CN101496870B

CN101496870B - Chinese medicinal composition for resolving phlegm and suppressing cough as well as preparation method and quality control method thereof

Info

Publication number: CN101496870B
Application number: CN2008100569267A
Authority: CN
Inventors: 付立家; 付建家
Original assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Current assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Priority date: 2008-01-28
Filing date: 2008-01-28
Publication date: 2011-12-07
Anticipated expiration: 2028-01-28
Also published as: CN101496870A

Abstract

The present invention relates to a Chinese traditional medicine composition for relieving cough and reducing sputum, a method for preparing the same and a method for controlling the quality. The Chinese traditional medicine composition comprises bulk drugs of pummelo peel, dried orange peel, rhizoma pinellinae praeparata, coltsfoot flower, balloon flower, bitter apricot kernel (fried without peel), tuckahoe, trichosanthes bark, liquorice, aster, dwarf lilyturf root, rhizoma anemarrhenae, radix rehmanniae, gypsum, perilla seed (fried), and the like which are added with auxiliary materials and are prepared into clinically acceptable dosage forms including capsules, soft capsules, pills, tablets, powder, injections, granules, mixtures, and the like. The quality control method for the Chinese traditional medicine composition identifies the dwarf lilyturf root, the liquorice, the bitter apricot kernel, the dried orange peel, the balloon flower and the trichosanthes bark, performs content determination on the pummelo peel, and ensures the curative effect of the medicine. The Chinese traditional medicine composition has the efficacy of relieving cough and reducing sputum, and is applied to treating cough with more sputum, fullness sensation in chest with short breath, and dry pharynx and tickling throat caused by phlegm heat lung obstruction.

Description

A kind of detection method of relieving cough and reducing sputum Chinese medicine composition

Technical field

The present invention relates to a kind of Chinese medicine composition and preparation method thereof and method of quality control, particularly a kind of relieving cough and reducing sputum Chinese medicine composition and preparation method thereof and method of quality control.

Background technology

From physiological angle, cough is a kind of body protective sexuality, and it can be discharged the sputum in the respiratory tract, foreign matter, thereby the cleaning of maintenance respiratory tract and unobstructed is beneficial to healthy.But more frequent and violent cough will have influence on people's work, living and studying, at this moment just should select the relieving cough and reducing sputum medicine of suiting the medicine to the illness for use according to symptom.Some more serious diseases usually also are attended by the cough symptom, as pleurisy, spontaneous pneumothorax, pulmonary tuberculosis, lung cancer, heart failure etc., cough all can occur.In the medicine of expelling phlegm and arresting coughing, Western medicine and Chinese patent drug have his own strong points: Western medicine is directly suited the medicine to the illness, instant effect now, but habituation is arranged, need the complex treatment of associating other drug more, need use antibiotics simultaneously as respiratory tract infection, flu then need be share anti-cold medicine; Chinese patent drug commonly used uses as suiting the medicine to the illness, determined curative effect, though take effect not as Western medicine rapid, take stopgap measures and effect a permanent cure.

Summary of the invention

First purpose of the present invention is to provide a kind of relieving cough and reducing sputum Chinese medicine composition; Second purpose of the present invention is to provide the preparation method of this Chinese medicine composition; The 3rd purpose of the present invention is to provide the method for quality control of this Chinese medicine composition.

Content of the present invention is to be achieved through the following technical solutions:

A kind of relieving cough and reducing sputum Chinese medicine composition is to be made by the bulk drug of following weight ratio:

Exocarpium Citri Grandis 50-100 weight portion, dried orange peel 50-100 weight portion, rhizoma arisaematis 50-100 weight portion, loguat leaf 50-100 weight portion, balloonflower root 30-70 weight portion, semen armeniacae amarae (peeling is fried) 30-70 weight portion, Poria cocos 30-70 weight portion, PERICARPIUM TRICHOSANTHIS 30-70 weight portion, Radix Glycyrrhizae 20-50 weight portion, aster 20-50 weight portion, the tuber of dwarf lilyturf 20-50 weight portion, wind-weed 20-50 weight portion, root of purple-flowered peucedanum 10-35 weight portion, gypsum 10-35 weight portion, banksia rose 10-35 weight portion.

Rhizoma arisaematis in the relieving cough and reducing sputum Chinese medicine composition of the present invention can be substituted by rhizoma typhonii, semen brassicae, rhizoma pinellinae praeparata, Inula britannica chinensis.

Loguat leaf in the relieving cough and reducing sputum Chinese medicine composition of the present invention can be substituted by the tuber of stemona, tussilago, lepidium seed, the root bark of white mulberry.

The banksia rose in the relieving cough and reducing sputum Chinese medicine composition of the present invention can be substituted by perilla seed (stir-fry).

The root of purple-flowered peucedanum in the relieving cough and reducing sputum Chinese medicine composition of the present invention can be substituted by caulis bambusae in taenian, bamboo juice, glutinous rehmannia, chlorite schist.

This Chinese medicine composition is to be made by the bulk drug of following weight ratio:

Exocarpium Citri Grandis 66 weight portions, dried orange peel 66 weight portions, rhizoma arisaematis 66 weight portions, loguat leaf 66 weight portions, balloonflower root 44 weight portions, semen armeniacae amarae (peeling is fried) 44 weight portions, Poria cocos 44 weight portions, PERICARPIUM TRICHOSANTHIS 44 weight portions, Radix Glycyrrhizae 33 weight portions, aster 33 weight portions, the tubers of dwarf lilyturf 33 weight portion, the wind-weed 33 weight portions, the root of purple-flowered peucedanum 22 weight portions, gypsum 22 weight portions, the banksia rose 22 weight portions.

Chinese medicine composition preparation method of the present invention is: gesso is broken into meal, and boiling 1-3 time each 0.5-2 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, semen armeniacae amarae four flavor steam distillations are collected distillate 200-300 parts by volume; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 50-70%, stirs evenly, and leaves standstill 16-36 hour, filters filtrate for later use; Ten flavors such as all the other rhizoma arisaematis, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 50-70% ethanol, flood after 12-36 hour diacolation in accordance with the law, the collection liquid 2500-3000 parts by volume of filtering, merge with above-mentioned reserve liquid, decompression recycling ethanol is to there not being the alcohol flavor, merge with the parget water decocting liquid, be concentrated into the clear cream of relative density 1.06 (50 ℃), add conventional auxiliary material again, through conventional method, make the preparation of clinical acceptance.

Chinese medicine composition preparation method of the present invention is preferably: gesso is broken into meal, and the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, semen armeniacae amarae four flavor steam distillations are collected distillate 250 parts by volume; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; Ten flavors such as all the other rhizoma arisaematis, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect liquid 2700 parts by volume of filtering, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add conventional auxiliary material again,, make the oral solid formulation of clinical acceptance through conventional method.

The preparation method of Chinese medicine composition mixture of the present invention is: gesso is broken into meal, and the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, semen armeniacae amarae four flavor steam distillations are collected distillate 250 parts by volume; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; Ten flavors such as all the other rhizoma arisaematis, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect the liquid 2700ml that filters, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add sucrose 80 weight portions, boil, left standstill 24 hours, filter, add ethyl hydroxy benzoate 0.3 weight portion, benzoic acid 0.5 weight portion (both use an amount of hot water dissolving earlier) and distillate, add water and adjust total amount to 950 parts by volume, stir evenly, refrigerate 48 hours, get supernatant, embedding, sterilization, promptly.

