CN103604901B - Thin-layer identification method of compound andrographis tablet - Google Patents
Thin-layer identification method of compound andrographis tablet Download PDFInfo
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Abstract
The invention discloses a thin-layer identification method of a compound andrographis tablet. The thin-layer identification method comprises the followings steps: a step of pre-treating a compound andrographis tablet sample to be identified to obtain liquor of test products; a step of pre-treating negative reference substances to obtain negative liquor of test products; a step of pre-treating the reference substances to obtain reference substance liquor; and a step of carrying out thin-layer chromatographic identification analysis onto the liquor of test products, wherein ethyl acetate and methyl alcohol in a volume ratio of 9:1-99:1 are used as a developer while developing is carried out in the thin-layer chromatographic identification analysis step, a developed thin-layer plate is taken out, dried and sprayed with sulfuric acid liquor of vanillic aldehyde for developing during viewing; the sulfuric acid liquor of vanillic aldehyde is prepared from the vanillic aldehyde and the sulfuric acid with concentration of 60%-70%; mass percent of the vanillic aldehyde in the sulfuric acid liquor of vanillic aldehyde is 1%-2.5%. The identification method disclosed by the invention is high in identification efficiency, high in identification precision and convenient to operate.
Description
Technical field
The present invention relates to the discrimination method of FUFANG CHUANXINLIAN PIAN, particularly relate to a kind of thin-layer identification method of FUFANG CHUANXINLIAN PIAN.
Background technology
FUFANG CHUANXINLIAN PIAN is made up of Herba Andrographitis and yellow avens two kinds of raw materials, the standard of current execution be " the Sanitation Ministry medicine standard " (Pharmacopoeia Commission of the Ministry of Public Health. Drug Standard of Ministry of Public Health of the Peoples Republic of China. Traditional Chinese medicine historical preparation (19) .1998.116), standard number is WS3-B-3624-98.FUFANG CHUANXINLIAN PIAN method for making is: get Herba Andrographitis 60g, be ground into fine powder, remaining Herba Andrographitis 540g and yellow avens 1400g boiling secondary, 2 hours first times, second time 1 hour, collecting decoction, leave standstill, filter, filtrate is condensed into the clear cream that relative density is 1.26 ~ 1.32 (70 DEG C), add Herba Andrographitis fine powder, mixing, adds appropriate amount of auxiliary materials, make particle, be pressed into 1000, sugar coating, obtain final product.
The primary standard of FUFANG CHUANXINLIAN PIAN embodies the discriminating to two kinds of medicinal materials respectively by two TLC distinguish, utilize Dehydro and drographolide standard items to differentiate (1) Herba Andrographitis medicinal material respectively, differentiate (2) yellow avens medicinal material with yellow avens medicinal material standard items, details are as follows:
1, this product 20 is got, removing sugar-coat, porphyrize, add sherwood oil (60 ~ 90 DEG C) 30ml, put in water-bath and add hot reflux 30 minutes, discard sherwood oil liquid, residue adds chloroform 30ml, puts in water-bath and adds hot reflux 30 minutes, let cool, filter, filtrate is decoloured with activated charcoal 2g, and filter, filtrate puts evaporate to dryness in water-bath, residue adds chloroform 0.5ml makes dissolving, as need testing solution.Separately get Dehydro and drographolide reference substance, the solution of every 1ml containing 2mg copied into by chlorination, product solution in contrast.Test according to thin-layered chromatography, draw each 10 ~ 15 μ l of above-mentioned two kinds of solution, put respectively in same with sodium carboxymethyl cellulose be bonding agent silica gel g thin-layer plate on, with chloroform-ethyl acetate-absolute ethyl alcohol (9:6:1) for developping agent, launch, take out, dry, spray the equivalent mixed liquor with 2%3,5-dinitrobenzoic acid ethanolic solution and 5% potassium hydroxide-ethanol solution.In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
2, get this product 10, removing sugar-coat, porphyrize, adds sherwood oil (60 ~ 90 DEG C) 20ml, puts in water-bath and add hot reflux 30 minutes, discard sherwood oil liquid, residue adds chloroform 20ml, puts water-bath and refluxes 30 minutes, let cool, filter, filtrate evaporate to dryness, residue adds chloroform 0.5ml makes dissolving, as need testing solution.Separately get yellow avens control medicinal material 14g, add suitable quantity of water, decoct 30 minutes, filter, filtrate is concentrated into dry, and residue adds ethanol 10ml makes dissolving, and filter, filtrate evaporate to dryness, residue adds chloroform 0.5ml makes dissolving, medicinal material solution in contrast.Test according to thin-layered chromatography, draw each 10 ~ 15 μ l of above-mentioned two kinds of solution, put respectively in same with sodium carboxymethyl cellulose be bonding agent silica gel g thin-layer plate on, with chloroform-ethyl acetate-absolute ethyl alcohol (9:6:1) for developping agent, launch, take out, dry, spray 70% sulfuric acid solution with 0.2% vanillic aldehyde.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, an aobvious same color.
