CN100372563C

CN100372563C - Compound preparation for treating bronchitis, its preparation method and quality control method

Info

Publication number: CN100372563C
Application number: CNB2005100596538A
Authority: CN
Inventors: 张友生; 张思宁
Original assignee: Individual
Current assignee: Individual
Priority date: 2005-03-30
Filing date: 2005-03-30
Publication date: 2008-03-05
Anticipated expiration: 2025-03-30
Also published as: CN1840118A

Abstract

The present invention discloses a preparation method and a quality controlling method for a compound granule. The compound preparation is prepared from a liquid extract of thunberg fritillary bulb, a liquid extract of liquorice, a liquid extract of platycodi liquidum, stemona root, peucedanum root, ginger pinellia, tangerine peel, ammonium chloride, ephedrine hydrochloride and menthol by specific technology. The quality controlling method of the compound granule adds an identifying method for the thin-layer chromatography of the tangerine peel, the peucedanum roots and the liquorice on the basis of the original syrup agent, and also adds a measuring method for the content of ephedrine hydrochloride; thereby, the quality control standard is further perfected and enhanced.

Description

Treat bronchitic compound preparation and preparation method thereof, method of quality control

Technical field

The present invention relates to a kind of compound preparation, particularly relate to bronchitic compound granular agent of a kind of treatment and preparation method thereof, method of quality control.

Background technology

Bronchitis belongs to Chinese medicine " cough ", and " asthma " sick category is common clinical, frequently-occurring disease.The peace of the Xiao Shouing syrup of coughing is the compound preparation that Chinese medicine and Western medicine are formed in the market, and said preparation is with Bulbus Fritillariae Thunbergii, the Radix Stemonae, Radix Platycodonis, Radix Peucedani, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens), Chinese medicine nourishing the lung to arrest cough such as Pericarpium Citri Reticulatae, Herba Menthae, Radix Glycyrrhizae cooperate and reduce phlegm with Western medicine ammonium chloride, the ephedrine hydrochloride cough-relieving, the Chinese and western drugs organic assembling is the agent of moistening the lung and resolving phlegm, relieving cough and asthma, altogether for accumulation of phlegm-heat in the lung, pant and breathe hard, cough with ropy sputum, the bronchitis of performance such as thirsty and dry pharynx has better curative effect.But because existing market has only a kind of dosage form of syrup, dosage form is single, can not satisfy different patients' needs, therefore develop a kind of new dosage form and seemed that it is necessary compeling, and former syrupy quality standard is only differentiated ephedrine hydrochloride, only detect the content of ammonium chloride in the medicine, and other Chinese medicine ingredients in the medicine such as Pericarpium Citri Reticulatae, Radix Peucedani, Radix Glycyrrhizae etc. are not set up quality control standard, therefore need further to improve and improve.

Summary of the invention

The object of the invention is to provide a kind of treatment bronchitic compound granular agent; The object of the invention also is to provide a kind of preparation method for the treatment of bronchitic compound granular agent; The object of the invention also is to provide a kind of method of quality control for the treatment of bronchitic compound granular agent.

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

Compound granular agent of the present invention is that the crude drug by following w/v (in the ratio of g/ml) is prepared from:

Bulbus Fritillariae Thunbergii fluid extract 20-25 parts by volume, Radix Glycyrrhizae fluidextract 20-25 parts by volume, Radix Stemonae 20-25 weight portion, Radix Platycodonis fluidextract 15-20 parts by volume, Radix Peucedani 20-25 weight portion, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 20-25 weight portion, Pericarpium Citri Reticulatae 10-15 weight portion, ammonium chloride 20-25 weight portion, ephedrine hydrochloride 0.3-0.4 weight portion, Mentholum 0.1-0.2 weight portion; Wherein Bulbus Fritillariae Thunbergii fluid extract standard is recorded in " one one of Chinese pharmacopoeia version in 1977; Radix Glycyrrhizae fluidextract records in " under an appendix fluid extract of Chinese pharmacopoeia version in 2000 the general rule item; The method for making of Radix Platycodonis fluidextract is recorded in " pharmaceutical preparation note ".

The preferential proportioning of above-mentioned raw materials medicine is:

Bulbus Fritillariae Thunbergii fluid extract 25 parts by volume, Radix Glycyrrhizae fluidextract 20 parts by volume, the Radix Stemonae 25 weight portions, Radix Platycodonis fluidextract 18 parts by volume, Radix Peucedani 22 weight portions, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 22 weight portions, Pericarpium Citri Reticulatae 12 weight portions, ammonium chloride 20 weight portions, ephedrine hydrochloride 0.365 weight portion, Mentholum 0.15 weight portion.

Compound granular agent of the present invention is prepared by following method:

Get above-mentioned ten flavors crude drug of the present invention, Radix Peucedani adds water in 85～90 ℃ of warm macerating 2-3 time, and each 2-3 hour, filter, filtrate merges, and is concentrated in right amount, puts coldly, adds ethanol and makes and contain alcohol and measure and reach 25% approximately, and is standby; The Radix Stemonae, Pericarpium Citri Reticulatae pulverize separately become coarse powder, and appendix 1 O of Chinese Pharmacopoeia version in 2000 is according to the percolation under fluid extract and the extractum item, the Radix Stemonae with 55% ethanol make solvent, Pericarpium Citri Reticulatae is made solvent with 80% ethanol, carry out percolation respectively, collect the Radix Stemonae liquid of filtering, the Pericarpium Citri Reticulatae liquid of filtering, standby; Rhizoma Pinelliae powder is broken into coarse powder, with 50% ethanol in 70～80 ℃ of warm macerating 2-3 time, each 2-3 hour, merge leachate, standby; Above four kinds of reserve liquids are merged, decompression recycling ethanol is to there not being the alcohol flavor, with Bulbus Fritillariae Thunbergii, Radix Glycyrrhizae, three kinds of fluid extract mixings of Radix Platycodonis, leave standstill, filter, be concentrated into relative density under 22 ℃ of temperature and be 1.29～1.35 clear paste, add ephedrine hydrochloride, get the clear paste of above-mentioned mixing, add ammonium chloride, an amount of dextrin and sucrose, mixing; It is an amount of to add 40% ethanol, makes granule, drying, and spray adds 90% alcoholic solution of Mentholum, and mixing is distributed into bag, and every bag of 5g promptly gets compound granular agent of the present invention.Oral, a 5-7.5g, 3 times on the one.

