CN100370253C

CN100370253C - Method for quality control of Qianbai biyan solid prepn. for treating rhinitis

Info

Publication number: CN100370253C
Application number: CNB2005100032709A
Authority: CN
Inventors: 叶湘武; 王泽坤
Original assignee: Guizhou Yibai Pharmaceutical Co Ltd
Current assignee: Guizhou Yibai Pharmaceutical Co Ltd
Priority date: 2005-11-10
Filing date: 2005-11-10
Publication date: 2008-02-20
Anticipated expiration: 2025-11-10
Also published as: CN1785232A

Abstract

The present invention discloses a quality control method for solid preparation 'Qianbai' for treating rhinitis. The method adopts thin-layer chromatography to carry out qualitative identification to groundsel, ephedra and notopterygium root and other medicine in solid preparation, and an efficient liquid-phase chromatography is used as the index to measure the content of ephedrin in medicine. The quality control method of the present invention has the advantages of stable quality control to products, convenience and accuracy and good repeatability in results.

Description

The method of quality control of thousand cypress rhinitis solid pharmaceutical preparations

Technical field:

The present invention relates to a kind of method of quality control of the thousand cypress rhinitis solid pharmaceutical preparations of making by climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, particularly differentiate Chinese crude drug climbing groundsel, Chinese ephedra and notopterygium root and to thousand cypress rhinitis quality control of solid dosage forms methods of the qualitative identification of other drug in the solid pharmaceutical preparation with the content of this medicine epheday intermedia alkali of high effective liquid chromatography for measuring with thin-layered chromatography.

Background technology:

Treat acute and chronic rhinitis in " Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Chinese patent drug prescribed preparation) the 5th (WS3-B-0884-91), the qianbai biyan tablets of disease such as nasosinusitis and pharyngitis, this pharmaceutical formulation is made up of climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, and above-mentioned raw materials is made 1000 altogether; And issued its quality control standard, but in the existing qianbai biyan tablets method of quality control of Ministry of Public Health promulgation, only with the climbing groundsel medicinal material as qualitative identification, its specificity is not strong, limitation is bigger, lacks assay in addition, is difficult to control the quality of qianbai biyan tablets; And " 2000 editions one qianbai biyan tablets method of quality control that records of Chinese pharmacopoeia, with the existing qianbai biyan tablets method of quality control of Ministry of Public Health promulgation much at one, and none microscopical identification does not have and contains the survey index, is difficult to control the quality of qianbai biyan tablets.

Chinese medicine preparation for a long time, difficult quality is effectively controlled, the quality of effectively controlling tcm product is a great problem, the method for available technology adopting is imperfect, the minority specific aim is not strong, uses these methods to be difficult to reach the purpose of real control of quality.Above qianbai biyan tablets preparation is difficult to control the quality of said preparation owing to lack method of quality control; Brought difficulty for normal production and operation, utilize some known technology to be difficult to the method for quality control that the said preparation product is provided, the present invention is directed to the deficiencies in the prior art, adopt new means that the quality of qianbai biyan tablets is controlled, particularly increase assay detection method in the preparation, content with this medicine epheday intermedia alkali of high effective liquid chromatography for measuring, increase Chinese ephedra, the thin-layered chromatography discrimination method of notopterygium root two flavor medicinal materials and to the qualitative identification method of other drug in the solid pharmaceutical preparation, make sample preparation easier, guarantee this compound preparation higher quality standard level.

Summary of the invention:

The object of the present invention is to provide a kind of method of quality control of thousand cypress rhinitis solid pharmaceutical preparations, in process of production product quality is monitored, improve the accuracy of quality monitoring standard to guarantee said preparation.

The present invention be directed to thousand cypress rhinitis solid pharmaceutical preparations and propose method of quality control, this solid pharmaceutical preparation specifically comprises capsule, granule and tablet etc.

Described thousand cypress rhinitis solid pharmaceutical preparations of the present invention are to be made by the Chinese medicine material of following weight/part:

Climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g.

The preparation method of the present invention's thousand cypress rhinitis solid pharmaceutical preparations can adopt following method:

More than seven flavors, climbing groundsel, Selaginella tamariscina, Cassia occidentalis, Chinese ephedra boiling secondary, collecting decoction filters, filtrate is condensed into thick paste; Three flavors such as all the other notopterygium roots are ground into fine powder, add above-mentioned thick paste, stir evenly, and drying is ground into fine powder, mixing, and formulation method adds different auxiliary materials routinely, can make capsule respectively, sheet system or 1000/sheet/g of granule, promptly.

Thousand cypress rhinitis solid pharmaceutical preparations of the present invention, in its Chinese medicine preparation, part Chinese medicine can be replaced with the Chinese medicine of equivalent effect, replaces the back curative effect and can not change.

The present invention is achieved by following technical proposals: the one content of high effective liquid chromatography for measuring thousand cypress rhinitis solid pharmaceutical preparation epheday intermedia alkali; Its dual-purpose thin-layered chromatography is differentiated thousand cypress rhinitis solid pharmaceutical preparation Chinese crude drug climbing groundsels, Chinese ephedra and notopterygium root; Its three couple carries out qualitative identification to thousand cypress rhinitis solid pharmaceutical preparations

The method of quality control of thousand concrete cypress rhinitis solid pharmaceutical preparations can be achieved by following steps:

Step with the content of high effective liquid chromatography for measuring thousand cypress rhinitis solid pharmaceutical preparation epheday intermedia alkali:

The preparation of ephedrine reference substance solution,

The preparation of thousand cypress rhinitis solid pharmaceutical preparation sample solutions,

Measure above solution with high performance liquid chromatograph, get chromatogram,

Draw the reference substance typical curve, according to thousand cypress rhinitis solid pharmaceutical preparation sample peak areas, with typical curve, calculate the content of this sample epheday intermedia alkali.

Concrete high performance liquid chromatography step is as follows:

The content of measuring ephedrine according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D) may further comprise the steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine; Set flow velocity; Column temperature; Detect wavelength.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

B. it is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and make solution, promptly.

C. this product content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol, close plug claims to decide weight, sonicated is put coldly, claims to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate, regulates the pH value with sodium hydroxide test solution, uses chloroform extraction, get chloroform solution, add the hydrochloric acid solution jolting and extract, divide and get hydrochloric acid solution, chloroform solution washes with water, merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, be transferred in the measuring bottle, be diluted to scale, shake up with methyl alcohol, filter membrane filters, promptly.

D. accurate respectively reference substance solution and the need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;

Every of this product contains Chinese ephedra with ephedrine (C ₁₀H ₁₅NO) meter must not be less than 0.65mg.