Described parts by volume/weight portion is corresponding with g/ml.

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

[discriminating]

(1) get pharmaceutical composition of the present invention and be equivalent to crude drug 20-30g,, add hydrochloric acid 2-4ml, put in the water-bath heating 0.5-2 hour, to put coldly, the 20-40ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 20-40 minute, filter, filtrate is concentrated into 30-50ml, shines medicinal material solution in pairs with legal system.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (3.5-4.5: 0.5-1.5) be developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 100～110 ℃ of heating 4-6 minute.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

(2) get pharmaceutical composition of the present invention and be equivalent to crude drug 10-15g, add hydrochloric acid 2-4ml, reflux 0.5-2 hour, put cold, add the methenyl choloride jolting and extract 1-3 time, each 10-20ml merges the methenyl choloride extract, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (8-12: 15-25: 5-10: 0-1) be developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 100～110 ℃ of heating 4-6 minute.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(3) get pharmaceutical composition of the present invention and be equivalent to crude drug 10-15g, adding the water-saturated n-butanol jolting extracts 1-3 time, each 15-25ml, merge n-butanol extracting liquid, use ammonia scrubbing 1-3 time, each 15-25ml, discard ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, (10-15: 5-10: 1-3) lower floor's solution of the layering of placement below 10 ℃ is developping agent with chloroform-methanol-water, launch, take out, dry, (phosphomolybdic acid 2g adds water 20ml and makes dissolving with the phosphomolybdic acid sulfuric acid solution in spray, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 100～110 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(4) get pharmaceutical composition of the present invention and be equivalent to crude drug 10-15g, adding sherwood oil (60 ℃～90 ℃) 15-25ml jolting extracts 1 time, discard sherwood oil, adding the ethyl acetate jolting extracts 1-3 time, each 15-25ml merges the ethyl acetate extract, evaporate to dryness, residue adds methyl alcohol 2ml makes dissolving, as need testing solution.Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, (10-15: 5-10: 1-3) lower floor's solution of the layering of placement below 10 ℃ is developping agent with chloroform-methanol-water, launch about 10-20cm, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(5) get pharmaceutical composition of the present invention and be equivalent to crude drug 10-15g, add 10% ethanol solution of sulfuric acid 4-6ml, reflux 4-6h, put coldly, extract 1-3 time with the chloroform jolting, at every turn 15-25ml, combined chloroform liquid, add water 20-40ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ether (45-55: 45-55) be developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing at 100～110 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(6) get pharmaceutical composition of the present invention and be equivalent to crude drug 15-20g, add water saturated normal butyl alcohol and extract 1-3 time, each 25-40ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃)-ethyl acetate (35-45: 5-15) be developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show mutually the spot of color just.

[assay]

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

Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With methanol-water-acetic acid (30-40: 60-70: 0-1) be moving phase; The detection wavelength is 283nm; Column temperature is 40 ℃.Theoretical cam curve is calculated by the aurantiin peak should be not less than 3000.

The preparation of reference substance solution is taken at 100～120 ℃ of about 15mg of aurantiin reference substance that are dried to constant weight, and accurate the title decides, and puts in the 100ml measuring bottle, adds methyl alcohol and makes dissolving in right amount and be diluted to scale, shakes up; Precision is measured 3ml, puts in the 10ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets (every 1ml contains aurantiin 45 μ g).

The preparation precision of need testing solution is measured this product 1ml, puts in the 25ml measuring bottle, adds methyl alcohol and is diluted to scale, shakes up, and filters, and gets subsequent filtrate, promptly.

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

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

[discriminating]

(1) get pharmaceutical composition mixture 40ml of the present invention, add hydrochloric acid 3ml, put in the water-bath heating 1 hour, put coldly, the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 30 minutes filters, and filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (4: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

(2) get pharmaceutical composition mixture 20ml of the present invention, add hydrochloric acid 3ml, reflux 1 hour is put coldly, adds the methenyl choloride jolting and extracts 2 times, and each 15ml merges the methenyl choloride extract, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) be developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(3) get pharmaceutical composition mixture 20ml of the present invention, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch, take out, dry, (phosphomolybdic acid 2g adds water 20ml and makes dissolving with the phosphomolybdic acid sulfuric acid solution in spray, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(4) get pharmaceutical composition mixture 20ml of the present invention, add sherwood oil (60～90 ℃) 20ml jolting and extract 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml merges the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch about 15cm, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(5) get pharmaceutical composition mixture 20ml of the present invention, add 10% ethanol solution of sulfuric acid 5ml, reflux 3h, put coldly, extract 2 times each 20ml with the chloroform jolting, combined chloroform liquid, add water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 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 developping agent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(6) get pharmaceutical composition mixture 30ml of the present invention, add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃)-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color.

[assay]

Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With methanol-water-acetic acid (38: 62: 0.5) is moving phase; The detection wavelength is 283nm; Column temperature is 40 ℃.Theoretical cam curve is calculated by the aurantiin peak should be not less than 3000.

The preparation of reference substance solution is taken at 110 ℃ of about 15mg of aurantiin reference substance that are dried to constant weight, and accurate the title decides, and puts in the 100ml measuring bottle, adds methyl alcohol and makes dissolving in right amount and be diluted to scale, shakes up; Precision is measured 3ml, puts in the 10ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets (every 1ml contains aurantiin 45 μ g).

Get the every 1ml of pharmaceutical composition mixture of the present invention and contain Exocarpium Citri Grandis with aurantiin (C ₂₇H ₃₂O ₁₄) meter, must not be less than 0.80mg.

Pharmaceutical composition clearing lung-heat of the present invention, cough-relieving is reduced phlegm, and can fundamentally treat because coughing with a lot of sputum, the fullness sensation in chest that pulmonary retention of phlegmopyrexia causes breathed hard, the dry throat larynx is itched.The present composition is compared existing preparation and is possessed good drug effect, and scope of the present invention through screening, finds in some scope of composition, to possess more outstanding drug effect unexpectedly when can realizing drug effect of the present invention.Rhizoma arisaematis in the Chinese medicine composition, rhizoma typhonii, semen brassicae, rhizoma pinellinae praeparata, Inula medicine for warming and resolving cold-phlegm mainly are applicable to the cough and asthma that cold phlegm, wet phlegm cause, excessive dilute and white sputum; Loguat leaf, the tuber of stemona, tussilago, lepidium seed, the root bark of white mulberry belong to that the hot phlegm medicine in Qinghua is applicable to mainly that hot phlegm causes coughs and breathes heavily uncomfortable in chestly, and the yellow thickness of phlegm is difficult for bringing up; The banksia rose, perilla seed (stir-fry) belong to the strengthening the spleen and reducing phlegm medicine and mainly are applicable to chest gastral cavity turgor, and food is few vomits and diarrhoea coughing with a lot of sputum; The root of purple-flowered peucedanum, caulis bambusae in taenian, bamboo juice, glutinous rehmannia, chlorite schist belong to that the hot phlegm medicine in Qinghua is applicable to mainly that hot phlegm causes coughs and breathes heavily uncomfortable in chestly, and the yellow thickness of phlegm is difficult for bringing up.

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 developping agent, make and differentiate that specificity is fine, and method is economic and practical, the result is quick, and can both use different thin layer plates.Pass through screening in the content assaying method to sample, test sample disposal route, the selection of developping agent, make content assaying method effectivelyly to carry out quality control, and compare more stable that product that additive method measures shows on drug effect with the product that this method is measured to product.