The method of quality control of compound granular agent of the present invention comprises one or more in following discriminating and/or the content assaying method.

Differentiate:

A. get compound granular agent 5g of the present invention, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is 15～25: 4～6: chloroform-methanol-strong ammonia solution of 0.5 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

B. get compound granular agent 20g of the present invention, add methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml, merge n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, add neutral alumina 1g and mix thoroughly, evaporate to dryness is contained in neutral alumina pillar (100～120 orders, 3g, internal diameter 1cm) on, is ethyl acetate-methanol 30ml eluting of 1～2: 1～2, collects the eluent evaporate to dryness with ratio, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Citri Reticulatae control medicinal material 0.3g, adds methanol 30ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that normal hexane-ethyl acetate of 1～3: 2～4 is developing solvent, launch, take out, dry, put under the wavelength 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

C. get Radix Peucedani control medicinal material 0.8g, decoct with water 30 minutes, filter, filtrate is concentrated into about 20ml, puts coldly, adds ethyl acetate 30ml jolting extraction, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw need testing solution and each 5 μ l of control medicinal material solution of differentiating under the B item, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is 1～2: 1～2 normal hexane-ethyl acetate is developing solvent, launch, take out, dry, spray is put under the wavelength 365nm ultra-violet lamp and is inspected with 1% sodium hydroxide alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

D. get and differentiate the pillar behind the eluting under the B item, add 40% methanol 30ml eluting, collect the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds methanol 30ml in addition, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ratio is 10～20: ethyl acetate-formic acid of 1: 1: 2-glacial acetic acid-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Assay:

E. ammonium chloride

Get the compound granular agent of the present invention under the content uniformity item, grind well, get 2.5g, it is fixed to claim, put in the 500ml kjeldahl flask, add water 250ml, jolting makes dissolving, adds bead number, add 40% sodium hydroxide solution 5ml, with the nitrogen ball kjeldahl flask is connected with condensing tube immediately, the tip of condensing tube is immersed under the liquid level of 4% boric acid solution 50ml (adding 10 of 0.1% methyl-0.2% bromocresol green mixing indicator solutions in the solution), adds thermal distillation, when the cumulative volume that receives liquid is about 200ml, with the most advanced and sophisticated liquid level that proposes of condensing tube, made steam washing 1 minute, with stopping distillation behind the water wash tip; Distillate becomes gray purple with sulphuric acid volumetric solution (0.05mol/L) titration to solution by aeruginous, and titrating result is proofreaied and correct with blank assay, promptly; The sulphuric acid volumetric solution (0.05mol/L) of every 1ml is equivalent to the ammonium chloride NH4Cl of 5.349mg;

Compound granular agent of the present invention contains ammonium chloride (NH4Cl) for every bag and should be 170～230mg.

F. ephedrine hydrochloride

Measure according to high performance liquid chromatography (Chinese Pharmacopoeia appendix III in 2000);

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With ratio be 5～15: 95～85 to regulate pH value with phosphoric acid be that methanol-0.05mol/L potassium dihydrogen phosphate of 3.0 is a mobile phase; The detection wavelength is 209nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 2800;

It is an amount of to take by weighing ephedrine hydrochloride, adds 0.1% methanol hydrochloride solution and makes the solution that every 1ml contains 30 μ g, promptly gets reference substance solution; Get the compound preparation of the present invention under the content uniformity item, grind well, get 1g, it is fixed to claim, puts in the tool plug conical flask, adds 0.1% methanol hydrochloride solution 25ml, claim to decide weight, supersound process 30 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 0.1% methanol hydrochloride solution, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly get need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly; Every bag of hydrochloric ephedrine of compound granular agent of the present invention (C10H15NOHCl) should be 3.10～4.20mg.

Compound granular agent of the present invention has moistening the lung and resolving phlegm, and the effect of relieving cough and asthma is used for accumulation of phlegm-heat in the lung, and pant and breathe hard, cough with ropy sputum, mouth is drunk dry pharynx, and bronchitis belongs to above-mentioned patient; Oral, a 5～7.5g, 3 times on the one.

The quality control standard of compound granular agent of the present invention is on the basis of former " peace cough syrup " quality standard, and the thin layer chromatography that has increased Pericarpium Citri Reticulatae, Radix Peucedani, Radix Glycyrrhizae is differentiated; After in addition the discriminating of ephedrine hydrochloride being launched with former developing solvent, the ephedrine hydrochloride spot and R f value is higher, adopt developing solvent chloroform-methanol-strong ammonia solution of the present invention after spot and R f value moderate.

In addition, the present invention also increases the Determination of Ephedrine Hydrochloride assay method, and this law adopts high performance liquid chromatography, is filler with octadecylsilane chemically bonded silica; Methanol-0.05mol/L potassium dihydrogen phosphate (regulating pH value with phosphoric acid is 3.0) (10: 90) is mobile phase; The detection wavelength is 209nm.The range of linearity 0.09664～0.5134 μ g, r=1.000, average recovery rate 99.59%6, RSD%=0.72.

After granule of the present invention increases thin layer chromatography discriminating and Determination of Ephedrine Hydrochloride mensuration, at ambient temperature, carried out stability test, the content that the result checked (comprising character, discriminating, inspection, limit test of microbe, assay) does not all have obviously change, shows that granule of the present invention has good stability.

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

Experimental example 1. granules of the present invention (peace cough granule) treatment bronchitis 30 routine observation of curative effect

Case is all from outpatient service.

One, diagnostic criteria and observational technique

Diagnostic criteria is all with reference to " the clinical guidance principle of new Chinese medicine treatment chronic bronchitis ".

Medication: the oral peace granule of coughing, one time 1 bag, 3 times on the one; 20 days is a course of treatment.

Two, physical data

1, sex: male 19 examples, women 11 examples.

2, the age: 15～30 years old 10 example, 31～50 years old 11 example, 51～65 years old 9 example.Minimal ages 16 years old, maximum 61 years old age, 44.1 years old mean age.

3, different pattern of syndrome simple type 12 examples, the type of panting 18 examples.

4, the course of disease :≤7 days 8 examples, 7～15 days 12 examples, 15 days 10 examples.

5, the state of an illness: slight 9 examples, moderate 12 examples, severe 9 examples.

