CN102520113B - Detection method for fifteen Rupeng preparation - Google Patents

Abstract

The invention relates to a detection method for a fifteen Rupeng preparation. The detection method comprises one or several of detection of Common Vladimiria Root, Semen Cassiae and Rhizoma Acori Calami in the preparation through thin layer chromatography, and detection of the content of aconitine in the preparation.

Description

The detection method of ten five tastes breast roc preparations

Technical field

The invention belongs to technical field of traditional Chinese medicines, be specifically related to a kind of detection method of Chinese medicine preparation.

Background technology

Ten five tastes breast roc balls are Tibetan medicine's tradition proved recipes, and prescription is comprised of shellfish main officer of Tibet, wide muscle rattan, cassia seed, the tame and docile cream of slag, Semen seu folium abelmoschi moschati, Rhizoma Acori Calami, BAXIAGA, catechu, myrobalan's (stoning), styrax, terminaliae billericae,fructus, Aconitum Szechenyianum Gay (system), the banksia rose, emblic, Moschus ten gomi herbs.Can anti-inflammatory analgetic, dry yellow water, for joint congestion and swelling pain, itches, gout, yellow water gathers.

Ten five tastes breast roc balls that use are at present recorded in first of < < Drug Standard of Ministry of Public Health of the Peoples Republic of China nineteen ninety-five version Tibetan medicine:

Tibetan medicine name: Bi Qiong Ai Bu of a specified duration

Phonetic name: Shiwuwei Rupeng Wan

Book page number: C1-199 standard number: WS3-BC-0196-95

[prescription] shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax 60g, terminaliae billericae,fructus 150g, Aconitum Szechenyianum Gay 75g, banksia rose 150g, Moschus 1.5g, emblic 150g.

[method for making] above ten five tastes, slagging-off is tamed and dociled outside cream, Moschus, and all the other are ground into fine powder, sieve, and add muscone's fine powder, mix, and tame and docile cream add appropriate water pill with slag, dry in the shade and get final product.

[proterties] this product is the sepia water-bindered pill; The micro-perfume (or spice) of gas, bitter.

[inspection] should meet every regulation relevant under pill item (8 pages of appendix).

[function with cure mainly] anti-inflammatory analgetic, dry yellow water.For joint congestion and swelling pain, itch, gout, yellow water gathers.

[usage and consumption] 2～4 balls, 2 times on the one.

The heavy 3g of [specification] every 10 balls.

[storage] is airtight, puts shady and cool dry place

Not yet there is at present the detection method of ten five tastes breast roc preparations, also do not have toxic component in the other side to carry out the method for content detection.

Summary of the invention

The object of the present invention is to provide a kind of detection method of ten five tastes breasts roc preparations, described detection method has that specificity is strong, favorable reproducibility, the advantage such as easy and simple to handle.

For achieving the above object, technical scheme of the present invention is:

A detection method for ten five tastes breast roc preparations, said preparation bulk drug consists of: frankincense 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g; The method comprises one or more of following discriminating and/or inspection method: cross thin-layer chromatography the radix jurineae in preparation, cassia seed, Rhizoma Acori Calami are carried out to qualitative discriminating, and check preparation mesaconitine content.

Particularly, the qualitative discriminating of radix jurineae comprises:

1a) prepare radix jurineae need testing solution, radix jurineae control medicinal material solution, not containing the negative sample solution of radix jurineae;

2a), according to 2010 editions one appendix VI B of thin-layered chromatography < < Chinese Pharmacopoeia > > test, draw respectively step 1a) need testing solution, control medicinal material solution, negative sample solution the point sample that obtain be on same silica gel g thin-layer plate;

3a) take cyclohexane-methylene chloride-ethyl acetate as developping agent, by step 2a) thin layer plate that obtains be placed on saturated in expansion cylinder, launch, take out, dry, spray is with vanillic aldehyde sulfuric acid solution, it is clear that hot blast blows to spot colour developing, in thin-layer chromatography, with the corresponding position of radix jurineae control medicinal material chromatogram on, the spot of the aobvious same color of test sample chromatogram;

The qualitative discriminating of cassia seed comprises:

1b) prepare cassia seed need testing solution, cassia seed control medicinal material solution, not containing the negative sample solution of cassia seed;

2b), according to 2010 editions one appendix VI B of thin-layered chromatography < < Chinese Pharmacopoeia > > test, draw respectively step 1b) need testing solution, control medicinal material solution, negative sample solution the point sample that obtain be on same silica gel g thin-layer plate;

3b) take the upper strata liquid of sherwood oil-ethyl formate-formic acid as developping agent, by step 2b) thin layer plate that obtains be placed on saturated in expansion cylinder, launch, take out, dry, under ultraviolet lamp, inspect, in thin-layer chromatography, with the corresponding position of cassia seed control medicinal material chromatogram on, the fluorescence spot of the aobvious same color of test sample chromatogram;

The qualitative discriminating of Rhizoma Acori Calami comprises:

1c) prepare Rhizoma Acori Calami need testing solution, Rhizoma Acori Calami control medicinal material solution, not containing the negative sample solution of Rhizoma Acori Calami;

2c), according to 2010 editions one appendix VI B of thin-layered chromatography < < Chinese Pharmacopoeia > > test, draw respectively rapid 1c) need testing solution, control medicinal material solution, negative sample solution the point sample that obtain be in same silica G F ₂₅₄on thin layer plate;

3c) take petroleum ether-ethyl acetate as developping agent, by step 2c) thin layer plate that obtains be placed on saturated in expansion cylinder, launch, take out, dry, under ultraviolet lamp, inspect; In thin-layer chromatography, with the corresponding position of Rhizoma Acori Calami control medicinal material chromatogram on, the blackening point of the aobvious same color of test sample chromatogram;

Content of Aconitine inspection comprises:

1d) prepare aconitine need testing solution, aconitine reference substance solution;

2d) according to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw respectively step 1d) need testing solution that obtains, aconitine reference substance solution point sample are on same silica gel g thin-layer plate;