Embodiment

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

Experimental example 1 pharmaceutical composition pharmacodynamic study of the present invention

1 material

1.1 medicine medicine group of the present invention I makes according to embodiment 6 methods; Medicine group II of the present invention makes according to embodiment 7 methods; Medicine group III of the present invention makes according to embodiment 8 methods; The Chinese medicine positive controls, 'Baikejing ' syrup; Codeine phosphate tablets; Ammonium chloride; Ammoniacal liquor; It is pure that agents useful for same is analysis.

1.2 animal NIH mouse, frog.

1.3 statistics is selected for use, and t checks relatively between group.

2 methods

2.1 antitussive effect

2.1.1 ammoniacal liquor is drawn the influence of coughing

Select 18～22g mouse (spraying 20s, making mouse cough is qualified more than 25 times/30s) for use, male and female half and half are divided into 5 groups at random, and the ig administration (first group, equal-volume distilled water; Second and third group, medicine group I of the present invention, II are equivalent to crude drug 0.3g/kg; The 4th group, Chinese medicine positive controls 0.3g/kg, the 5th group, codeine 10ml), every day 1 time, continuous 3d, 1h behind the last medicine draws the method for coughing by mouse ammoniacal liquor and experimentizes, and compares between the work group, the results are shown in Table 1.

Table 1 pair ammoniacal liquor draw the influence of coughing number of times (x ± s, n=15)

Compare with the distilled water group ^*P＜0.05, ^*P＜0.01; Compare △ P＜0.05, △ △ P＜0.01 with the Chinese medicine positive controls;

Medicine group I of the present invention, II can significantly reduce the cough number of times of mouse, and relatively there were significant differences with control group, and relatively there were significant differences with the Chinese medicine positive controls.

2.1.2 sulphuric dioxide is drawn the influence of coughing

Select 90 of the NIH mouse of cough latent period 15s for use, body weight 18～22g, male and female half and half.Be divided into 5 groups at random, medication is the same, and 1h behind the last medicine experimentizes, and compares between the work group, the results are shown in Table 2.

Table 2 pair sulphuric dioxide draw cough preclinical influence (x ± s, n=18)

Cough latent period but medicine group I of the present invention, II significant prolongation sulphuric dioxide draw, relatively there were significant differences with control group, and relatively there were significant differences with the Chinese medicine positive controls.

2.2 phlegm-dispelling functions

2.2.1 influence to the phenol red discharge rate of mouse tracheae

Select 55 of 18～22gNIH mouse for use, male and female half and half are divided into 5 groups at random, and medication is the same, and 1h behind the last medicine by the phenol red expelling phlegm method experiment of mouse, measures phenol red discharge rate, the results are shown in Table 3.

The influence of the table 3 pair phenol red discharge rate of mouse tracheae (x ± s, n=11)

[0073]?

Medicine group I of the present invention, II and Chinese medicine positive controls can significantly increase the phenol red discharge rate of mouse tracheae, and relatively there were significant differences with control group, and relatively there were significant differences for medicine group I of the present invention, II and Chinese medicine positive controls.

2.2.2 in body frog oral mucosa effect on ciliary movement

Select 50～100g frog for use, destroy spinal cord with pin, be fixed on the frog board, fully expose upper jaw mucosal surface, local medicine group I of the present invention, II, Chinese medicine positive controls, 0.0125% isoprel, the equivalent physiological saline of dripping is surveyed the required time from the start line to the finishing line of 3min wood chip before and after the administration with stopwatch, write down 3 times, average, carry out statistical treatment, the results are shown in Table 4.

The influence of table 4 pair frog oral mucosa ciliary movement speed (x ± s, n=11)

Every kg body weight animals administer 0.1ml, ↑ expression is accelerated, and ↓ expression is slowed down

Compare with the physiological saline group ^*P＜0.05, ^*P＜0.01; Compare △ P＜0.05, △ △ P＜0.01 with the Chinese medicine positive controls;

Medicine group I of the present invention, II can significantly increase the ciliary movement of frog oral mucosa, and relatively there were significant differences with physiological saline, and relatively there were significant differences with the Chinese medicine positive controls, illustrate that said preparation has the effect that promotes expectoration.

3 brief summaries

Medicine group of the present invention can significantly reduce the number of times that ammoniacal liquor causes mouse cough, energy significant prolongation sulphuric dioxide causes the latent period of mouse cough, can significantly increase the phenol red discharge rate of mouse tracheae, can significantly increase the ciliary movement of frog oral mucosa, point out this medical instrument that relieving cough and reducing sputum effect is arranged.

The inhibiting research of experimental example 2 medicine group pair cell cytochrome p 450s of the present invention (CYP3A4 and CYP1A2)

Medicine group of the present invention is a kind of antitussive that is prepared from by plurality of Chinese composition such as Exocarpium Citri Grandis, rhizoma arisaematis, dried orange peel etc., because it has good expelling phlegm and arresting coughing effect and is able to widespread use clinically.Medicine group of the present invention contains number of chemical compositions such as flavone compound aurantiamarin, citral and monoterpenes compound.CYP3A4 is important Cytochrome P450, the metabolism of the clinical medicine of catalysis 60%.

In experimentation, our surprised discovery medicine group of the present invention has inhibiting effect to CYP3A4 and CYP1A2, causes the medicine of catalysis metabolism to produce bad drug interaction.

Materials and methods

1. material

Medicine group of the present invention, Dormicum and 1-hydroxyl Dormicum; 1, the 7-dimethyl xanthine; Caffeine.Chloroform, formic acid and 2-propyl alcohol are analytical reagent, and methyl alcohol is chromatographically pure reagent.

2. method

Tried: the experimenter is 10 healthy male volunteers, 23 ± 2 years old mean age, body weight standard body weight ± 20% in, all experimenters are through health check-up, and are healthy, tried not take in preceding 2 weeks other any medicines.

Medication adopts cross-over design, 10 experimenters are divided into two groups at random, the medicine group of the present invention (according to technology preparation among the embodiment 6) that first three day A organizes oral clinical dosage is equivalent to crude drug 15g/ day, B organizes the placebo of oral isodose, and 4d begins two groups of all oral 7.5mg Dormicums of A, B and 100mg caffeine.In the 0th, 0.25,0.5,1,2,3,4,5,6 and 8h gather venous blood 3ml, two groups of intersection behind the wash-out 10d repeat previous stage and test.Blood sample is centrifugal, separated plasma, and-40 ℃ of preservations are to be measured.

The mensuration of Dormicum and caffeine and metabolic product thereof: Dormicum and caffeine and metabolic product thereof are measured with HPLC-MS, chromatographic column is Xterra TM MS C18 (3.9 * 150mm, 5um), moving phase is that water (contains 0.01% formic acid, pH=4.1)-and methyl alcohol, adopt linear gradient elution method, preceding 10min ratio is 52: 48, ratio becomes 95: 5 then, and flow velocity is 0.2mlmin ^-1, column temperature is 40 ℃.Detecting device is ESI, and voltage is set to capillary voltage: 3.2KV, taper hole voltage: 45v, extraction voltage: 6v.

Sample preparation: get blood plasma 200 μ l and interior mark 10 μ l (alprazolam, 2.03mgL ^-1) adding 2ml extraction solvent after the centrifuge tube mesoscale eddies of 15ml mixes (chloroform: 2-propyl alcohol=9: 1), vortex 3min, it is centrifugal that (3000rmin 5min), separates organic phase, and organic phase dries up with nitrogen under 40 ℃ of conditions, and is residual to be measured with dissolve with methanol.

