol 15ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml medical material solution in contrast.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) test, draw above-mentioned need testing solution, negative sample solution 5 μ l, control medicinal material solution 2 μ l put in same silica gel G F respectively ₂₅₄On the lamellae, be developing solvent, launch, take out, dry, put under the ultra-violet lamp (254nm) and inspect with lower floor's solution of chloroform-methanol-water (8: 2.5: 1).In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, and the speckle colour developing is clear, negative noiseless.

Method two: get this drug combination preparation content and be equivalent to 40g crude drug amount, add methanol 30ml, supersound process 30 minutes filters, and filtrate is concentrated into 1.5ml, as need testing solution.The negative sample solution that lacks Rhizoma Belamcandae according to test sample preparation and the preparation of sample solution preparation method; Other gets Rhizoma Belamcandae control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 1 μ l of above-mentioned three kinds of solution, put respectively on same polyamide film, with chloroform-butanone-methanol (3:1:1) is developing solvent, launches, and takes out, dry, spray is put under the ultra-violet lamp (365nm) and is inspected with the aluminum chloride test solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, but colour developing is more shallow.

In sum, method for optimizing one is as the method for quality control of Rhizoma Belamcandae in this pharmaceutical composition.

Pheretima

Method one: get this drug combination preparation content and be equivalent to 60g crude drug amount, add chloroform 30ml, supersound process 30 minutes filters, and filtrate is concentrated into 1ml, as need testing solution.The negative sample solution that lacks Pheretima according to test sample preparation and the preparation of sample solution preparation method; Other gets Pheretima control medicinal material 1g, adds chloroform 30ml, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned control medicinal material solution 5 μ l, need testing solution, negative sample solution 10 μ l put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (9: 1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color, the speckle colour developing is clear, and negative noiseless.

Method two: get this drug combination preparation content and be equivalent to 60g crude drug amount, add water 30ml, be heated to and boil, put coldly, centrifugal, get supernatant as need testing solution.The negative sample solution that lacks Pheretima according to test sample preparation and the preparation of sample solution preparation method; Other gets lysine reference substance, leucine reference substance, valine reference substance and adds water and make the solution that every 1ml contains 1mg, 1mg and 0.5mg, in contrast product solution.According to thin layer chromatography (appendix VI B) test, draw each 3 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (4:1:1) is developing solvent, launches, and takes out, dry, spray is with ninhydrin solution, and it is clear to be heated to speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color, but the sample separation effect is bad.

Method three: get this drug combination preparation content and be equivalent to 60g crude drug amount, add chloroform 30ml, supersound process 20 minutes filters, and filtrate is put evaporate to dryness in the water-bath, and residue adds chloroform 1ml makes dissolving, as need testing solution.The negative sample solution that lacks Pheretima according to test sample preparation and the preparation of sample solution preparation method; Other gets Pheretima control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-acetone (9:1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, but sample point is unintelligible.

Method four: get this drug combination preparation content and be equivalent to 60g crude drug amount, porphyrize adds chloroform 20ml, close plug, and supersound process 20 minutes filters, and filtrate is concentrated into about 1ml, as need testing solution.The negative sample solution that lacks Pheretima according to test sample preparation and the preparation of sample solution preparation method; Other gets Pheretima control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VIB) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with benzene-ethyl acetate (9:1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, but spot colors is more shallow.

In sum, method for optimizing one is as the method for quality control of Pheretima in this pharmaceutical composition.

The Radix Astragali

Method one: get this drug combination preparation content and be equivalent to 120g crude drug amount, add methanol 60ml, reflux 1 hour, filter, filtrate is added on neutral alumina post (100～120 orders, 5g, on the internal diameter 10～15mm), with 40% methanol 100ml eluting, collect eluent, evaporate to dryness, residue adds water 30ml makes dissolving, with water saturated n-butanol extraction 2 times, each 20ml merges n-butyl alcohol liquid; Wash each 20ml with water 2 times; Discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution.The negative sample solution that lacks the Radix Astragali according to test sample preparation and the preparation of sample solution preparation method; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VI B), draw each 2 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, 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 reference substance chromatograph on, daylight shows down identical sepia speckle, ultra-violet lamp (365nm) shows identical orange-yellow fluorescence speckle down, but that sample point separates is bad.

Method two: get this drug combination preparation content and be equivalent to 120g crude drug amount, porphyrize adds 2% potassium hydroxide methanol solution 50ml, claim to decide weight, reflux keeps little and boiled 1 hour, filter, filtrate adds chloroform-n-butyl alcohol (2: 1) mixed solution extracts 2 times, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 2 times, each 40ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.The negative sample solution that lacks the Radix Astragali according to test sample preparation and the preparation of sample solution preparation method; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, negative sample solution 10 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution of placing below 10 ℃ with chloroform-methanol-water (15: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color, the speckle colour developing is clear, and negative noiseless.

Method three: get this drug combination preparation content and be equivalent to 60g crude drug amount, add ethanol 60ml, reflux 20 minutes filters, the filtrate evaporate to dryness, residue adds 0.3% sodium hydroxide solution 15ml makes dissolving, filters, and filtrate is regulated pH value to 5 ~ 6 with dilute hydrochloric acid, extract with ethyl acetate 15ml jolting, divide and get acetic acid ethyl fluid, filter the filtrate evaporate to dryness with the filter paper that is covered with an amount of anhydrous sodium sulfate.Residue adds ethyl acetate 1ml makes dissolving, as need testing solution.The negative sample solution that lacks the Radix Astragali according to test sample preparation and the preparation of sample solution preparation method; Other gets Radix Astragali control medicinal material, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw above-mentioned three kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate,, launch as developing solvent with chloroform-methanol (10:1), take out, dry, put in the ammonia steam and inspect under the smoked rearmounted ultra-violet lamp (365nm).In the test sample chromatograph, spreading on the corresponding position, show the fluorescence principal spot of same color, but sample point is being more shallow with the control medicinal material color.