Preferable methods is:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine (88-96: 4.6-9: 0.05-0.13); Flow velocity: 1.2ml/min; Column temperature: 36-45 ℃; Detect wavelength 207 ± 2nm.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

B. it is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds that methyl alcohol is made, solution, promptly.

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 20-35 minute, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulates the pH value to 10-13 with sodium hydroxide test solution, with chloroform extraction 4-6 time (each 20ml or 10ml), combined chloroform liquid adds hydrochloric acid solution (1 → 100) 30ml jolting and extracts, and divides and gets hydrochloric acid solution, chloroform solution washes with water and merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter with filter membrane (0.45 μ m), promptly.

D. accurate respectively reference substance solution and each the 5-10 μ l of need testing solution of drawing of determination method injects liquid chromatograph, measures, promptly;

Preferred method is:

Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D):

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine (93: 7: 0.1); Flow velocity: 1.2ml/min; Column temperature: 40 ℃; Detect wavelength 207nm.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

B. it is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and make the solution that every 1ml contains 0.1mg (amounting to ephedrine 0.08190mg), promptly.

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 30 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulate pH value to 11 with sodium hydroxide test solution, with 4 (20ml of chloroform extraction, 20ml, 10ml, 10ml), combined chloroform liquid, adding hydrochloric acid solution (1 → 100) 30ml jolting extracts, divide and get hydrochloric acid solution, chloroform solution washes with water 2 times, each 10ml, merge hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter with filter membrane (0.45 μ m), promptly.

D. accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly;

In the method for quality control of the present invention, differentiate that with thin-layered chromatography the step of Chinese crude drug climbing groundsel, Chinese ephedra and notopterygium root in the thousand cypress rhinitis solid pharmaceutical preparations is as follows:

Thin-layered chromatography is differentiated the step of Chinese crude drug Chinese ephedra (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of need testing solution: get this product content, porphyrize adds strong ammonia solution respectively, and chloroform is put in the water-bath and refluxed, filter, filtrate evaporate to dryness, residue add methyl alcohol make molten, as need testing solution.

B. the preparation of reference substance solution: get the Chinese ephedra control medicinal material, shine medicinal material solution in pairs with legal system.

C. get the ephedrine hydrochloride reference substance again, add methyl alcohol and make solution, in contrast product solution.

D. according to the thin-layered chromatography test, drawing above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-methanol-strong ammonia solution, launches, and takes out, and dries, and spray is dried with ninhydrin solution.

The processing of above-mentioned this Chinese ephedra thin layer discriminating sample can adopt strong ammonia solution to extract, chloroform removal of impurities, developping agent are chloroform-methanol-strong ammonia solution, and chloroform-methanol-strong ammonia solution ratio (4: 1: 0.1) is best, developer is ninhydrin solution the best, and 105 ℃ of baking colour developings in about 5 minutes down.

Thin-layered chromatography is differentiated the step of Chinese crude drug notopterygium root (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of need testing solution: get this product content, porphyrize adds sherwood oil, and sonicated filters, filtrate evaporate to dryness, residue add ethyl acetate make molten, as need testing solution.

B. the preparation of control medicinal material solution: get the notopterygium root control medicinal material, shine medicinal material solution in pairs with legal system.

C. according to the thin-layered chromatography test, drawing need testing solution, control medicinal material solution respectively, put respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate, launches, and takes out, and dries, and spray dries up with 5% vanillic aldehyde sulfuric acid solution.

The processing of above-mentioned this notopterygium root thin layer discriminating sample can be adopted Petroleum ether extraction, the ethyl acetate removal of impurities, developping agent is normal hexane-benzene-ethyl acetate, normal hexane-benzene-ethyl acetate ratio (2: 1: 1) is best, developer 5% vanillic aldehyde sulfuric acid solution is best, and blow to the spot colour developing with hot blast clear.

Thin-layered chromatography is differentiated the step of Chinese crude drug climbing groundsel (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of need testing solution: get this product content, porphyrize adds ethanol, puts reflux in the water-bath, filter, filtrate evaporate to dryness, residue add ethanol make molten, as need testing solution.

B. the preparation of control medicinal material solution: get the climbing groundsel control medicinal material, boiling filters, and filtrate is condensed into thick paste, adds ethanol, heats in the water-bath, filter, filtrate evaporate to dryness, residue add ethanol make approximately molten, medicinal material solution in contrast.

C. according to the thin-layered chromatography test, drawing above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with toluene-ethyl acetate-formic acid, launches, and takes out, and dries, and spray is with the ferric trichloride test solution.

The processing of above-mentioned this climbing groundsel thin layer discriminating sample can be adopted alcohol extract, and developping agent is toluene-ethyl acetate-formic acid, and toluene-ethyl acetate-formic acid (5: 4: 1) is best, and developer 1-3% ferric trichloride test solution is best.

Preferred Chinese crude drug Chinese ephedra thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of this product 35-45 grain, porphyrize adds strong ammonia solution 1-5ml, adds chloroform 10-30ml, puts in the water-bath backflow 0.5-2 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1-3ml makes dissolving, as need testing solution.

B. the preparation of reference substance solution: get Chinese ephedra control medicinal material 1-3g, shine medicinal material solution in pairs with legal system.

C. get the ephedrine hydrochloride reference substance again, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution.

D. according to the thin-layered chromatography test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-strong ammonia solution (2-6: 0.9-1.2: 0.08-0.12) be developping agent, launch, take out, dry, spray is with ninhydrin solution, 105 ℃ of bakings about 3-7 minute.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation.

Preferred Chinese crude drug notopterygium root thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of this product 5-15 grain, porphyrize adds sherwood oil (60～90 ℃) 15-30ml, and sonicated 25-35 minute, filter, filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get notopterygium root control medicinal material 0.5-1g, shine medicinal material solution in pairs with legal system.

C. test according to thin-layered chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, with normal hexane-benzene-ethyl acetate (1.1-2.4: 0.8-1. 1: 0.8-1.1) be developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

Preferred Chinese crude drug climbing groundsel thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of this product 3-8 grain, porphyrize adds ethanol 15-30ml, puts in the water-bath reflux 1-2 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1-3ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 20-30g, during boiling 1-3, filter, filtrate is condensed into thick paste, add ethanol 30-50ml, in the water-bath reflux 1-2 hour, filter, filtrate evaporate to dryness, residue add the about 1-3ml of ethanol makes dissolving, in contrast medicinal material solution.

C. according to the thin-layered chromatography test, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (3.8-6: 3.5-5: 0.9-1.2) be developping agent, launch, take out that dry, spray is with the ferric trichloride test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

More preferably Chinese crude drug Chinese ephedra thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of 40 of this product, porphyrize adds strong ammonia solution 1ml, adds chloroform 20ml, puts in the water-bath and refluxes 1 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.

B. the preparation of reference substance solution: get Chinese ephedra control medicinal material 1g, shine medicinal material solution in pairs with legal system.