3d) take cyclohexane-ethyl acetate-diethylamine as developping agent, by step 2d) thin layer plate that obtains be placed on saturated in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, in thin-layer chromatography, test sample chromatogram with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

The preparation method of the need testing solution of described radix jurineae, cassia seed, Rhizoma Acori Calami and aconitine, control medicinal material solution, negative sample solution comprises:

(1) radix jurineae

Get described ten five tastes breast roc preparations, porphyrize, adds acetone, and ultrasonic processing filters, and filtrate is concentrated, as the need testing solution of radix jurineae; Separately get radix jurineae control medicinal material, be made in the same way of radix jurineae control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution;

(2) cassia seed

Get described ten five tastes breast roc preparations, porphyrize, adds methyl alcohol, adds hot reflux, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution, ultrasonic processing, then add methenyl choloride, add hot reflux, cooling, in dislocation separating funnel, divide and get methenyl choloride layer, acid solution is extracted with methenyl choloride, adds anhydrous sodium sulfate and dewaters in right amount, filters, filtrate volatilizes, and residue adds methyl alcohol to be made to dissolve, as the need testing solution of cassia seed; Separately depend on pine torch control medicinal material, be made in the same way of cassia seed control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution;

(3) Rhizoma Acori Calami

Get described ten five tastes breast roc preparations, add methyl alcohol, ultrasonic, filter, filtrate is concentrated, as the need testing solution of Rhizoma Acori Calami; Separately get Rhizoma Acori Calami control medicinal material, be made in the same way of Rhizoma Acori Calami control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution;

(4) aconitine

Get described ten five tastes breast roc preparations, porphyrize, adds ammonia solution, stirs evenly, and places, and adds diethyl ether, and jolting, places, filter, and filtrate evaporate to dryness, residue adds absolute ethyl alcohol to be made to dissolve, the need testing solution detecting as Content of Aconitine; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make solution, as aconitine reference substance solution.

Preferably, said method comprising the steps of:

A) the qualitative discriminating of radix jurineae

Get described ten five tastes breast roc preparation 1～3 weight portions, porphyrize, adds acetone 10～30 parts by volume, and ultrasonic processing 10～30 minutes filters, and filtrate concentrates approximately 1～3 parts by volume, as the need testing solution of radix jurineae; Separately get radix jurineae control medicinal material 0.1～0.8 weight portion, be made in the same way of radix jurineae control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution; According to 2010 editions appendix VIB tests of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw each 0.003～0.007 parts by volume of above-mentioned three kinds of solution, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as 10～20: 2～8: cyclohexane-methylene chloride-ethyl acetate of 0.5～2 is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the spot of the aobvious same color of test sample chromatogram;

B) the qualitative discriminating of cassia seed

Get described ten five tastes breast roc preparation 1～3 weight portions, porphyrize, add methyl alcohol 10～30 parts by volume, add hot reflux 20～45 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10～30 parts by volume, ultrasonic processing 3～8 minutes, add again methenyl choloride 10～30 parts by volume, add hot reflux 20～45 minutes, cooling, in dislocation separating funnel, divide and get methenyl choloride layer, acid solution is extracted 1～3 time with methenyl choloride, each 5～15 parts by volume, merge methenyl choloride liquid, adding anhydrous sodium sulfate dewaters in right amount, filter, filtrate volatilizes, residue adds methyl alcohol 1～3 parts by volume to be made to dissolve, as the need testing solution of cassia seed, separately depend on pine torch control medicinal material 0.1～0.8g, be made in the same way of cassia seed control medicinal material solution, in prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution, according to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 0.005～0.02 parts by volume, control medicinal material solution 0.003～0.007 parts by volume and negative sample solution 0.005～0.02 parts by volume, point sample is on same silica gel g thin-layer plate respectively, with volume ratio 10～20: 2～8: 0.5～2, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect, in thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of the aobvious same color of test sample chromatogram,

C) the qualitative discriminating of Rhizoma Acori Calami

Get described ten five tastes breast roc preparation 1～3 weight portions, add methyl alcohol 10～30 parts by volume, ultrasonic 10～30 minutes, filter, filtrate is concentrated into approximately 1～3 parts by volume, as the need testing solution of Rhizoma Acori Calami; Separately get Rhizoma Acori Calami control medicinal material 0.1～0.8 weight portion, be made in the same way of Rhizoma Acori Calami control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 0.005～0.02 parts by volume, control medicinal material solution 0.003～0.007 parts by volume and negative sample solution 0.005～0.02 parts by volume, point sample is in same silica G F respectively ₂₅₄on thin layer plate, take volume ratio as 5～15: 0.5～4, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the blackening point of the aobvious same color of test sample chromatogram;

D) Content of Aconitine inspection

Get described ten five tastes breast roc preparation 3～8 weight portions, porphyrize, adds ammonia solution 0.5～2 parts by volume, stirs evenly, place 1～3 hour, 10～20 parts by volume that add diethyl ether, jolting 0.5～2 hour, places 18～30 hours, filter, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 1～3 parts by volume to be made to dissolve, as the need testing solution of aconitine; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every parts by volume containing 0.0005～0.002 weight portion, as aconitine reference substance solution; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw need testing solution 0.005～0.03 parts by volume, aconitine reference substance solution 0.0015～0.015 parts by volume, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as 2～8: 1～3: cyclohexane-ethyl acetate-diethylamine of 0.5～2 is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In thin layer test sample chromatogram, test sample chromatogram with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

More preferably, said method comprising the steps of:

I) the qualitative discriminating of radix jurineae

Get described ten five tastes breast roc preparation 2g, porphyrize, adds acetone 20ml, and ultrasonic processing 20 minutes filters, and filtrate concentrates about 2ml, as the need testing solution of radix jurineae; Separately get radix jurineae control medicinal material 0.5g, be made in the same way of radix jurineae control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 5 μ l of above-mentioned three kinds of solution, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as the cyclohexane-methylene chloride-ethyl acetate of 15: 5: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the spot of the aobvious same color of test sample chromatogram;