Data processing and statistical study: AUC _0-shCalculate C with trapezoidal method _MaxAnd t _MaxCan directly read from drug-time curve.Ke and t _1/2Utilization pharmacokinetics special software 3P87 calculates.The difference of pharmacokinetic parameter is added up with SPSS (10.0version) before and after the administration, and paired t-test is adopted in significance test, and P＜0.05 thinks that there were significant differences.

The result

Dormicum, 1-hydroxyl Dormicum, caffeine and 1 before and after the administration, the blood concentration and the pharmacokinetic parameter of 7-dimethyl xanthine see Table 5-8.

The influence of table 5 couple Dormicum and 1-hydroxyl Dormicum's blood concentration (μ gL)

Table 6 pair caffeine and 1, the influence of the blood concentration of 7-dimethyl xanthine (μ gL)

[0103]?

Table 7 Dormicum pharmacokinetic parameter

Table 8 caffeine pharmacokinetic parameter

[0108]?

The medicine group of the present invention that gives clinical dosage is after three days, Dormicum's Cmax (C _Max) and medicine area under curve raising for the moment (116.8 ± 64.32vs127.3 ± 54.96, P＞0.05; 54.20 ± 33.15vs60.00 ± 32.76, P＞0.05), same, peak time (T) slightly improves, but there was no significant difference (0.45 ± 0.10vs0.58 ± 0.35, P＞0.05).Dormicum's elimination half life period (t _1/2) significantly improve (1.86 ± 0.35vs2.07 ± 0.36, P＜0.05).Before and after the oral medicine group of the present invention, the pharmacokinetic parameter of caffeine, (10.40 ± 4.23vs12.83 ± 6.00, P=0.163), other parameter all was irregular variation area slightly improved under drug-time curve.

The result shows: give medicine group of the present invention after three days, Dormicum's AUC, C _Max, t _MaxAll increase t _1/2Significantly improve, point out medicine group of the present invention can suppress Dormicum's metabolism.Because Dormicum's 1 monohydroxy metabolism by CYP3A4 catalysis, can be represented the activity of CYP3A4 with Dormicum's 1 monohydroxy metabolism in the research.Medicine group of the present invention is because it is inhibited to CYP3A4 to Dormicum's inhibiting effect.The CYP3A4 that suppresses in the intestines and stomach causes C _MaxAnd t _MaxMain cause, the CYP3A4 that suppresses in the liver causes t _1/2The main cause that prolongs, two kinds of factors all can cause the increase of AUC.

The screening of experimental example discrimination test 3 tuber of dwarf lilyturf

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 24.08g, porphyrize adds hydrochloric acid 3ml, puts in the water-bath heating 0.5 hour, puts coldly, and the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 24.08g, porphyrize adds hydrochloric acid 3ml, puts in the water-bath heating 1 hour, puts coldly, and the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 24.08g, porphyrize adds hydrochloric acid 3ml, puts in the water-bath and heats 1.5 hours, put coldly, add sherwood oil 30ml jolting and extract, divide and get sherwood oil liquid, evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.

Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 20-40 minute, filter, filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system.Get to lack and write out a prescription the tuber of dwarf lilyturf, press the technology preparation scarce tuber of dwarf lilyturf of negative preparation among the embodiment 6, prepare by the test sample preparation method and lack the negative controls tuber of dwarf lilyturf.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (4: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The preparation of table 9. need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: toluene-methyl alcohol-glacial acetic acid (80: 5: 0.1)

Developping agent two: methenyl choloride-acetone (4: 1)

Developping agent three: methenyl choloride-acetone (4.5: 0.5)

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 24.08g, add hydrochloric acid 3ml, put in the water-bath heating 1 hour, put coldly, the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 30 minutes filters, and filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system.Get to lack and write out a prescription the tuber of dwarf lilyturf, press the technology preparation scarce tuber of dwarf lilyturf of negative preparation among the embodiment 6, prepare by the test sample preparation method and lack the negative controls tuber of dwarf lilyturf.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 5 μ 1 of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch respectively, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

Table 10. developping agent proportioning preferred

When developping agent is methenyl choloride-acetone (4: 1) as can be seen from the above table, launch effectively on thin layer plate, principal spot is clear, does not have hangover, and is identical with the speckle displacement and the color of reference substance, is fit to testing requirements.

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 24.08g, add hydrochloric acid 3ml, put in the water-bath heating 1 hour, put coldly, the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 30 minutes filters, and filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system.Get to lack and write out a prescription the tuber of dwarf lilyturf, press the technology preparation scarce tuber of dwarf lilyturf of negative preparation among the embodiment 6, prepare by the test sample preparation method and lack the negative controls tuber of dwarf lilyturf.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw above-mentioned two kinds of solution each 1 μ l, 3 μ l, 5 μ l, 10 μ l, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (4: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

Table 11. sample solution point sample amount optimization experiment is table as a result

Test sample point sample amount is when 5 μ l as can be seen from the above table, and color developing effect is good on thin layer plate, is fit to testing requirements.The screening of experimental example 4 Radix Glycyrrhizae discrimination tests

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds hydrochloric acid 3ml, reflux 0.5 hour, put coldly, add the methenyl choloride jolting and extract 2 times, each 10ml merges the methenyl choloride extract, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds hydrochloric acid 3ml, reflux 1 hour, put coldly, add the methenyl choloride jolting and extract 2 times, each 15ml merges the methenyl choloride extract, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds hydrochloric acid 3ml, reflux 2 hours, put coldly, add the methenyl choloride jolting and extract 2 times, each 20ml merges the methenyl choloride extract, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.

Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.Get scarce Radix Glycyrrhizae prescription, press the negative preparation of the scarce Radix Glycyrrhizae of technology preparation among the embodiment 6, prepare scarce Radix Glycyrrhizae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 8: 0.5) be developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The preparation of table 12. need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 8: 1.2)

Developping agent two: sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 8: 0.5)

Developping agent three: ethyl acetate-methanol-water (7: 2.5: 2.5)

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add hydrochloric acid 3ml, reflux 1 hour is put coldly, adds the methenyl choloride jolting and extracts 2 times, and each 15ml merges the methenyl choloride extract, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.Get scarce Radix Glycyrrhizae prescription, press the negative preparation of the scarce Radix Glycyrrhizae of technology preparation among the embodiment 6, prepare scarce Radix Glycyrrhizae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 15 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch respectively, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 13. developping agent proportioning preferred

Developping agent is sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 8: in the time of 0.5) as can be seen from the above table, launch effectively on thin layer plate, principal spot is clear, does not have hangover, identical with the speckle displacement and the color of reference substance, be fit to testing requirements.

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add hydrochloric acid 3ml, reflux 1 hour is put coldly, adds the methenyl choloride jolting and extracts 2 times, and each 15ml merges the methenyl choloride extract, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.Get scarce Radix Glycyrrhizae prescription, press the negative preparation of the scarce Radix Glycyrrhizae of technology preparation among the embodiment 6, prepare scarce Radix Glycyrrhizae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw above-mentioned two kinds of solution each 10 μ l, 15 μ l, 20 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 8: 0.5) be developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 14. sample solution point sample amount optimization experiment is table as a result

Test sample point sample amount is when 15 μ l as can be seen from the above table, and color developing effect is good on thin layer plate, is fit to testing requirements.The screening of experimental example 5 semen armeniacae amarae discrimination tests

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, the n-butanol layer evaporate to dryness, the residue 5ml that adds diethyl ether makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, the n-butanol layer evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, the methyl alcohol jolting is extracted 2 times, each 20ml, merge methanol extract liquid, with normal hexane washing 2 times, each 20ml discards normal hexane liquid, normal hexane layer evaporate to dryness, the residue 5ml that adds diethyl ether makes dissolving, as need testing solution.

Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.Get scarce semen armeniacae amarae prescription, press the negative preparation of the scarce semen armeniacae amarae of technology preparation among the embodiment 6, prepare scarce semen armeniacae amarae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch, take out, dry, (phosphomolybdic acid 2g adds water 20ml and makes dissolving with the phosphomolybdic acid sulfuric acid solution in spray, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The preparation of table 15. need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: methenyl choloride-ethyl acetate-methanol-water (15: 40: 22: 10) 10 ℃ of lower floor's solution of placing 12 hours

Developping agent two: chloroform-methanol-water (10: 10: 1) is placed lower floor's solution of layering below 10 ℃

Developping agent three: chloroform-methanol-water (13: 7: 2) is placed lower floor's solution of layering below 10 ℃

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.Get scarce semen armeniacae amarae prescription, press the negative preparation of the scarce semen armeniacae amarae of technology preparation among the embodiment 6, prepare scarce semen armeniacae amarae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned 3 kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch respectively, take out, dry, (phosphomolybdic acid 2g adds water 20ml and makes dissolving with the phosphomolybdic acid sulfuric acid solution in spray, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 100～110 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 16 developping agent proportioning preferred

Developping agent is chloroform-methanol-water (13: 7: 2) when placing the lower floor solution of layering below 10 ℃ as can be seen from the above table, launches effectively on thin layer plate, and principal spot is clear, does not have hangover, and is identical with the speckle displacement and the color of reference substance, is fit to testing requirements.

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.Get scarce semen armeniacae amarae prescription, press the negative preparation of the scarce semen armeniacae amarae of technology preparation among the embodiment 6, prepare scarce semen armeniacae amarae negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 3 μ l of above-mentioned 3 kinds of solution, 5 μ l, 10 μ l, 15 μ l, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch, take out, dry, (phosphomolybdic acid 2g adds water 20ml and makes dissolving with the phosphomolybdic acid sulfuric acid solution in spray, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 17 sample solution point sample amount optimization experiment

Test sample point sample amount is when 10 μ l as can be seen from the above table, and color developing effect is good on thin layer plate, is fit to testing requirements.

The screening of experimental example 6 PERICARPIUM TRICHOSANTHIS discrimination tests

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 18.06g, porphyrize adds methanol extraction 2 times, each 30ml at every turn, methanol extract liquid steam in, residue adds ethanol 1ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 18.06g, porphyrize adds water saturated normal butyl alcohol and extracts 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 18.06g, porphyrize adds alcohol extract 2 times, each 30ml at every turn, ethanol extract steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.

Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, press the negative preparation of the scarce PERICARPIUM TRICHOSANTHIS of technology preparation among the embodiment 6, prepare scarce PERICARPIUM TRICHOSANTHIS negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃)-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color.

The preparation of table 18 need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: sherwood oil (60～90 ℃)-ethyl acetate (5: 1)

Developping agent two: sherwood oil (60～90 ℃)-ethyl acetate (4: 1)

Developping agent three: sherwood oil (60～90 ℃)-ethyl acetate (3: 1)

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 18.06g, add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, press the negative preparation of the scarce PERICARPIUM TRICHOSANTHIS of technology preparation among the embodiment 6, prepare scarce PERICARPIUM TRICHOSANTHIS negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned 3 kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch respectively, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color.

Table 19. developping agent proportioning preferred

When developping agent is a sherwood oil (60～90 ℃)-ethyl acetate (4: 1) as can be seen from the above table, launch effectively on thin layer plate, principal spot is clear, does not have hangover, and is identical with the speckle displacement and the color of reference substance, is fit to testing requirements.

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 18.06g, add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, press the negative preparation of the scarce PERICARPIUM TRICHOSANTHIS of technology preparation among the embodiment 6, prepare scarce PERICARPIUM TRICHOSANTHIS negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw above-mentioned 3 kinds of solution each 3 μ l, 5 μ l, 10 μ l, 15 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃)-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color.

Sample solution point sample amount optimization experiment is table as a result

Table 20 sample solution point sample amount preferred

Test sample point sample amount is when 10 μ l as can be seen from the above table, and color developing effect is good on thin layer plate, is fit to testing requirements.The screening of experimental example 7 dried orange peel discrimination tests

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, the 20ml jolting that adds diethyl ether is extracted 1 time, discards ether, add the ethyl formate jolting and extract 2 times, each 20ml merges the ethyl formate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, adding sherwood oil (60～90 ℃) 20ml jolting extracts 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml, merge the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize, adding sherwood oil (60～90 ℃) 20ml jolting extracts 1 time, discard sherwood oil, add the ethyl formate jolting and extract 2 times, each 20ml, merge the ethyl formate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.

Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.Get scarce dried orange peel prescription, press the negative preparation of the scarce dried orange peel of technology preparation among the embodiment 6, prepare scarce dried orange peel negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 6 μ l of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch about 15cm, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

The preparation of table 21. need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: chloroform-methanol-water (10: 10: 2) is placed lower floor's solution of layering below 10 ℃

Developping agent two: chloroform-methanol (19: 1)

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add sherwood oil (60～90 ℃) 20ml jolting and extract 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml merges the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.Get scarce dried orange peel prescription, press the negative preparation of the scarce dried orange peel of technology preparation among the embodiment 6, prepare scarce dried orange peel negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 6 μ l of above-mentioned 3 kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch about 15cm respectively, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

Table 22 developping agent proportioning preferred

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add sherwood oil (60～90 ℃) 20ml jolting and extract 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml merges the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.Get scarce dried orange peel prescription, press the negative preparation of the scarce dried orange peel of technology preparation among the embodiment 6, prepare scarce dried orange peel negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw above-mentioned 3 kinds of solution each 1 μ l, 3 μ l, 6 μ l, 10 μ l, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch about 15cm, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

Table 23 sample solution point sample amount optimization experiment result

Test sample point sample amount is when 6 μ l as can be seen from the above table, and color developing effect is good on thin layer plate, is fit to testing requirements.The screening of experimental example 8 balloonflower root discrimination tests

(1) preparation of need testing solution

Method one: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds 10% ethanol solution of sulfuric acid 5ml, reflux 2h puts coldly, extracts 2 times with the chloroform jolting, each 20ml, combined chloroform liquid adds water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds 10% ethanol solution of sulfuric acid 5ml, reflux 3h puts coldly, extracts 2 times with the chloroform jolting, each 20ml, combined chloroform liquid adds water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.

Method three: get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, porphyrize adds ethanolic solution 5ml, reflux 4h puts coldly, extracts 2 times with the chloroform jolting, each 20ml, combined chloroform liquid adds water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration filters the filtrate evaporate to dryness, residue adds ethanol 5ml makes dissolving, as need testing solution.

Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.Get scarce balloonflower root prescription, press the negative preparation of the scarce balloonflower root of technology preparation among the embodiment 6, prepare scarce balloonflower root negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, drawing each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-ether (1: 1), launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The preparation of table 24 need testing solution

(2) the developping agent proportioning is preferred:

Developping agent one: chloroform-methanol (20: 1)

Developping agent two: chloroform-methanol-formic acid (16: 10: 1)

Developping agent three: chloroform-ether (1: 1)

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add 10% ethanol solution of sulfuric acid 5ml, reflux 3h, put coldly, extract 2 times each 20ml with the chloroform jolting, combined chloroform liquid, add water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.Get scarce balloonflower root prescription, press the negative preparation of the scarce balloonflower root of technology preparation among the embodiment 6, prepare scarce balloonflower root negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, developping agent one, two, three, launch respectively, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 25. developping agent proportioning preferred

When developping agent is chloroform-ether (1: 1) as can be seen from the above table, launch effectively on thin layer plate, principal spot is clear, does not have hangover, and is identical with the speckle displacement and the color of reference substance, is fit to testing requirements.

(3) sample solution point sample amount is preferred:

Get pharmaceutical preparation of the present invention and be equivalent to crude drug 12.04g, add 10% ethanol solution of sulfuric acid 5ml, reflux 3h, put coldly, extract 2 times each 20ml with the chloroform jolting, combined chloroform liquid, add water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.Get scarce balloonflower root prescription, press the negative preparation of the scarce balloonflower root of technology preparation among the embodiment 6, prepare scarce balloonflower root negative controls by the test sample preparation method.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, drawing above-mentioned 3 kinds of solution each 5 μ l, 10 μ l, 15 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-ether (1: 1), launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

Table 26 sample solution point sample amount optimization experiment is table as a result

The content assaying method test of experimental example 9 Exocarpium Citri Grandises

Monarch drug in a prescription in the assay Exocarpium Citri Grandis side of being, aurantiin are its effective constituent, for ensuring drug quality, are the assay object so select aurantiin, adopt high performance liquid chromatography, and aurantiin is carried out assay.

1. preparation method's precision of test sample is measured this product 1ml, puts in the 25ml measuring bottle, adds methyl alcohol and is diluted to scale, shakes up, and filters, and gets subsequent filtrate, promptly.

2. measure the selection of wavelength and draw aurantiin reference substance solution 10 μ l, need testing solution 10 μ l, inject high performance liquid chromatograph, through diode array detector the aurantiin absorption peak is carried out absorption spectromtry in 190nm～370nm, the ultra-violet absorption spectrum basically identical of test sample and reference substance as a result, all strong absorption is arranged, be decided to be 283nm so this product is measured wavelength at the 283nm place.

3. linear relationship is investigated precision and is taken by weighing the aurantiin reference substance, adds dissolve with methanol, makes the reference substance solution that every 1ml contains 49.8 μ g, and sample introduction 2.5,5,7.5,10,12.5,15 μ l by above-mentioned chromatographic condition, measure aurantiin peak area integrated value respectively.With the reference substance sample size is horizontal ordinate, is ordinate with the aurantiin peak area.The result shows, in 0.1245 μ g～0.747 μ g scope, the amount of aurantiin and peak area integrated value are good linear relationship, and be a straight line that was close to initial point, therefore can adopt in test one point external standard method to measure, regression equation Y=1.83556 * 106X-2623.08, γ=0.9995.

4. the precision test prepares need testing solution according to technology among the embodiment 6, replication 5 times, and record aurantiin absorption peak peak area, mean value is 870942.8, RSD=1.37% (n=5).

5. to get be example 6 samples for reappearance test, according to 5 parts of test liquids of the parallel preparation of [1] method, carries out assay respectively, and mean value is=1.0252 (mg/ml), RSD=1.49% (n=5).

6. stability test is got embodiment 6 samples according to above-mentioned chromatographic condition, and the preparation need testing solution is respectively at 0,1,2,4,6,8h carries out assay, record aurantiin absorption peak peak area, RSD=1.24% (n=6).Stable in the need testing solution 8h as a result, can satisfy the mensuration needs.

7. the negative control test is got and is lacked the Exocarpium Citri Grandis prescription, press the negative preparation of the scarce Exocarpium Citri Grandis of technology preparation among the embodiment 6,, record aurantiin negative control chromatogram according to the text content assaying method, as a result in the side other composition under this condition determination, noiseless to the assay of aurantiin.

8. embodiment 6 samples (naringin content is 1.0252mg/ml) 2.5ml is got in the average recovery test, gets 5 parts altogether, adds the aurantiin reference substance respectively, according to content assaying method among the embodiment 10, measures in accordance with the law, and calculate recovery rate the results are shown in Table 1.

The test of table 27 average recovery

9. sample size is measured according to [1] method and is prepared need testing solution, measures the content of aurantiin in 3 batch samples, the results are shown in Table 2.

Naringin content in table 28 sample

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

Embodiment 1

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, loguat leaf 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, banksia rose 22g

This pharmaceutical composition adds conventional auxiliary material, makes capsule, soft capsule, pill, tablet, powder, parenteral solution, granule, mixture by common process.

Embodiment 2

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma pinellinae praeparata 55g, loguat leaf 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, banksia rose 22g

Embodiment 3

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, tussilago 66g, root bark of white mulberry 22g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, banksia rose 22g

Add conventional auxiliary material again,, make capsule, soft capsule, pill, tablet, powder, parenteral solution, granule, the mixture of clinical acceptance through conventional method.

Embodiment 4

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, loguat leaf 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, perilla seed (stir-fry) 22g

Embodiment 5

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, loguat leaf 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, glutinous rehmannia 22g, gypsum 22g, banksia rose 22g

Embodiment 6

More than ten five tastes, gesso is broken into meal, the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, semen armeniacae amarae four flavor steam distillations are collected distillate 250ml; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; Ten flavors such as all the other rhizoma arisaematis, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect the liquid 2700ml that filters, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add sucrose 80g, boil, left standstill 24 hours, filter, add ethyl hydroxy benzoate 0.3g, benzoic acid 0.5g (both use an amount of hot water dissolving earlier) and distillate, add water and adjust total amount, stir evenly, refrigerate 48 hours, get supernatant to 950ml, embedding, sterilization, promptly.

Embodiment 7

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, loguat leaf 66g, lepidium seed 66g, root bark of white mulberry 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, banksia rose 22g

More than 17 flavors, gesso is broken into meal, the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, lepidium seed, semen armeniacae amarae five tastes steam distillation are collected distillate 250ml; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; All the other rhizoma arisaematis etc. ten simply, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect the liquid 2700ml that filters, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add sucrose 80g, boil, left standstill 24 hours, filter, add ethyl hydroxy benzoate 0.3g, benzoic acid 0.5g (both use an amount of hot water dissolving earlier) and distillate, add water and adjust total amount, stir evenly, refrigerate 48 hours, get supernatant to 950ml, embedding, sterilization, promptly.

Embodiment 8

Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma pinellinae praeparata 66g, tussilago 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, glutinous rehmannia 22g, gypsum 22g, perilla seed (stir-fry) 22g

More than ten five tastes, gesso is broken into meal, the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, tussilago, semen armeniacae amarae four flavor steam distillations are collected distillate 250ml; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; Ten flavors such as all the other rhizoma pinellinae praeparata, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect the liquid 2700ml that filters, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add sucrose 80g, boil, left standstill 24 hours, filter, add ethyl hydroxy benzoate 0.3g, benzoic acid 0.5g (both use an amount of hot water dissolving earlier) and distillate, add water and adjust total amount, stir evenly, refrigerate 48 hours, get supernatant to 950ml, embedding, sterilization, promptly.