D. according to the thin-layered chromatography test, drawing each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-methanol-strong ammonia solution (4: 1: 0.1), launches, and takes out, and dries, and spray is with ninhydrin solution, about 5 minutes of 105 ℃ of bakings.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation.

More preferably Chinese crude drug notopterygium root thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of 10 of this product, porphyrize adds sherwood oil (60～90 ℃) 20ml, and sonicated 30 minutes filters, and filtrate steams thousand, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get notopterygium root control medicinal material 0.5g, shine medicinal material solution in pairs with legal system.

C. test according to thin-layered chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate (2: 1: 1), launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

More preferably Chinese crude drug climbing groundsel thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of 6 of this product, porphyrize adds ethanol 25ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 25g, boiling 1 hour filters, and filtrate is condensed into thick paste, add ethanol 40ml, reflux is 1 hour in the water-bath, filters, filtrate evaporate to dryness, residue add the about 2ml of ethanol makes dissolving, in contrast medicinal material solution.

C. according to the thin-layered chromatography test, drawing each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with toluene-ethyl acetate-formic acid (5: 4: 1), launches, and takes out, and dries, and spray is with the ferric trichloride test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

Thousand cypress rhinitis solid pharmaceutical preparations are carried out qualitative identification, and step is as follows: (is example with the capsule)

(1) microscopic character of powder of notopterygium root, the root of Dahurain angelica, Ligusticum wallichii is differentiated and is got this product content, porphyrize, put microscopically and observe: amylum body is numerous, simple grain ellipse, Long Circle, similar round, oval or kidney shape etc., diameter 5～16 μ m, omphalion point-like, slit shape or herringbone shape; Accidental composite grain is made up of 2～4 gradation.Bunch shape crystallization is present in the parenchyma cell, is similar round agglomerate or class cluster crystal shape, diameter 10～25 μ m.The pale brown look of cork cell is the class polygon.How broken the grease chamber is, the visible fragment of idol, and the secretory cell wall is thin, contains more oil droplet.How cataclasm secretory canal is, and common golden yellow or yellowish-brown strip secretion.Conduit is mainly reticulate vessel and spiral duct, diameter 14～50 μ m.

(2) content of 8 of this product is got in the discriminating of alkaloids, add acidic ethanol 30ml, put in the water-bath reflux 20 minutes, and filtered, get filtrate 20ml, put and fling to ethanol in the water-bath, residue adds watery hydrochloric acid 1ml, water 10ml makes dissolving, filters, and gets each 1ml of filtrate, add each 1～2 of potassium mercuric iodide, bismuth potassium iodide and silico-tungstic acid test solution respectively, all generate precipitation.

(3) content of 5 of this product is got in the discriminating of volatile oil, and porphyrize adds ether 20ml, and jolting filters, and gets filtrate 2ml, puts in the porcelain dish, and low temperature is flung to ether, adds 1 of 1% vanillic aldehyde sulfuric acid solution, shows bluish violet to aubergine.

(4) discriminating of Cassia occidentalis is got [discriminating] (3) residual filtrate down and is concentrated into about 2ml, hydro-oxidation sodium test solution 1ml, and jolting is left standstill, and adds 1 of superoxol, puts in the water-bath and heats, and is apparent orange red, after the acidifying of usefulness watery hydrochloric acid, orange red should taking off.

The TCL that the invention provides Chinese ephedra, notopterygium root and climbing groundsel differentiates.The qualitative identification method of other drug also is provided, increased quality determining method to preparation epheday intermedia alkali content, revision test repeatedly, confirm that this method of quality control is comparatively stable, easy, the result is accurate, the degree of separation of the developping agent of selecting for use is good, the spot colour developing is clear, and negative control is noiseless, favorable reproducibility.Can be used as the index of the stability of said preparation quality control and investigation technology.

Further specify the present invention by the following examples, but not as limitation of the present invention.

The used preparation of experimental example test sample of the present invention all adopts the embodiment of the invention 1 described capsule, can adopt the embodiment of the invention 2 or 3 described tablets, granule, also available have other preparations that same materials is formed with the solid pharmaceutical preparation described in each embodiment.

The research of experimental example 1,000 cypress rhinitis solid pharmaceutical preparation quality assurance of drug of the present invention

1. differentiate: (1) gets thousand cypress rhinitis solid pharmaceutical preparation contents, and put microscopically and observe: amylum body is numerous, simple grain ellipse, Long Circle, similar round, oval or kidney shape etc., diameter 5～16 μ m, omphalion point-like, slit shape or herringbone shape; Accidental composite grain is formed (for the microscopic character of powder of the root of Dahurain angelica, Ligusticum wallichii in the side) by 2～4 gradation.Bunch shape crystallization is present in the parenchyma cell, is similar round agglomerate or class cluster crystal shape, diameter 10～25 μ m (root of Dahurain angelica, Ligusticum wallichii).The pale brown look of cork cell is class polygon (for the microscopic character of powder of notopterygium root, the root of Dahurain angelica, Ligusticum wallichii in the side).How broken the grease chamber is, the visible fragment of idol, and the secretory cell wall is thin, contains more oil droplet (for the microscopic features of rhizoma chuanxiong volatile oil).How cataclasm secretory canal is.And common golden yellow or yellowish-brown strip secretion (being the microscopic features of Rhizoma Et Radix Notopterygii volatile oil).Conduit is mainly reticulate vessel and spiral duct, diameter 14～50 μ m (microscopic character of powder of notopterygium root, the root of Dahurain angelica in the side of being).

(2) get the content of 8 of thousand cypress rhinitis solid pharmaceutical preparations, add acidic ethanol 30ml, put in the water-bath reflux 20 minutes, and filtered, get filtrate 20ml, put and fling to ethanol in the water-bath, residue adds watery hydrochloric acid 1ml, water 10ml makes dissolving, filters, and gets each 1ml of filtrate, add each 1～2 of potassium mercuric iodide, bismuth potassium iodide and silico-tungstic acid test solution respectively, all generate precipitation.The discriminating of alkaloids in this side of being.Through test of many times, the result all is positive.

(3) get the content of 5 of thousand cypress rhinitis solid pharmaceutical preparations, porphyrize adds ether 20ml, and jolting filters, and gets filtrate 2ml, puts in the porcelain dish, and low temperature is flung to ether, adds 1 of 1% vanillic aldehyde sulfuric acid solution, shows bluish violet to aubergine.The discriminating of volatile oil in this side of being.Through test of many times, the result all is positive.

The residual filtrate of (4) getting under the item of [discriminating] (3) is concentrated into about 2ml, hydro-oxidation sodium test solution 1ml, and jolting is left standstill, and adds 1 of superoxol, puts in the water-bath and heats, and shows orange red, after the watery hydrochloric acid acidifying, orange red should taking off.The discriminating of Cassia occidentalis in this side of being.Through test of many times, the result all is positive.