Ii) the qualitative discriminating of cassia seed

Get described ten five tastes breast roc preparation 2g, porphyrize, adds methyl alcohol 20ml, add hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 20ml, ultrasonic processing 5 minutes, add again methenyl choloride 20ml, add hot reflux 30 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 2 times for acid solution, each 10ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, residue adds methyl alcohol 2ml to be made to dissolve, as the need testing solution of cassia seed; Separately depend on pine torch control medicinal material 0.5g, be made in the same way of cassia seed control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, point sample is on same silica gel g thin-layer plate respectively, with volume ratio 15: 5: 1, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of the aobvious same color of test sample chromatogram;

Iii) the qualitative discriminating of Rhizoma Acori Calami

Get described ten five tastes breast roc preparation 2g, add methyl alcohol 20ml, ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as the need testing solution of Rhizoma Acori Calami; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of Rhizoma Acori Calami control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5～10 μ l, control medicinal material solution 5 μ l and negative sample solution 5～10 μ l, point sample is in same silica G F respectively ₂₅₄on thin layer plate, take volume ratio as 10: 2, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, and thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the blackening point of the aobvious same color of test sample chromatogram;

Iv) Content of Aconitine inspection

Get described ten five tastes breast roc preparation 5g, porphyrize, adds ammonia solution 1ml, stirs evenly, place 2 hours, and the 15ml that adds diethyl ether, jolting 1 hour, places 24 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 2ml to be made to dissolve, as the need testing solution of aconitine; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1mg, as aconitine reference substance solution; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw need testing solution 20 μ l, 10 μ l, aconitine reference substance solution 10 μ l, 5 μ l, 2.5 μ l, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as the cyclohexane-ethyl acetate-diethylamine of 5: 2: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In thin-layer chromatography, test sample chromatogram with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

The formulation of above-mentioned ten five tastes breast roc preparations can be tablet, capsule, granule, pill or powder.Preparation method is: get described ten five tastes breast roc preparations, be ground into fine powder, sieve, mix, add suitable quantity of water pill, dry in the shade and get final product; Or get described ten five tastes breast roc preparations, and be ground into fine powder, sieve, mix, mix, by pharmacy conventional method, add conventional auxiliary material, make the oral formulations of accepting clinically.

The constituent that the present invention is directed to ten five tastes breast roc preparations, detects the existence of index composition radix jurineae, cassia seed, Rhizoma Acori Calami, and detects the content of the toxic component aconitine in preparation, to reach the object that the quality of preparation is controlled.Detection method of the present invention detects needs according to reality, and the testing conditions of existing detection method is screened, and has determined the preferably detection method of ten five tastes breast roc preparations, and that this detection method has is sensitive, favorable reproducibility and specificity strong feature.

Accompanying drawing explanation

Fig. 1 is radix jurineae thin-layer chromatography detection figure in ten five tastes breast roc preparations;

Wherein, from left to right sample order is: the need testing solution 3 of the need testing solution 1 of radix jurineae, the need testing solution 2 of radix jurineae, radix jurineae, the need testing solution 4 of radix jurineae, radix jurineae control medicinal material solution, not containing the negative sample solution of radix jurineae;

Wherein, developping agent is cyclohexane-methylene chloride-ethyl acetate (15: 5: 1), and developer is 5% vanillic aldehyde sulfuric acid.

Fig. 2 is cassia seed thin-layer chromatography detection figure in ten five tastes breast roc preparations;

Wherein, from left to right sample order is: the need testing solution 3 of the need testing solution 1 of cassia seed, the need testing solution 2 of cassia seed, cassia seed, the need testing solution 4 of cassia seed, cassia seed control medicinal material solution, cassia seed control medicinal material solution, not containing the negative sample solution of cassia seed;

Wherein, developping agent is the upper strata liquid of sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (15: 5: 1), under ultraviolet lamp (365nm), inspects.

Fig. 3 is Rhizoma Acori Calami thin-layer chromatography detection figure in ten five tastes breast roc preparations;

Wherein from left to right sample order is: the need testing solution 3 of the need testing solution 1 of Rhizoma Acori Calami, the need testing solution 2 of Rhizoma Acori Calami, Rhizoma Acori Calami, the need testing solution 4 of Rhizoma Acori Calami, Rhizoma Acori Calami control medicinal material solution, Rhizoma Acori Calami control medicinal material solution, not containing the negative controls of Rhizoma Acori Calami;

Wherein, developping agent is sherwood oil (60 ℃～90 ℃)-ethyl acetate (10: 2), under ultraviolet lamp (254nm), inspects.

Fig. 4 is ten five tastes breast roc preparation mesaconitine content detection figure;

Wherein from left to right sample order is: (point sample amount 10 μ l) for the need testing solution 1 that Content of Aconitine detects, (point sample amount 10 μ l) for the need testing solution 2 that Content of Aconitine detects, (point sample amount 10 μ l) for the need testing solution 3 that Content of Aconitine detects, (point sample amount 5 μ l) for aconitine reference substance solution 1, (point sample amount 10 μ l) for aconitine reference substance solution 2, (point sample amount 20 μ l) for the need testing solution 4 that Content of Aconitine detects, (point sample amount 20 μ l) for the need testing solution 5 that Content of Aconitine detects, (point sample amount 20 μ l) for the need testing solution 6 that Content of Aconitine detects, (point sample amount 2.5 μ l) for aconitine reference substance solution, (point sample amount 2.5 μ l) for aconitine reference substance solution,

Wherein, developping agent is cyclohexane-ethyl acetate-diethylamine (5: 2: 1), and developer is rare bismuth iodide test solution; Under daylight, inspect.

Embodiment

The following preparation for each composition detection all referred to as " test sample ", is interpreted as the test sample that it refers to require according to each detection method preparation under each detection method, for example, at the test sample described in radix jurineae method, be " test sample of radix jurineae ".

The preparation of [embodiment 1] ten five tastes breast roc preparations

Frankincense 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled to cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g, slagging-off is tamed and dociled outside cream, Moschus or muscone, all the other are ground into fine powder, sieve, add Moschus or muscone's fine powder, mix, with slag, tame and docile cream and add suitable quantity of water pill, dry in the shade and get final product.Or above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, add Moschus or muscone's fine powder, mix, by pharmacy conventional method, add conventional auxiliary material, make the oral formulations of accepting clinically: granule, pill, capsule, tablet or powder.