Method four: get this drug combination preparation content and be equivalent to 60g crude drug amount, add methanol 50ml, reflux 1 hour, put coldly, filter the filtrate evaporate to dryness, residue adds water 30ml makes dissolving, in order to water saturated n-butanol extraction 2 times, and each 30ml, merge n-butyl alcohol liquid, add 1% sodium hydroxide solution washing 2 times, each 20ml, get n-butyl alcohol liquid in order to n-butyl alcohol saturated be washed to neutrality, get n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.The negative sample solution that lacks the Radix Astragali according to test sample preparation and the preparation of sample solution preparation method; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography, (appendix VI B) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-ethyl acetate-methanol-water (20:40:22:10) is developing solvent, launches, 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 reference substance chromatograph on, show the speckle of same color, but the sample point color is more shallow.

Conclusion: in sum, method for optimizing two is as one of the method for quality control of the Radix Astragali in this pharmaceutical composition.

Herba Epimedii

Method one: get this drug combination preparation content and be equivalent to 40g crude drug amount, porphyrize, the accurate title, decide, put in the 100ml tool plug conical flask accurate Diluted Alcohol 50ml, the close plug of adding, claim to decide weight, supersound process (power 250W, frequency 33kHz) 1 hour, put coldly, supply the weight that subtracts mistake, shake up with Diluted Alcohol, filter, get subsequent filtrate 10ml, evaporate to dryness, residue adds methanol 2ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.The negative sample solution that lacks Herba Epimedii according to test sample preparation and the preparation of sample solution preparation method; Test according to thin layer chromatography (appendix VI B), draw each 10 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica HF 254 lamellae of adhesive with the sodium carboxymethyl cellulose, be developing solvent with ethyl acetate-acetone-methanol-water (10:1:1.5:1), launch, take out, dry, put under the ultra-violet lamp (254nm) and inspect, in the test sample chromatograph, on the corresponding position of reference substance chromatograph, show the speckle of same color, but speckle is unintelligible.

Method two: get this drug combination preparation content and be equivalent to 40g crude drug amount, porphyrize adds 50% ethanol 30ml, reflux 30 minutes is taken out, and puts cold, filter, filtrate is steamed near and is done, and adds water 3ml and makes dissolving, be added on polyamide column (5g, on the post footpath 10～15mm), water 100ml eluting, discard eluent, reuse ethanol 100ml eluting is collected eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.The negative sample solution that lacks Herba Epimedii according to test sample preparation and the preparation of sample solution preparation method; Other gets the icariin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of the yellow version in 2000 of middle traditional Chinese medicines B) test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively on same polyamide film, with dehydrated alcohol-water (55: 44) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical kermesinus speckle, clear spot, and negative noiseless.

Conclusion: in sum, the mode of priority two is as one of the discrimination method of Herba Epimedii in this pharmaceutical composition.

Herba Ephedrae

Method one: get this drug combination preparation content and be equivalent to 40g crude drug amount, porphyrize adds strong ammonia solution 4ml and makes moisteningly, adds chloroform 40ml, and reflux 1 hour filters, and filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.The negative sample solution that lacks Herba Ephedrae according to test sample preparation and the preparation of sample solution preparation method; Other gets the ephedrine hydrochloride reference substance, and chlorination is copied into the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with isopropyl alcohol-n-butyl alcohol-strong ammonia solution (15: 2: 2.5) is developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation, the speckle colour developing is clear and negative noiseless.

Method two: get this drug combination preparation content and be equivalent to 40g crude drug amount, add strong ammonia solution number droplet, add chloroform 10ml again, reflux 1 hour filters, and filtrate evaporate to dryness, residue add methanol 2ml shake well, filter, and filtrate is as need testing solution.The negative sample solution that lacks Herba Ephedrae according to test sample preparation and the preparation of sample solution preparation method; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VI B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-strong ammonia solution (20:5:0.5) is developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation, but separating effect is bad.

Method three: get this drug combination preparation content and be equivalent to 40g crude drug amount, lack the negative sample solution of Herba Ephedrae according to test sample preparation and the preparation of sample solution preparation method; Test according to thin layer chromatography (appendix VI B).Draw each the 3 μ l of need testing solution, negative sample solution and reference substance solution under [assay] item, put respectively on same silica gel g thin-layer plate, with ethanol-ammonia (10:0.5) is developing solvent, launches, and takes out, dry, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show the speckle of same color, but the speckle colour developing is unintelligible, but the speckle colour developing is more shallow.

Conclusion: in sum, method for optimizing one is as the discriminating of this pharmaceutical composition epheday intermedia.

Radix Platycodonis

Method one: get this drug combination preparation content and be equivalent to 40g crude drug amount, inclining content, adds 7% sulphuric acid alcohol-water (1: 3) mixed solution 40ml, reflux 3 hours is put coldly, adds the chloroform jolting and extracts 2 times, each 20ml, combined chloroform liquid adds water 30ml washing, discard water liquid, the chloroform solution anhydrous sodium sulfate dehydration filters, filtrate evaporate to dryness, residue add methanol 2ml makes dissolving, adds activated carbon and handles in right amount, filter, filtrate is as need testing solution.The negative sample solution that lacks Radix Platycodonis according to test sample preparation and the preparation of sample solution preparation method; Other gets Radix Platycodonis control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 6 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (1: 1) 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 principal spot of same color, the speckle colour developing is clear, and negative noiseless.

