CN1813973A

CN1813973A - Quality control method of keke oral preparation for relieving cough

Info

Publication number: CN1813973A
Application number: CN 200510200745
Authority: CN
Inventors: 叶湘武; 汤琼; 张梅
Original assignee: Guizhou Yibai Pharmaceutical Co Ltd
Current assignee: Guizhou Yibai Pharmaceutical Co Ltd
Priority date: 2005-11-25
Filing date: 2005-11-25
Publication date: 2006-08-09
Anticipated expiration: 2025-11-25
Also published as: CN100510744C

Abstract

The present invention relates to a quality control method of Keke oral preparation for relieving cough and asthma. It includes trait, examination, identification and content determination. The described trait identification includes: soft extract identification, said product is a dark brown thick semifluid, its taste is sweet and slight bitter; syrup identification, said product is a brownish yellow thick liquid, its taste is sweet and slight bitter; and oral liquor identification, said product is a brownish yellow liquid, its taste is sweet and light bitter. The described identification includes thin-layer identification of ephedra root, poppy husk and licorice, the described content determination includes the content determination of ephedrine hydrochloride and codeine phosphate. Said invention can effectively control the quality of Keke soft extract, Keke syrup and Keke oral liquor preparation.

Description

The method of quality control of KEKE KOUFU ZHIJI

Technical field: the present invention relates to a kind of method of quality control of KEKE KOUFU ZHIJI, belong to technical field of medicaments.

Background technology: cough is a kind of commonly encountered diseases in our daily life, cough stopping formulation is to develop on the MAXINGSHIGAN TANG basis of China's traditional Chinese medical science famous classic treatise on Febrile Diseases, is total to seven flavor medicine by Herba Ephedrae, Pericarpium Papaveris, Radix Glycyrrhizae, Semen Armeniacae Amarum, Semen Raphani, Radix Platycodonis, Gypsum Fibrosum and forms; Have the lung qi dispersing heat clearing away, cough-relieving, the effect of relievining asthma, to upper respiratory tract infection, tracheitis, pneumonia, diseases such as cough have definite curative effect." KEKE JIAONANG " publication is the 17 of Drug Standard of Ministry of Public Health of the Peoples Republic of China, this medicine is used for many years clinically, obtain satisfied therapeutic effect at treatment cough, asthma fermentation, though this product is being obtained certain achievement aspect the diseases such as treatment cough, but, existing dosage form on the market only has capsule, is difficult to satisfy patient's needs.And because cough, asthma occurs in winter mostly since winter dry weather, patient is except cough, also often with dry throat and mouth, dry cough expectorant is few or do not have symptom such as expectorant, need take and have the medicine of moisturizing the function of promoting the production of body fluid, the therapeutic effect that can play this moment.General solid preparation is not moisturized the effect of promoting the production of body fluid with regard to the characteristics of its dosage form itself, can not reach optimum therapeuticing effect.Therefore the applicant has made soft extract, oral liquid or syrup with the cough stopping formulation of routine, takes in exsiccant season, just can play a multiplier effect.But how effectively to control the quality of cough-relieving soft extract, oral liquid or syrup, also do not have relevant quality standard can supply reference at present.

Summary of the invention:

The objective of the invention is to: the method for quality control that a kind of KEKE KOUFU ZHIJI is provided.This oral formulations comprises soft extract, oral liquid and syrup, by method of quality control of the present invention, effectively controls the quality of this KEKE KOUFU ZHIJI, guarantees the clinical efficacy of said preparation.

KEKE KOUFU ZHIJI of the present invention is to constitute like this: calculate according to composition by weight: it mainly is prepared from by Herba Ephedrae 43.2-240, Pericarpium Papaveris 43.2-240, Radix Glycyrrhizae 43.2-240, Semen Armeniacae Amarum 43.2-240, Semen Raphani 13.6-75, Radix Platycodonis 13.6-75, Gypsum Fibrosum 13.6-75 and adjuvant.

Cough-relieving soft extract of the present invention and syrup are preparations like this: Herba Ephedrae, Pericarpium Papaveris are pulverized, and add 3-30 and doubly measure acidic aqueous solution, soak after 0.5-5 hour to decoct 1-5 time, and each 0.5-5 hour, filter, merging filtrate, medicinal liquid transfer PH to 6-8 standby; Semen Armeniacae Amarum steamed 20-60 minute earlier, Radix Glycyrrhizae, Semen Raphani, Radix Platycodonis, Gypsum Fibrosum four Chinese medicine add 3-15 times of water gaging and soaked 0.5-5 hour, decoct 1-5 time with Semen Armeniacae Amarum then, each 0.5-5 hour, filter, filtrate merges, with Herba Ephedrae, the Pericarpium Papaveris extracting solution mixes, concentrating under reduced pressure, add different adjuvants again, can make syrup or soft extract with conventional method.

Cough-relieving oral liquid of the present invention is preparation like this: Herba Ephedrae, Pericarpium Papaveris are pulverized, and add 3-30 and doubly measure acidic aqueous solution, soak after 0.5-5 hour to decoct 1-5 time, and each 0.5-5 hour, filter, merging filtrate, medicinal liquid transfer PH to 6-8 standby; Semen Armeniacae Amarum steamed 20-60 minute earlier, and Radix Glycyrrhizae, Semen Raphani, Radix Platycodonis, Gypsum Fibrosum four Chinese medicine add 3-15 times of water gaging and soaked 0.5-5 hour, decocted 1-5 time each 0.5-5 hour then with Semen Armeniacae Amarum, filter, filtrate merges, with Herba Ephedrae, the Pericarpium Papaveris extracting solution mixes, and concentrating under reduced pressure is put cold, add the ethanol precipitate with ethanol, leave standstill, filtrate recycling ethanol also continues to be condensed into thick paste, add water-cooled again and hid 30-150 hour, filter, filtrate adds the adjuvant dissolving, add water to 1000ml, transfer PH to 3-7, promptly.

The method of quality control of KEKE KOUFU ZHIJI of the present invention is such: it comprises character, inspection, discriminating and assay, and described character comprises: for soft extract, product is the stiff semifluid of sepia, and it is sweet to distinguish the flavor of, little hardship; For syrup, product is pale brown color thick liquid, and it is sweet to distinguish the flavor of, little hardship; For oral liquid, product is pale brown color liquid, is equipped with the easily diffusing precipitation of jog for a long time, and it is sweet to distinguish the flavor of, little hardship; Described inspection comprises: relative density, and the soft extract relative density should be between 1.30～1.50, and the syrup relative density is not less than 1.20; PH value, the oral liquid pH value should be 3.5～6.5; Other should meet every regulation relevant under Chinese Pharmacopoeia syrup, soft extract or the mixture item; Described discriminating comprises the epheday intermedia with the ephedrine hydrochloride reference substance side of discriminating, with Pericarpium Papaveris in codeine phosphate reference substance and the Pericarpium Papaveris control medicinal material discriminating side, with the thin layer of Radix Glycyrrhizae in the enoxolone reference substance discriminating side differentiate partly or entirely; Described assay comprise to the assay of ephedrine hydrochloride, codeine phosphate partly or entirely.

Specifically, the discrimination method of this preparation epheday intermedia comprises that with the ephedrine hydrochloride reference substance be contrast, and with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is the thin layer discrimination method of developing solvent.