6, symptom and sign see Table 1.

Table 1 clinical symptoms and sign distribute

Symptom and sign	The example number	Do not have	Gently	In	Heavy
Symptom and sign	The example number	Do not have	Gently	In	Heavy	The cough wheezing sound of panting of coughing up phlegm	30 30 30 30	0 0 9 10	10 9 9 6	11 12 7 8	9 9 5 6

7, picture of the tongue: yellow and thin fur 18 examples, yellow and greasy fur 12 examples; Smooth pulse 21 examples, sliding several 9 examples.

8, leukocyte, Chest X-rays, pulmonary's sign see Table 2.

Table 2 leukocyte, Chest X-rays, pulmonary's sign situation

Project leukocyte Chest X-rays pulmonary sign	Example several 30 30 30	The treatment group
		The treatment group	Normal 9 10 9	Unusual 21 20 21

Three, curative effect determinate standard:

1, clinical symptoms efficacy evaluation

Clinical cure: disappearances such as coughing, cough up phlegm, pant.

Produce effects: (+++→+) the pulmonary's wheezing sound of coughing, cough up phlegm, pant etc. and be clearly better obviously alleviates.

Effectively: cough, cough up phlegm to some extent and to take a turn for the better (+++→ ++ or ++ →+) pulmonary's wheezing sound obviously alleviates.

Invalid: each disease does not all have improvement or increases the weight of.

2, comprehensive therapeutic effect criterion:

According to the standards of grading of every observation index, calculate the integrated value of treatment front and back earlier, calculate therapeutic index then and determined curative effect.

Clinical cure: clinical cardinal symptom, sign disappears, and unusual physical and chemical index recovers normal, therapeutic index 〉=90%.

Produce effects: cardinal symptom, sign is obviously improved, and unusual physical and chemical index is approaching normal, therapeutic index 〉=70%＜90%.

Effectively: cardinal symptom, sign alleviate, and unusual physical and chemical index makes moderate progress, and treat and produce index 〉=30%70%.

Four, result:

1, total effects: clinical cure 9 examples (30%), produce effects 12 examples (40%), effective 8 examples (26.67%), invalid 1 example (3.33%).

2, symptom, sign and curative effect see Table 3.

Table 3 clinical symptoms and sign curative effect

Symptom, sign	The example number	Clinical cure	Produce effects	Effectively	Invalid
Symptom, sign	The example number	Clinical cure	Produce effects	Effectively	Invalid	The cough wheezing sound of panting of coughing up phlegm	30 30 30 30	11 10 12 18	10 11 10 10	8 9 7 1	1 0 1 1

3, the state of an illness and curative effect see Table 4.

Table 4 state of an illness curative effect

The state of an illness	The example number	Clinical cure	Produce effects	Effectively	Invalid
The state of an illness	The example number	Clinical cure	Produce effects	Effectively	Invalid	Slight moderate severe	9 12 9	7 7 2	1 2 6	1 2 1	0 1 0

4, the course of disease and curative effect see Table 5.

Table 5 course of disease curative effect

The course of disease (my god)	The example number	Clinical cure	Produce effects	Effectively	Invalid
The course of disease (my god)	The example number	Clinical cure	Produce effects	Effectively	Invalid	≤7 ～15 >15	8 12 10	6 2 1	1 4 7	1 6 1	0 0 1

5, the curative effect of different pattern of syndrome sees Table 6.

The curative effect of the different pattern of syndrome of table 6

Pattern of syndrome	The example number	Clinical cure	Produce effects	Effectively	Invalid
Pattern of syndrome	The example number	Clinical cure	Produce effects	Effectively	Invalid	The simple type type of panting	12 18	3 6	4 8	4 4	1 0

6, leukocyte, Chest X-rays, pulmonary's sign situation of change see Table 7.

Table 7 leukocyte, Chest X-rays, pulmonary's sign change

Project	The example number	Change normal	Improve	Improvement rate %
Project	The example number	Change normal	Improve	Improvement rate %	Leukocyte (very total) Chest X-rays pulmonary sign	21 20 21	14 15 15	6 4 5	92.24 95.00 95.24

7, tongue, pulse condition situation of change see Table 8.

Table 8 tongue pulse condition changes

The tongue pulse condition	The example number	Change normal	Improve	Improvement rate %
The tongue pulse condition	The example number	Change normal	Improve	Improvement rate %	Yellow and thin fur yellow and greasy fur rolling pulse arteries and veins bone number	18 12 21 9	7 3 3 4	5 3 13 1	66.67 50.00 76.19 55.56

Experimental example 2: the thin layer discrimination method of granule ephedrine hydrochloride of the present invention

Instrument and reagent: KQ-100 type ultrasonic cleaner; 101-1-BS electric heating constant temperature air dry oven; Ephedrine hydrochloride reference substance (0090-9801 is for assay Nat'l Pharmaceutical ﹠ Biological Products Control Institute); Lamellae; Hands bed board (thick 0.3mm); Tlc silica gel (lot number: 010503, Qingdao Marine Chemical Co., Ltd. makes) methanol etc.: be analytical pure.

1, the preparation of need testing solution: get this product 5g, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution.

2, the preparation of reference substance solution: get the ephedrine hydrochloride reference substance, add the liquor-saturated solution that every 1ml contains 1mg, the product solution in contrast made of first.

3, the preparation of negative control solution:, make the negative control sample that lacks ephedrine hydrochloride in the ratio and the method for making of the described prescription of technical scheme.Get 5g, make negative control solution by the preparation method of need testing solution.

4, thin layer chromatography: according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000) test, draw each 4 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (20: 5: 0.5) is developing solvent, and room temperature is launched, and exhibition is apart from about 8cm, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.The negative control chromatograph is noiseless on the relevant position.

Because ephedrine hydrochloride is a crude drug, is solubilized so directly add methanol by the described method of technical scheme, simplify the extracting method of proper mass standard.After former in addition developing solvent launched, the ephedrine hydrochloride spot and R f value was higher, and now changing developing solvent is that spot and R f value is moderate behind chloroform-methanol-strong ammonia solution.