Method two: get this drug combination preparation content and be equivalent to 40g crude drug amount, extract 2 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butanol extraction and annotates, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.The negative sample solution that lacks Radix Platycodonis according to test sample preparation and the preparation of sample solution preparation method; Other gets Radix Platycodonis control medicinal material 0.5g, adds methanol 10ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 2ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VI B), draw each 2 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-formic acid (16:10:1) 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 principal spot of same color, but that speckle separates is bad.

Method three: get this drug combination preparation content and be equivalent to 40g crude drug amount, porphyrize adds the mixed solution 30ml of chloroform-water-hydrochloric acid (10:10:3), put in the water-bath reflux 1 hour, and divided and get chloroform liquid, evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.The negative sample solution that lacks Radix Platycodonis according to test sample preparation and the preparation of sample solution preparation method; Other gets Radix Platycodonis control medicinal material 2g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VI B), draw need testing solution, negative sample solution 10 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-methanol (20:1) 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, but the speckle colour developing is more shallow.

Conclusion: in sum, method for optimizing one is as the method for quality control of this drug regimen species Radix Platycodonis.

The screening of experimental example 3. content assaying methods

Adopt the content Determination of Icariin in the high-efficient liquid phase color popularize law mensuration medicine of the present invention, to improve quality determining method of the present invention, part test the results are shown in down:

1. the preparation of need testing solution

Get 20 in this product tablet, remove film-coat, porphyrize, get about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 20ml that adds claims to decide weight, supersound process (power 250W, frequency 50kHz), puts coldly, claim to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.Compared different ultrasonic times, content Determination of Icariin in the need testing solution, result such as following table:

The selection of table 10 ultrasonic time

Ultrasonic time	20 minutes	40 minutes	60 minutes	80 minutes
Ultrasonic time	20 minutes	40 minutes	60 minutes	80 minutes	Icariin content (mg/g)	0.4307	0.6232	0.7118	0.7151

As can be seen from the above table, supersound extraction can be extracted the icariin in the medicine of the present invention fully in 60 minutes, so the preparation of need testing solution selects supersound extraction to get final product in 60 minutes.

2. the selection of mobile phase

Get test sample molten night, respectively with the mobile phase of acetonitrile-water with methanol-different proportionings of 0.05mol/L potassium dihydrogen phosphate, the separating effect at each peak in the need testing solution chromatogram relatively the results are shown in following table:

The selection of table 11. mobile phase

Proportion of mobile phase	Methanol-water (40: 60)	Methanol-water (53:47)	Methanol-water (60: 40)	Methanol-0.05mol/L potassium dihydrogen phosphate (50: 55)	Methanol-0.05mol/L potassium dihydrogen phosphate (40: 65)	Methanol-0.05mol/L potassium dihydrogen phosphate (30:75)
Proportion of mobile phase	Methanol-water (40: 60)	Methanol-water (53:47)	Methanol-water (60: 40)	Methanol-0.05mol/L potassium dihydrogen phosphate (50: 55)	Methanol-0.05mol/L potassium dihydrogen phosphate (40: 65)	Methanol-0.05mol/L potassium dihydrogen phosphate (30:75)	The chromatographic peak separating effect	Inferior separating effect has interference	Good separating effect, noiseless	Inferior separating effect has interference	Inferior separating effect has interference	Inferior separating effect has interference	Inferior separating effect has interference

As can be seen from the above table, disturbing does not appear in the good separating effect at each peak in test sample chromatogram when mobile phase is methanol-water (53:47), main peak.

3. the methodological study of content detection

To the detection method of content that medicine of the present invention adopted, carried out related side's science of law from aspects such as linear relationship, stability, precision, repeatability, the response rate and investigated, concrete grammar and result are as follows:

Detecting instrument (room temperature detection): Agilent 1100 type High Performance Liquid Chromatography posts: (Zorbax C18 4.6 * 150mm, 5 μ m) producer: Agilent Techologies Anjelen Sci. ﹠ Tech. Inc (China)

Mobile phase: methanol-water (53:47) detects wavelength: 270nm

Flow velocity: 1.0ml/min column temperature: room temperature

The reference substance source: icariin is purchased lot number: the 0638-0613 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

Assay method: accurate respectively each the 10 μ l of negative controls, reference substance liquid and need testing solution that draw, inject chromatograph of liquid, measure, promptly.

(1) stability test reference substance solution respectively at preparing the back 0,2,4,6,12,24 hour, is measured in accordance with the law, and the result shows that it is basicly stable in 24 hours, the results are shown in following table:

The stability test of table 12 content assaying method

(2) linear relationship is investigated and to be got reference substance solution (52.3 μ g/ml) and shake up, accurate respectively 2,4,6,8,10, the 12 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve, show that icariin is linear between 0.097 μ g-0.582 μ g, its regression equation is:

Area＝13562714*Amt+77626(r＝0.9998)

Table 13 linear relationship is investigated

(3) the accurate need testing solution 10 μ l that draw of precision test repeat sample introduction 5 times, try to achieve relative standard deviation＜2%, the results are shown in following table:

The test of table 14. precision

(4) the text method is pressed in repeatability test, gets five parts of same bolus of drug of the present invention, and every part is measured, and tries to achieve relative standard deviation＜2%, the results are shown in following table:

The test of table 15. repeatability

(5) the recovery test precision take by weighing known content medicinal tablet 9.0g same of the present invention more respectively precision take by weighing icariin reference substance 50mg, be configured to the solution of 0.5mg/ml, precision is measured 10ml, preparation method operation by above-mentioned need testing solution, measure its content, and calculate its response rate, measurement result sees the following form:

Table 16 recovery test

Tested number	Sampling amount (g)	Peoniflorin amount mg in the sample)	Add peoniflorin amount (mg)	Measure peoniflorin amount (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)
Tested number	Sampling amount (g)	Peoniflorin amount mg in the sample)	Add peoniflorin amount (mg)	Measure peoniflorin amount (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)	1	9.0023	6.5579	5.00	11.3209	95.26
2	9.0321	6.5212	5.00	11.1807	93.19			1	9.0023	6.5579	5.00	11.3209	95.26
2	9.0321	6.5212	5.00	11.1807	93.19			3	9.0983	6.5475	5.00	11.3790	96.63	95.13	1.3732
4	9.0432	6.6651	5.00	11.3991	94.68			3	9.0983	6.5475	5.00	11.3790	96.63	95.13	1.3732
4	9.0432	6.6651	5.00	11.3991	94.68			5	9.0119	6.5831	5.00	11.3781	95.90

(6) blank assay

Ratio according to drug prescription taste of Chinese medicine of the present invention, press oral liquid formulations technology, preparation does not contain the negative sample of Herba Epimedii, according to preparation of need testing solution preparation method and detection, negative sample solution is that the identical retention time of peoniflorin reference substance place does not have chromatographic peak as a result, so negative noiseless.

By above methodology examination result as can be seen, its linear relationship of the content assaying method that medicine of the present invention adopted, stability, precision, repeatability etc. are all good, can effectively control drug quality of the present invention.

Experimental example 3. pharmacodynamic experiments

1 material

1.1 medicine and reagent:

Chinese medicinal composition preparation of the present invention is made according to embodiment 6 methods;

HUANGLONGKECHUAN JIAONAN, Yangling Dongke Maidisen Pharmaceutical Co., Ltd produces, the accurate word Z20025060 of traditional Chinese medicines, lot number: 060809; Acecoline (Ach), chemical reagent three factories in Shanghai produce, lot number 031025; Histamine phosphate (Hist), Shanghai Inst. of Biochemistry, Chinese Academy of Sciences provides, lot number 031025; Phenol red, chemical reagent factory in Beijing's produces, lot number 040507; Dimethylbenzene, chemical reagent three factories in Shanghai produce, lot number 031018.

1.2 key instrument YSD-5 type pharmacology, physiology are used instrument more, radio two factories in Bangbu produce; DC-001 type isolated organ analyzer, analytical tool factory in Nanjing produces; 721 type spectrophotometers, Shanghai the 3rd analytical tool factory produces.

1.3 the animal Kunming mouse, SD rat, Cavia porcellus, domestic cat.Above animal male and female dual-purpose, body weight is seen below and is stated.Provide by China Academy of TCM's Experimental Animal Center.

2 methods and result

2.1 antitussive experiment

2.1.1 strong aqua ammonia is caused the influence of mouse cough reaction

60 of mices, body weight 20 ± 2g is divided into 4 groups at random.Of the present invention group and the every d of HUANGLONGKECHUAN JIAONAN group press dosage shown in the table 1 respectively and irritate the continuous 5d of stomach (ig) administration; Normal saline (NS) group and codeine filling with etc. capacity NS, 5d codeine group is irritated with codeine 0.03gKg ^-1Each the group all after the last administration 1h begin the experiment.Mice is put in the airtight glass box respectively, utilize the constant-pressure atomization device in case, to spray people's strong aqua ammonia aerosol, cause the mouse cough reaction.Spray time 10S, spouting liquid 3ml.The record spraying finishes mouse cough number of times in the 2min of back.The results are shown in Table 17.

Table 17 the present invention causes the influence (x ± S) of mouse cough reaction to strong aqua ammonia

Compare * * P＜0.01 with the NS group, with HUANGLONGKECHUAN JIAONAN group ratio, ※ P＜0.05

2.1.2 the present invention draws the influence of coughing threshold value to electricity irritation cat superior laryngeal nerve

32 of domestic cats, body weight 2.5 ± 0.3kg.With pentobarbital sodium 25～30mgKg ^-1Intraperitoneal injection of anesthesia, back of the body position is fixed on the operating-table.Near thyroid cartilage,, find out and separate superior laryngeal nerve, set up shield electrode in order to stimulating along neck midline incision skin.One special glass sleeve pipe, one end is inserted people cat bottleneck throat from the oral cavity, the other end joins with latex tubing and agate king Li Shi tympanites and is connected on the tracing device, so as to the record status of cough.Press the ig of grouping shown in the table 2 administration.30min begins behind the medicine, at regular intervals, stimulates superior laryngeal nerve with rectangular pulse stimulator.Stimulus parameter: the wide 0.5ms of 40 times/S. of frequency ripple, each continued stimulus 5S, adjacent twice stimulation time be 2～5min at interval, and stimulation voltage is ascending progressively to be increased progressively.Cough threshold value with the minimum voltage value that causes cough as drawing, the results are shown in Table 18.Of the present invention group and HUANGLONGKECHUAN JIAONAN group and codeine group are drawn and are coughed threshold value and all obviously improve, and can keep more than the 150min.

Table 18 the present invention draws the influence of coughing threshold value (n=8, x ± s) to electricity irritation cat superior laryngeal nerve

Compare with the NS group ^*P＜0.01 is with HUANGLONGKECHUAN JIAONAN group ratio, ※ P＜0.05

The experiment 2.2 relieving asthma

2.2.1 drawing Cavia porcellus, the present invention breathes heavily preclinical influence

Cavia porcellus is put in people's special glass case, and constant voltage sprays into capacity mixed liquors such as 1% Ach and 0.5% Hist in case.Spray time 20S, spouting liquid 6ml.Finishing rapid breathing time fall of twitching to occur to Cavia porcellus with spraying breathes heavily incubation period as drawing.Select body weight 180 ± 20g Cavia porcellus, 1d rejects responsive person (draw and breathe heavily incubation period〉120S) before the experiment.Get 32 of qualified Cavia porcelluss next day, press table 3 grouping ig administration, 40min opens to draw as spraying and breathes heavily behind the medicine.The results are shown in Table 19 of the present invention groups, HUANGLONGKECHUAN JIAONAN group and aminophylline group draws and breathes heavily equal significant prolongation incubation period.