The discrimination method of Pericarpium Papaveris comprises that with codeine phosphate reference substance and Pericarpium Papaveris control medicinal material be contrast in this preparation, and with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is the thin layer discrimination method of developing solvent.

The discrimination method of Radix Glycyrrhizae comprises that with the enoxolone reference substance be contrast in this preparation, and with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is the thin layer discrimination method of developing solvent.

In particular, the discriminating of this preparation is selected from one or more in the following method:

(1) Herba Ephedrae: get soft extract, syrup or oral liquid, wherein soft extract adds the water mixing earlier, and syrup needs thin up, and medicinal liquid is transferred pH8～13 with ammonia, with chloroform extraction 1-5 time, the combined chloroform extracting solution extracts with hydrochloric acid solution, the acid solution evaporate to dryness, residue is with methanol or dissolve with ethanol, as need testing solution; Other gets the ephedrine hydrochloride reference substance, with methanol or dissolve with ethanol, and product solution in contrast; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract, syrup or oral liquid, add ethyl acetate and ammonia water mixture, supersound extraction, extracting solution filters, and filtrate evaporate to dryness, residue add methanol or dissolve with ethanol, as need testing solution; Other gets the Pericarpium Papaveris control medicinal material, adds the mixed liquor of ethyl acetate and ammonia, supersound extraction, and extracting solution filters, and filtrate evaporate to dryness, residue add methanol or ethanol makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, uses dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract, syrup or oral liquid add methanol or ethanol ultrasonic extraction, and extracting solution filters, the filtrate evaporate to dryness, residue add water make the dissolving and be transferred in the separatory funnel, add dilute sulfuric acid, shake up, with ether extraction 1-5 time, discard ether solution, water liquid closes n-butanol extracting liquid with water saturated n-butanol extraction 1-5 time, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, discard water lotion, n-butanol extracting liquid evaporate to dryness, residue with dissolve with methanol to flask, evaporate to dryness, add hydrochloric acid and chloroform, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds methanol or alcoholic solution makes dissolving, as need testing solution; In addition extracting liquorice subacid reference substance is an amount of, with methanol or dissolve with ethanol, and product solution in contrast; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Say that accurately the above-mentioned discriminating of this preparation is selected from one or more in the following method:

(1) Herba Ephedrae: get soft extract 2g, add water 10-50ml mixing; Or get syrup 5ml, add water 10-50ml mixing; Or get oral liquid 20ml.Transfer pH8～13 with ammonia, use chloroform extraction 1-5 time, each 10-50ml, the combined chloroform extracting solution extracts 1-5 time with 10-50ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5-1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract 5g or syrup 5ml or oral liquid 20ml, add ethyl acetate 10-100ml and ammonia 0.5-5ml, supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds the mixed liquor of ethyl acetate 10-100ml and ammonia 0.5-5ml, and supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH8～13 with ammonia solution, use ethyl acetate extraction 1-5 time, each 10-100ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 10-100ml, and supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=5-25: 1-9: 0.2-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

The another kind of discrimination method of this preparation is selected from one or more in the following method:

(2) Pericarpium Papaveris: get soft extract, syrup or oral liquid, transfer PH8～13 with ammonia solution, use ethyl acetate extraction 1-5 time, merge ethyl acetate liquid, water bath method, residue add methanol or dissolve with ethanol, as need testing solution; Other gets the Pericarpium Papaveris control medicinal material, adds ethyl acetate, supersound extraction, and extracting solution filters, and filtrate evaporate to dryness, residue add methanol or ethanol makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, uses dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) Radix Glycyrrhizae: get soft extract, syrup or oral liquid, thin up also is transferred in the separatory funnel, add dilute sulfuric acid, shake up, use ether extraction 1-5 time, discard ether solution, water liquid is with water saturated n-butanol extraction 1-5 time, close n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time discards water lotion, the n-butanol extracting liquid evaporate to dryness, to flask, evaporate to dryness adds hydrochloric acid and chloroform to residue with dissolve with methanol, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds methanol or ethanol makes dissolving, as need testing solution; In addition extracting liquorice subacid reference substance is an amount of, with methanol or dissolve with ethanol, and product solution in contrast; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with petroleum ether (30～60 ℃): chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Say that accurately the above-mentioned another kind of discrimination method of this preparation is selected from one or more in the following method:

(1) gets soft extract 2g, add water 10-50ml mixing; Or get syrup 5ml, add water 10-50ml mixing; Or get oral liquid 20ml.Transfer pH8～13 with ammonia, use chloroform extraction 1-5 time, each 10-50ml, the combined chloroform extracting solution extracts 1-5 time with 10-50ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5-1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(2) get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH8～13 with ammonia solution, use ethyl acetate extraction 1-5 time, each 10-100ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 10-100ml, and supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 10-50ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 1-5ml, shake up, with ether extraction 1-5 time, each 10-100ml discards ether solution, water liquid is with water saturated n-butanol extraction 1-5 time, each 10-100ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, each 10-100ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 5-20ml, evaporate to dryness, add hydrochloric acid 1-10ml and chloroform 20-100ml, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds ethanol 0.5-2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 0.5-2mg with dissolve with ethanol, in contrast product solution.According to the test of Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with petroleum ether (30～60 ℃): chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Specifically, the Determination of Ephedrine Hydrochloride assay method comprises that adopting chromatographic column is C18 or C4 or C8 post in this preparation, with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and detecting wavelength is the high performance liquid chromatography of 200-500nm.

The content assaying method of codeine phosphate comprises that adopting chromatographic column is C18 or C4 or C8 post in this preparation, with 0.01-1mol/L sodium acetate solution: methanol=1-9: 9-1 is mobile phase, detecting wavelength is the high performance liquid chromatography of 200-500nm, and wherein the 0.01-1mol/L sodium acetate solution needs to transfer pH=2-5 with glacial acetic acid.

In particular, the content assaying method of this preparation is selected from one or more in the following method:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and the detection wavelength is 200-500nm; Get the ephedrine hydrochloride reference substance, the accurate title, decide, and uses dissolve with methanol, shakes up, and promptly gets reference substance solution; Get soft extract, syrup or oral liquid, with the mobile phase dilution, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; Contain Herba Ephedrae in the soft extract in ephedrine hydrochloride, must not be less than 0.5mg/g, contain Herba Ephedrae in the syrup, must not be less than 0.5mg/ml, contain Herba Ephedrae in the oral liquid, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with 0.01-1mol/L sodium acetate solution (glacial acetic acid is transferred pH=2-5): methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get the codeine phosphate reference substance, the methanol-water solution dissolving with 0.5-5 doubly measures shakes up, and precision is measured 1ml, puts in the 5-25ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, and promptly gets reference substance solution; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add silicon dried soil and mix thoroughly, and be transferred in the round-bottomed flask, with ethanol water-bath reflux, extract,, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue is dissolved in water and is transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 1-8 time, combined chloroform extracting solution, water-bath is waved to 10-30ml, quantitatively be transferred in the separatory funnel, extract 1-5 time, merge acid solution with hydrochloric acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, cross chloroform extraction 2-8 time with water saturation, combined chloroform extracting solution, water bath method, residue dissolves with mobile phase, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Say that accurately the content assaying method of above-mentioned preparation is selected from one or more in the following method:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and the detection wavelength is 200-500nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.05-0.15mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and each 5-20 μ l of need testing solution of drawing injects hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with 0.01-1mol/L sodium acetate solution (glacial acetic acid is transferred pH=2-5): methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5-50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5-25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, adding silicon dried soil mixes thoroughly, and be transferred in the round-bottomed flask, use 10-100ml ethanol water-bath reflux, extract, 0.5-5 hour, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue add the 5-50ml water dissolution and are transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 1-8 time, each 5-50ml, combined chloroform extracting solution, water-bath is waved to 10-30ml, quantitatively be transferred in the separatory funnel, extract 1-5 time, each 5-50ml with hydrochloric acid solution 1 → 200, merge acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, crosses chloroform extraction 2-8 time with water saturation, each 5-50ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 5-25ml measuring bottle, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