Also have researcher to improve above-mentioned two kinds of methods, but employ the larger organic reagent of toxicity " chloroform " equally, this is intended to be avoided in new edition pharmacopeia as far as possible; In addition, yellow spotting is difficult to obtain in the developping agent having water, and moisture-sensitive wet environment affects.In addition, for the discriminating of medicinal material yellow avens, employ the larger organic reagent of toxicity " chloroform " equally, this is intended to be avoided in new edition pharmacopeia as far as possible.
The discrimination method of the FUFANG CHUANXINLIAN PIAN of prior art has following shortcoming:
1, existing technical method is comparatively loaded down with trivial details to test sample pre-treating method, and this method directly carries out ultrasonic extraction to medicinal material, safety easy to operate relative to water-bath with the organic reagent of the safety such as sherwood oil, ethyl acetate.
2, the thin-layer developing condition of prior art all employ " chloroform ", and this reagent is volatile, easily causes body hepatic injury, and toxicity is comparatively large, for the reagent that do not use is advised by the Chinese Pharmacopoeia council.This method adopts acetate-methanol as developping agent, and security is high.
3, prior art method therefor, the colour developing spot for yellow avens is yellow, and this point is usually yellow band, fuzzyyer, develops the color not obvious, easily causes error.
4, certain methods of the prior art requires the anhydrous existence of developping agent, in humidity higher season and area, operates and more difficultly reaches requirement.Therefore its processing ease is restricted.
As can be seen here, how to provide a kind of easy and simple to handle, reduce toxicity, identify the thin-layer identification method of FUFANG CHUANXINLIAN PIAN accurately, this is the current technical issues that need to address in this area.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of thin-layer identification method of FUFANG CHUANXINLIAN PIAN, simplify the operation course, improve the security of operation and the accuracy of discriminating.
For solving above technical matters, technical scheme of the present invention is:
A kind of thin-layer identification method of FUFANG CHUANXINLIAN PIAN, comprise respectively FUFANG CHUANXINLIAN PIAN sample to be identified is carried out pre-treatment obtain need testing solution step, reference substance is carried out the step that pre-treatment obtains reference substance solution, also comprise the step of need testing solution being carried out to indentification by TLC analysis; Wherein, volume ratio is adopted to be that the ethyl acetate of 9:1 ~ 99:1 and methyl alcohol are as developping agent when launching in the step of indentification by TLC analysis, take out launching complete thin layer plate when inspecting, dry, spray develops the color with the sulfuric acid solution of vanillic aldehyde, wherein, the sulfuric acid that the sulfuric acid solution of described vanillic aldehyde is 60% ~ 70% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of described vanillic aldehyde, the mass percent of vanillic aldehyde is 1% ~ 2.5%.
Preferably, FUFANG CHUANXINLIAN PIAN sample to be identified is carried out the step that pre-treatment obtains need testing solution to be specially: get sample powder to be identified and add the first organic solvent process, filter, obtain the first filtrate and the first residue; First filtrate is discarded, the first residue is added the second organic solvent process again, filter, obtain the second filtrate and the second residue; Second residue is discarded, by the second filtrate evaporate to dryness, obtains the 3rd residue, the 3rd residue is added organic solvent dissolution, as need testing solution.