Usage and consumption: a day dose is 1.2-2.4g crude drug amount, 2 times on the one.

The screening experiment of the thin-layer chromatography condition of radix jurineae in [embodiment 2] ten five tastes breast roc preparations

(1) instrument

Mortar, electronic scales, tool plug conical flask, transfer pipet, ultrasound wave extraction apparatus, funnel, filter paper, evaporating dish, sample applicator, silica G plate, chromatography cylinder, spray bottle, hair dryer.

(2) control medicinal material

Radix jurineae control medicinal material (lot number: 1091-200001), purchased from Nat'l Pharmaceutical & Biological Products Control Institute.

(3) reagent

Ether, cyclohexane, acetone, sherwood oil (60 ℃～90 ℃), ethyl formate, formic acid, ethyl acetate, methylene chloride, 5% vanillic aldehyde sulfuric acid.

(4) method of inspection:

Extract the selection of solvent: adopt respectively ether and acetone for extracting solvent;

The selection of extracting method: adopt respectively ultrasonic processing and add hot reflux;

The selection of developping agent: respectively take cyclohexane-acetone (10: 3), sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (15: 5: 0.5), sherwood oil (60 ℃～90 ℃)-ethyl acetate (9: 1), cyclohexane-methylene chloride-ethyl acetate (15: 5: 1) as developping agent.

(5) select respectively above different solvents, extracting method and developping agent to test, result is as follows

Repetition test in the above conditions, finally determines preferred detection method, specific as follows:

Get described ten five tastes breast roc preparation 2g, porphyrize, adds acetone 20ml, and ultrasonic processing 20 minutes filters, and filtrate concentrates about 2ml, as need testing solution; Separately get radix jurineae control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take volume ratio as the cyclohexane-methylene chloride-ethyl acetate of 15: 5: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color, as shown in Figure 1.The method operates through many people test of many times, favorable reproducibility, and specificity is strong, can be used as the detection method of radix jurineae in ten five tastes breast roc preparations.

The screening experiment of the thin-layer chromatography condition of cassia seed in [embodiment 3] ten five tastes breast roc preparations

(1) instrument

Mortar, electronic scales, tool plug conical flask, transfer pipet, ultrasound wave extraction apparatus, funnel, filter paper, evaporating dish, sample applicator, silica G plate, chromatography cylinder, spray bottle, hair dryer.

(2) control medicinal material

Cassia seed control medicinal material (lot number: 1211011-200403), purchased from Nat'l Pharmaceutical & Biological Products Control Institute.

(3) reagent

Methyl alcohol, 95% ethanol, 2.5mol/L sulfuric acid solution, methenyl choloride, anhydrous sodium sulfate, sherwood oil (60 ℃～90 ℃), ethyl formate, formic acid.

(4) method of inspection:

Extract the selection of solvent: adopt respectively methyl alcohol and 95% ethanol for extracting solvent;

The selection of extracting method: adopt respectively dipping, add hot reflux, ultrasonic extraction;

The selection of developping agent: respectively take the upper strata liquid of sherwood oil (30～60 ℃)-acetone (2: 1), sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (15: 5: 1) as developping agent.

(5) select respectively above different solvents, extracting method and developping agent to test, result is as follows:

Repetition test in the above conditions, has finally determined cassia seed preferred detection method, specific as follows:

Get described ten five tastes breast roc preparation 2g, porphyrize, adds methyl alcohol 20ml, adds hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 20ml, ultrasonic processing 5 minutes, add again methenyl choloride 20ml, add hot reflux 30 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 2 times for acid solution, each 10ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 2ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, put respectively on same silica gel g thin-layer plate, with volume ratio 15: 5: 1, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.As shown in Figure 2.The method operates through many people test of many times, favorable reproducibility, and specificity is strong, can be used as the detection method of cassia seed in ten five tastes breast roc preparations.

The screening experiment of the thin-layer chromatography condition of Rhizoma Acori Calami in [embodiment 4] ten five tastes breast roc preparations

(1) instrument

Mortar, electronic scales, tool plug conical flask, transfer pipet, ultrasound wave extraction apparatus, funnel, filter paper, evaporating dish, sample applicator, silica G and GF254 thin layer plate, chromatography cylinder, spray bottle, hair dryer.

(2) control medicinal material

Rhizoma Acori Calami control medicinal material (lot number: 121084-200502), purchased from Nat'l Pharmaceutical & Biological Products Control Institute.

(3) reagent

Methyl alcohol, ether, sherwood oil (60 ℃～90 ℃), ethyl acetate, normal hexane, formic acid.

(4) method of inspection:

Extract the selection of solvent: adopt respectively methyl alcohol and ether for extracting solvent;

The selection of thin layer plate: adopt respectively silica gel g thin-layer plate and silica GF254 thin layer plate;

The selection of developping agent: respectively take the upper strata liquid of sherwood oil (30～60 ℃)-ethyl acetate (10: 2), normal hexane-ethyl acetate-formic acid (9: 1: 1) as developping agent.

(5) select respectively above different solvents, thin layer plate and developping agent to test, result is as follows:

Repetition test in the above conditions, has finally determined the preferred detection method of Rhizoma Acori Calami, specific as follows:

Get described ten five tastes breast roc preparation 2g, porphyrize, adds methyl alcohol 20ml, and ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5～10 μ l, control medicinal material solution 5 μ l and negative sample solution 5～10 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take volume ratio as 10: 2, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, and thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.As shown in Figure 3.The method operates through many people test of many times, favorable reproducibility, and specificity is strong, can be used as the detection method of Rhizoma Acori Calami in ten five tastes breast roc preparations.

The screening experiment of the thin-layer chromatography condition of [embodiment 5] ten five tastes breast roc preparation mesaconitine content

(1) instrument

Mortar, electronic scales, tool plug conical flask, transfer pipet, ultrasound wave extraction apparatus, funnel, filter paper, evaporating dish, sample applicator, silica gel g thin-layer plate, chromatography cylinder, spray bottle, hair dryer.