Experimental example 3: the thin layer discrimination method of granule Pericarpium Citri Reticulatae of the present invention

Instrument and reagent: KQ-100 type ultrasonic cleaner; Pericarpium Citri Reticulatae control medicinal material (0969-9803 Nat'l Pharmaceutical ﹠ Biological Products Control Institute); Pericarpium Citri Reticulatae (feed intake and use crude drug); The chromatography neutral alumina (lot number: 010412, Shanghai the May 4th chemical reagent company limited); Lamellae: hands bed board (thick 0.3mm); Tlc silica gel (lot number: 010503, Qingdao Marine Chemical Co., Ltd. makes) methanol etc.: be analytical pure.

1, the preparation of need testing solution; Get this product 20g, add methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml, merge n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, add neutral alumina 1g and mix thoroughly, evaporate to dryness is contained in neutral alumina pillar (100～120 orders, 3g, internal diameter 1cm) on,, collects the eluent evaporate to dryness with ethyl acetate-methanol (1: 1) 30ml eluting, residue adds methanol 1ml makes dissolving, as need testing solution.

2, the preparation of control medicinal material solution: get Pericarpium Citri Reticulatae control medicinal material 0.3g, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.

3, the preparation of negative control solution:, make the negative control sample that lacks Pericarpium Citri Reticulatae in the ratio and the method for making of the described prescription of technical scheme.Get 20g, make negative control solution by the preparation method of need testing solution.

4, thin layer chromatography: according to thin layer chromatography (the about allusion quotation of China appendix VIU in 2000) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate (2: 3) is developing solvent, room temperature is launched, and exhibition is taken out apart from about 8cm, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.Negative control medical material chromatograph with corresponding 4 the principal spot positions of control medicinal material chromatograph on, Rf value has interference by last two, by under two noiseless, can be used as the discriminating of this product Pericarpium Citri Reticulatae.

Experimental example 4: the thin layer discrimination method of granule Radix Peucedani of the present invention

Instrument and reagent: Radix Peucedani control medicinal material (951-9201, Nat'l Pharmaceutical ﹠ Biological Products Control Institute); Radix Peucedani (feed intake and use crude drug); All the other are with experimental example 3.

1, the preparation of need testing solution: with experimental example 3.

2, the preparation of control medicinal material solution: get Radix Peucedani control medicinal material 0.8g, decoct with water 30 minutes, filter, filtrate is concentrated into about 20ml, puts coldly, adds ethyl acetate 30ml.Jolting is extracted, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.

3, the preparation of negative control solution:, make the negative control sample that lacks Radix Peucedani in the ratio and the method for making of the described prescription of technical scheme.Get 20g, make negative control solution by the preparation method of need testing solution.

4, thin layer chromatography: according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate (1: 1) is developing solvent, and room temperature is launched, and exhibition is apart from about 8cm, take out, dry, spray is put under the ultra-violet lamp (365nm) and is inspected with 1% sodium hydroxide alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.The negative control chromatograph is noiseless.

Experimental example 5: the thin layer discrimination method of granule Radix Glycyrrhizae of the present invention

Instrument and reagent: 101-1-BS electric heating constant temperature air dry oven; Radix Glycyrrhizae control medicinal material (904-8801 Nat'l Pharmaceutical ﹠ Biological Products Control Institute); Radix Glycyrrhizae (feed intake and use crude drug); All the other are with experimental example 3.

1, the preparation of need testing solution: get the pillar behind the eluting under the described discriminating of the technical scheme B item, add 40% methanol 30ml eluting, collect the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution.

2, the preparation of control medicinal material solution: extracting liquorice control medicinal material 0.3g, add methanol 30ml, make control medicinal material solution with the test sample method.

3, the preparation of negative control solution:, make the negative control sample that lacks Radix Glycyrrhizae in the ratio and the method for making of the described prescription of technical scheme.Get 20g, make negative control solution by the preparation method of need testing solution.

4, thin layer chromatography: according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000) test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, (15: 1: 1: 2) be developing solvent, room temperature was launched, and exhibition is apart from about 8cm with ethyl acetate-formic acid-glacial acetic acid-water, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to speckle colour developing clear.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.The negative control chromatograph is noiseless.

Experimental example 6: the content assaying method of granule ammonium chloride of the present invention

As follows with reference to former " peace cough syrup " content assaying method result of the test:

1, test method: get this product under the content uniformity item, grind well, get 2.5g respectively, the accurate title, decide, put in the 500ml kjeldahl flask, add water 250ml, jolting makes dissolving, adds bead number, add 40% sodium hydroxide solution 5ml, with the nitrogen ball kjeldahl flask is connected with condensing tube immediately, the tip of condensing tube is immersed under the liquid level of 4% boric acid solution 50ml (adding 10 of 0.1% C.I. 13020 .-0.2% bromocresol green mixing indicator solutions in the solution), adds thermal distillation, when the cumulative volume that receives liquid is about 200ml, with the most advanced and sophisticated liquid level that proposes of condensing tube, made steam washing 1 minute, with stopping distillation behind the water wash tip.Distillate becomes gray purple with sulphuric acid volumetric solution (0.05mol/L) titration to solution by aeruginous, and titrating result is proofreaied and correct with blank assay, gets promptly that [the sulphuric acid volumetric solution (0.05mol/L) of every 1ml is equivalent to the ammonium chloride NH of 5.35mg ₄Cl].

The result proofreaies and correct with blank assay, promptly gets [the ammonium chloride NH4Cl that the sulphuric acid volumetric solution (0.05mol/L) of every 1ml is equivalent to 5.35mg].

2, result of the test: get 3 batches of this product (lot number: 20030801,20030802,20030803), the scarce ammonium chloride negative control of making in prescription ratio and method for making is measured ammonium chloride content as stated above, average 192.6mg/ bag, be equivalent to 96.3% of labelled amount, the results are shown in Table 9.

Table 9

Test sample	030801	030802	030803	Negative control
Test sample	030801	030802	030803	Negative control	Content (mg/ bag)	192.2	193.3	192.3	0.68

3, ammonium chloride assay: get the ammonium chloride 0.1g of the usefulness that feeds intake, the accurate title, decide, and measures as stated above, and content is pressed dry product and calculated, and the results are shown in Table 10.

Check 3 batches, all meet two " ammonium chloride " items of (Chinese Pharmacopoeia) version in 2000 regulation (press dry product and calculate, content must not be less than 99.5g) down.