The another kind of content assaying method of this preparation is selected from one or more in the following method:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and the detection wavelength is 200-500nm; Get the ephedrine hydrochloride reference substance, the accurate title, decide, and uses dissolve with methanol, shakes up, and promptly gets reference substance solution; Get soft extract, syrup or oral liquid, with the mobile phase dilution, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride.

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with 0.01-1mol/L sodium acetate solution (glacial acetic acid is transferred pH=2-5): methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get the codeine phosphate reference substance, the methanol-water solution dissolving with 0.5-5 doubly measures shakes up, and precision is measured 1ml, puts in the 5-25ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, and promptly gets reference substance solution; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, thin up also is transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 2-8 time, combined chloroform extracting solution, water-bath are waved to 10-30ml, quantitatively are transferred in the separatory funnel, extract 1-5 time with hydrochloric acid solution, merge acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, crosses chloroform extraction 2-8 time with water saturation, the combined chloroform extracting solution, water bath method, residue dissolves with mobile phase, shakes up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Say that accurately the another kind of content assaying method of above-mentioned preparation is selected from one or more in the following method:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and the detection wavelength is 200-500nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.05-0.15mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and each 5-20 μ l of need testing solution of drawing injects hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride.

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with 0.01-1mol/L sodium acetate solution (glacial acetic acid is transferred pH=2-5): methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5-50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5-25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 5-50ml water dilution and be transferred in the separatory funnel, transfer pH8～13, chloroform extraction 2-8 time with sodium hydroxide test solution, each 5-50ml, combined chloroform extracting solution, water-bath are waved to 10-30ml, quantitatively are transferred in the separatory funnel, extract 1-5 time with hydrochloric acid solution 1 → 200, each 5-50ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH8～13, cross chloroform extraction 2-8 time with water saturation, each 5-50ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 5-25ml measuring bottle, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

In a word, the method for quality control of KEKE KOUFU ZHIJI of the present invention comprises character, inspection, discriminating and assay, and wherein character comprises:

For soft extract, product is the stiff semifluid of sepia, and it is sweet to distinguish the flavor of, little hardship;

For syrup, product is pale brown color thick liquid, and it is sweet to distinguish the flavor of, little hardship;

For oral liquid, product is pale brown color liquid, is equipped with the easily diffusing precipitation of jog for a long time, and it is sweet to distinguish the flavor of, little hardship;

Inspection comprises:

Relative density: the soft extract relative density should be between 1.30～1.50, and the syrup relative density is not less than 1.20;

PH value: the oral liquid pH value should be 3.5～6.5;

Other: should meet every regulation relevant under Chinese Pharmacopoeia syrup, soft extract or the mixture item;

Discriminating comprises:

(1) Herba Ephedrae: get soft extract 2g, add water 20ml mixing; Or get syrup 5ml, add water 20ml mixing; Or get oral liquid 20ml, and transfer pH10～11 with ammonia, use chloroform extraction 2 times, each 20ml, the combined chloroform extracting solution extracts 1 time with 20ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=5: 5: 1 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract 5g or syrup 10ml or oral liquid 20ml, add ethyl acetate 40ml and ammonia 1ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds the mixed liquor of ethyl acetate 40ml and ammonia 1ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; Other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=20: 20: 3: 1 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 30 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml add methanol 50ml, supersound extraction 5 minutes, filter, filtrate evaporate to dryness, residue add 20ml water to be made dissolving and is transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, use ether extraction 2 times, each 30ml discards ether solution, and water liquid is with water saturated n-butanol extraction 3 times, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 1mg with dissolve with ethanol, in contrast product solution; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution and reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=10: 5: 1 is developing solvent, launches, and takes out, and dries, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

Assay comprises:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is the C18 post, and with 0.1% phosphoric acid solution: methanol or acetonitrile: triethylamine is a mobile phase at 94: 6: 0.1, and the detection wavelength is 207nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.08mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane filtration with 0.45 μ m promptly gets need testing solution; Accurate respectively reference substance solution and each 5-20 μ l of need testing solution of drawing injects hplc determination content; Contain Herba Ephedrae in the soft extract in ephedrine hydrochloride, must not be less than 0.5mg/g, contain Herba Ephedrae in the syrup, must not be less than 0.5mg/ml, contain Herba Ephedrae in the oral liquid, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C18 post, transfers the 0.03mol/L sodium acetate solution of pH=3.5 with glacial acetic acid: methanol=8: 2 is mobile phase, and the detection wavelength is 238nm; Get codeine phosphate reference substance 10mg, accurate claim surely, put in the 10ml measuring bottle, dissolve and be diluted to scale with 50% solution, shake up, precision is measured 1ml, puts in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, the silicon dried soil that adds 1 times of amount is mixed thoroughly, and is transferred in the round-bottomed flask, with 50ml ethanol water-bath reflux, extract, 1 hour, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue add the 20ml water dissolution and are transferred in the separatory funnel, transfer pH10～11 with sodium hydroxide test solution, chloroform extraction 4 times, each 20ml, combined chloroform extracting solution, water-bath is waved to 20ml, quantitatively be transferred in the separatory funnel, extract 3 times, each 20ml with hydrochloric acid solution 1 → 200, merge acid solution, the reuse sodium hydroxide test solution is transferred pH10～11, crosses chloroform extraction 4 times with water saturation, each 20ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 5ml measuring bottle, microporous filter membrane with 0.45 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; Contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than 0.05mg/g, contain Pericarpium Papaveris in the syrup, must not be less than 0.05mg/ml, contain Pericarpium Papaveris in the oral liquid, must not be less than 0.025mg/ml in codeine phosphate in codeine phosphate.