(2) reference substance

Aconitine reference substance (lot number: 320756035164), purchased from Nat'l Pharmaceutical & Biological Products Control Institute.

(3) reagent

Ammonia solution, ether, absolute ethyl alcohol, cyclohexane, benzene, ethyl acetate, diethylamine, methylene chloride, acetone, methyl alcohol, rare bismuth potassium iodide, improvement bismuth potassium iodide test solution.

(4) extracting method

The selection of developping agent: respectively take benzene-ethyl acetate-ethylenediamine (14: 4: 1), cyclohexane-ethyl acetate-ethylenediamine (14: 4: 1), cyclohexane-ethyl acetate-diethylamine (5: 2: 1), methylene chloride-acetone-methyl alcohol (6: 1: 1) as developping agent;

The selection of developer: respectively take rare bismuth potassium iodide test solution and improvement bismuth potassium iodide test solution as colour developing.

(5) select respectively above different developping agents and developer to test, result is as follows:

Repetition test in the above conditions, finally determined the preferred detection method of Content of Aconitine:

Get described ten five tastes breast roc preparation 5g, porphyrize, adds ammonia solution 1ml, stirs evenly, place 2 hours, and the 15ml that adds diethyl ether, jolting 1 hour, places 24 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 2ml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast; According to thin-layered chromatography test described in 2010 editions one appendix VI B of < < Chinese Pharmacopoeia > >, accurate need testing solution 20 μ l, the 10 μ l of drawing, aconitine reference substance solution 10 μ l, 5 μ l, 2.5 μ l, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate-diethylamine (5: 2: 1) as developping agent, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot, as shown in Figure 4.The method operates through many people test of many times, favorable reproducibility, and specificity is strong, can be used as the detection method of ten five tastes breast roc preparation mesaconitine.

[test example 1]: the detection of ten five tastes breast roc balls

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, slagging-off is tamed and dociled outside cream, Moschus or muscone, and all the other are ground into fine powder, sieve, and add Moschus or muscone's fine powder, mix, and tame and docile cream add suitable quantity of water pill with slag, dry in the shade and get final product.

A. the thin layer of radix jurineae is differentiated

Get described ten five tastes breast roc ball 2g, porphyrize, adds acetone 20ml, and ultrasonic processing 20 minutes filters, and filtrate concentrates about 2ml, as need testing solution; Separately get radix jurineae control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take volume ratio as the cyclohexane-methylene chloride-ethyl acetate of 15: 5: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

B. the thin layer of cassia seed is differentiated

Get described ten five tastes breast roc ball 2g, porphyrize, adds methyl alcohol 20ml, adds hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 20ml, ultrasonic processing 5 minutes, add again methenyl choloride 20ml, add hot reflux 30 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 2 times for acid solution, each 10ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 2ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, put respectively on same silica gel g thin-layer plate, with volume ratio 15: 5: 1, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.

C. the thin layer of Rhizoma Acori Calami is differentiated

Get described ten five tastes breast roc ball 2g, add methyl alcohol 20ml, ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5～10 μ l, control medicinal material solution 5 μ l and negative sample solution 5～10 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take volume ratio as 10: 2, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, and thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.

D. the content inspection of aconitine

Get described ten five tastes breast roc ball 5g, porphyrize, adds ammonia solution 1ml, stirs evenly, place 2 hours, and the 15ml that adds diethyl ether, jolting 1 hour, places 24 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 2ml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw need testing solution 10 μ l and aconitine reference substance solution 2.5 μ l, negative sample solution 10 μ l, put respectively on same silica gel g thin-layer plate, take volume ratio as the cyclohexane-ethyl acetate-diethylamine of 5: 2: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

[test example 2]: the detection of ten five tastes breast roc sheets

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, and add Moschus or muscone's fine powder, mix, and by pharmacy conventional method, add conventional auxiliary material, make acceptable ten five tastes breast roc sheets clinically.

A. the thin layer of radix jurineae is differentiated

Get above-mentioned ten five tastes breast roc sheet 1g, porphyrize, adds acetone 10ml, and ultrasonic processing 10 minutes filters, and filtrate is waved to about 1ml, as need testing solution; Separately get radix jurineae control medicinal material 0.3g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-methylene chloride-ethyl acetate (12: 3: 0.8) as developping agent, thin layer plate is put saturated 10min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

B. the thin layer of cassia seed is differentiated

Get above-mentioned ten five tastes breast roc sheet 3g, porphyrize, adds methyl alcohol 30ml, adds hot reflux 45 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 30ml, ultrasonic processing 8 minutes, add again methenyl choloride 30ml, add hot reflux 45 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 3 times for acid solution, each 5ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 3ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.8g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 15 μ l, control medicinal material solution 7 μ l and negative sample solution 15 μ l, put respectively on same silica gel g thin-layer plate, take the upper strata liquid of sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (17: 6: 1.5) as developping agent, thin layer plate is put saturated 30min in expansion cylinder, launch, take out, dry, put under ultraviolet lamp (365nm) and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color, and spot separates better, without hangover, negative without obviously disturbing.C. the limit examine of aconitine

C. the content inspection of aconitine

Get above-mentioned ten five tastes breast roc sheet 8g, porphyrize, adds ammonia solution 2ml, stirs evenly, place 3 hours, and the 20ml that adds diethyl ether, jolting 2 hours, places 30 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 3ml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1.5mg, product solution in contrast; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, accurate need testing solution 15 μ l and the aconitine reference substance solution 5 μ l of drawing, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate-diethylamine (7: 2.4: 1.3) as developping agent, thin layer plate is put saturated 30min in expansion cylinder, launches, and takes out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

[test example 3]: the detection of ten five tastes breast roc hard shell capsules

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, and add Moschus or enter muscone's fine powder, mix, and by pharmacy conventional method, add conventional auxiliary material, make acceptable ten five tastes breast roc hard shell capsules clinically.