(5) get the content of 6 of thousand cypress rhinitis solid pharmaceutical preparations, porphyrize adds ethanol 25ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 2ml, make dissolving, as need testing solution.Other gets climbing groundsel control medicinal material 25g, and boiling 1 hour filters, and filtrate is condensed into thick paste, adds ethanol 40ml, and reflux is 1 hour in the water-bath, filters, and filtrate evaporate to dryness, residue add the about 2ml of ethanol makes dissolving, in contrast medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 3 μ l of above-mentioned two kinds of solution, on the same silica gel g thin-layer plate of idea, be developping agent respectively, launch with toluene-ethyl acetate-formic acid (5: 4: 1), take out, dry, spray is with the ferric trichloride test solution, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.This method is the thin layer discrimination method of climbing groundsel in the primary standard, the result of reproduction, and the spot colour developing is clear, and negative control is noiseless, so continue to use primary standard, lists it in quality standard.

(6) get the content of 40 of thousand cypress rhinitis solid pharmaceutical preparations, porphyrize adds strong ammonia solution 1ml, adds chloroform 20ml, puts in the water-bath and refluxes 1 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets Chinese ephedra control medicinal material 1g, shines medicinal material solution in pairs with legal system.Get the ephedrine hydrochloride reference substance again, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-strong ammonia solution (4: 1: 0.1) is developping agent, launch, take out, dry, spray is with ninhydrin solution, about 5 minutes of 105 ℃ of bakings.In the test sample chromatogram, respectively with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show identical punctation.This method is the thin layer discrimination method of ephedrine hydrochloride in the primary standard, the result of reproduction, and the spot colour developing is clear, and negative control is noiseless, so continue to use primary standard, lists it in quality standard text.

(7) with notopterygium root medicinal material in the notopterygium root control medicinal material discriminating side.Method one: get this product content 5g, the 20ml that adds diethyl ether, cold soaking spends the night, and filters, and filtrate evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Method two: get this product content 5g, porphyrize adds sherwood oil (60～90 ℃) 20ml, and sonicated 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, in contrast product solution.Be developping agent with benzene-ethyl acetate (4: 1), normal hexane-benzene-ethyl acetate (2: 1: 1) respectively, launch.The result shows, selecting method two is that the degree of separation launched of developping agent is good with normal hexane-benzene-ethyl acetate (2: 1: 1), and the spot colour developing is clear, and negative control is noiseless, thus selection its list the quality standard text in.

(8) in the quality research process, we once adopted the medicinal materials such as Selaginella tamariscina, Cassia occidentalis, the root of Dahurain angelica and Ligusticum wallichii among several different methods the other side to carry out test of many times, all because of feminine gender have disturb or spot unintelligible, factors such as method poor reproducibility fail to find ripe discrimination method, so exclude the quality standard text.

3. check: press the regulation under capsule (the appendix I L) item in appendix rules of preparations of Chinese Pharmacopoeia version in 2000, carry out moisture, content uniformity, disintegration time limited, heavy metal inspection, the inspection of arsenic salt, limit test of microbe respectively.

The methodological study of experimental example 2 assay methods of the present invention

1. the research of Chinese ephedra assay

1.1 it is reported, mainly contain compositions such as organic amine alkaloid such as ephedrine, pseudoephedrine, ephedine etc. and a small amount of volatility, flavonoids in the Chinese ephedra.The assay of chemical constitution in the Chinese ephedra, what bibliographical information was more is the assay of ephedrine, method for measuring mostly is high performance liquid chromatography.With reference to relevant documents and materials, the Content of Ephedrine With in the preparation is measured, and method for measuring is studied.

1.1.1 Tianjin, instrument island LC-10ATvp high performance liquid chromatograph, the LC-10ATvp infusion pump; SPD-M10Avp secondary array detector; The SCL-10Avp system controller; CTO-10ASvp thermocolumn case; The CLASS-VP6.1 workstation data processing system; Microsyringe (25 μ L), HAMILTON company; Supersonic wave cleaning machine (250W, Beijing's Medical Devices two factories).

1.1.2 reagent methyl alcohol (chromatographically pure), phosphoric acid (analyzing pure), triethylamine (analyzing pure), NaOH (analyzing pure), hydrochloric acid (analyzing pure), ethanol (analyzing pure), water are the secondary redistilled water; (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the ephedrine hydrochloride reference substance, for assay usefulness, lot number: 714-0202).

1.1.3 chromatographic condition chromatographic column: Hypersil ODS2 (5.0 * 200mm) Dalian Yi Lite; Moving phase: 0.1% phosphoric acid solution-methyl alcohol-triethylamine (93: 7: 0.1); Detect wavelength: 207nm; Flow velocity: 1.2ml/min; Column temperature: 40 ℃; Sample size: 5 μ l.

1.1.4 system suitability test

The negative need testing solution of getting ephedrine hydrochloride reference substance solution, need testing solution and scarce Chinese ephedra medicinal material respectively injects liquid chromatograph, the record chromatogram.Retention time (the t of ephedrine _R) being about 11.0 minutes, the negative sample chromatogram is at non-false positive peak, ephedrine position, and ephedrine separates fully (degree of separation＞1.5) with close impurity peaks, and promptly ephedrine separates with other components fully under this test condition.Number of theoretical plate is calculated as 3500 with ephedrine, and the degree of separation of ephedrine and other component peaks is greater than 1.5.Calculate with the ephedrine peak so decide number of theoretical plate, should be not less than 2000.

1.1.5 linear relationship is investigated

1.1.5.1 it is an amount of that the preparation precision of standard solution takes by weighing the ephedrine hydrochloride reference substance, adds dissolve with methanol, makes the solution of the hydrochloric ephedrine 0.1060mg of every 1ml (amounting to ephedrine 0.0868mg).

1.1.5.2 accurate ephedrine hydrochloride reference substance solution (amounting to ephedrine 0.0868mg/ml) 2,4,6,8, the 10 μ l that draw of the drafting of typical curve, inject liquid chromatograph respectively, the record chromatogram, with peak area A mass number (μ g) being carried out linear regression calculates, getting equation of linear regression is: A=2042331.80X-3057.00, r=0.9999.Precision is measured reference substance solution 5 μ l and is injected liquid chromatograph, the gained peak area is 883440, one need testing solution sample introduction is analyzed the gained peak area calculate with regression equation and one point method respectively, result's relative deviation is 0.05%, so text adopts one point method to carry out assay.The range of linearity: 0.1736 μ g～0.868 μ g.

1.1.6 the selective extraction method of need testing solution extraction time: ultrasonic Extraction, measurement result sees Table 1.