Table 10

Lot number	Criticize 1	Criticize 2	Criticize 3
Lot number	Criticize 1	Criticize 2	Criticize 3	Content (%)	99.7	99.8	99.7

Above-mentioned result of the test shows, by former " peace cough syrup " sampling amount conversion, gets this product 2.5g and tests, and method is feasible.Because this product is a preparation, by " appendix a granular preparation of Chinese pharmacopoeia version in 2000 general rule requirement, moisture must not cross 5.0%, the content uniformity limit for ± 7%, add in the preparation process factor such as weighing error, determine that it is 85～115% of recipe quantity that this product contains ammonium chloride, promptly every bag of content is 170～230mg.

Experimental example 7: granule Determination of Ephedrine Hydrochloride assay method of the present invention

1, instrument and reagent

The SP-8810 high performance liquid chromatograph; The Spectra UV-detector; Sepu 3000P chromatographic work station; Unicam UV530 ultraviolet-uisible spectrophotometer; KQ-100 type ultrasonic cleaner.Ephedrine hydrochloride reference substance (0090-9801 is for assay Nat'l Pharmaceutical ﹠ Biological Products Control Institute).Pacify the granule (lot number: 20030801,20030802,20030803) of coughing.Methanol: chromatographically pure; Water is ultra-pure water; Other reagent: be analytical pure.

2, high-efficient liquid phase chromatogram condition and system suitability test

C ₁₈Stainless steel column (Hypersil ODS2 4.6 * 200mm 5 μ m); Mobile phase: methanol-0.05mol/L potassium dihydrogen phosphate (regulating pH value with phosphoric acid is 3.0) (10: 90); Flow velocity: 1.0ml/min, 1.2ml/min; Detect wavelength: 209nm column temperature: room temperature.Number of theoretical plate is pressed the ephedrine hydrochloride peak and is calculated, and should be not less than 2800.

3, standard curve

It is an amount of that precision takes by weighing the ephedrine hydrochloride reference substance, adds methanol and make the solution that contains 9.664,20.536,30.200,41.072,51.340 μ g among every 1ml.Draw 10 μ l respectively, inject high performance liquid chromatograph, measure peak area, the results are shown in Table 11 by above-mentioned condition.

Table 11

C(μg/ml)	9.664	20.536	30.200	41.072	51.340
C(μg/ml)	9.664	20.536	30.200	41.072	51.340	Peak area	175728	374744	520988	716887	889397

As abscissa, the peak area that records carries out linear regression as vertical coordinate with reference substance concentration, and getting linear equation is Y=17039X+14783, r=1.000.The range of linearity is 0.09664～0.5134 μ g.

4, the preparation of need testing solution

Get this product under the content uniformity item, grind well, get 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 0.1% methanol hydrochloride solution 25ml that adds, claim to decide weight, supersound process 30 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 0.1% methanol hydrochloride solution, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.

1. extract sample solvent; Select for use methanol, 0.1% methanol hydrochloride solution to test, observe from the HPLC collection of illustrative plates, extract sample with 0.1% methanol hydrochloride solution, the ephedrine hydrochloride peak shape is good than methanol, thereby with its solvent as the extraction sample.

2. extracting method; As extracting solvent, select supersound process 30 minutes, 60 minutes with 0.1% methanol hydrochloride solution; Reflux was extracted in 1 hour, and extracting solution is pressed content assaying method and measured ephedrine hydrochloride, the results are shown in Table 12.

Table 12

Extracting method	Supersound process 30 minutes	Supersound process 60 minutes	Reflux 1 hour
Extracting method	Supersound process 30 minutes	Supersound process 60 minutes	Reflux 1 hour	Content (mg/ bag)	3.756	3.739	3.798

Berberine hydrochloride is soluble in methanol, and result of the test is represented above-mentioned extracting method, and it is basic identical to measure the ephedrine hydrochloride content results, all can be used as the extracting method of ephedrine hydrochloride assay.The present invention is in conjunction with recovery test, selects for use comparatively easily supersound process 30 minutes as the described method of technical scheme.

5, the preparation of reference substance solution

It is an amount of that precision takes by weighing the ephedrine hydrochloride reference substance, adds 0.1% methanol hydrochloride solution and make the solution that every 1ml contains 30 μ g, in contrast product solution.

6, algoscopy

(1) selection of mensuration wavelength: get the ephedrine hydrochloride reference substance solution in the interscan of 200～360nm wave-length coverage, the result has absorption maximum at 209nm wavelength place, is λ=209nm so select to measure wavelength.

(2) selection of mobile phase: select methanol-water (1: 1), acetonitrile-0.01% phosphoric acid solution (5: 95), methanol-0.05mol/L potassium dihydrogen phosphate (8: 92) and methanol-0.05mol/L.Potassium dihydrogen phosphate (regulating pH value with phosphoric acid is 3.0) (10: 90) waits to be tested, and the result shows last a kind of mobile phase separating effect, peak shape is best and as the mobile phase of the described method of technical scheme.

Assay method such as technical scheme, accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure by above-mentioned condition.

7, blank assay

Get scarce ephedrine hydrochloride negative sample, handle and measure by the preparation and the assay method of need testing solution in prescription ratio and method for making preparation.The result does not see that blank assay has interference.

8, precision test

1. reference substance; Accurate ephedrine hydrochloride reference substance solution (30 μ g/ml) the 10 μ l that draw repeat sample introduction 5 times by above-mentioned condition.Measurement result sees Table 13.

Table 13

Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)
Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)	Peak area	520960	522046	525358	521795	520988	522229	0.34

2. sample: accurate draw same test sample (lot number: 20030801) solution 10 μ l, repeat sample introduction 5 times by above-mentioned condition.The results are shown in Table 14.

Table 14

Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)
Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)	Peak area	513596	502937	499503	508298	513062	507479	1.21

9, replica test

(lot number: 20030802) grind well, precision takes by weighing 5 parts, handles and measures by content assaying method, the results are shown in Table 15 to get same test sample.

Table 15

Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)
Sequence number	1	2	3	4	5	Meansigma methods	RSD(％)	Content (mg/ bag)	3.611	3.584	3.400	3.614	3.437	3.529	2.92

10, stability test

20030801) and each 10 μ l of reference substance solution (30.2 μ g/ml) accurate draw same need testing solution (lot number:, after start was waited to stablize in 1 hour, every 2 hours injection high performance liquid chromatographs, measurement result saw Table 16.