Table 19 the present invention draws Cavia porcellus and breathes heavily preclinical influence (X ± S)

Compare * * P＜0.01 with the NS group

2.2.2 the present invention is to the influence of isolated helical strips of guinea contraction movement

Get 300 ± 30g Cavia porcellus, the back of hitting unconsciously is taken out trachea rapidly, makes spiral bar, insert the bath that contains Kreds-Henseleit liquid 25ml poor in, logical people mixes oxygen.Tracheal strip one end is fixed in the bath bottom, and the other end links to each other with monitor by tonotransducer.1. add people Hist (final concentration 5.2 * 10 in the bath ^-3MolL ^-1, down together); 2. add Hist in the bath, treat that the tracheal smooth muscle contraction adds the flat electuary of people's cough with asthma (final concentration 8mgml when reaching the peak ^-1, down together); 3. add the flat electuary of people's cough with asthma in the bath, add people Hist behind the 2min; Add Hist in the bath, treat that the tracheal smooth muscle contraction adds people's aminophylline (final concentration 0.5mgml when reaching the peak ^-1).The results are shown in Table 20.Guinea pig tracheal smooth muscle is shunk due to the remarkable antagonism Hist of the flat electuary of cough with asthma, acts on more than the lasting 30min, and is close with aminophylline.

Table 20 the present invention causes influence (n=10, the X ± S) of guinea pig tracheal smooth muscle contraction movement to histamine

The experiment 2.3 eliminate the phlegm

2.3.1 the present invention is to the phenol red influence of mice trachea section excretion

Get 30 of mices, body weight 21 ± 2g.Press the ig of grouping shown in the table 5 administration, continuously 4d.Behind the last administration 30min, every Mus lumbar injection 0.25% phenol red normal saline solution 0.5ml.Animal is put to death in the cervical vertebra dislocation behind the 30min, uses 5% NaHCO ₃Lavation mice trachea 3 times, each 0.5ml.Collect the about altogether 1.5ml of irrigating solution, centrifugal back is sentenced 5% NaHCO with 721 type spectrophotometers at 520nm ₃For the blank pipe is measured absorption value.Carry out rectilinear regression with concentration and absorption value, draw the standard curve equation.Obtain the phenol red concentration of each measured tube according to standard curve, the results are shown in Table 21.

Table 21 the present invention influence phenol red to mice trachea section excretion (X ± S)

Compare * P＜0.05 with the NS group

2.3.2 the flat influence of cough with asthma to rat expectoration amount

32 of rats, body weight 200 ± 20g presses the ig of grouping shown in the table 6 administration, continuously 6d.Water is can't help in the 8-12h fasting before the last administration, and 1h is with 3% pentobarbital sodium intraperitoneal injection of anesthesia behind the medicine, and back of the body position is fixing, cuts off skin along the neck center line, separates trachea.Hit exactly two cartilaginous rings at the thyroid cartilage lower edge and interleave one of people's capillary glass-tube (the about 0.8mm of internal diameter), draw intra-tracheal sputum liquid.Sputum length the results are shown in Table 22 as the expectoration amount in people's capillary glass-tube to inhale.Of the present invention group and chlorination all can significantly increase the expectoration amount by group.

Table 22 the present invention is to the influence of rat expectoration amount (x ± s)

Compare * P＜0.05 with the NS group

2.4 antiinflammatory experiment

2.4.1 the present invention causes the influence of ankle swelling in rat to Ovum Gallus domesticus album

Get 40 of rats, body weight 200 ± 7.6g presses the ig of grouping shown in the table 7 administration.Each all surveys the following position of right ankle joint volume as normal value by the capillary tube amplifying method before organizing medicine.Behind the successive administration 5d, cause inflammation to sufficient sole of the foot injection fresh albumen 0.1ml in the right ankle of every Mus.With the right ankle of different time behind the medicine with lower volume and medicine before the difference of normal value as the inflammatory swelling degree, the results are shown in Table 23, high low dose group of the flat electuary of cough with asthma and indometacin group swelling degree significantly alleviate, and act on more than the lasting 3h.

Table 23 the present invention causes the influence (X ± S) of ankle swelling in rat to Ovum Gallus domesticus album

2.4.2 xylol of the present invention causes the influence of mice auricular concha inflammation

Get 40 of body weight 18 ± 2g mices, press the ig of grouping shown in the table 8 administration.Behind the successive administration 7d, 2 of hard of hearing melted paraxylenes cause inflammation in an every Mus left side.Put to death mice after causing scorching 30min, with diameter 9mm card punch respectively at left ear swelling position and the Umbillcaria esculenta corresponding site respectively lay an auricle and weigh, as the inflammatory swelling degree, the results are shown in Table 24 with left and right sides auricle weight difference.Of the present invention group and indometacin group swelling degree significantly alleviate.

Table 24 xylol of the present invention causes the influence of mice auricular concha inflammation

Experimental result shows: present composition tablet can significantly suppress strong aqua ammonia and cause the mouse cough reaction, obviously improves drawing of electricity irritation cat superior laryngeal nerve and coughs threshold value; Obviously prolonged guinea pig is drawn and is breathed heavily incubation period and antagonism Host and cause guinea pig tracheal smooth muscle and shrink; To little, rat trachea excretion is phenol red and the expectoration amount has remarkable promotion; Ovum Gallus domesticus album caused rat is stamped swelling and mice caused by dimethylbenzene xylene auricular concha inflammation all has remarkable inhibition.Prompting this product has obvious antitussive, relievings asthma, eliminates the phlegm, antiinflammatory action, and except that the phlegm-dispelling functions advantage was not obvious, tablet of the present invention all was better than the HUANGLONGKECHUAN JIAONAN group above-mentioned various using.