The investigation (n=2) of the different ultrasonic times of table 1 Diluted Alcohol

Ultrasonic time (min)	10	20	30	45	60
Ultrasonic time (min)	10	20	30	45	60	Content (%) RSD (%)	0.1557 2.43	0.1591 2.55	0.1795 1.01	0.1796 2.91	0.1789 2.71

As seen from Table 1, ultrasonic Extraction can be extracted the ephedrine in the preparation fully in 30 minutes, so select sonicated 30 minutes.

Get this product 1.1.7 the extraction time of extract and Content of Ephedrine With measurement result are investigated, the accurate title, decide, and adds Diluted Alcohol, close plug, weigh, sonicated 30 minutes is put cold, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, subsequent filtrate is regulated pH value to 11 with sodium hydroxide test solution, use chloroform extraction, the assay of its extraction time and ephedrine the results are shown in Table 2.

The assay result (n=2) of table 2 extraction time and ephedrine

Extraction time	2	3	4	5	6
Extraction time	2	3	4	5	6	Content (%) RSD (%)	0.1548 3.04	0.1695 2.79	0.1793 0.95	0.1791 1.69	0.1790 1.91

Extract as seen from the table 4 times with 5 times, 6 times there was no significant difference as a result, therefore in quality standard, extraction times is decided to be 4 times.

1.1.8 precision test

Get ephedrine hydrochloride reference substance solution (amounting to ephedrine 0.0868mg/ml), repeat sample introduction 5 times, measure peak area, average peak area is 885140, and RSD is 0.27%.

1.1.9 reappearance test

Get this product (lot number: 20021124) content, prepare 5 parts of test liquids by the preparation method of test liquid under the assay item in the quality standard, sample introduction is measured peak area respectively, result of calculation is listed table 3 in, average content is 0.1793%, RSD is 0.67%.

The reappearance test of table 3 preparation test sample epheday intermedia alkali

The sample introduction number of times	1	2	3	4	5	Mean value	RSD(％)
The sample introduction number of times	1	2	3	4	5	Mean value	RSD(％)	Content (%)	0.1785	0.1790	0.1800	0.1780	0.1810	0.1793	0.67

1.1.10 stability test

Get this product (lot number: 20021124) content, preparation method by test liquid under the assay item in the quality standard prepares test liquid, respectively at 0,1,2,4,8 hour mensuration ephedrine peak area, the result lists table 4 in, average peak area is 921884, RSD is 0.44%, illustrates that test liquid is good at 8 hours internal stabilities.

The stability test of table 4 preparation test sample epheday intermedia alkali

Minute (h)	0	1	2	4	8	Mean value	RSD(％)
Minute (h)	0	1	2	4	8	Mean value	RSD(％)	Peak area	921225	918637	928141	918165	923251	921884	0.44

1.1.11 recovery test

Adopt the application of sample absorption method, the accurate respectively 5 parts of samples of having measured content (lot number: 20021124, average content is 0.1793%) that take by weighing are an amount of, put in the tool plug conical flask, (with the Diluted Alcohol is solvent to accurate adding ephedrine hydrochloride reference substance solution, 0.0225mg/ml, amount to ephedrine 0.0184mg/ml) 50ml, sonicated 30 minutes filters, the accurate filtrate 25ml that draws, with sodium hydroxide solution adjust pH to 11, put in the separating funnel, with 4 (20ml of chloroform extraction, 20ml, 10ml, 10ml), combined chloroform liquid, adding hydrochloric acid solution (1 → 100) 30ml jolting extracts, divide and get hydrochloric acid solution, chloroform solution washes with water 2 times, each 10ml, merge hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter, make test liquid with filter membrane (0.45 μ m).The accurate respectively 5 μ l of absorption inject liquid chromatograph, and the record chromatogram is measured content, calculate recovery rate (the results are shown in Table 5), and average recovery rate is 98.82%, RSD is 0.58%.

Table 5 thousand cypress capsule for treating rhinitis epheday intermedia alkali are measured recovery test

Test number (TN)	Sample size (g)	Contain ephedrine (mg)	Add ephedrine (mg)	The amount of recording (mg)	The recovery (%)	Average recovery rate (%)	RSD (％)
Test number (TN)	Sample size (g)	Contain ephedrine (mg)	Add ephedrine (mg)	The amount of recording (mg)	The recovery (%)	Average recovery rate (%)	RSD (％)	1 2 3 4 5	0.5210 0.5046 0.4992 0.5042 0.5254	0.9342 0.9047 0.8951 0.9040 0.9420	0.920	1.8396 1.8205 1.8049 1.8161 1.8445	98.41 99.54 98.89 99.14 98.10	98.82	0.58

1.1.12 sample determination

Prepare test liquid and reference substance solution by quality standard, sample introduction 5 μ l write down chromatogram respectively, measure peak area, are calculated as follows content:

In the formula: Ai: the need testing solution peak area

As: reference substance solution peak area

Cs: reference substance solution concentration (mg/ml)

W: test sample sample weighting amount (g)

W ₁: average particle heavy (g)

3 batch sample epheday intermedia alkali content measurement results (seeing Table 6)

Table 63 batch sample epheday intermedia alkali content measurement results

Lot number	Content (mg/ grain)		Average content (mg/ grain)
	Content (mg/ grain)			1	2
	20030107 20030112 20030117	0.96 0.98 0.98		1	2	0.93 0.98 0.96	0.95 0.98 0.97

According to 3 batch sample measurement results, the yield of sample epheday intermedia alkali is all more than 50%, so preparation epheday intermedia alkali content limit is calculated as follows:

Content of Ephedrine With limit=preparation contains Chinese ephedra medicinal material (g/g) * medicinal material and contains heavy (mg)=162/500 * 0.8% * 50% * 500=0.648mg/ grain of ephedrine limit (pharmacopeia regulation) * yield * grain.

So deciding every of this product contains Chinese ephedra with ephedrine (C ₁₀H ₁₅NO) meter must not be less than 0.65mg.

Embodiment of the present invention

The quality control of embodiment 1 capsule of the present invention

Get Chinese crude drug climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, above-mentioned raw materials, climbing groundsel, Selaginella tamariscina, Cassia occidentalis, Chinese ephedra boiling secondary, collecting decoction filters, and filtrate is condensed into thick paste; Three flavors such as all the other notopterygium roots are ground into fine powder, add above-mentioned thick paste, stir evenly, and drying is ground into fine powder, and mixing incapsulates, and makes 1000, promptly.

1, measure the content of ephedrine with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D):

Test apparatus, reagent, reagent

Tianjin, island LC-10ATvp high performance liquid chromatograph, the LC-10ATvp infusion pump; SPD-M10Avp secondary array detector; The SCL-10Avp system controller; CTO-10ASvp thermocolumn case; The CLASS-VP6.1 workstation data processing system; Microsyringe (25 μ L), HAMILTON company; Supersonic wave cleaning machine (250W, Beijing's Medical Devices two factories).

(Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the ephedrine hydrochloride reference substance, for assay usefulness, lot number: 714-0202).

Methyl alcohol (chromatographically pure), phosphoric acid (analyzing pure), triethylamine (analyzing pure), glacial acetic acid (analyzing pure), water are the secondary redistilled water.

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase: 0.1% phosphoric acid-methyl alcohol-triethylamine (93: 7: 0.1); Flow velocity: 1.2ml/min; Column temperature: 40 ℃; Detect wavelength 207nm.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

2, differentiate this medicine Chinese crude drug Chinese ephedra with thin-layered chromatography

3, differentiate this medicine Chinese crude drug notopterygium root with thin-layered chromatography

A. the preparation of need testing solution: get the content of 10 of this product, porphyrize adds sherwood oil (60～90 ℃) 20ml, and sonicated 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

C. test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw need testing solution 10 μ l, control medicinal material solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate (2: 1: 1), launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

4, differentiate this medicine Chinese crude drug climbing groundsel with thin-layered chromatography

C. according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (5: 4: 1) is developping agent, launches, and takes out, dry, spray is with the ferric trichloride test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

5, the qualitative identification of other drug in this capsule

The quality control of embodiment 2 tablets of the present invention

Get Chinese crude drug climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, above-mentioned raw materials, climbing groundsel, Selaginella tamariscina, Cassia occidentalis, Chinese ephedra boiling secondary, collecting decoction filters, and filtrate is condensed into thick paste; Three flavors such as all the other notopterygium roots are ground into fine powder, add above-mentioned thick paste, stir evenly, and drying is ground into fine powder, and mixing adds appropriate amount of auxiliary materials, and compressing tablet is made 1000, promptly.

Test apparatus, reagent, reagent

C. this product 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 30 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulate pH value to 11 with sodium hydroxide test solution, with 4 (20ml of chloroform extraction, 20ml, 10ml, 10ml), combined chloroform liquid, adding hydrochloric acid solution (1 → 100) 30ml jolting extracts, divide and get hydrochloric acid solution, chloroform solution washes with water 2 times, each 10ml, merge hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter with filter membrane (0.45 μ m), promptly.

A. the preparation of need testing solution: get the content of 20 of this product, porphyrize adds strong ammonia solution 1ml, adds chloroform 20ml, puts in the water-bath and refluxes 1 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.

A. the preparation of need testing solution: get 10 of this product, porphyrize adds sherwood oil (60～90 ℃) 20ml, and sonicated 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

A. the preparation of need testing solution: get 6 of this product, porphyrize adds ethanol 25ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution.

5, the sufficient property discriminating of other drug in this tablet

(1) microscopic character of powder of notopterygium root, the root of Dahurain angelica, Ligusticum wallichii is differentiated and is got this product, porphyrize, put microscopically and observe: amylum body is numerous, simple grain ellipse, Long Circle, similar round, oval or kidney shape etc., diameter 5～16 μ m, omphalion point-like, slit shape or herringbone shape; Accidental composite grain is made up of 2～4 gradation.Bunch shape crystallization is present in the parenchyma cell, is similar round agglomerate or class cluster crystal shape, diameter 10～25 μ m.The pale brown look of cork cell is the class polygon.How broken the grease chamber is, the visible fragment of idol, and the secretory cell wall is thin, contains more oil droplet.How cataclasm secretory canal is, and common golden yellow or yellowish-brown strip secretion.Conduit is mainly reticulate vessel and spiral duct, diameter 14～50 μ m.

(2) 8 of this product are got in the discriminating of alkaloids, add acidic ethanol 30ml, put in the water-bath reflux 20 minutes, and filtered, get filtrate 20ml, put and fling to ethanol in the water-bath, residue adds watery hydrochloric acid 1ml, water 10ml makes dissolving, filters, and gets each 1ml of filtrate, add each 1～2 of potassium mercuric iodide, bismuth potassium iodide and silico-tungstic acid test solution respectively, all generate precipitation.

(3) 5 of this product are got in the discriminating of volatile oil, and porphyrize adds ether 20ml, and jolting filters, and gets filtrate 2ml, puts in the porcelain dish, and low temperature is flung to ether, adds 1 of 1% vanillic aldehyde sulfuric acid solution, show bluish violet to aubergine.

The quality control of embodiment 3 granules of the present invention

Get Chinese crude drug climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, above-mentioned raw materials, climbing groundsel, Selaginella tamariscina, Cassia occidentalis, Chinese ephedra boiling secondary, collecting decoction filters, and filtrate is condensed into thick paste; Three flavors such as all the other notopterygium roots are ground into fine powder, add above-mentioned thick paste, stir evenly, and the system particle, drying, whole grain, mixing, packing, every bag of capacity 10g, promptly.

Test apparatus, reagent, reagent

The every 10g of this product contains Chinese ephedra with ephedrine (C ₁₀H ₁₅NO) meter must not be less than 0.65mg.

A. the preparation of need testing solution: get this product 30g, porphyrize adds strong ammonia solution 1ml, adds chloroform 20ml, puts in the water-bath and refluxes 1 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.

A. the preparation of need testing solution: get this product 30g, porphyrize adds sherwood oil (60～90 ℃) 20ml, and sonicated 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

A. the preparation of need testing solution: get this product 20g, porphyrize adds ethanol 25ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution.

5, the qualitative identification of other drug in this granule

(2) this product 25g is got in the discriminating of alkaloids, add acidic ethanol 30ml, put in the water-bath reflux 20 minutes, and filtered, get filtrate 20ml, put and fling to ethanol in the water-bath, residue adds watery hydrochloric acid 1ml, water 10ml makes dissolving, filters, and gets each 1ml of filtrate, add each 1～2 of potassium mercuric iodide, bismuth potassium iodide and silico-tungstic acid test solution respectively, all generate precipitation.

(3) content of this product 10g is got in the discriminating of volatile oil, and porphyrize adds ether 20ml, and jolting filters, and gets filtrate 2ml, puts in the porcelain dish, and low temperature is flung to ether, adds 1 of 1% vanillic aldehyde sulfuric acid solution, shows bluish violet to aubergine.

The quality control of embodiment 4 capsules of the present invention

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine (88: 4.6: 0.05); Flow velocity: 1.2ml/min; Column temperature: 36 ℃; Detect wavelength 205nm.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 20 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulates pH value to 10 with sodium hydroxide test solution, with 3 (20ml of chloroform extraction, 20ml, 20ml), combined chloroform liquid adds hydrochloric acid solution (1 → 100) 30ml jolting and extracts, and divides and gets hydrochloric acid solution, chloroform solution washes with water and merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter with filter membrane (0.45 μ m), promptly.