In a word, the method for quality control of KEKE KOUFU ZHIJI of the present invention also can be that this is such, and it comprises character, inspection, discriminating and assay, and wherein character is:

Inspection comprises:

PH value: the oral liquid pH value should be 3.5～6.5;

Discriminating is: (1) Herba Ephedrae: get soft extract 2g, add water 20ml mixing; Or get syrup 5ml, add water 20ml mixing; Or get oral liquid 20ml, and transfer pH10～11 with ammonia, use chloroform extraction 2 times, each 20ml, the combined chloroform extracting solution extracts 1 time with 20ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=5: 5: 1 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH9～10 with ammonia solution, use ethyl acetate extraction 2 times, each 20ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 20ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=20: 20: 3: 1 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 30 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 20ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, with ether extraction 2 times, each 30ml discards ether solution, water liquid is with water saturated n-butanol extraction 3 times, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is made the solution that every 1ml contains 1mg with dissolve with ethanol, in contrast product solution in addition; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution and reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=10: 5: 1 is developing solvent, launches, and takes out, and dries, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

Assay is:

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C18 post, transfers the 0.03mol/L sodium acetate solution of pH=3.5 with glacial acetic acid: methanol=8: 2 is mobile phase, and the detection wavelength is 238nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 10ml measuring bottle, with 50% dissolve with methanol solution and be diluted to scale, shake up, precision is measured 1ml, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 20ml water dilution and be transferred in the separatory funnel, transfer pH10～11 with sodium hydroxide test solution, chloroform extraction 4 times, each 20ml, combined chloroform extracting solution, water-bath are waved to 20ml, quantitatively are transferred in the separatory funnel, extract 3 times with hydrochloric acid solution 1 → 200, each 20ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH10～11, cross chloroform extraction 4 times with water saturation, each 20ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 5ml measuring bottle, shake up, the microporous filter membrane filtration with 0.45 μ m promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; Contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate in codeine phosphate.

KEKE KOUFU ZHIJI comprises soft extract, oral liquid and syrup, Herba Ephedrae and Pericarpium Papaveris are the medicines that plays a major role in the KEKE KOUFU ZHIJI, owing to codeine phosphate in ephedrine hydrochloride in the Herba Ephedrae and the Pericarpium Papaveris is not only the active substance that plays a major role in the KEKE KOUFU ZHIJI, also be malicious numb material simultaneously, so among the present invention the content of ephedrine hydrochloride and codeine phosphate is set up assay method, so that can better control drug quality; And the present invention also carries out the thin layer Study on Identification to Herba Ephedrae, Pericarpium Papaveris and three medical materials of Radix Glycyrrhizae.

Method of quality control of the present invention is the preferred plan that obtains through a large amount of screenings, and following experimentation is a preferred process of the present invention.

One, ephedrine hydrochloride content assaying method research

1, need testing solution preparation method research:

Method 1: get soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, with methanol water-bath reflux, extract,, extracting solution filters, and filtrate volatilizes, and adds dissolve with methanol, and is diluted to 25ml, shakes up, with the microporous filter membrane filtration of 0.45 μ m, promptly.

Method 2: get soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, put in the 25ml measuring bottle, mobile phase is diluted to scale, shakes up, with the microporous filter membrane filtration of 0.45 μ m, promptly.

Sample	Method 1 (mg/g)	Method 2 (mg/g)
Sample	Method 1 (mg/g)	Method 2 (mg/g)	1	0.5642	0.7295
2	0.5037	0.7152	1	0.5642	0.7295
2	0.5037	0.7152	3	0.5127	0.7224

According to result of the test as can be known, employing method 2 preparation need testing solutions, ephedrine hydrochloride extracts more complete.

2, the selection of mobile phase:

Mobile phase 1: the mixed solution with acetonitrile and 0.2 phosphoric acid solution is a mobile phase.

Mobile phase 2: the mixed solution with 0.01mol/L potassium dihydrogen phosphate and methanol is a mobile phase.

Mobile phase 3: the mixed solution with phosphoric acid solution, methanol or acetonitrile and triethylamine is a mobile phase.

Result: with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase; The negative sample chromatogram is at non-false positive peak, ephedrine hydrochloride position, and ephedrine hydrochloride separates fully (separating degree＞1.5) with close impurity peaks, and promptly ephedrine hydrochloride separates with other components fully under this condition.Optimal flow is mutually: 0.1% phosphoric acid solution: methanol or acetonitrile: triethylamine=94: 6: 0.1.

Two, codeine phosphate content assaying method research

1, need testing solution preparation method research:

Method one: get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, the silicon dried soil that adds 1 times of amount is mixed thoroughly and is transferred in the round-bottomed flask, add 50ml methanol, ethanol or dehydrated alcohol water-bath reflux, extract, 1 hour, filter, and use methanol, ethanol or dehydrated alcohol are washed and are dripped a flask and a filter, and filtrate water bath method, residue add the 20ml water dissolution and are transferred to branch and move in the funnel, transfer pH 10～11 with sodium hydroxide test solution, chloroform extraction 4 times, each 20ml, combined chloroform extracting solution, water-bath is waved to about 20ml, quantitatively be transferred to branch and move in the funnel, extract 3 times, each 20ml with hydrochloric acid solution (1 → 200), merge acid solution, the reuse sodium hydroxide test solution is transferred pH10～11, crosses chloroform extraction 4 times with water saturation, each 20ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 5ml measuring bottle, filter (0.45 μ m), promptly.

The result: methanol and ethanol are for extracting solvent, and is suitable to the codeine phosphate leaching effect in the preparation, but methanol is in the reflux, extract, process, the saccharide impurity that leaches is a lot of, and emulsifying degree when the chloroform extraction is strengthened, and is unfavorable for operation, so among the present invention, extract solvent and select ethanol for use.

Method two: get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 20ml water dilution and be transferred in the separatory funnel, transfer pH10～11, add chloroform or extracted with diethyl ether 4 times with sodium hydroxide test solution, each 20ml, combined chloroform extracting solution, water-bath are waved to about 20ml, quantitatively are transferred in the separatory funnel, extract 3 times with hydrochloric acid solution (1 → 200), each 20ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH10～11, cross chloroform extraction 4 times with water saturation, each 20ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 5ml measuring bottle, shake up, filter (0.45 μ m), promptly.

The result: select for use chloroform as extractant, codeine phosphate extracts comparatively complete.

2, the selection of mobile phase:

Mobile phase 1: the mixed solution with acetonitrile, ammonium acetate solution and triethylamine is a mobile phase.

Mobile phase 2: the mixed solution with potassium dihydrogen phosphate, methanol and oxolane is a mobile phase.

Mobile phase 3: the mixed solution with sodium acetate solution and methanol is a mobile phase.

Result: with 0.01-1mol/L sodium acetate solution (glacial acetic acid transfer pH=2-5): methanol=1-9: 9-1 is a mobile phase; The negative sample chromatogram is at non-false positive peak, codeine phosphate position, and codeine phosphate separates fully (separating degree＞1.5) with close impurity peaks, and promptly codeine phosphate separates with other components fully under this condition.Optimal flow is mutually: 0.03mol/L sodium acetate solution (glacial acetic acid is transferred pH=3.5): methanol=80: 20.

Three, Herba Ephedrae thin layer Study on Identification

Need testing solution preparation method one: get soft extract 2g, add water 20ml mixing; Or get syrup 5ml, add water 20ml mixing; Or get oral liquid 20ml.Transfer PH10～11 with ammonia, use chloroform extraction 2 times, each 20ml, the combined chloroform extracting solution extracts 1 time with 20ml (1 → 200) hydrochloric acid solution, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Lack the Herba Ephedrae negative controls with the method preparation.

Need testing solution preparation method two: get soft extract 2g, syrup 2ml or get oral liquid 5ml.Add methanol 20ml, dissolving filters, as need testing solution; Lack the Herba Ephedrae negative controls with the method preparation.

Developing solvent is selected: respectively with the mixed solution of ethyl acetate, acetone and ammonia; The mixed solution of chloroform, methanol and ammonia is developing solvent.