Experimental example 5. clinical trials

1. case screening

110 routine cough with asthma patients are divided into 2 groups at random.56 examples are organized in treatment, male's 36 examples, women's 20 examples, age 18-78 year, average 45.5 years old.Acute bronchitis 18 examples wherein, chronic bronchitis 20 examples, bronchial asthma 10 examples, COPD 8 examples.Matched group 54 examples, man's 35 examples, woman's 19 examples, age 19-78 year, average 46.2 years old, acute bronchitis 19 examples wherein, chronic bronchitis 20 examples, bronchial asthma 8 examples, COPD 7 examples, all cases all belong to stage of attack, and data such as the sick kind of two groups of cases, the course of disease, sex, age and Chinese medical discrimination typing are learned by statistics and handled no significant difference (P〉0.05) and have comparability.

2. therapeutic evaluation

1. curative effect judging standard cures: cough, expectorant, asthma shape disappear substantially, pulmonary breathes heavily, wheezing sound disappears.2. produce effects: cough, expectorant, asthma shape is clearly better, pulmonary breathes heavily, wheezing sound obviously alleviates.3. effective: cough, expectorant, asthma shape takes a turn for the better, pulmonary breathes heavily, wheezing sound alleviates.4. invalid: symptom and pulmonary's sign do not have the improvement or the person of increasing the weight of.

3. Therapeutic Method

Therapeutic Method matched group routine is taken HUANGLONGKECHUAN JIAONAN, and Yangling Dongke Maidisen Pharmaceutical Co., Ltd produces, and the accurate word Z20025060 of traditional Chinese medicines is oral, one time 4,3 times on the one.The treatment group gives Chinese medicine composition tablet of the present invention on the basis of above-mentioned conventional therapy, one time 3; 3 times on the one.In the drug administration process, the person gives infection and the treatment of eliminating the phlegm to cough up the purulent sputum, and a week is a course of treatment.One course of treatment of acute bronchitis medication, chronic bronchitis asthma, COPD medication 2-3 course of treatment.

4. therapeutic outcome

Table 25 treatment group and matched group observation of curative effect table

Learn by statistics and handle, there were significant differences for treatment group and matched group curative effect (P＜0.05), further specifies, and medicinal tablet curative effect of the present invention is better than HUANGLONGKECHUAN JIAONAN.

Following embodiment all can realize above invention effect

Following embodiment all can realize the described effect of above-mentioned experimental example

Embodiment 1.

Rhizoma Belamcandae 150g Semen Lepidii (Semen Descurainiae) 150g Herba Epimedii 150g

Fructus Crataegi (life) 375g Radix Platycodonis 75g Herba Houttuyniae 500g

Radix Astragali 225g Herba Ephedrae (processing) 75g Pheretima 75g

Rhizoma dioscoreae 150g Rhizoma Anemarrhenae 75g Gecko 50g

More than 12 flavors, make the preparation of clinical acceptance according to this area routine techniques, as granule, capsule, tablet, soft capsule, pill.

Embodiment 2. capsules

Radix Astragali 225g Herba Ephedrae (processing) 75g Pheretima 75g

Rhizoma dioscoreae 150g Radix Platycodonis 75g

More than eight flavors decoct with water 2 times, each 2 hours, collecting decoction, being concentrated into relative density is 115～1.20, adds dextrin 500g, mixing is made granule, drying, granulate is made 1000 promptly.

[discriminating]

Get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, porphyrize adds methanol 50ml, ultrasonic place 30 minutes, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 2ml methanol makes dissolving, as need testing solution.Other gets Rhizoma Belamcandae control medicinal material 1g, adds methanol 15ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml medical material solution in contrast.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) test, draw above-mentioned need testing solution 5 μ l, control medicinal material solution 2 μ l put in same silica gel G F respectively ₂₅₄On the lamellae, be developing solvent, launch, take out, dry, put under the ultra-violet lamp (254nm) and inspect with lower floor's solution of chloroform-methanol-water (8: 2.5: 1).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 methanol-water (53:47) is mobile phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

Algoscopy

Embodiment 3. tablets

Radix Astragali 225g Pheretima 75g Herba Epimedii 250g

Fructus Crataegi (life) 375g Radix Platycodonis 75g Herba Houttuyniae 500g

Rhizoma Belamcandae 150g Herba Ephedrae (processing) 75g Semen Lepidii (Semen Descurainiae) 150g

More than nine flavor medical materials, decoct with water secondary, each 2 hours, collecting decoction filtered, filtrate is concentrated into the extractum that relative density is 1.15～1.20 (50 ℃), adds starch 140g, granulates drying, the magnesium stearate mix homogeneously of adding 0.5%, tabletting, the bag film-coat is made 1000, promptly.

[discriminating]

[assay]

The assay of Herba Epimedii

Algoscopy

Embodiment 4. soft capsules

Radix Astragali 225g Herba Ephedrae (processing) 75g Pheretima 75g

Rhizoma dioscoreae 150g Rhizoma Anemarrhenae 75g

More than eight flavor decoctings boil time, each hour, collecting decoction, being concentrated into relative density is 1.15～1.20, drying under reduced pressure is pulverized, it is an amount of to add vegetable oil, stirs evenly, and makes 600 of soft capsules, promptly.