A. the preparation of need testing solution: get the content of 35 of this product, porphyrize adds strong ammonia solution 1ml, adds chloroform 10ml, puts in the water-bath and refluxes 0.5 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.

D. according to the thin-layered chromatography test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-strong ammonia solution (2: 0.9: 0.08) is developping agent, launches, and takes out, dry, spray is with ninhydrin solution, about 3 minutes of 105 ℃ of bakings.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation.

A. the preparation of need testing solution: get the content of 5 of this product, porphyrize adds sherwood oil (60～90 ℃) 15ml, and sonicated 25 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

C. test according to thin-layered chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate (1.1: 0.8: 0.8), launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

A. the preparation of need testing solution: get the content of 3 of this product, porphyrize adds ethanol 15ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 20g, boiling 1 hour filters, and filtrate is condensed into thick paste, add ethanol 30ml, reflux is 1 hour in the water-bath, filters, filtrate evaporate to dryness, residue add the about 1ml of ethanol makes dissolving, in contrast medicinal material solution.

C. according to the thin-layered chromatography test, drawing each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with toluene-ethyl acetate-formic acid (3.8: 3.5: 0.9), launches, and takes out, and dries, and spray is with the ferric trichloride test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

5, the qualitative identification of other drug in this capsule

With the 5th of embodiment 1

The quality control of embodiment 5 capsules of the present invention

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine (96: 9: 0.13); Flow velocity: 1.2ml/min; Column temperature: 45 ℃; Detect wavelength 209nm.Number of theoretical plate is with ephedrine (C ₁₀H ₁₅NO) peak meter should be not less than 2000.

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 35 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulates pH value to 13 with sodium hydroxide test solution, with 6 (20ml of chloroform extraction, 20ml, 10 ml, 10ml 10ml, 10ml), combined chloroform liquid adds hydrochloric acid solution (1 → 100) 30ml jolting and extracts, and divides and gets hydrochloric acid solution, chloroform solution washes with water and merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, is transferred in the 10ml measuring bottle, be diluted to scale with methyl alcohol, shake up, filter with filter membrane (0.45 μ m), promptly.

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

A. the preparation of need testing solution: get the content of 45 of this product, porphyrize adds strong ammonia solution 5ml, adds chloroform 30ml, puts in the water-bath and refluxes 2 hours, filters, and filtrate evaporate to dryness, residue add methyl alcohol 3ml makes dissolving, as need testing solution.

B. the preparation of reference substance solution: get Chinese ephedra control medicinal material 3g, shine medicinal material solution in pairs with legal system.

D. according to the thin-layered chromatography test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-strong ammonia solution (6: 1.2: 0.12) is developping agent, launches, and takes out, dry, spray is with ninhydrin solution, about 7 minutes of 105 ℃ of bakings.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation.

A. the preparation of need testing solution: get the content of 15 of this product, porphyrize adds sherwood oil (60～90 ℃) 30ml, and sonicated 35 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get notopterygium root control medicinal material 1g, shine medicinal material solution in pairs with legal system.

C. test according to thin-layered chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate (2.4: 1.1: 1.1), launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

A. the preparation of need testing solution: get the content of 8 of this product, porphyrize adds ethanol 30ml, puts in the water-bath reflux 2 hours, filters, and filtrate evaporate to dryness, residue add ethanol 3ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 30g, boiling 3 hours filters, and filtrate is condensed into thick paste, add ethanol 50ml, reflux is 2 hours in the water-bath, filters, filtrate evaporate to dryness, residue add the about 3ml of ethanol makes dissolving, in contrast medicinal material solution.

C. according to the thin-layered chromatography test, drawing each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with toluene-ethyl acetate-formic acid (6: 5: 1.2), launches, and takes out, and dries, and spray is with the ferric trichloride test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

5, the qualitative identification of other drug in this capsule

With the 5th of embodiment 1

Claims

1. cypress rhinitis quality control of solid dosage forms method, described thousand cypress rhinitis solid pharmaceutical preparations are made by climbing groundsel 4848g, Selaginella tamariscina 808g, Cassia occidentalis 484g, Chinese ephedra 162g, notopterygium root 32g, root of Dahurain angelica 16g and Ligusticum wallichii 16g, described method of quality control may further comprise the steps: with the content of high effective liquid chromatography for measuring thousand cypress rhinitis solid pharmaceutical preparation epheday intermedia alkali; Differentiate thousand cypress rhinitis solid pharmaceutical preparation Chinese crude drug climbing groundsels, Chinese ephedra and notopterygium root with thin-layered chromatography; Thousand cypress rhinitis solid pharmaceutical preparations are carried out qualitative identification; Wherein said content with high effective liquid chromatography for measuring thousand cypress rhinitis solid pharmaceutical preparation epheday intermedia alkali; The process following steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine of 88-96: 4.6-9: 0.05-0.13; Flow velocity: 1.2ml/min; Column temperature: 36-45 ℃; Detect wavelength 207 ± 2nm; Number of theoretical plate is with ephedrine C ₁₀H ₁₅NO peak meter should be not less than 2000;

B. it is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds that methyl alcohol is made, solution, promptly;

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 20-35 minute, put coldly, claim to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulate the pH value to 10-13 with sodium hydroxide test solution, with chloroform extraction 4-6 time, each 20ml or 10ml, combined chloroform liquid, adding hydrochloric acid solution 30ml jolting by 1 → 100 extracts, divide and get hydrochloric acid solution, chloroform solution washes with water and merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, be transferred in the 10ml measuring bottle, be diluted to scale, shake up with methyl alcohol, filter with 0.45 μ m filter membrane, promptly;

D. accurate respectively reference substance solution and each the 5-10 μ l of need testing solution of drawing of determination method injects liquid chromatograph, measures, promptly; Every of this product contains Chinese ephedra with ephedrine C ₁₀H ₁₅The NO meter must not be less than 0.65mg; Describedly differentiate the method for thousand cypress rhinitis solid pharmaceutical preparation Chinese crude drug climbing groundsels, Chinese ephedra and notopterygium root with thin-layered chromatography, through following steps:

Chinese ephedra is differentiated and follows these steps to carry out:

A. the preparation of need testing solution: get the content of this product 35-45 grain, porphyrize adds strong ammonia solution 1-5ml, adds chloroform 10-30ml, puts in the water-bath backflow 0.5-2 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1-3ml makes dissolving, as need testing solution;

B. the preparation of reference substance solution: get Chinese ephedra control medicinal material 1-3g, shine medicinal material solution in pairs with legal system;

C. get the ephedrine hydrochloride reference substance again, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution;

D. according to the thin-layered chromatography test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, chloroform-methanol-strong ammonia solution with 2-6: 0.9-1.2: 0.08-0.12 is a developping agent, launches, and takes out, dry, spray is with ninhydrin solution, 105 ℃ of bakings 3-7 minute; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation;