Table 16

Minute (h)	1	3	5	7	9	Meansigma methods	RSD(％)
Minute (h)	1	3	5	7	9	Meansigma methods	RSD(％)	The reference substance peak area	531961	530265	515572	513751	534252	525160	1.83
The sample peak area	523674	539642	541145	520989	537944	532679	1.79	The reference substance peak area	531961	530265	515572	513751	534252	525160	1.83

11, application of sample recovery test

Get 5 parts of each about 1g of same test sample (lot number: 20030801, content is 0.7278mg/g), accurate claim fixed, accurate respectively berberine hydrochloride reference substance solution (0.630mg/ml) 1ml that adds, heating in water bath is flung to methanol, presses the content assaying method sample introduction respectively and measures calculate recovery rate.The results are shown in Table 17.

Table 17

Sequence number	Test sample (g)	Content of berberine hydrochloride (mg)	Add reference substance amount (mg)	The amount of recording (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)
Sequence number	Test sample (g)	Content of berberine hydrochloride (mg)	Add reference substance amount (mg)	The amount of recording (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)	1 2 3 4 5	0.9463 0.9502 0.9509 0.9506 0.9519	0.6887 0.6916 0.6921 0.6918 0.6928	0.630 0.630 0.630 0.630 0.630	1.2994 1.3132 1.3192 1.3186 1.3293	98.54 99.36 99.78 99.76 100.49	99.59	0.72

12, assay: get 3 batches of this product, measure ephedrine hydrochloride content, the results are shown in Table 18 by the content assaying method of drafting.

Table 18

Lot number	20030801	20030802	20030803
Lot number	20030801	20030802	20030803	Content (mg/ bag)	3.71	3.74	3.71

13, the ephedrine hydrochloride material content is measured: get the ephedrine hydrochloride of the usefulness that feeds intake, measure by the content assaying method of drafting, the results are shown in Table 19.

Table 19

Lot number	Criticize 1	Criticize 2	Criticize 3
Lot number	Criticize 1	Criticize 2	Criticize 3	Content (%)	99.6	99.9	99.7

Check 3 batches, all meet " regulation (press dry product and calculate, content must not be less than 99.0%) under two " ephedrine hydrochloride " items of Chinese pharmacopoeia version in 2000.

Because this product is a preparation, by " appendix a granular preparation of Chinese pharmacopoeia version in 2000 general rule requirement, moisture must not cross 5.0%, the content uniformity limit for ± 7%, add in the preparation process factor such as weighing error, determine that the hydrochloric ephedrine of this product is 85～115% of a recipe quantity, promptly every bag of content is 3.10～4.20mg.

The following embodiment of the present invention all can reach the effect of above-mentioned experimental example.

Embodiment 1. granule of the present invention

Bulbus Fritillariae Thunbergii fluid extract 25ml, Radix Glycyrrhizae fluidextract 20ml, Radix Stemonae 25g, Radix Platycodonis fluidextract 18ml, Radix Peucedani 22g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 22g, Pericarpium Citri Reticulatae 12g, ammonium chloride 20g, ephedrine hydrochloride 0.365g, Mentholum 0.15g;

More than ten flavors, Radix Peucedani adds water in 85～90 ℃ of warm macerating three times, first and second time each 3 hours, 2 hours for the third time, filter, filtrate merges, and is concentrated in right amount, puts coldly, adds ethanol and makes and contain the alcohol amount and reach 25% approximately, and is standby; The Radix Stemonae, Pericarpium Citri Reticulatae pulverize separately become coarse powder, according to the percolation (Chinese Pharmacopoeia appendix 1 O in 2000) under fluid extract and the extractum item, the Radix Stemonae with 55% ethanol make solvent, Pericarpium Citri Reticulatae is made solvent with 80% ethanol, carry out percolation respectively, collect the Radix Stemonae liquid 25ml that filters, the Pericarpium Citri Reticulatae liquid 5ml that filters, standby.Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) is ground into coarse powder, with 50% ethanol in 70～80 ℃ of warm macerating three times, each 3 hours, merge leachate, standby.Above four kinds of reserve liquids are merged, decompression recycling ethanol is to there not being the alcohol flavor, with Bulbus Fritillariae Thunbergii, Radix Glycyrrhizae, three kinds of fluid extract mixings of Radix Platycodonis, leave standstill, filter, being concentrated into relative density is the clear paste of 1.29～1.35 (22 ℃), add ephedrine hydrochloride, get 1 part of the clear paste of above-mentioned mixing, add ammonium chloride 20g and dextrin 34g, sucrose 425g, mixing; It is an amount of to add 40% ethanol, makes granule, and drying is made 500g, and spray adds 90% alcoholic solution of Mentholum, mixing, and 100 bags of packing, every bag of 5g, promptly.Oral, a 5-7.5g, 3 times on the one.

Embodiment 2. the method for quality control of granule of the present invention

A. get compound granular agent 5g of the present invention, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (20: 5: 0.5) is developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

B. get compound granular agent 20g of the present invention, add methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml, merge n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, add neutral alumina 1g and mix thoroughly, evaporate to dryness is contained in neutral alumina pillar (100～120 orders, 3g, internal diameter 1cm) on,, collects the eluent evaporate to dryness with ethyl acetate-methanol (1: 1) 30ml eluting, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Citri Reticulatae control medicinal material 0.3g, adds methanol 30ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VI B in 2000), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate (2: 3) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

C. get Radix Peucedani control medicinal material 0.8g, decoct with water 30 minutes, filter, filtrate is concentrated into about 20ml, puts coldly, adds ethyl acetate 30ml jolting extraction, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw need testing solution and each 5 μ l of control medicinal material solution of differentiating under the B item, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is put under the ultra-violet lamp (365nm) and is inspected with 1% sodium hydroxide alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

D. get and differentiate the pillar behind the eluting under the B item, add 40% methanol 30ml eluting, collect the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds methanol 30ml in addition, shines medical material solution in pairs with legal system; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ethyl acetate-formic acid-glacial acetic acid-water (15: 1: 1: 2) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Embodiment 3. the method for quality control of granule of the present invention

B. get compound granular agent 20g of the present invention, add methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml, merge n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes dissolving, add neutral alumina 1g and mix thoroughly, evaporate to dryness is contained in neutral alumina pillar (100～120 orders, 3g, internal diameter 1cm) on,, collects the eluent evaporate to dryness with ethyl acetate-methanol (1: 1) 30ml eluting, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Citri Reticulatae control medicinal material 0.3g, adds methanol 30ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIB in 2000), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate (2: 3) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color.