A. the thin layer of radix jurineae is differentiated

Get above-mentioned ten five tastes breast roc hard shell capsules content 2g, porphyrize, adds acetone 20ml, and ultrasonic processing 20 minutes filters, and filtrate is waved to about 2ml, as need testing solution; Separately get radix jurineae control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-methylene chloride-ethyl acetate (15: 5: 1) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

B. the thin layer of Rhizoma Acori Calami is differentiated

Get above-mentioned ten five tastes breast roc hard shell capsules content 1g, add methyl alcohol 120ml, ultrasonic 10 minutes, filter, filtrate is concentrated into about 1ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.3g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5 μ l, control medicinal material solution 3 μ l and negative sample solution 5 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take sherwood oil (60 ℃～90 ℃)-ethyl acetate (7: 0.8) as developping agent, thin layer plate is put saturated 10min in expansion cylinder, launches, and takes out, and dries, and puts under ultraviolet lamp (254nm) and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.

C. the limit examine of aconitine

Get above-mentioned ten five tastes breast roc hard shell capsules 3g, porphyrize, adds ammonia solution 0.5ml, stirs evenly, place 1 hour, and the 10ml that adds diethyl ether, jolting 0.5 hour, places 18 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 1ml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 0.5mg, product solution in contrast; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, accurate need testing solution 5 μ l and the aconitine reference substance solution 1.5 μ l of drawing, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate-diethylamine (3: 1.2: 0.7) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

[test example 4]: the detection of ten five tastes breast roc particles

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, and add Moschus or muscone's fine powder, mix, and by pharmacy conventional method, add conventional auxiliary material, make acceptable ten five tastes breast roc particles clinically.

A. the thin layer of radix jurineae is differentiated

Get above-mentioned ten five tastes breast roc particle 3g, porphyrize, adds acetone 30ml, and ultrasonic processing 30 minutes filters, and filtrate is waved to about 3ml, as need testing solution; Separately get radix jurineae control medicinal material 0.8g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 7 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-methylene chloride-ethyl acetate (18: 7: 1.5) as developping agent, thin layer plate is put saturated 30min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

B. the thin layer of cassia seed is differentiated

Get above-mentioned ten five tastes breast roc particle 1g, porphyrize, adds methyl alcohol 10ml, adds hot reflux 20 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml, ultrasonic processing 3 minutes, add again methenyl choloride 10ml, add hot reflux 20 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 1 time for acid solution, each 15ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 1ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.3g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5 μ l, control medicinal material solution 3 μ l and negative sample solution 5 μ l, put respectively on same silica gel g thin-layer plate, take the upper strata liquid of sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (12: 2: 0.8) as developping agent, thin layer plate is put saturated 10min in expansion cylinder, launch, take out, dry, put under ultraviolet lamp (365nm) and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color, and spot separates better, without hangover, negative without obviously disturbing.

C. the thin layer of Rhizoma Acori Calami is differentiated

Get above-mentioned ten five tastes breast roc particle 2g, add methyl alcohol 20ml, ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take sherwood oil (60 ℃～90 ℃)-ethyl acetate (10: 2) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under ultraviolet lamp (254nm) and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.

[test example 5]: the loose detection of ten five tastes breast rocs

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, the banksia rose/radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, and add Moschus or muscone's fine powder, mix, and by pharmacy conventional method, add conventional auxiliary material, make acceptable ten five tastes breast rocs clinically and fall apart.

A. the thin layer of cassia seed is differentiated

Get the loose 2g of above-mentioned ten five tastes breast roc, porphyrize, adds methyl alcohol 20ml, adds hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 20ml, ultrasonic processing 5 minutes, add again methenyl choloride 20ml, add hot reflux 30 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 2 times for acid solution, each 10ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 2ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes not containing cassia seed negative sample solution by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, put respectively on same silica gel g thin-layer plate, take the upper strata liquid of sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (15: 5: 1) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, put under ultraviolet lamp (365nm) and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color, and spot separates better, without hangover, negative without obviously disturbing.

B. the thin layer of Rhizoma Acori Calami is differentiated

Get the loose 1g of above-mentioned ten five tastes breast roc, add methyl alcohol 120ml, ultrasonic 10 minutes, filter, filtrate is concentrated into about 1ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.3g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5 μ l, control medicinal material solution 3 μ l and negative sample solution 5 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take sherwood oil (60 ℃～90 ℃)-ethyl acetate (7: 0.8) as developping agent, thin layer plate is put saturated 10min in expansion cylinder, launches, and takes out, and dries, and puts under ultraviolet lamp (254nm) and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.

C. the limit examine of aconitine

Get the loose 3g of above-mentioned ten five tastes breast roc, porphyrize, adds ammonia solution 0.5ml, stirs evenly, place 1 hour, and the 10ml that adds diethyl ether, jolting 0.5 hour, places 18 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol lml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 0.5mg, product solution in contrast; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, accurate need testing solution 20 μ l and the aconitine reference substance solution 10 μ l of drawing, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate-diethylamine (3: 1.2: 0.7) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

[test example 6]: the detection of ten five tastes breast roc soft capsules

Shellfish main officer of Tibet 150g, wide muscle rattan 150g, cassia seed 120g, slag are tamed and dociled cream 75g, Semen seu folium abelmoschi moschati 120g, Rhizoma Acori Calami 120g, BAXIAGA 110g, catechu 75g, myrobalan 150g, styrax/bitter Kui Nabu 60g, the terminaliae billericae,fructus 150g of stoning, Aconitum Szechenyianum Gay (system) 75g, banksia rose radix jurineae 150g, Moschus/muscone 1.5g, emblic 150g;

Above ten five tastes, except Moschus or muscone, all the other are ground into fine powder, sieve, and add Moschus or muscone's fine powder, mix, and by pharmacy conventional method, add conventional auxiliary material, make acceptable ten five tastes breast roc soft capsules clinically.