Notopterygium root is differentiated and follows these steps to carry out:

A. the preparation of need testing solution: get the content of this product 5-15 grain, porphyrize adds 60～90 ℃ sherwood oil 15-30ml, and sonicated 25-35 minute, filter, filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get notopterygium root control medicinal material 0.5-1g, shine medicinal material solution in pairs with legal system;

C. test according to thin-layered chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, putting respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-benzene-ethyl acetate of 1.1-2.4: 0.8-1.1: 0.8-1.1, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

Climbing groundsel is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of this product 3-8 grain, porphyrize adds ethanol 15-30ml, puts in the water-bath reflux 1-2 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1-3ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 20-30g, during boiling 1-3, filter, filtrate is condensed into thick paste, add ethanol 30-50ml, in the water-bath reflux 1-2 hour, filter, filtrate evaporate to dryness, residue add ethanol 1-3ml makes dissolving, in contrast medicinal material solution;

C. according to thin-layered chromatography test, drawing each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with toluene-ethyl acetate-formic acid of 3.8-6: 3.5-5: 0.9-1.2, launches, and takes out, and dries, and spray is with the ferric trichloride test solution; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

Described thousand cypress rhinitis solid pharmaceutical preparations are carried out qualitative identification, may further comprise the steps:

A. the microscopic character of powder of notopterygium root, the root of Dahurain angelica, Ligusticum wallichii is differentiated, gets this product content, porphyrize, and put microscopically and observe: amylum body is numerous, simple grain ellipse, Long Circle, similar round, oval or kidney shape; , diameter 5～16 μ m, omphalion point-like, slit shape or herringbone shape; Accidental composite grain is made up of 2～4 gradation; Bunch shape crystallization is present in the parenchyma cell, is similar round agglomerate or class cluster crystal shape, diameter 10～25 μ m; The pale brown look of cork cell is the class polygon; How broken the grease chamber is, the visible fragment of idol, and the secretory cell wall is thin, contains more oil droplet; How cataclasm secretory canal is, and common golden yellow or yellowish-brown strip secretion; Conduit is mainly reticulate vessel and spiral duct, diameter 14～50 μ m,

B. the content of 8 of this product is got in the discriminating of alkaloids, adds acidic ethanol 30ml, put in the water-bath reflux 20 minutes, filter, get filtrate 20ml, put and fling to ethanol in the water-bath, residue adds watery hydrochloric acid 1ml, water 10ml makes dissolving, filter, get each 1ml of filtrate, add each 1～2 of potassium mercuric iodide, bismuth potassium iodide and silico-tungstic acid test solution respectively, all generate precipitation

C. the content of 5 of this product is got in the discriminating of volatile oil, and porphyrize adds ether 20ml, and jolting filters, and gets filtrate 2ml, puts in the porcelain dish, and low temperature is flung to ether, adds 1 of 1% vanillic aldehyde sulfuric acid solution, shows bluish violet to aubergine,

D. the discriminating of Cassia occidentalis, the residual filtrate of getting under [discriminating] c item is concentrated into 2ml, hydro-oxidation sodium test solution 1ml, jolting is left standstill, and adds 1 of superoxol, puts in the water-bath and heats, and shows orange red, after the watery hydrochloric acid acidifying, orange red should taking off.

2. the method for quality control of claim 1 is characterized in that: the described high performance liquid chromatography of using, and step is as follows:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filling agent; Moving phase is 0.1% phosphoric acid-methyl alcohol-triethylamine of 93: 7: 0.1; Flow velocity: 1.2ml/min; Column temperature: 40 ℃; Detect wavelength 207nm; Number of theoretical plate is with ephedrine C ₁₀H ₁₅NO peak meter should be not less than 2000;

B. it is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and make every 1ml and contain 0.1mg, amounts to the solution of ephedrine 0.08190mg, promptly;

C. this product content 1.0g under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug conical flask, add Diluted Alcohol 50ml, close plug claims to decide weight, sonicated 30 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, precision is measured subsequent filtrate 25ml, regulates pH value to 11 with sodium hydroxide test solution, uses chloroform extraction 4 times, be followed successively by 20ml, 20ml, 10ml, 10ml, combined chloroform liquid adds hydrochloric acid solution 30ml jolting by 1 → 100 and extracts, and divides and gets hydrochloric acid solution, chloroform solution washes with water 2 times, each 10ml merges hydrochloric acid solution and water liquid, evaporate to dryness, residue adds dissolve with methanol, be transferred in the 10ml measuring bottle, be diluted to scale, shake up with methyl alcohol, filter with 0.45 μ m filter membrane, promptly;

D. accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly; Every of this product contains Chinese ephedra with ephedrine C ₁₀H ₁₅The NO meter must not be less than 0.65mg.

3. the method for quality control of claim 1 is characterized in that: describedly differentiate the method for thousand cypress rhinitis solid pharmaceutical preparation Chinese crude drug climbing groundsels, Chinese ephedra and notopterygium root with thin-layered chromatography, through following steps:

Chinese ephedra is differentiated and follows these steps to carry out:

A. the preparation of need testing solution: get the content of 40 of this product, porphyrize adds strong ammonia solution 1ml, adds chloroform 20ml, puts in the water-bath and refluxes 1 hour, filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution;

B. the preparation of reference substance solution: get Chinese ephedra control medicinal material 1g, shine medicinal material solution in pairs with legal system;

D. according to the thin-layered chromatography test, drawing each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with chloroform-methanol-strong ammonia solutions of 4: 1: 0.1, launches, and takes out, and dries, and spray is with ninhydrin solution, 105 ℃ of bakings 5 minutes; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical punctation;

Notopterygium root is differentiated and follows these steps to carry out:

A. the preparation of need testing solution: get the content of 10 of this product, porphyrize adds 60～90 ℃ sherwood oil 20ml, and sonicated 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get notopterygium root control medicinal material 0.5g, shine medicinal material solution in pairs with legal system;

C. according to the thin-layered chromatography test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l put respectively on same silica gel g thin-layer plate, with 2: 1: 1 normal hexane-benzene-ethyl acetates was developping agent, launched, and took out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

Climbing groundsel is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get the content of 6 of this product, porphyrize adds ethanol 25ml, puts in the water-bath reflux 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get climbing groundsel control medicinal material 25g, boiling 1 hour filters, and filtrate is condensed into thick paste, adds ethanol 40ml, and reflux is 1 hour in the water-bath, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, in contrast medicinal material solution;

C. according to the thin-layered chromatography test, drawing each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with 5: 4: 1 toluene-ethyl acetate-formic acid, launches, and takes out, and dries, and spray is with the ferric trichloride test solution; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.