The method of quality control of embodiment 9 preparations of the present invention

Getting embodiment 6 contents differentiates:

[discriminating]

(1) get this product 40ml, add hydrochloric acid 3ml, put in the water-bath heating 1 hour, put coldly, the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 30 minutes filters, and filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (4: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

(2) get this product 20ml, add hydrochloric acid 3ml, reflux 1 hour is put coldly, adds the methenyl choloride jolting and extracts 2 times, and each 15ml merges the methenyl choloride extract, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (30～60 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) be developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(3) get this product 20ml, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, uses ammonia scrubbing 2 times, and each 20ml discards ammoniacal liquor liquid, and n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution.Get scarce semen armeniacae amarae prescription, lack semen armeniacae amarae feminine gender preparation, prepare by the test sample preparation method and lack the semen armeniacae amarae negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch, take out, dry, spray is with phosphomolybdic acid sulfuric acid solution (phosphomolybdic acid 2g, add water 20ml and make dissolving, slowly adding sulfuric acid 30ml, mixing), it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color, lacking semen armeniacae amarae negative control liquid chromatography does not have corresponding spot.

(4) get this product 20ml, add sherwood oil (60～90 ℃) 20ml jolting and extract 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml merges the ethyl acetate extract, and evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution.Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution.Get scarce dried orange peel prescription, lack dried orange peel feminine gender preparation, prepare by the test sample preparation method and lack the dried orange peel negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 6 μ l of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with chloroform-methanol-water (13: 7: 2) below 10 ℃ is developping agent, launch about 15cm, take out, dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.Lack dried orange peel negative control liquid chromatography and do not have corresponding spot.

(5) get this product 20ml, add 10% ethanol solution of sulfuric acid 5ml, reflux 3h, put coldly, extract 2 times each 20ml with the chloroform jolting, combined chloroform liquid, add water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution.Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method.Get scarce balloonflower root prescription, lack balloonflower root feminine gender preparation, prepare by the test sample preparation method and lack the balloonflower root negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 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 developping agent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.Lack balloonflower root negative control liquid chromatography and do not have corresponding spot.

(6) get this product 30ml and add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, lack PERICARPIUM TRICHOSANTHIS feminine gender preparation, prepare by the test sample preparation method and lack the PERICARPIUM TRICHOSANTHIS negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, [one is developping agent with sherwood oil (60～90 ℃) ethyl acetates (4: 1), launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show mutually the spot of color just; Negative controls does not have this spot,

The method of quality control of embodiment 10 preparations of the present invention

Get embodiment 6 contents and carry out assay:

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

The every 1ml of this product contains Exocarpium Citri Grandis with aurantiin (C ₂₇H ₃₂O ₁₄) meter, must not be less than 0.80mg.

The method of quality control of embodiment 11 preparations of the present invention

Getting embodiment 6 contents differentiates and assay:

[discriminating]

(6) get this product 30ml and add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, lack PERICARPIUM TRICHOSANTHIS feminine gender preparation, prepare by the test sample preparation method and lack the PERICARPIUM TRICHOSANTHIS negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃) ethyl acetates (4: 1) is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show mutually the spot of color just; Negative controls does not have this spot,

The preparation method and the method for quality control of the oral liquid of embodiment 12 medicine groups of the present invention

[prescription] Exocarpium Citri Grandis 66g, dried orange peel 66g, rhizoma arisaematis 66g, loguat leaf 66g, balloonflower root 44g, semen armeniacae amarae (peeling is fried) 44g, Poria cocos 44g, PERICARPIUM TRICHOSANTHIS 44g, Radix Glycyrrhizae 33g, aster 33g, the tuber of dwarf lilyturf 33g, wind-weed 33g, root of purple-flowered peucedanum 22g, gypsum 22g, banksia rose 22g

[method for making] above ten five tastes, gesso is broken into meal, and the boiling secondary each 1 hour, filters filtrate for later use; Exocarpium Citri Grandis, dried orange peel, loguat leaf, semen armeniacae amarae four flavor steam distillations are collected distillate 250ml; The distiller inner liquid medicine filters, and filtrate adds ethanol to be made and contain the alcohol amount and reach 60%, stirs evenly, and leaves standstill 24 hours, filters filtrate for later use; Ten flavors such as all the other rhizoma arisaematis, be ground into meal and above-mentioned dregs of a decoction mixing, according to the percolation under liquid extract and the extract item (" appendix IO of Chinese pharmacopoeia version in 2005), make solvent with 60% ethanol, flood after 24 hours diacolation in accordance with the law, collect the liquid 2700ml that filters, merge with above-mentioned reserve liquid, decompression recycling ethanol merges with the parget water decocting liquid to there not being the alcohol flavor, is concentrated into the clear cream of relative density 1.06 (50 ℃).Add sucrose 80g, boil, left standstill 24 hours, filter, add ethyl hydroxy benzoate 0.3g, benzoic acid 0.5g (both use an amount of hot water dissolving earlier) and distillate, add water and adjust total amount, stir evenly, refrigerate 48 hours, get supernatant to 950ml, embedding, sterilization, promptly.

[discriminating]

(6) get this product 30ml and add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid steam in, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution.Get scarce PERICARPIUM TRICHOSANTHIS prescription, lack PERICARPIUM TRICHOSANTHIS feminine gender preparation, prepare by the test sample preparation method and lack the PERICARPIUM TRICHOSANTHIS negative controls by the technology preparation.According to thin-layered chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 10 μ L of above-mentioned 3 kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil (60～90 ℃) ethyl acetates (4: 1) is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing.In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show mutually the spot of color just; Negative controls does not have this spot.

[function with cure mainly] clearing lung-heat, cough-relieving is reduced phlegm.Be used for that coughing with a lot of sputum, fullness sensation in chest that pulmonary retention of phlegmopyrexia causes are breathed hard, the dry throat larynx is itched.

[usage and consumption] is oral, a 10ml, 2～3 times on the one; Children's consumption is followed the doctor's advice.

[attention] avoids pungent food greasy.

[specification] every dress 10ml.

Shady and cool place is put in [storage] sealing.

Claims

1. the detection method of a relieving cough and reducing sputum Chinese medicine composition is characterized in that this method is:

Described Chinese medicine composition is to be made by the bulk drug of following weight ratio:

Semen armeniacae amarae 30-70 weight portion, Poria cocos 30-70 weight portion, PERICARPIUM TRICHOSANTHIS 30-70 weight portion, Radix Glycyrrhizae 20-50 weight portion, aster 20-50 weight portion, tuber of dwarf lilyturf 20-50 weight portion, wind-weed 20-50 weight portion, root of purple-flowered peucedanum 10-35 weight portion, gypsum 10-35 weight portion, banksia rose 10-35 weight portion are fried in Exocarpium Citri Grandis 50-100 weight portion, dried orange peel 50-100 weight portion, rhizoma arisaematis 50-100 weight portion, loguat leaf 50-100 weight portion, balloonflower root 30-70 weight portion, peeling;

Discrimination method is following steps (1)-(6):

(1) get this pharmaceutical composition and be equivalent to crude drug 20-30g, add hydrochloric acid 2-4ml, put in the water-bath heating 0.5-2 hour, put coldly, the 20-40ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution; Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 20-40 minute, filter, filtrate is concentrated into 30-50ml, shines medicinal material solution in pairs with legal system; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 3.5-4.5: the methenyl choloride-acetone of 0.5-1.5 proportioning is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 100～110 ℃ of heating 4-6 minute; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