Embodiment 4. the method for quality control of granule of the present invention

E. ammonium chloride

Get the compound granular agent of the present invention under the content uniformity item, grind well, get 2.5g, it is fixed to claim, put in the 500ml kjeldahl flask, add water 250ml, jolting makes dissolving, adds bead number, add 40% sodium hydroxide solution 5ml, with the nitrogen ball kjeldahl flask is connected with condensing tube immediately, the tip of condensing tube is immersed under the liquid level of 4% boric acid solution 50ml (adding 10 of 0.1% C.I. 13020 .-0.2% bromocresol green mixing indicator solutions in the solution), adds thermal distillation, when the cumulative volume that receives liquid is about 200ml, with the most advanced and sophisticated liquid level that proposes of condensing tube, made steam washing 1 minute, with stopping distillation behind the water wash tip; Distillate becomes gray purple with sulphuric acid volumetric solution (0.05mol/L) titration to solution by aeruginous, and titrating result is proofreaied and correct with blank assay, promptly; The sulphuric acid volumetric solution (0.05mol/L) of every 1ml is equivalent to the ammonium chloride NH4Cl of 5.349mg;

F. ephedrine hydrochloride

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-0.05mol/L potassium dihydrogen phosphate (regulating pH value with phosphoric acid is 3.0) (10: 90) is mobile phase; The detection wavelength is 209nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 2800;

It is an amount of to take by weighing ephedrine hydrochloride, adds 0.1% methanol hydrochloride solution and makes the solution that every 1ml contains 30 μ g, promptly gets reference substance solution;

Get the compound preparation of the present invention under the content uniformity item, grind well, get 1g, it is fixed to claim, puts in the tool plug conical flask, the accurate 0.1% methanol hydrochloride solution 25ml that adds, claim to decide weight, supersound process 30 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 0.1% methanol hydrochloride solution, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly get need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly; Every bag of hydrochloric ephedrine of compound granular agent of the present invention (C10H15NOHCl) should be 3.10～4.20mg.

Embodiment 5. the method for quality control of granule of the present invention

Ephedrine hydrochloride:

Embodiment 6. the method for quality control of granule of the present invention

Differentiate:

A. get compound granular agent 5g of the present invention, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (Chinese Pharmacopoeia appendix VIF in 2000), draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol-strong ammonia solution (20: 5: 0.5) is developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay:

E. ammonium chloride

F. ephedrine hydrochloride

Embodiment 7. the method for quality control of granule of the present invention

Differentiate:

Assay:

F. ephedrine hydrochloride

Embodiment 8. the preparation of granule of the present invention and quality control standard thereof

More than ten flavors, Radix Peucedani adds water in 85～90 ℃ of warm macerating three times, first and second time each 3 hours, 2 hours for the third time, filter, filtrate merges, and is concentrated in right amount, puts coldly, adds ethanol and makes and contain the alcohol amount and reach 25% approximately, and is standby.The Radix Stemonae, Pericarpium Citri Reticulatae pulverize separately become coarse powder, according to the percolation (Chinese Pharmacopoeia appendix l O in 2000) under fluid extract and the extractum item, the Radix Stemonae with 55% ethanol make solvent, Pericarpium Citri Reticulatae is made solvent with 80% ethanol, carry out percolation respectively, collect the Radix Stemonae liquid 25ml that filters, the Pericarpium Citri Reticulatae liquid 5ml that filters, standby.Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) is ground into coarse powder, with 50% ethanol in 70～80 ℃ of warm macerating three times, each 3 hours, merge leachate, standby.Above four kinds of reserve liquids are merged, decompression recycling ethanol, leaves standstill with Bulbus Fritillariae Thunbergii, Radix Glycyrrhizae, three kinds of fluid extract mixings of Radix Platycodonis to there not being the alcohol flavor, filter, being concentrated into relative density is the clear paste of 1.29～1.35 (22 ℃), adds ephedrine hydrochloride, gets 1 part of the clear paste of above-mentioned mixing, add 1 part in dextrin, ammonium chloride 20g, sucrose is an amount of, mixing; It is an amount of to add 40% ethanol, makes granule, and drying is made 500g, and spray adds 90% alcoholic solution of Mentholum, mixing, and 100 bags of packing, promptly.

Quality control standard by the granule of the present invention of above-mentioned preparation method preparation is:

Differentiate:

Assay:

E. ammonium chloride

F. ephedrine hydrochloride

Claims

One kind the treatment bronchitic compound granular agent method of quality control, it is characterized in that the content assaying method in this method is:

Ephedrine hydrochloride shines high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; To regulate pH value with phosphoric acid be that methanol-0.05mol/L potassium dihydrogen phosphate of 3.0 is a mobile phase, and the mobile phase ratio of methanol-0.05mol/L potassium dihydrogen phosphate is 5～15: 95～85; The detection wavelength is 209nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 2800;

It is an amount of to take by weighing ephedrine hydrochloride, adds 0.1% methanol hydrochloride solution and makes the solution that every 1ml contains 30 μ g, promptly gets reference substance solution; Get the compound granular agent under the content uniformity item, grind well, get 1g, it is fixed to claim, puts in the tool plug conical flask, the accurate 0.1% methanol hydrochloride solution 25ml that adds, claim to decide weight, supersound process 30 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 0.1% methanol hydrochloride solution, shake up, with the microporous filter membrane filtration of 0.45 μ m, get subsequent filtrate, promptly get need testing solution; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure, promptly; Every bag of hydrochloric ephedrine of compound granular agent should be 3.10～4.20mg;