Preferably, described first organic solvent is sherwood oil or the compound close with sherwood oil polarity, and described second organic solvent is any one or two or more potpourris in methyl alcohol, ethanol, normal butyl alcohol, ethyl acetate.
Preferably, the processing time is 10 ~ 45 minutes.
Preferably, the processing time is 20 ~ 30 minutes.
Preferably, the method for process be ultrasonic, backflow, decoct, dipping in any one or two or more.
Preferably, described reference substance solution comprises: the yellow avens reference substance solution be made up through pre-treatment separately of yellow avens medicinal material, Dehydro and drographolide respectively and Dehydro and drographolide reference substance solution.
Preferably, in the step that described indentification by TLC is analyzed, inspect and be specially: first the thin layer plate having sprayed developer is developed the color at normal temperatures, until the spot characterizing Herba Andrographitis feature occurs, and then heating blankets plate occurs to yellow avens unique point, carries out the colour developing of yellow avens feature.
Preferably, volume ratio is adopted to be that the ethyl acetate of 18:1 ~ 80:1 and methyl alcohol are as developping agent when launching in the step of indentification by TLC analysis.
Compared with prior art, the thin-layer identification method of FUFANG CHUANXINLIAN PIAN of the present invention, thin-layered chromatography is adopted to differentiate, developping agent due to thin-layered chromatography selects ethyl acetate and methyl alcohol, ethyl acetate compares with the chloroform used in prior art, there is hypotoxicity, substantially increase the security of operation; In developping agent, the volume ratio of ethyl acetate and methyl alcohol is set to the so appropriate scope of 9:1 ~ 99:1, clear spot can be made to distinguish, make easy and simple to handle, improve identification efficiency.Adopt the sulfuric acid solution of vanillic aldehyde as developer during owing to inspecting, wherein the concentration of vanillic aldehyde controls, in 1% ~ 2.5% such scope, can effectively develop the color to the point of the distinctive blue cyan of yellow avens, makes discriminating more quick and easy.
Embodiment
In order to make those skilled in the art understand technical scheme of the present invention better, below by specific embodiment, the present invention is described in further detail.
The thin-layer identification method of FUFANG CHUANXINLIAN PIAN of the present invention comprises the steps:
Sample to be identified and reference substance are carried out pre-treatment respectively, then adopts thin-layered chromatography to differentiate.Wherein, the step of pre-treatment can select pre-treatment means of the prior art.
Specifically, discrimination method of the present invention specifically comprises the steps:
1, the preparation of need testing solution:
The powder getting test sample (sample namely to be identified) adds organic solvent, ultrasonic 5 ~ 60 minutes, and filter, filtrate is abandoned, and residue adds organic solvent supersonic 5 ~ 60 minutes, as need testing solution after filtrate is concentrated;
2, the preparation of reference substance and control medicinal material solution: get Dehydro and drographolide reference substance, adds organic solvent and makes reference substance solution; Get yellow avens control medicinal material, make control medicinal material solution according to need testing solution preparation method.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution point sample 10 μ l ~ 15 μ l respectively;
Developping agent: volume ratio is the ethyl acetate of 9:1 ~ 99:1: methyl alcohol;
Expansion mode: ascending development;
Inspect: taking out launching complete thin layer plate, drying, spraying 60% ~ 70% sulfuric acid solution with 1% ~ 2.5% vanillic aldehyde.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, then illustrate that test sample is certified products.
Wherein, the present invention's test material used can select following material, but is not limited to following material during actual enforcement:
Reagent: silica G (lot number 051007) Haiyang Chemical Plant, Qingdao produces, self-control plate gauge lattice 10 × 20mm, thickness 0.5mm; Prefabricated board is that Haiyang Chemical Plant, Qingdao produces, and specification is 10 × 20mm, thickness 0.2 ~ 0.25mm.Other agents useful for same is pure for analyzing.
Control medicinal material: Dehydro and drographolide (lot number: 110854-201007), yellow avens control medicinal material (lot number: 121227-200804), Herba Andrographitis control medicinal material (lot number: 121082-201004) is purchased from Nat'l Pharmaceutical & Biological Products Control Institute.