(2) get this pharmaceutical composition and be equivalent to crude drug 10-15g, add hydrochloric acid 2-4ml, reflux 0.5-2 hour, put cold, add the methenyl choloride jolting and extract 1-3 time, each 10-20ml merges the methenyl choloride extract, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 8-12: 15-25: 5-10: 30～60 ℃ of sherwood oil-benzene-ethyl acetate-glacial acetic acid of 0-1 proportioning are developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 100～110 ℃ of heating 4-6 minute; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) get this pharmaceutical composition and be equivalent to crude drug 10-15g, adding the water-saturated n-butanol jolting extracts 1-3 time, each 15-25ml, merge n-butanol extracting liquid, use ammonia scrubbing 1-3 time, each 15-25ml, discard ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10-15: 5-10: the chloroform-methanol-water of 1-3 proportioning is placed layering below 10 ℃ lower floor's solution is developping agent, launch, take out, dry, spray is with the phosphomolybdic acid sulfuric acid solution, it is clear to be heated to spot colour developing at 100～110 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(4) get this pharmaceutical composition and be equivalent to crude drug 10-15g, add 60～90 ℃ of sherwood oil 15-25ml joltings and extract 1 time, discard sherwood oil, add the ethyl acetate jolting and extract 1-3 time, each 15-25ml merges the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10-15: 5-10: the chloroform-methanol-water of 1-3 proportioning is placed layering below 10 ℃ lower floor's solution is developping agent, launch about 10-20cm, take out, dry, spray is put under the 365nm ultraviolet lamp and is inspected with the aluminium choride test solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

(5) get this pharmaceutical composition and be equivalent to crude drug 10-15g, add 10% ethanol solution of sulfuric acid 4-6ml, reflux 4-6h, put coldly, extract 1-3 time with the chloroform jolting, at every turn 15-25ml, combined chloroform liquid, add water 20-40ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution; Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 45-55: the chloroform-ether of 45-55 proportioning is a developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing at 100～110 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(6) get this pharmaceutical composition and be equivalent to crude drug 15-20g, add water saturated normal butyl alcohol and extract 1-3 time, each 25-40ml at every turn, n-butanol extracting liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 35-45: 60～90 ℃ of sherwood oil-ethyl acetates of 5-15 proportioning are developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing; In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color;

Assay: chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With 30-40: 60-70: the methanol-water-acetic acid of 0-1 proportioning is moving phase; The detection wavelength is 283nm; Column temperature is 40 ℃; Theoretical cam curve is calculated by the aurantiin peak should be not less than 3000; The preparation of reference substance solution: be taken at 100-120 ℃ of aurantiin reference substance 15mg that is dried to constant weight, the accurate title, decide, and puts in the 100ml measuring bottle, adds methyl alcohol and make dissolving in right amount and be diluted to scale, shakes up; Precision is measured 3ml, puts in the 10ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets the reference substance solution that every 1ml contains aurantiin 45 μ g; The preparation of need testing solution: precision is measured pharmaceutical composition of the present invention and is equivalent to crude drug 0.3-1.0g, puts in the 25ml measuring bottle, adds methyl alcohol and is diluted to scale, shakes up, and filters, and gets subsequent filtrate, promptly; Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly; The every 1ml of this product contains Exocarpium Citri Grandis in aurantiin, must not be less than 0.80mg.

2. the detection method of Chinese medicine composition as claimed in claim 1 is characterized in that this method is:

(1) get this pharmaceutical composition mixture 40ml, add hydrochloric acid 3ml, put in the water-bath heating 1 hour, put coldly, the 30ml jolting that adds diethyl ether is extracted, and divides and gets ether solution, and evaporate to dryness, residue add methenyl choloride 1ml makes dissolving, as need testing solution; Other gets the control medicinal material 2g tuber of dwarf lilyturf, and boiling 30 minutes filters, and filtrate is concentrated into 40ml, shines medicinal material solution in pairs with legal system; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, methenyl choloride-acetone with 4: 1 proportionings is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, 105 ℃ of heating 5 minutes; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

(2) get this pharmaceutical composition mixture 20ml, add hydrochloric acid 3ml, reflux 1 hour is put coldly, adds the methenyl choloride jolting and extracts 2 times, and each 15ml merges the methenyl choloride extract, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice hypo acid reference substance adds absolute ethyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution in addition; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10: 20: 7: 30～60 ℃ of sherwood oil-benzene-ethyl acetate-glacial acetic acid of 0.5 proportioning were developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(3) get this pharmaceutical composition mixture 20ml, add the water-saturated n-butanol jolting and extract 2 times, each 20ml merges n-butanol extracting liquid, with ammonia scrubbing 2 times, each 20ml discards ammoniacal liquor liquid, n-butanol layer evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that 1ml contains 2mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with the chloroform-methanol-water of 13: 7: 2 proportionings below 10 ℃ is developping agent, launch, take out, dry, spray is with the phosphomolybdic acid sulfuric acid solution, it is clear to be heated to spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(4) get this pharmaceutical composition mixture 20ml, the sherwood oil 20ml jolting that adds 60～90 ℃ is extracted 1 time, discards sherwood oil, add the ethyl acetate jolting and extract 2 times, each 20ml merges the ethyl acetate extract, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets dried orange peel glycosides reference substance, adds methyl alcohol and makes saturated solution, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing layering with the chloroform-methanol-water of 13: 7: 2 proportionings below 10 ℃ is developping agent, launch 15cm, take out, dry, spray is put under the 365nm ultraviolet lamp and is inspected with the aluminium choride test solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

(5) get this pharmaceutical composition mixture 20ml, add 10% ethanol solution of sulfuric acid 5ml, reflux 3h, put coldly, extract 2 times each 20ml with the chloroform jolting, combined chloroform liquid, add water 30ml washing, discard washing lotion, the chloroform solution anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution; Other gets balloonflower root control medicinal material 1g, makes reference substance solution with method; According to the test of an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, chloroform-ether with 1: 1 proportioning is a developping agent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

(6) get this pharmaceutical composition mixture 30ml, add water saturated normal butyl alcohol and extract 2 times, each 30ml at every turn, n-butanol extracting liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Get PERICARPIUM TRICHOSANTHIS control medicinal material 3g, add 60% ethanol 20ml cold soaking 2h after, sonicated 30min filters, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, makes control medicinal material solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin-layered chromatography, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, 60～90 ℃ of sherwood oil-ethyl acetates with 4: 1 proportionings are developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing; In the test sample chromatogram, with PERICARPIUM TRICHOSANTHIS control medicinal material chromatogram relevant position on, show the spot of same color.

3. detection method as claimed in claim 1 is characterized in that what described composition was made by the bulk drug of following weight ratio: semen armeniacae amarae 44 weight portions, Poria cocos 44 weight portions, PERICARPIUM TRICHOSANTHIS 44 weight portions, Radix Glycyrrhizae 33 weight portions, aster 33 weight portions, the tubers of dwarf lilyturf 33 weight portion, the wind-weed 33 weight portions, the root of purple-flowered peucedanum 22 weight portions, gypsum 22 weight portions, the banksia rose 22 weight portions are fried in Exocarpium Citri Grandis 66 weight portions, dried orange peel 66 weight portions, rhizoma arisaematis 66 weight portions, loguat leaf 66 weight portions, balloonflower root 44 weight portions, peeling.