Described compound granular agent prepares by the following method: Bulbus Fritillariae Thunbergii fluid extract 20-25 parts by volume, Radix Glycyrrhizae fluidextract 20-25 parts by volume, Radix Stemonae 20-25 weight portion, Radix Platycodonis fluidextract 15-20 parts by volume, Radix Peucedani 20-25 weight portion, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 20-25 weight portion, Pericarpium Citri Reticulatae 10-15 weight portion, ammonium chloride 20-25 weight portion, ephedrine hydrochloride 0.3-0.4 weight portion, Mentholum 0.1-0.2 weight portion; Get above-mentioned ten flavor crude drug, Radix Peucedani adds water in 85～90 ℃ of warm macerating 2-3 time, and each 2-3 hour, filter, filtrate merges, and is concentrated in right amount, puts coldly, adds ethanol and makes and contain alcohol and measure and reach 25%, and is standby; The Radix Stemonae, Pericarpium Citri Reticulatae pulverize separately become coarse powder, and appendix of Chinese Pharmacopoeia version in 2000 10 is according to the percolation under fluid extracts and the extractum item, the Radix Stemonae with 55% ethanol make solvent, Pericarpium Citri Reticulatae is made solvent with 80% ethanol, carry out percolation respectively, collect the Radix Stemonae liquid of filtering, the Pericarpium Citri Reticulatae liquid of filtering, standby; Rhizoma Pinelliae powder is broken into coarse powder, with 50% ethanol in 70～80 ℃ of warm macerating 2-3 time, each 2-3 hour, merge leachate, standby; Above four kinds of reserve liquids are merged, decompression recycling ethanol is to there not being the alcohol flavor, with Bulbus Fritillariae Thunbergii, Radix Glycyrrhizae, three kinds of fluid extract mixings of Radix Platycodonis, leave standstill, filter, be concentrated into relative density under 22 ℃ of temperature and be 1.29～1.35 clear paste, add ephedrine hydrochloride, get the clear paste of above-mentioned mixing, add ammonium chloride, an amount of dextrin and sucrose, mixing; It is an amount of to add 40% ethanol, makes granule, drying, and spray adds 90% alcoholic solution of Mentholum, and mixing is distributed into bag, and every bag of 5g promptly gets the compound granular agent.
2. the method for quality control of compound granular agent as claimed in claim 1 is characterized in that the ratio of methanol-0.05mol/L potassium dihydrogen phosphate mobile phase in the described Determination of Ephedrine Hydrochloride mensuration of this method is 10: 90.
3. the method for quality control of compound granular agent as claimed in claim 1 or 2 is characterized in that this method comprises one or more in the following discriminating:

A. get described compound granular agent 5g, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is 15～25: 4～6: chloroform-methanol-strong ammonia solution of 0.5 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get described compound granular agent 20g, add methanol 50ml, supersound process 15 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml merges n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, adds neutral alumina 1g and mixes thoroughly, evaporate to dryness, being contained on the neutral alumina pillar, is ethyl acetate-methanol 30ml eluting of 1～2: 1～2 with ratio, collects the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Citri Reticulatae control medicinal material 0.3g, adds methanol 30ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that normal hexane-ethyl acetate of 1～3: 2～4 is developing solvent, launches, and takes out, dry, put under the wavelength 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color;

C. get Radix Peucedani control medicinal material 0.8g, decoct with water 30 minutes, filter, filtrate is concentrated into 20ml, puts coldly, adds ethyl acetate 30ml jolting and extracts, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution of differentiating under the B item, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that normal hexane-ethyl acetate of 1～2: 1～2 is developing solvent, launch, take out, dry, spray is put under the wavelength 365nm ultra-violet lamp and is inspected with 1% sodium hydroxide alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color;

D. get and differentiate the pillar behind the eluting under the B item, add 40% methanol 30ml eluting, collect the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds methanol 30ml in addition, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ratio is 10～20: ethyl acetate-formic acid of 1: 1: 2-glacial acetic acid-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
4. the method for quality control of compound granular agent as claimed in claim 3 is characterized in that this method comprises one or more in the following differential method:

A. get described compound granular agent 5g, add methanol 30ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, filters, and filtrate is as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that chloroform-methanol-strong ammonia solution of 20: 5: 0.5 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get described compound granular agent 20g, add methanol 50ml, supersound process 15 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, with water saturated n-butanol extraction 2 times, each 30ml merges n-butanol extracting liquid, add water 15ml washing, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, adds neutral alumina 1g and mixes thoroughly, evaporate to dryness, being contained on the neutral alumina pillar, is ethyl acetate-methanol 30ml eluting of 1: 1 with ratio, collects the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Citri Reticulatae control medicinal material 0.3g, adds methanol 30ml, and supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that normal hexane-ethyl acetate of 2: 3 is developing solvent, launches, and takes out, dry, put under the wavelength 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color;

C. get Radix Peucedani control medicinal material 0.8g, decoct with water 30 minutes, filter, filtrate is concentrated into 20ml, puts coldly, adds ethyl acetate 30ml jolting and extracts, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution and each 5 μ l of control medicinal material solution of differentiating under the B item, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ratio is that normal hexane-ethyl acetate of 1: 1 is developing solvent, launch, take out, dry, spray is put under the wavelength 365nm ultra-violet lamp and is inspected with 1% sodium hydroxide alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence principal spot of same color;

D. get and differentiate the pillar behind the eluting under the B item, add 40% methanol 30ml eluting, collect the eluent evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.3g adds methanol 30ml in addition, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ratio is 15: 1: 1: ethyl acetate-formic acid of 2-glacial acetic acid-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
5. the method for quality control of compound granular agent as claimed in claim 4 is characterized in that this method also comprises following assay:

Get the described compound granular agent under the content uniformity item, grind well, get 2.5g, it is fixed to claim, put in the 500ml kjeldahl flask, add water 250ml, jolting makes dissolving, add bead number, add 40% sodium hydroxide solution 5ml, with the nitrogen ball kjeldahl flask is connected with condensing tube immediately, the tip of condensing tube is immersed under the liquid level of 4% boric acid solution 50ml, add 0.1% C.I. 13020 .-0.2% bromocresol green in the solution and mix 10 of indicator solutions, add thermal distillation, when the cumulative volume that receives liquid is 200ml, with the most advanced and sophisticated liquid level that proposes of condensing tube, made steam washing 1 minute, with stopping distillation behind the water wash tip; Distillate becomes gray purple with 0.05mol/L sulphuric acid volumetric solution titration to solution by aeruginous, and titrating result is proofreaied and correct with blank assay, promptly; The 0.05mol/L sulphuric acid volumetric solution of every 1ml is equivalent to the ammonium chloride NH4Cl of 5.349mg; Described compound granular agent contains ammonium chloride for every bag and should be 170～230mg.