A. the thin layer of radix jurineae is differentiated

Get above-mentioned ten five tastes breast roc soft capsule 3g, porphyrize, adds acetone 30ml, and ultrasonic processing 30 minutes filters, and filtrate is waved to about 3ml, as need testing solution; Separately get radix jurineae control medicinal material 0.8g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 7 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-methylene chloride-ethyl acetate (18: 7: 1.5) as developping agent, thin layer plate is put saturated 30min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

B. the thin layer of cassia seed is differentiated

Get above-mentioned ten five tastes breast roc soft capsule 1g, porphyrize, adds methyl alcohol 10ml, adds hot reflux 20 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml, ultrasonic processing 3 minutes, add again methenyl choloride 10ml, add hot reflux 20 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 1 time for acid solution, each 15ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, and residue adds methyl alcohol 1ml to be made to dissolve, as need testing solution; Separately depend on pine torch control medicinal material 0.3g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5 μ l, control medicinal material solution 3 μ l and negative sample solution 5 μ l, put respectively on same silica gel g thin-layer plate, take the upper strata liquid of sherwood oil (60 ℃～90 ℃)-ethyl formate-formic acid (12: 2: 0.8) as developping agent, thin layer plate is put saturated 10min in expansion cylinder, launch, take out, dry, put under ultraviolet lamp (365nm) and inspect; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color, and spot separates better, without hangover, negative without obviously disturbing.

C. the thin layer of Rhizoma Acori Calami is differentiated

Get above-mentioned ten five tastes breast roc soft capsule 2g, add methyl alcohol 20ml, ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of control medicinal material solution; In prescription ratio and preparation technology, configuration does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution.According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, put respectively in same silica G F ₂₅₄on thin layer plate, take sherwood oil (60 ℃～90 ℃)-ethyl acetate (10: 2) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under ultraviolet lamp (254nm) and inspects; In test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the blackening point of aobvious same color.

D. the limit examine of aconitine

Get above-mentioned ten five tastes breast roc soft capsule 5g, porphyrize, adds ammonia solution 1ml, stirs evenly, place 2 hours, and the 15ml that adds diethyl ether, jolting 1 hour, places 24 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 2ml to be made to dissolve, as need testing solution; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, accurate need testing solution 10 μ l and the aconitine reference substance solution 2.5 μ l of drawing, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate-diethylamine (5: 2: 1) as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, dry, spray is with rare bismuth potassium iodide test solution; In test sample chromatogram, with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

Claims (4)

1. a detection method for ten five tastes breast roc preparations, said preparation bulk drug consists of: frankincense 150 weight portions, wide muscle rattan 150 weight portions, cassia seed 120 weight portions, slag are tamed and dociled cream 75 weight portions, Semen seu folium abelmoschi moschati 120 weight portions, Rhizoma Acori Calami 120 weight portions, BAXIAGA 110 weight portions, catechu 75 weight portions, myrobalan's 150 weight portions of stoning, styrax/bitter Kui Nabu 60 weight portions, terminaliae billericae,fructus 150 weight portions, Aconitum Szechenyianum Gay processed 75 weight portions, the banksia rose/radix jurineae 150 weight portions, Moschus/muscone's 1.5 weight portions, emblic 150 weight portions;

The method comprises following discriminating and/or inspection method, comprises the following steps:

A) the qualitative discriminating of radix jurineae

Get described ten five tastes breast roc preparation 1～3 weight portions, porphyrize, adds acetone 10～30 parts by volume, and ultrasonic processing 10～30 minutes filters, and filtrate concentrates approximately 1～3 parts by volume, as the need testing solution of radix jurineae; Separately get radix jurineae control medicinal material 0.1～0.8 weight portion, be made in the same way of radix jurineae control medicinal material solution; In prescription ratio and preparation technology, preparation does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution; According to 2010 editions appendix VIB tests of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw each 0.003～0.007 parts by volume of above-mentioned three kinds of solution, point sample is on same silica gel g thin-layer plate respectively, cyclohexane-methylene chloride-ethyl acetate take volume ratio as 10～20:2～8:0.5～2 is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the spot of the aobvious same color of test sample chromatogram;

B) the qualitative discriminating of cassia seed

Get described ten five tastes breast roc preparation 1～3 weight portions, porphyrize, add methyl alcohol 10～30 parts by volume, add hot reflux 20～45 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10～30 parts by volume, ultrasonic processing 3～8 minutes, add again methenyl choloride 10～30 parts by volume, add hot reflux 20～45 minutes, cooling, in dislocation separating funnel, divide and get methenyl choloride layer, acid solution is extracted 1～3 time with methenyl choloride, each 5～15 parts by volume, merge methenyl choloride liquid, adding anhydrous sodium sulfate dewaters in right amount, filter, filtrate volatilizes, residue adds methyl alcohol 1～3 parts by volume to be made to dissolve, as the need testing solution of cassia seed, separately depend on pine torch control medicinal material 0.1～0.8g, be made in the same way of cassia seed control medicinal material solution, in prescription ratio and preparation technology, preparation does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution, according to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 0.005～0.02 parts by volume, control medicinal material solution 0.003～0.007 parts by volume and negative sample solution 0.005～0.02 parts by volume, point sample is on same silica gel g thin-layer plate respectively, with volume ratio 10～20: 2～8: 0.5～2, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect, in thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of the aobvious same color of test sample chromatogram,

C) the qualitative discriminating of Rhizoma Acori Calami

Get described ten five tastes breast roc preparation 1～3 weight portions, add methyl alcohol 10～30 parts by volume, ultrasonic 10～30 minutes, filter, filtrate is concentrated into approximately 1～3 parts by volume, as the need testing solution of Rhizoma Acori Calami; Separately get Rhizoma Acori Calami control medicinal material 0.1～0.8 weight portion, be made in the same way of Rhizoma Acori Calami control medicinal material solution; In prescription ratio and preparation technology, preparation does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 0.005～0.02 parts by volume, control medicinal material solution 0.003～0.007 parts by volume and negative sample solution 0.005～0.02 parts by volume, point sample is in same silica G F respectively ₂₅₄on thin layer plate, take volume ratio as 5～15:0.5～4, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, and thin layer plate is put saturated 0～40min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the blackening point of the aobvious same color of test sample chromatogram;