Test sample: FUFANG CHUANXINLIAN PIAN three batches (01,02,03), for Guangzhou Baiyunshan Heji Huangpu Chinese Medicine Co., Ltd. produces.
Embodiment one
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 20, removing sugar-coat, porphyrize, adds organic solvent---and sherwood oil 30ml, ultrasonic 30 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 30 minutes, filtrate is concentrated into about 2ml, as need testing solution.In other embodiments, also filtrate can be concentrated into any value in 1 ~ 2ml.In other embodiments, as any value that the FUFANG CHUANXINLIAN PIAN of test sample can be in 10 ~ 20.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 30 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution is point sample 15 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 95:5;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 70% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 2.5%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
Embodiment two
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 15, removing sugar-coat, porphyrize, adds organic solvent---sherwood oil 30ml, ultrasonic 40 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 40 minutes, filtrate is concentrated into about 1.4ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 40 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: silica G is from making sheet;
Point sample: need testing solution and control medicinal material solution is point sample 12 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 9:1;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 60% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 2%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
Embodiment three
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 10, removing sugar-coat, porphyrize, adds organic solvent---sherwood oil 30ml, ultrasonic 50 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 50 minutes, filtrate is concentrated into about 1.0ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 50 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution is point sample 13 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 20:1;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 60% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 1%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
Embodiment four
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 12, removing sugar-coat, porphyrize, adds organic solvent---sherwood oil 30ml, ultrasonic 40 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 38 minutes, filtrate is concentrated into about 1.2ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 38 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: silica G is from making sheet;
Point sample: need testing solution and control medicinal material solution is point sample 10 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 35:1;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 70% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 1%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
Embodiment five
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 16, removing sugar-coat, porphyrize, adds organic solvent---and sherwood oil 30ml, ultrasonic 40 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 45 minutes, filtrate is concentrated into about 2ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 45 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution is point sample 12 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 55:1;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 70% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 1.5%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
Embodiment six
The discrimination method of the present embodiment comprises the steps:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 18, removing sugar-coat, porphyrize, adds organic solvent---and sherwood oil 30ml, ultrasonic 60 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 42 minutes, filtrate is concentrated into about 2ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 45 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution is point sample 15 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 99:1;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 60% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 2.3%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point, this illustrates that test sample is certified products.
The thin-layer identification method tool of FUFANG CHUANXINLIAN PIAN of the present invention has the following advantages:
Because unfolding condition developping agent used is that acetate-methanol mixes with any one ratio in (9:1) ~ (99:1), this reagent is compared to the safer environmental protection of the chloroform of prior art, easy and simple to handle.
In sample pretreatment process, the so safer solvent of selection ethyl acetate carries out sample preparation and the disposal route of sample is more simple and convenient, the safer environmental protection of reagent, easy and simple to handle.
In colour developing mode, select concentration be 60% ~ 70% sulfuric acid solution of 1% ~ 2.5% vanillic aldehyde as developer, develop the color all right.And concentration is talked about improperly, as selected 70% sulfuric acid solution of 0.2% vanillic aldehyde, then can not effectively develop the color to the point of the distinctive blue cyan of yellow avens.
In colour developing mode, compared with prior art, although one block of plate can be utilized to differentiate Herba Andrographitis and yellow avens two kinds of medicinal materials simultaneously, but the present invention changes development step, namely, after having sprayed 60% ~ 70% sulfuric acid solution of developer 1% ~ 2.5% vanillic aldehyde, employed the coloration method do not heated for Herba Andrographitis, namely developed the color at normal temperatures, the spot impurity making Herba Andrographitis develop the color like this is less, more can highlight the characteristic of principal spot.And employ heating means for yellow avens, and namely after Herba Andrographitis colour developing, heating blankets plate, carry out yellow avens colour developing, make the discriminating of yellow avens more be added with characteristic, namely in case of heating, there is the point of distinctive blue cyan, make identification result more accurate like this.
Specificity is verified:
The specificity of discrimination method of the present invention:
1, the preparation of need testing solution:
Get test sample-FUFANG CHUANXINLIAN PIAN 15, removing sugar-coat, porphyrize, adds organic solvent---and sherwood oil 30ml, ultrasonic 40 minutes, discard sherwood oil liquid, residue adds ethyl acetate 30ml, ultrasonic 30 minutes, filtrate is concentrated into about 1ml, as need testing solution.
2, the preparation of reference substance solution: get yellow avens medicinal material 10g, add 200ml water, ultrasonic 30 minutes, filter, use equal-volume extraction into ethyl acetate, ethyl acetate portion was concentrated into 2ml, as yellow avens reference substance solution.Get Dehydro and drographolide reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.
3, the preparation of negative control medicinal material solution: Herba Andrographitis 600g, according to the method for making of FUFANG CHUANXINLIAN PIAN, makes not containing the negative controls of yellow avens medicinal material; Yellow avens 1400g, according to the method for making of FUFANG CHUANXINLIAN PIAN, makes not containing the negative controls of Herba Andrographitis medicinal material; Again according to the preparation method of need testing solution, be prepared into respectively not containing the negative need testing solution of Herba Andrographitis with not containing the negative need testing solution of yellow avens.
4, tlc identification method is adopted to differentiate test sample:
Thin layer condition is as follows:
Thin layer plate: common silica G prefabricated board;
Point sample: need testing solution and control medicinal material solution is point sample 4 μ l respectively;
Developping agent: volume ratio is the acetate-methanol of 95:5;
Expansion mode: ascending development;
Inspect: take out launching complete thin layer plate, dry, spray the sulfuric acid solution with vanillic aldehyde, the sulfuric acid that the sulfuric acid solution of this vanillic aldehyde is 60% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of this vanillic aldehyde, the mass percent of vanillic aldehyde is 2%.
Chromatogram identification: in test sample chromatogram, on the position corresponding with Dehydro and drographolide reference substance chromatogram to Herba Andrographitis control medicinal material, aobvious identical spot, not containing on the negative corresponding position of test sample chromatogram of Herba Andrographitis, does not show identical spot; Be heated to spot development with electric-heating air blower or electric hot plate more clear, observe under natural light, on the position corresponding to yellow avens control medicinal material chromatogram, an aobvious identical blue blue spot point; Not containing on the corresponding position of negative controls chromatogram of yellow avens medicinal material, there is no blue cyan spot.This illustrates that test sample is certified products.
Compliance test result:
Verify that discrimination method durability of the present invention is good below by way of testing:
The comparison of different thin layer plate: choose hand paving G-CMC.Na and prefabricated silica G plate, respectively by the test method test drafted.Result shows, all can reach discriminating requirement from making sheet and prefabricated board.
The comparison of different relative humidity: get the thin layer plate after point sample, regulate humidity to be launch in the chromatography cylinder of 32% and 72% with sulfuric acid respectively, colour developing, inspects.Result shows, in relative humidity 32% and 72%, to have no significant effect chromatogram.
The comparison of different temperatures: the chromatography cylinder of existing developping agent is placed in refrigerator (5 DEG C), after placing a period of time, puts into chromatography cylinder by the thin layer plate after point sample, launches, and colour developing, inspects.Result shows, cryogenic conditions
Under thin layer clear spot, degree of separation is better, similar to the thin layer situation under normal temperature condition, shows that low temperature has no significant effect this thin-layer identification method.
Through above-mentioned test, show that the thin-layer identification method of FUFANG CHUANXINLIAN PIAN of the present invention is in thin layer plate, hand bed board and prefabricated board all can reach separating effect, and the change of temperature, humidity is little to expansion influential effect, therefore the method good tolerance.
Below be only the preferred embodiment of the present invention, it should be pointed out that above-mentioned preferred implementation should not be considered as limitation of the present invention, protection scope of the present invention should be as the criterion with claim limited range.For those skilled in the art, without departing from the spirit and scope of the present invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (9)
1. the thin-layer identification method of a FUFANG CHUANXINLIAN PIAN, it is characterized in that, comprise respectively FUFANG CHUANXINLIAN PIAN sample to be identified is carried out pre-treatment obtain need testing solution step, reference substance is carried out the step that pre-treatment obtains reference substance solution, also comprise the step of need testing solution being carried out to indentification by TLC analysis; Wherein, volume ratio is adopted to be that the ethyl acetate of 9:1 ~ 99:1 and methyl alcohol are as developping agent when launching in the step of indentification by TLC analysis, take out launching complete thin layer plate when inspecting, dry, spray develops the color with the sulfuric acid solution of vanillic aldehyde, wherein, the sulfuric acid that the sulfuric acid solution of described vanillic aldehyde is 60% ~ 70% by vanillic aldehyde and concentration is made, and in the sulfuric acid solution of described vanillic aldehyde, the mass percent of vanillic aldehyde is 1% ~ 2.5%.
2. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 1, it is characterized in that, FUFANG CHUANXINLIAN PIAN sample to be identified is carried out the step that pre-treatment obtains need testing solution to be specially: get sample powder to be identified and add the first organic solvent process, filter, obtain the first filtrate and the first residue; First filtrate is discarded, the first residue is added the second organic solvent process again, filter, obtain the second filtrate and the second residue; Second residue is discarded, by the second filtrate evaporate to dryness, obtains the 3rd residue, the 3rd residue is added organic solvent dissolution, as need testing solution.
3. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 2, it is characterized in that, described first organic solvent is sherwood oil or the compound close with sherwood oil polarity, and described second organic solvent is any one or two or more potpourris in methyl alcohol, ethanol, normal butyl alcohol, ethyl acetate.
4. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 2, it is characterized in that, the processing time is 10 ~ 45 minutes.
5. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 4, it is characterized in that, the processing time is 20 ~ 30 minutes.
6. the thin-layer identification method of the FUFANG CHUANXINLIAN PIAN as described in any one of claim 2 ~ 5, is characterized in that, the method for process is ultrasonic, backflow, decoct, in dipping any one or two or more.
7. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 1, it is characterized in that, described reference substance solution comprises: the yellow avens reference substance solution be made up through pre-treatment separately of yellow avens medicinal material, Dehydro and drographolide respectively and Dehydro and drographolide reference substance solution.
8. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 1, it is characterized in that, in the step that described indentification by TLC is analyzed, inspect and be specially: first the thin layer plate having sprayed developer is developed the color at normal temperatures, until the spot characterizing Herba Andrographitis feature occurs, and then heating blankets plate occurs to yellow avens unique point, carries out the colour developing of yellow avens feature.
9. the thin-layer identification method of FUFANG CHUANXINLIAN PIAN as claimed in claim 1, it is characterized in that, when launching in the step of indentification by TLC analysis, adopt volume ratio to be that the ethyl acetate of 95:5 or 9:1 or 20:1 or 35:1 or 55:1 or 99:1 and methyl alcohol are as developping agent.
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| CN201310578033.XA CN103604901B (en) | 2013-11-18 | 2013-11-18 | Thin-layer identification method of compound andrographis tablet |
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| CN201310578033.XA CN103604901B (en) | 2013-11-18 | 2013-11-18 | Thin-layer identification method of compound andrographis tablet |
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| CN1088834A (en) * | 1993-10-27 | 1994-07-06 | 黎红 | " Yiliwan " pill for treating poultry diseases |
| US8084495B2 (en) * | 2004-02-03 | 2011-12-27 | Herbal Powers Corporation | Composition of labdane diterpenes extracted from andrographis paniculata, useful for the treatment of autoimmune diseases, and alzheimer disease by activation for PPR-gamma receptors |
| CN1733105B (en) * | 2005-08-18 | 2011-04-20 | 贵阳云岩西创药物科技开发有限公司 | Detection method of Golden Preparation for treating gynecological disease |
| CN100369614C (en) * | 2005-11-10 | 2008-02-20 | 贵州益佰制药股份有限公司 | Quality control method of compound gallblader freeflow solid preparation |
| CN101002859A (en) * | 2006-01-16 | 2007-07-25 | 陈茜 | Compounding Herba Andrographitis dripping pills, and its preparing method |
| CN101690752B (en) * | 2008-06-06 | 2013-04-10 | 广西灵峰药业有限公司 | Quality control method of medicine composition |
| CN101700262A (en) * | 2009-10-26 | 2010-05-05 | 四川禾邦阳光制药股份有限公司 | Quality control method of andrographis paniculata dropping pills |
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