[discriminating]

[assay]

The assay of Herba Epimedii

Algoscopy

Embodiment 5. granules

Rhizoma Belamcandae 150g Radix Platycodonis 75g Herba Epimedii 250g

Radix Astragali 225g Herba Ephedrae (processing) 75g Pheretima 75g

Fructus Crataegi (life) 375g Gecko 50g Herba Houttuyniae 500g

More than nine flavor decoctings boil 2 times, each 2 hours, collecting decoction, being concentrated into relative density is 1.15～1.20, adds sucrose 350g, dextrin 150g, mixing is made granule, drying, granulate is made 1000g promptly.

[discriminating]

[assay]

The assay of Herba Epimedii

Algoscopy

Embodiment 6. pills

Rhizoma Belamcandae 150g Radix Platycodonis 75g Herba Epimedii 150g

Radix Astragali 225g Herba Ephedrae (processing) 75g Pheretima 75g

Rhizoma dioscoreae 150g Fructus Crataegi (life) 375g

More than nine the flavor medical materials, Rhizoma dioscoreae, the Radix Astragali, Pheretima, Fructus Crataegi powder are broken into 100 order fine powders, all the other flavour of a drug decoct with water secondary, each 2 hours, collecting decoction filters, and filtrate is concentrated into the concentrated solution that relative density is 1.05～1.10 (50 ℃), with the general ball of above-mentioned medicated powder, make 600, promptly.

[discriminating]

C. get and be equivalent to crude drug 56g drug combination preparation content of the present invention, porphyrize adds 2% potassium hydroxide methanol solution 50ml, claim to decide weight, reflux keeps little and boiled 1 hour, filter, filtrate adds chloroform-n-butyl alcohol (2: 1) mixed solution extracts 2 times, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 2 times, each 40ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution 10 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution of placing below 10 ℃ with chloroform-methanol-water (15: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

[assay]

The assay of Herba Epimedii

Algoscopy

Claims

1. Chinese medicine composition for the treatment of cough illness is characterized in that said composition is to be made by the crude drug of following weight ratio:

Radix Astragali 30-50 weight portion Herba Ephedrae (processing) 10-20 weight portion Pheretima 10-20 weight portion.

2. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

Rhizoma dioscoreae 20-40 weight portion Semen Lepidii (Semen Descurainiae) 25-40 weight portion.

3. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

Rhizoma dioscoreae 20-40 weight portion Rhizoma Anemarrhenae 10-30 weight portion.

4. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

Rhizoma dioscoreae 20-40 weight portion Fructus Crataegi (life) 60-80 weight portion.

5. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

Fructus Crataegi (life) 60-80 weight portion Semen Lepidii (Semen Descurainiae) 25-40 weight portion.

6. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

Fructus Crataegi (life) 60-80 weight portion Gecko 5-15 weight portion.

7. as the described Chinese medicine composition of the arbitrary claim of claim 2-6, it is characterized in that said composition can be to add Herba Houttuyniae 80-120 weight portion in its crude drug.

8. Chinese medicine composition as claimed in claim 1 is characterized in that said composition can be to be made by the crude drug of following weight ratio:

The Rhizoma dioscoreae 30 weight portion Rhizoma Anemarrhenaes 15 weight portion Geckos 10 weight portions.

9. as the preparation method of claim 7 or 8 described Chinese medicine compositions, it is characterized in that this preparation method is:

10. the preparation method of Chinese medicine composition as claimed in claim 9 is characterized in that this method is:

11., it is characterized in that this method of quality control comprises one or more in following discrimination method and/or the assay as the method for quality control of claim 7 or 8 described Chinese medicine compositions:

[discriminating]

A. get and be equivalent to crude drug 10-50g drug combination preparation content of the present invention, porphyrize adds methanol 40-80ml, supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add water 20-60ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 1-3ml methanol makes dissolving, as need testing solution; Other gets Rhizoma Belamcandae control medicinal material 0.5-2g, adds methanol 10-20ml, and supersound process 20-60 minute, filter, filtrate is concentrated into 1ml medical material solution in contrast; According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) test, draw above-mentioned need testing solution 3-10 μ l, control medicinal material solution 1-5 μ l puts in same silica gel G F respectively ₂₅₄On the lamellae, (6-10: 1-5: lower floor's solution 0.5-2) is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp (254nm) and inspects with chloroform-methanol-water; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

B. get and be equivalent to crude drug 40-70g drug combination preparation content of the present invention, porphyrize adds chloroform 10-40ml, and supersound process 10-60 minute, filter, filtrate is concentrated into 0.5-2ml, as need testing solution; Other gets Pheretima control medicinal material 0.5-2g, adds chloroform 10-40ml, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned control medicinal material solution 3-10 μ l, need testing solution 5-20 μ l put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (8-12: 0.5-2) be developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

C. get and be equivalent to crude drug 40-70g drug combination preparation content of the present invention, porphyrize, add 1-3% potassium hydroxide methanol solution 40-60ml, claim to decide weight, reflux, keep little and boiled 0.5-2.0 hour, filter, filtrate adds chloroform-n-butyl alcohol, and (1-3: 0.5-2) mixed solution extracts 1-3 time, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 1-3 time, each 30-50ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 0.5-2.0ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.1-1.0mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution 10 μ l, reference substance solution 1-5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, (10-20: 5-10: 1-5) lower floor's solution of placing below 10 ℃ is developing solvent with chloroform-methanol-water, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

D. get and be equivalent to crude drug 5-20g drug combination preparation content of the present invention, porphyrize adds 30-70% ethanol 30ml, reflux 10-50 minute, take out, put cold, filter, filtrate is steamed near and is done, and adds water 1-5ml and makes dissolving, be added on polyamide column (5g, on the post footpath 10～15mm), water 100ml eluting, discard eluent, reuse ethanol 100ml eluting is collected eluent, evaporate to dryness, residue add ethanol 0.5-2.0ml makes dissolving, as need testing solution; Other gets the icariin reference substance, adds ethanol and makes the solution that every 1ml contains 0.5-2.0mg, in contrast product solution; According to thin layer chromatography (an appendix VI of the yellow version in 2005 of middle traditional Chinese medicines B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with dehydrated alcohol-water (55: 44) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical kermesinus speckle;

E. get and be equivalent to crude drug 20-40g drug combination preparation content of the present invention, porphyrize adds strong ammonia solution 2-8ml and makes moistening, add chloroform 20-60ml, reflux 0.5-2.0 hour, filter, filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, and chlorination is copied into the solution that every 1ml contains 0.5-2.0mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 3-10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with isopropyl alcohol-n-butyl alcohol-strong ammonia solution (10-20: 1-3: 1-5) be developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation;

F. get and be equivalent to crude drug 20-50g drug combination preparation content of the present invention, add 7% sulphuric acid alcohol-water (0.5-2.0: 1-5) mixed solution 40ml, reflux 1-5 hour, put coldly, add the chloroform jolting and extract 1-3 time, at every turn 10-30ml, combined chloroform liquid adds water 20-40ml washing, discards water liquid, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methanol 1-3ml makes dissolving, add activated carbon and handle in right amount, filter, filtrate is as need testing solution; Other gets Radix Platycodonis control medicinal material 0.5-2.0g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 2-10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (0.5-2.0: 0.5-2.0) be 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 principal spot of same color;

[assay]

The assay of Herba Epimedii

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol-water (40-60:60-40) is mobile phase; The detection wavelength is 200-300nm; Number of theoretical plate calculates by the icariin peak should be not less than 2000;

It is an amount of that the icariin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 0.05-0.5mg, promptly;

The preparation of need testing solution is got and is equivalent to crude drug 30-50g drug combination preparation content of the present invention, and porphyrize is got about 0.5-2.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 10-20ml that adds, claim to decide weight, supersound process (power 250W, frequency 50kHz) 0.5-2.0 hour, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 2-10 μ l of need testing solution of drawing of algoscopy injects chromatograph of liquid, measures, promptly;

12. the method for quality control of Chinese medicine composition as claimed in claim 11 is characterized in that this method of quality control comprises one or more in following discrimination method and/or the assay:

[discriminating]

A. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, porphyrize adds methanol 50ml, supersound process 30 minutes, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, adding the n-butyl alcohol jolting extracts three times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds 2ml methanol makes dissolving, as need testing solution; Other gets Rhizoma Belamcandae control medicinal material 1g, adds methanol 15ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml medical material solution in contrast; According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) test, draw above-mentioned need testing solution 5 μ l, control medicinal material solution 2 μ l put in same silica gel G F respectively ₂₅₄On the lamellae, be developing solvent, launch, take out, dry, put under the ultra-violet lamp (254nm) and inspect with lower floor's solution of chloroform-methanol-water (8: 2.5: 1); In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

B. get and be equivalent to crude drug 56g drug combination preparation content of the present invention, porphyrize adds chloroform 30ml, and supersound process 30 minutes filters, and filtrate is concentrated into 1ml, as need testing solution; Other gets Pheretima control medicinal material 1g, adds chloroform 30ml, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned control medicinal material solution 5 μ l, need testing solution 10 μ l put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (9: 1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

C. get and be equivalent to crude drug 56g drug combination preparation content of the present invention, porphyrize adds 2% potassium hydroxide methanol solution 50ml, claim to decide weight, reflux keeps little and boiled 1 hour, filter, filtrate adds chloroform-n-butyl alcohol (2: 1) mixed solution extracts 2 times, each 50ml, merge extractive liquid,, the ammonia solution saturated with n-butyl alcohol washs 2 times, each 40ml, discard ammonia solution, chloroform-n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution 10 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution of placing below 10 ℃ with chloroform-methanol-water (15: 7: 2) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

D. get and be equivalent to crude drug 11g drug combination preparation content of the present invention, porphyrize adds 50% ethanol 30ml, reflux 30 minutes is taken out, and puts cold, filter, filtrate is steamed near and is done, and adds water 3ml and makes dissolving, be added on polyamide column (5g, on the post footpath 10～15mm), water 100ml eluting, discard eluent, reuse ethanol 100ml eluting is collected eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the icariin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography (an appendix VI of the yellow version in 2000 of middle traditional Chinese medicines B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with dehydrated alcohol-water (55: 44) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical kermesinus speckle;

E. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, porphyrize adds strong ammonia solution 4ml and makes moisteningly, adds chloroform 40ml, and reflux 1 hour filters, and filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, and chlorination is copied into 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 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with isopropyl alcohol-n-butyl alcohol-strong ammonia solution (15: 2: 2.5) is developing solvent, launch, take out, dry, spray is with ninhydrin 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 reference substance chromatograph on, show identical punctation;

F. get and be equivalent to crude drug 37.5g drug combination preparation content of the present invention, add 7% sulphuric acid alcohol-water (1: 3) mixed solution 40ml, reflux 3 hours, put coldly, add the chloroform jolting and extract 2 times, each 20ml, combined chloroform liquid adds water 30ml washing, discards water liquid, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methanol 2ml makes dissolving, add activated carbon and handle in right amount, filter, filtrate is as need testing solution; Other gets Radix Platycodonis control medicinal material 1g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (1: 1) 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 principal spot of same color;

[assay]

The assay of Herba Epimedii

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol-water (53:47) is mobile phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the icariin peak should be not less than 2000;

It is an amount of that the icariin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 0.1mg, promptly;

The preparation of need testing solution is got and is equivalent to crude drug 37.5g drug combination preparation content of the present invention, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 20ml that adds, claim to decide weight, supersound process (power 250W, frequency 50kHz) 1 hour, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

Algoscopy

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