D) Content of Aconitine inspection

Get described ten five tastes breast roc preparation 3～8 weight portions, porphyrize, adds ammonia solution 0.5～2 parts by volume, stirs evenly, place 1～3 hour, 10～20 parts by volume that add diethyl ether, jolting 0.5～2 hour, places 18～30 hours, filter, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 1～3 parts by volume to be made to dissolve, as the need testing solution of aconitine; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every parts by volume containing 0.0005～0.002 weight portion, as aconitine reference substance solution; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw need testing solution 0.005～0.03 parts by volume, aconitine reference substance solution 0.0015～0.015 parts by volume, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as 2～8: 1～3: cyclohexane-ethyl acetate-diethylamine of 0.5～2 is developping agent, thin layer plate is put saturated 0～40min in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In thin layer test sample chromatogram, test sample chromatogram with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

2. detection method as claimed in claim 1, comprises the following steps:

I) the qualitative discriminating of radix jurineae

Get described ten five tastes breast roc preparation 2g, porphyrize, adds acetone 20ml, and ultrasonic processing 20 minutes filters, and filtrate concentrates about 2ml, as the need testing solution of radix jurineae; Separately get radix jurineae control medicinal material 0.5g, be made in the same way of radix jurineae control medicinal material solution; In prescription ratio and preparation technology, preparation does not contain the negative sample of radix jurineae, and makes the not negative sample solution containing radix jurineae by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw the each 5 μ l of above-mentioned three kinds of solution, point sample is on same silica gel g thin-layer plate respectively, cyclohexane-methylene chloride-ethyl acetate take volume ratio as 15:5:1 is developping agent, and thin layer plate is put saturated 20min in expansion cylinder, launches, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to spot colour developing; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the spot of the aobvious same color of test sample chromatogram;

Ii) the qualitative discriminating of cassia seed

Get described ten five tastes breast roc preparation 2g, porphyrize, adds methyl alcohol 20ml, add hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 20ml, ultrasonic processing 5 minutes, add again methenyl choloride 20ml, add hot reflux 30 minutes, cooling, in dislocation separating funnel, point get methenyl choloride layer, methenyl choloride extracts 2 times for acid solution, each 10ml, merge methenyl choloride liquid, add anhydrous sodium sulfate and dewater in right amount, filter, filtrate volatilizes, residue adds methyl alcohol 2ml to be made to dissolve, as the need testing solution of cassia seed; Separately depend on pine torch control medicinal material 0.5g, be made in the same way of cassia seed control medicinal material solution; In prescription ratio and preparation technology, preparation does not contain the negative sample of cassia seed, and makes the not negative sample solution containing cassia seed by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 10 μ l, control medicinal material solution 5 μ l and negative sample solution 10 μ l, point sample is on same silica gel g thin-layer plate respectively, with volume ratio 15: 5: 1, boiling point is that the upper strata liquid of 60 ℃～90 ℃ of sherwood oil-ethyl formate-formic acid is developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, put under 365nm ultraviolet lamp and inspect; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of the aobvious same color of test sample chromatogram;

Iii) the qualitative discriminating of Rhizoma Acori Calami

Get described ten five tastes breast roc preparation 2g, add methyl alcohol 20ml, ultrasonic 20 minutes, filter, filtrate is concentrated into about 2ml, as the need testing solution of Rhizoma Acori Calami; Separately get Rhizoma Acori Calami control medicinal material 0.5g, be made in the same way of Rhizoma Acori Calami control medicinal material solution; In prescription ratio and preparation technology, preparation does not contain the negative sample of Rhizoma Acori Calami, and makes the not negative sample solution containing Rhizoma Acori Calami by the preparation method of described need testing solution; According to 2010 editions one appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > >, draw need testing solution 5～10 μ l, control medicinal material solution 5 μ l and negative sample solution 5～10 μ l, point sample is in same silica G F respectively ₂₅₄on thin layer plate, take volume ratio as 10:2, boiling point is that 60 ℃～90 ℃ petroleum ether-ethyl acetates are developping agent, and thin layer plate is put saturated 20min in expansion cylinder, launches, and takes out, and dries, and puts under 254nm ultraviolet lamp and inspects; In thin-layer chromatography, with the corresponding position of control medicinal material chromatogram on, the blackening point of the aobvious same color of test sample chromatogram;

Iv) Content of Aconitine inspection

Get described ten five tastes breast roc preparation 5g, porphyrize, adds ammonia solution 1ml, stirs evenly, place 2 hours, and the 15ml that adds diethyl ether, jolting 1 hour, places 24 hours, filters, filtrate evaporate to dryness, residue adds absolute ethyl alcohol 2ml to be made to dissolve, as the need testing solution of aconitine; Separately get aconitine reference substance appropriate, add absolute ethyl alcohol and make the solution of every 1ml containing 1mg, as aconitine reference substance solution; According to appendix VI B test of thin-layered chromatography < < Chinese Pharmacopoeia > > version in 2010, draw need testing solution 20 μ l, 10 μ l, aconitine reference substance solution 10 μ l, 5 μ l, 2.5 μ l, point sample is on same silica gel g thin-layer plate respectively, take volume ratio as the cyclohexane-ethyl acetate-diethylamine of 5: 2: 1 is as developping agent, thin layer plate is put saturated 20min in expansion cylinder, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In thin-layer chromatography, test sample chromatogram with the corresponding position of reference substance chromatogram on the spot that occurs should be less than the spot of reference substance or not occur spot.

3. the detection method as described in any one in claim 1 to 2, is characterized in that, the formulation of described ten five tastes breast roc preparations is tablet, capsule, granule, pill or powder.

4. the detection method as described in claim 1 to 2 any one, is characterized in that, gets the bulk drug of described weight portion, is ground into fine powder, sieves, and mixes, and adds suitable quantity of water pill, dries in the shade and get final product; Or get the bulk drug of described weight portion, and be ground into fine powder, sieve, mix, by pharmacy conventional method, add conventional auxiliary material, make the oral formulations of accepting clinically.

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