The result: employing method one preparation need testing solution, with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is developing solvent, and its separating degree is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is: ethyl acetate: acetone: ammonia=5: 5: 1.

Four, Pericarpium Papaveris Study on Identification

Need testing solution preparation method one: get soft extract 10g, syrup 10ml or oral liquid 20ml, add ethyl acetate 40ml and ammonia 1ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Lack the Pericarpium Papaveris negative controls with the method preparation.

Need testing solution preparation method two: get soft extract 10g, syrup 10ml or oral liquid 10ml, add methanol 50ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 5ml makes dissolving, as supplying need testing solution; Lack the Pericarpium Papaveris negative controls with the method preparation.

Need testing solution preparation method three: get soft extract 10g, syrup 10ml or oral liquid 20ml, transfer pH9～10 with ammonia solution, use ethyl acetate extraction 2 times, each 20ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Lack the Pericarpium Papaveris negative controls with the method preparation.

Need testing solution preparation method four: get soft extract 10g, syrup 10ml or oral liquid 10ml, use ethanol supersound process 30 minutes, filter, the filtrate evaporate to dryness adds methanol 2ml and makes dissolving, as need testing solution; Lack the Pericarpium Papaveris negative controls with the method preparation.

Developing solvent is selected: respectively with the mixed solution of toluene, acetone, dehydrated alcohol and ammonia; The mixed solution of chloroform, methanol and ammonia; The mixed solution of toluene, acetone and ammonia is developing solvent.

The result: employing method one or method three preparation need testing solutions, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, and its separating degree is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is: toluene: acetone: dehydrated alcohol: ammonia=20: 20: 3: 1.

Five, Radix Glycyrrhizae Study on Identification

Need testing solution preparation method one: get soft extract 5g, syrup 5ml or oral liquid 20ml add methanol 50ml, supersound extraction 5 minutes, filter, filtrate evaporate to dryness, residue add 20ml water make the dissolving and be transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, with ether extraction twice, each 30ml discards ether solution, water liquid is crossed n-butanol extraction 3 times with water saturation, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Need testing solution preparation method two: get soft extract 5g, syrup 5ml or oral liquid 20ml, add 30ml water make the dissolving and be transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, with ether extraction twice, each 30ml discards ether solution, water liquid is crossed n-butanol extraction 3 times with water saturation, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Need testing solution preparation method three: get soft extract 5g, syrup 5ml or oral liquid 20ml, add chloroform 30ml, hydrochloric acid 5ml, water-bath reflux, extract, 30 minutes is put coldly, filters, filtrate is transferred to and washes 3 times in the separatory funnel with water, each 20ml abandons water layer, and chloroform solution adds an amount of anhydrous sodium sulfate, filter, filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Need testing solution preparation method four: get soft extract 5g, syrup 5ml or oral liquid 20ml, add dilute hydrochloric acid 1ml, water 40ml shakes up, adding water-saturated n-butanol extracts 5 times, each 30ml merges n-butyl alcohol liquid, water bath method, residue dissolves with methanol 5ml gradation, be transferred in the 100ml round-bottomed flask, volatilization goes methanol solution, residue to add hydrochloric acid 5ml, chloroform 50ml, reflux 3h.Put coldly, divide and to get chloroform layer, filter, with 10ml chloroform gradation washing filter paper, merging filtrate, water bath method, residue add dissolve with methanol and are settled to 5ml, as test sample liquid; Lack the Radix Glycyrrhizae negative controls with the method preparation.

Developing solvent is selected: respectively with the mixed solution of petroleum ether (30～60 ℃), chloroform and glacial acetic acid; The mixed solution of petroleum ether (60～90 ℃), benzene, ethyl acetate and glacial acetic acid; The mixed solution of cyclohexane extraction, ethyl acetate and glacial acetic acid; The mixed solution of cyclohexane extraction, ethyl acetate, benzene and formic acid is developing solvent.

The result: employing method one or method two prepare need testing solution, and with petroleum ether (30～60 ℃): chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, and its separating degree is good, and the speckle colour developing is clear, and negative control is noiseless, the method favorable reproducibility.Best developing solvent is petroleum ether (30～60 ℃): chloroform: glacial acetic acid=10: 5: 1.

Compared with prior art, adopt method of quality control of the present invention effectively to control, thereby guarantee the clinical efficacy of KEKE KOUFU ZHIJI the quality of KEKE KOUFU ZHIJI.

The specific embodiment: further specify the present invention by the following examples, but not as limitation of the present invention.

Embodiments of the invention 1:

Character: for soft extract, product is the stiff semifluid of sepia, and it is sweet to distinguish the flavor of, little hardship; For syrup, product is pale brown color thick liquid, and it is sweet to distinguish the flavor of, little hardship; For oral liquid, product is pale brown color liquid, is equipped with the easily diffusing precipitation of jog for a long time, and it is sweet to distinguish the flavor of, little hardship;

Check: relative density: the soft extract relative density should be between 1.30～1.50, and the syrup relative density is not less than 1.20; PH value: the oral liquid pH value should be 3.5～6.5; Other: should meet every regulation relevant under Chinese Pharmacopoeia syrup, soft extract or the mixture item;

Differentiate:

Assay:

Embodiments of the invention 2:

Differentiate:

Assay:

Embodiments of the invention 3:

Differentiate:

(1) gets soft extract 2g, add water 10ml mixing; Or get syrup 5ml, add water 10ml mixing; Or get oral liquid 20ml.Transfer pH8 with ammonia, use chloroform extraction 1 time, each 10ml, the combined chloroform extracting solution extracts 1-5 time with 10ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2: 2: 0.3 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(2) get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH13 with ammonia solution, use ethyl acetate extraction 5 times, each 10ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 100ml, and supersound process 60 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 20 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10: 35: 0.5: 0.5 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 60 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 50ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 1-5ml, shake up, with ether extraction 5 times, each 10ml discards ether solution, water liquid is with water saturated n-butanol extraction 5 times, each 10ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 5 times, each 10ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 20ml, evaporate to dryness, add hydrochloric acid 10ml and chloroform 100ml, reflux 5 hours is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 2mg with dissolve with ethanol, in contrast product solution.According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 20 μ l of need testing solution and reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with petroleum ether (30～60 ℃): chloroform: glacial acetic acid=5: 9: 0.2 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Assay:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is the C8 post, and with 1% phosphoric acid solution: methanol or acetonitrile: triethylamine=115: 15: 2 is a mobile phase, and the detection wavelength is 500nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.05mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane filtration with 0.65 μ m promptly gets need testing solution; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride.

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C8 post, and with 1mol/L sodium acetate solution (glacial acetic acid is transferred pH=5): methanol=9: 1 is mobile phase, and the detection wavelength is 500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 50ml water dilution and be transferred in the separatory funnel, transfer pH13 with sodium hydroxide test solution, chloroform extraction 8 times, each 5ml, combined chloroform extracting solution, water-bath are waved to 30ml, quantitatively are transferred in the separatory funnel, extract 5 times with hydrochloric acid solution 1 → 200, each 5ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH13, cross chloroform extraction 8 times with water saturation, each 5ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 25ml measuring bottle, shake up, the microporous filter membrane filtration with 0.65 μ m promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Embodiments of the invention 4:

Differentiate:

(1) Herba Ephedrae: get soft extract 2g, add water 50ml mixing; Or get syrup 5ml, add water 50ml mixing; Or get oral liquid 20ml.Transfer pH13 with ammonia, use chloroform extraction 5 times, each 50ml, the combined chloroform extracting solution extracts 5 times with 50ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 25 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=10: 10: 2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract 5g or syrup 5ml or oral liquid 20ml, add ethyl acetate 10ml and ammonia 0.5ml, supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds the mixed liquor of ethyl acetate 10ml and ammonia 5ml, and supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=35: 10: 5: 3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH8 with ammonia solution, use ethyl acetate extraction 1 time, each 100ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 10ml, and supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=5: 9: 0.2 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is the C4 post, and with 0.05% phosphoric acid solution: methanol or acetonitrile: triethylamine=50: 3: 2 is a mobile phase, and the detection wavelength is 200nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.15mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane filtration with 0.1 μ m promptly gets need testing solution; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C4 post, and with 1mol/L sodium acetate solution (glacial acetic acid is transferred pH=5): methanol=9: 1 is mobile phase, and the detection wavelength is 200nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml add silicon dried soil and mix thoroughly, and be transferred in the round-bottomed flask, with 10ml ethanol water-bath reflux, extract, 0.5 hour, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue add the 5ml water dissolution and are transferred in the separatory funnel, transfer pH8 with sodium hydroxide test solution, chloroform extraction 8 times, each 5ml, combined chloroform extracting solution, water-bath is waved to 10ml, quantitatively be transferred in the separatory funnel, extract 1 time, each 50ml with hydrochloric acid solution 1 → 200, merge acid solution, the reuse sodium hydroxide test solution is transferred pH8, crosses chloroform extraction 2 times with water saturation, each 50ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 5ml measuring bottle, microporous filter membrane with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Embodiments of the invention 5:

Differentiate:

(1) Herba Ephedrae: get soft extract 2g, add water 10ml mixing; Or get syrup 5ml, add water 50ml mixing; Or get oral liquid 20ml.Transfer pH12 with ammonia, use chloroform extraction 3 times, each 30ml, the combined chloroform extracting solution extracts 1 time with 10ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=10: 2: 0.3 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) Pericarpium Papaveris: get soft extract 5g or syrup 5ml or oral liquid 20ml, add ethyl acetate 100ml and ammonia 5ml, supersound process 60 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds the mixed liquor of ethyl acetate 100ml and ammonia 0.5ml, and supersound process 60 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 20 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10: 35: 0.5: 0.5 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 60 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH13 with ammonia solution, use ethyl acetate extraction 5 times, each 10ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 100ml, and supersound process 60 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 20 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=25: 1: 3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 60 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is the C18 post, and with 0.05% phosphoric acid solution: methanol or acetonitrile: triethylamine=50: 3: 0.05 is a mobile phase, and the detection wavelength is 200nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.05mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane filtration with 0.65 μ m promptly gets need testing solution; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C8 post, and with 1mol/L sodium acetate solution (glacial acetic acid is transferred pH=2): methanol=1: 9 is mobile phase, and the detection wavelength is 500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml add silicon dried soil and mix thoroughly, and be transferred in the round-bottomed flask, with 100ml ethanol water-bath reflux, extract, 5 hours, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue add the 50ml water dissolution and are transferred in the separatory funnel, transfer pH13 with sodium hydroxide test solution, chloroform extraction 1 time, each 50ml, combined chloroform extracting solution, water-bath is waved to 30ml, quantitatively be transferred in the separatory funnel, extract 5 times, each 5ml with hydrochloric acid solution 1 → 200, merge acid solution, the reuse sodium hydroxide test solution is transferred pH13, crosses chloroform extraction 8 times with water saturation, each 5ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 25ml measuring bottle, microporous filter membrane with 0.1 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Embodiments of the invention 6:

Differentiate:

(1) gets soft extract 2g, add water 50ml mixing; Or get syrup 5ml, add water 10ml mixing; Or get oral liquid 20ml.Transfer pH13 with ammonia, use chloroform extraction 5 times, each 10ml, the combined chloroform extracting solution extracts 4 times with 10ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2: 10: 0.3 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

(2) get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH8 with ammonia solution, use ethyl acetate extraction 1 time, each 100ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 10ml, and supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution, control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=35: 10: 5: 3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10 minutes, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 10ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 1ml, shake up, with ether extraction 1 time, each 100ml discards ether solution, water liquid is with water saturated n-butanol extraction 1 time, each 100ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1 time, each 100ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 5ml, evaporate to dryness, add hydrochloric acid 1ml and chloroform 20ml, reflux 0.5 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 0.5ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 0.5mg with dissolve with ethanol, in contrast product solution.According to the test of Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put in same silica gel G F respectively ₂₅₄On the lamellae, with petroleum ether (30～60 ℃): chloroform: glacial acetic acid=25: 1: 3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Assay:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is the C8 post, and with 1% phosphoric acid solution: methanol or acetonitrile: triethylamine=115: 15: 2 is a mobile phase, and the detection wavelength is 500nm; Get the ephedrine hydrochloride reference substance, accurate claim surely, make the reference substance solution of the hydrochloric ephedrine reference substance of every 1ml 0.15mg, shake up, promptly with dissolve with methanol; Other gets soft extract 5.0g, syrup 2.0ml or oral liquid 5ml, puts in the 25ml measuring bottle, and mobile phase is diluted to scale, shakes up, and the microporous filter membrane filtration with 0.2 μ m promptly gets need testing solution; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride.

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is the C4 post, and with 0.01mol/L sodium acetate solution (glacial acetic acid is transferred pH=2): methanol=1: 9 is mobile phase, and the detection wavelength is 200nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 5ml water dilution and be transferred in the separatory funnel, transfer pH8 with sodium hydroxide test solution, chloroform extraction 2 times, each 50ml, combined chloroform extracting solution, water-bath are waved to 10ml, quantitatively are transferred in the separatory funnel, extract 1 time with hydrochloric acid solution 1 → 200, each 50ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH8, cross chloroform extraction 2 times with water saturation, each 50ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 5ml measuring bottle, shake up, the microporous filter membrane filtration with 0.1 μ m promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

Claims

1. the method for quality control of a KEKE KOUFU ZHIJI, it comprises character, inspection, discriminating and assay, it is characterized in that: described character comprises: for soft extract, product is the stiff semifluid of sepia, and it is sweet to distinguish the flavor of, little hardship; For syrup, product is pale brown color thick liquid, and it is sweet to distinguish the flavor of, little hardship; For oral liquid, product is pale brown color liquid, is equipped with the easily diffusing precipitation of jog for a long time, and it is sweet to distinguish the flavor of, little hardship; Described inspection comprises: relative density, and the soft extract relative density should be between 1.30～1.50, and the syrup relative density is not less than 1.20; PH value, the oral liquid pH value should be 3.5～6.5; Other should meet every regulation relevant under Chinese Pharmacopoeia syrup, soft extract or the mixture item; Described discriminating comprises the epheday intermedia with the ephedrine hydrochloride reference substance side of discriminating, with Pericarpium Papaveris in codeine phosphate reference substance and the Pericarpium Papaveris control medicinal material discriminating side, with the thin layer of Radix Glycyrrhizae in the enoxolone reference substance discriminating side differentiate partly or entirely; Described assay comprise to the assay of ephedrine hydrochloride, codeine phosphate partly or entirely.

2. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 1, it is characterized in that: the discrimination method of this preparation epheday intermedia comprises that with the ephedrine hydrochloride reference substance be contrast, and with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is the thin layer discrimination method of developing solvent.

3. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 1, it is characterized in that: the discrimination method of Pericarpium Papaveris comprises that with codeine phosphate reference substance and Pericarpium Papaveris control medicinal material be contrast in this preparation, and with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is the thin layer discrimination method of developing solvent.

4. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 1, it is characterized in that: the discrimination method of Radix Glycyrrhizae comprises that with the enoxolone reference substance be contrast in this preparation, and with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is the thin layer discrimination method of developing solvent.

5. according to the method for quality control of claim 1,2,3 or 4 described KEKE KOUFU ZHIJI, it is characterized in that: described discrimination method is selected from one or more in the following method:

(3) Radix Glycyrrhizae: get soft extract, syrup or oral liquid add methanol or ethanol ultrasonic extraction, and extracting solution filters, the filtrate evaporate to dryness, residue add water make the dissolving and be transferred in the separatory funnel, add dilute sulfuric acid, shake up, with ether extraction 1-5 time, discard ether solution, water liquid closes n-butanol extracting liquid with water saturated n-butanol extraction 1-5 time, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, discard water lotion, n-butanol extracting liquid evaporate to dryness, residue with dissolve with methanol to flask, evaporate to dryness, add hydrochloric acid and chloroform, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds methanol or alcoholic solution makes dissolving, as need testing solution; In addition extracting liquorice subacid reference substance is an amount of, with methanol or dissolve with ethanol, and product solution in contrast; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put respectively on same silica GF254 lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

6. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 5, it is characterized in that: described discrimination method is selected from one or more in the following method:

(1) Herba Ephedrae: get soft extract 2g, add water 10-50ml mixing; Or get syrup 5ml, add water 10-50ml mixing; Or get oral liquid 20ml; Transfer pH8～13 with ammonia, use chloroform extraction 1-5 time, each 10-50ml, the combined chloroform extracting solution extracts 1-5 time with 10-50ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5-1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2-10: 2-10: 0.3-2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

7. according to the method for quality control of claim 1,2,3 or 4 described KEKE KOUFU ZHIJI, it is characterized in that: described discrimination method is selected from one or more in the following method:

(2) Pericarpium Papaveris: get soft extract, syrup or oral liquid, transfer PH8～13 with ammonia solution, use ethyl acetate extraction 1-5 time, merge ethyl acetate liquid, water bath method, residue add methanol or dissolve with ethanol, as need testing solution; Other gets the Pericarpium Papaveris control medicinal material, adds ethyl acetate, supersound extraction, and extracting solution filters, and filtrate evaporate to dryness, residue add methanol or ethanol makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, uses dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract, syrup or oral liquid, thin up also is transferred in the separatory funnel, add dilute sulfuric acid, shake up, use ether extraction 1-5 time, discard ether solution, water liquid is with water saturated n-butanol extraction 1-5 time, close n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time discards water lotion, the n-butanol extracting liquid evaporate to dryness, to flask, evaporate to dryness adds hydrochloric acid and chloroform to residue with dissolve with methanol, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds methanol or ethanol makes dissolving, as need testing solution; In addition extracting liquorice subacid reference substance is an amount of, with methanol or dissolve with ethanol, and product solution in contrast; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put respectively on same silica GF254 lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

8. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 7, it is characterized in that: described discrimination method is selected from one or more in the following method: (1) gets soft extract 2g, adds water 10-50ml mixing; Or get syrup 5ml, add water 10-50ml mixing; Or get oral liquid 20ml; Transfer pH8～13 with ammonia, use chloroform extraction 1-5 time, each 10-50ml, the combined chloroform extracting solution extracts 1-5 time with 10-50ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 0.5-1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5-25 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=2-10: 2-10.: 0.3-2 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2) get soft extract 5g or syrup 5ml or oral liquid 20ml, transfer PH8～13 with ammonia solution, use ethyl acetate extraction 1-5 time, each 10-100ml merges ethyl acetate liquid, and water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Pericarpium Papaveris control medicinal material 1g, adds ethyl acetate 10-100ml, and supersound process 10-60 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, makes control medicinal material solution; It is an amount of that other gets the codeine phosphate reference substance, makes the solution that every lml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, each 5-20 μ l of control medicinal material solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene: acetone: dehydrated alcohol: ammonia=10-35: 10-35: 0.5-5: 0.5-3 is developing solvent, put a cuvette strong ammonia solution pre-equilibration in the chromatography cylinder in addition 10-60 minute, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 10-50ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 1-5ml, shake up, with ether extraction 1-5 time, each 10-100ml discards ether solution, water liquid is with water saturated n-butanol extraction 1-5 time, each 10-100ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 1-5 time, each 10-100ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 5-20ml, evaporate to dryness, add hydrochloric acid 1-10ml and chloroform 20-100ml, reflux 0.5-5 hour, divide and get the chloroform solution evaporate to dryness, residue adds ethanol 0.5-2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 0.5-2mg with dissolve with ethanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution and each 5-20 μ l of reference substance solution, put respectively on same silica GF254 lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=5-25: 1-9: 0.2-3 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 200-500nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

9. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 1, it is characterized in that: the Determination of Ephedrine Hydrochloride assay method comprises that adopting chromatographic column is C18 or C4 or C8 post in this preparation, with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and detecting wavelength is the high performance liquid chromatography of 200-500nm.

10. according to the method for quality control of the described KEKE KOUFU ZHIJI of claim 1, it is characterized in that: the content assaying method of codeine phosphate comprises that adopting chromatographic column is C18 or C4 or C8 post in this preparation, with 0.01-1mol/L sodium acetate solution: methanol=1-9: 9-1 is mobile phase, detecting wavelength is the high performance liquid chromatography of 200-500nm, and wherein the 0.01-1mol/L sodium acetate solution needs to transfer pH=2-5 with glacial acetic acid.

11. the method for quality control according to claim 1,9 or 10 described KEKE KOUFU ZHIJI is characterized in that: described content method is selected from one or more in the following method:

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and transfer the 0.01-1mol/L sodium acetate solution of pH=2-5 with glacial acetic acid: methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get the codeine phosphate reference substance, the methanol-water solution dissolving with 0.5-5 doubly measures shakes up, and precision is measured 1ml, puts in the 5-25ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, and promptly gets reference substance solution; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add silicon dried soil and mix thoroughly, and be transferred in the round-bottomed flask, with ethanol water-bath reflux, extract,, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue is dissolved in water and is transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 1-8 time, combined chloroform extracting solution, water-bath is waved to 10-30ml, quantitatively be transferred in the separatory funnel, extract 1-5 time, merge acid solution with hydrochloric acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, cross chloroform extraction 2-8 time with water saturation, combined chloroform extracting solution, water bath method, residue dissolves with mobile phase, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

12. the method for quality control according to the described KEKE KOUFU ZHIJI of claim 11 is characterized in that: described content method is selected from one or more in the following method:

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and transfer the 0.01-1mol/L sodium acetate solution of pH=2-5 with glacial acetic acid: methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5-50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5-25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, adding silicon dried soil mixes thoroughly, and be transferred in the round-bottomed flask, use 10-100ml ethanol water-bath reflux, extract, 0.5-5 hour, filter, and with washing with alcohol flask and filter, merging filtrate and washing liquid, evaporate to dryness, residue add the 5-50ml water dissolution and are transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 1-8 time, each 5-50ml, combined chloroform extracting solution, water-bath is waved to 10-30ml, quantitatively be transferred in the separatory funnel, extract 1-5 time, each 5-50ml with hydrochloric acid solution 1 → 200, merge acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, crosses chloroform extraction 2-8 time with water saturation, each 5-50ml, the combined chloroform extracting solution, water bath method, residue shakes up with mobile phase dissolving and fixed molten to the 5-25ml measuring bottle, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

13. the method for quality control according to claim 1,9 or 10 described KEKE KOUFU ZHIJI is characterized in that: described content method is selected from one or more in the following method:

Ephedrine hydrochloride: adopt high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and with the 0.05-1% phosphoric acid solution: methanol or acetonitrile: triethylamine=50-115: 3-15: 0.05-2 is a mobile phase, and the detection wavelength is 200-500nm; Get the ephedrine hydrochloride reference substance, the accurate title, decide, and uses dissolve with methanol, shakes up, and promptly gets reference substance solution; Get soft extract, syrup or oral liquid, with the mobile phase dilution, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects hplc determination content; In this cough stopping formulation, contain Herba Ephedrae in the soft extract, must not be less than in 0.5mg/g, the syrup and contain Herba Ephedrae, must not be less than in 0.5mg/ml, the oral liquid and contain Herba Ephedrae, must not be less than 0.25mg/ml in ephedrine hydrochloride in ephedrine hydrochloride in ephedrine hydrochloride;

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and transfer the 0.01-1mol/L sodium acetate solution of pH=2-5 with glacial acetic acid: methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get the codeine phosphate reference substance, the methanol-water solution dissolving with 0.5-5 doubly measures shakes up, and precision is measured 1ml, puts in the 5-25ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, and promptly gets reference substance solution; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, thin up also is transferred in the separatory funnel, transfer pH8～13 with sodium hydroxide test solution, chloroform extraction 2-8 time, combined chloroform extracting solution, water-bath are waved to 10-30ml, quantitatively are transferred in the separatory funnel, extract 1-5 time with hydrochloric acid solution, merge acid solution, the reuse sodium hydroxide test solution is transferred pH8～13, crosses chloroform extraction 2-8 time with water saturation, the combined chloroform extracting solution, water bath method, residue dissolves with mobile phase, shakes up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

14. the method for quality control according to the described KEKE KOUFU ZHIJI of claim 13 is characterized in that: described content method is selected from one or more in the following method:

Codeine phosphate: according to high performance liquid chromatography, chromatographic column is C18 or C4 or C8 post, and transfer the 0.01-1mol/L sodium acetate solution of pH=2-5 with glacial acetic acid: methanol=1-9: 9-1 is a mobile phase, and the detection wavelength is 200-500nm; Get codeine phosphate reference substance 10mg, the accurate title, decide, put in the 5-50ml measuring bottle, with methanol-water solution dissolving and be diluted to scale, shake up, precision is measured 1ml, put in the 5-25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, promptly get the reference substance solution of every 1ml phosphoric acid codeine reference substance 0.10mg; Get soft extract 5.0g, syrup 5.0ml or oral liquid 5ml, add 5-50ml water dilution and be transferred in the separatory funnel, transfer pH8～13, chloroform extraction 2-8 time with sodium hydroxide test solution, each 5-50ml, combined chloroform extracting solution, water-bath are waved to 10-30ml, quantitatively are transferred in the separatory funnel, extract 1-5 time with hydrochloric acid solution 1 → 200, each 5-50ml merges acid solution, and the reuse sodium hydroxide test solution is transferred pH8～13, cross chloroform extraction 2-8 time with water saturation, each 5-50ml, combined chloroform extracting solution, water bath method, residue is with the mobile phase dissolving and decide molten to the 5-25ml measuring bottle, shake up, the microporous filter membrane that reaches less than 0.65 μ m with 0.65 μ m filters, and promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn, inject hplc determination content, in this KEKE KOUFU ZHIJI, contain Pericarpium Papaveris in the soft extract in codeine phosphate, must not be less than in 0.05mg/g, the syrup and contain Pericarpium Papaveris in codeine phosphate, must not be less than in 0.05mg/ml, the oral liquid and contain Pericarpium Papaveris, must not be less than 0.025mg/ml in codeine phosphate.

15. the method for quality control according to claim 1,6 or 12 described KEKE KOUFU ZHIJI is characterized in that:

Character comprises:

Inspection comprises:

PH value: the oral liquid pH value should be 3.5～6.5;

Discriminating comprises:

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml add methanol 50ml, supersound extraction 5 minutes, filter, filtrate evaporate to dryness, residue add 20ml water to be made dissolving and is transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, use ether extraction 2 times, each 30ml discards ether solution, and water liquid is with water saturated n-butanol extraction 3 times, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is an amount of in addition, makes the solution that every 1ml contains 1mg with dissolve with ethanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ 1 of need testing solution and reference substance solution, put respectively on same silica GF254 lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=10: 5: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

Assay comprises:

16. the method for quality control according to claim 1,8 or 14 described KEKE KOUFU ZHIJI is characterized in that:

Described character is:

Inspection comprises:

PH value: the oral liquid pH value should be 3.5～6.5;

Other: should meet every regulation relevant under Chinese Pharmacopoeia syrup, soft extract or the mixture item; Discriminating is: (1) Herba Ephedrae: get soft extract 2g, add water 20ml mixing; Or get syrup 5ml, add water 20ml mixing; Or get oral liquid 20ml, and transfer pH10～11 with ammonia, use chloroform extraction 2 times, each 20ml, the combined chloroform extracting solution extracts 1 time with 20ml hydrochloric acid solution 1 → 200, the acid solution evaporate to dryness, residue makes dissolving with methanol 1ml, as need testing solution; Other gets the ephedrine hydrochloride reference substance, makes the solution that every 1ml contains 1mg with dissolve with methanol, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate: acetone: ammonia=5: 5: 1 is developing solvent, launch, take out, dry, spray is with ninhydrin solution, it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) Radix Glycyrrhizae: get soft extract 3g, syrup 5ml or oral liquid 20ml, add 20ml water make the dilution and be transferred in the separatory funnel, add dilute sulfuric acid 2ml, shake up, with ether extraction 2 times, each 30ml discards ether solution, water liquid is with water saturated n-butanol extraction 3 times, each 20ml closes n-butanol extracting liquid, the washing of the saturated mistake of reuse n-butyl alcohol 2 times, each 50ml, discard water lotion, the n-butanol extracting liquid evaporate to dryness, residue is dissolved in the flask with methanol 10ml, evaporate to dryness, add hydrochloric acid 4ml and chloroform 50ml, reflux 1 hour is divided and is got the chloroform solution evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance is made the solution that every 1ml contains 1mg with dissolve with ethanol, in contrast product solution in addition; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution and reference substance solution, put respectively on same silica GF254 lamellae, with 30～60 ℃ of petroleum ether: chloroform: glacial acetic acid=10: 5: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 254nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

Assay is: