CN101703728A - Quality detection method for stomach warming and soothing capsules - Google Patents
Quality detection method for stomach warming and soothing capsules Download PDFInfo
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Abstract
The invention discloses a quality detection method for stomach warming and soothing capsules. The method comprises the steps of identification of the stomach warming and soothing capsules, measurement of aconitine limit and contents of psoralen and isopsoralen, and the like. Through the measurement of the invention, each capsule contains not less than 0.30 milligram of psoralea in terms of the total amount of the psoralen (C11H6O3) and the isopsoralen (C11H6O3). The method of the invention has advanced technology and simple and convenient operation, and can effectively control the quality of a product.
Description
Technical field
The present invention relates to the quality determining method of a kind of quality determining method of Chinese patent medicine, particularly a kind of WENWEISHU JIAONANG.
Background technology
WENWEISHU JIAONANG is the capsules through being processed into such as Radix Codonopsis, Rhizoma Dioscoreae, Fructus Mume, Radix Aconiti Lateralis Preparata (system), Herba Cistanches (system), Pericarpium Citri Reticulatae, the Radix Astragali (processing), the Rhizoma Atractylodis Macrocephalae (stir-fry), Fructus Psoraleae, Cortex Cinnamomi, Fructus Crataegi (stir-fry), Fructus Amomi, kidney and spleen invigorating, the warming middle-JIAO nourishing the stomach, promoting the circulation of QI to relieve pain.Be used for the stomachache with cool feeling that deficiency of spleen-YANG and kidneyYANG causes, flatulence, belch, poor appetite, fear of cold unablely waits disease, and atrophic gastritis, chronic gastritis performance have above-mentioned patient.In existing WENWEISHU JIAONANG quality determining method, have only thin layer chromatography scanning that psoralen is carried out assay, have on the method that precision is not high, deviation appears in repeatability greatly, easily, be difficult to reflect really the shortcoming of product quality, and the limit handling method that does not have toxicity medical material Radix Aconiti Lateralis Preparata, therefore, can't guarantee the drug safety of product.
Summary of the invention
The objective of the invention is to overcome the deficiency of existing quality standard, a kind of drug safety that can effectively guarantee product is provided, the quality determining method of WENWEISHU JIAONANG controlled, stable and easy and simple to handle.
Its technical scheme is: a kind of quality determining method of WENWEISHU JIAONANG is characterized in that this method comprises
A, astragaloside are differentiated: get this product content, add the methanol supersound extraction, the extracting solution evaporate to dryness, residue adds water makes dissolving, extracts with water saturated n-butyl alcohol jolting, washs with ammonia solution, get n-butyl alcohol liquid evaporate to dryness, residue is dissolved in water, by D101 type macroporous adsorptive resins, and difference water, different concentration ethanol eluant solution, collect high concentration ethanol eluant solution liquid, evaporate to dryness, residue adds dissolve with methanol, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes solution, in contrast product solution.Test according to thin layer chromatography, draw above-mentioned need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts respectively to reach under the ultra-violet lamp (365nm) daylight under and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle of apparent same color under the daylight; Ultra-violet lamp (365nm) shows identical fluorescence speckle down.
B, Hesperidin are differentiated: get this product content, with the ethyl acetate heating and refluxing extraction, extracting solution is by polyamide column, and the water eluting is collected eluent, evaporate to dryness, and residue adds dissolve with methanol, leaves standstill, and gets supernatant as need testing solution.Other gets the Hesperidin reference substance, adds methanol and makes saturated solution, in contrast product solution.According to the thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-methanol-water, launch, take out, to dry, spray dries up with the aluminum chloride test solution, puts under the ultra-violet lamp (365nm) and inspects.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
C, Radix Codonopsis is differentiated: get this product content, add water and make dissolving, add ethanol again, leave standstill, filter the filtrate evaporate to dryness, residue adds dilute hydrochloric acid, the ether reflux is put coldly, divides and to get ether layer, the acid solution ether extraction merges ether solution, volatilizes, residue adds dissolve with ethanol, as need testing solution. and other gets the Radix Codonopsis control medicinal material, adds the water reflux, filter, filtrate is shone medical material solution in pairs with legal system. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, inspects under daylight. in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
D, psoralen and isopsoralen are differentiated: get this product content, the reflux that adds diethyl ether is put coldly, filters, and filtrate volatilizes, and residue adds dissolve with ethanol, as need testing solution.Other gets psoralen reference substance, isopsoralen reference substance, adds ethanol and makes mixed solution, in contrast product solution.According to the thin layer chromatography test, draw above-mentioned need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with normal hexane one ethyl acetate, launch, take out, dry, spray is dried with 10% sodium hydrate methanol solution, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
E, aconitine limit test: it is a certain amount of to get this product content, and porphyrize is put in the tool plug conical flask, and the jolting that adds diethyl ether is extracted, and adds the ammonia solution jolting again and extracts washing, places, and divide again and get the ether layer, evaporate to dryness, residue adds anhydrous alcohol solution, as need testing solution.Other gets the aconitine reference substance, adds dehydrated alcohol and makes certain density solution, in contrast product solution.Test according to thin layer chromatography, draw a certain amount of need testing solution and reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane-ethyl acetate-dehydrated alcohol is developing solvent, launch, take out, dry, spray is inspected under daylight with rare bismuth potassium iodide test solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on the speckle that occurs should be less than the speckle of reference substance, or speckle does not appear.
F, psoralen and isopsoralen assay: get a certain amount of this product content, with 1% hydrochloric acid methanol Soxhlet reflux, extract,, the extracting solution standardize solution filters, and gets subsequent filtrate as need testing solution; Other gets the psoralen reference substance, each is an amount of for the isopsoralen reference substance, with dissolve with methanol, makes mixed solution, in contrast product solution; According to high effective liquid chromatography for measuring, be filler with the octadecylsilane chemically bonded silica, methanol-0.4% phosphoric acid solution is a mobile phase, the detection wavelength is 246nm.Theoretical cam curve is calculated by the psoralen peak should be not less than 3000, and accurate respectively reference substance solution and the need testing solution drawn injects chromatograph of liquid, measures, and the result should be: every of WENWEISHU JIAONANG contains Fructus Psoraleae with psoralen (C
11H
6O
3) and isopsoralen (C
11H
6O
3) the total amount meter, must not be less than 0.30mg.
The quality determining method of WENWEISHU JIAONANG of the present invention, at first, content assaying method to Fructus Psoraleae medical material in the WENWEISHU JIAONANG improves, adopt that this method has that separating degree is good, precision is high, good reproducibility, easy and simple to handle, index components characteristics more fully, can carry out reliable quantified controlling to the quality of WENWEISHU JIAONANG; The second, increased the limit test of the aconitine of poisonous medical material Radix Aconiti Lateralis Preparata in the WENWEISHU JIAONANG,, can guarantee drug safety clinically by its limit is made control; The 3rd, increased qualitative identification method to astragaloside, Hesperidin, Radix Codonopsis and Fructus Psoraleae, the foundation of these qualitative identification methods has strengthened the control dynamics of WENWEISHU JIAONANG quality.Therefore, the quality determining method of WENWEISHU JIAONANG of the present invention has further improved the quality standard of WENWEISHU JIAONANG, has fully guaranteed the quality and the definite curative effect of product, and the foundation of science also is provided for the low-quality goods on the discriminating market.
The specific embodiment
WENWEISHU JIAONANG prescription composition: Radix Codonopsis, Rhizoma Dioscoreae, Fructus Mume, Radix Aconiti Lateralis Preparata (Radix Aconiti Lateralis Preparata), Herba Cistanches (wine steaming), Pericarpium Citri Reticulatae, Radix Astragali Preparata, the Rhizoma Atractylodis Macrocephalae (frying), Fructus Psoraleae, Cortex Cinnamomi, Fructus Crataegi cuneatae (parched), Fructus Amomi.
Preparation technology: get the recipe quantity medical material, wherein amomum powder is broken into fine powder; All the other Radix Codonopsis etc. ten decoct with water secondary simply, and 1.5 hours for the first time, 1 hour for the second time, collecting decoction filters, and filtrate is left standstill, get the thick paste that supernatant concentration to relative density is 1.35-1.38 (20 ℃), add Fructus Amomi fine powder and an amount of Pulvis Talci, starch and silicon dioxide, mixing, system granule, dry, incapsulate, make 1000, promptly.
WENWEISHU JIAONANG is differentiated:
1, astragaloside is differentiated: get this product content 5g, porphyrize adds methanol 30ml, supersound process 30 minutes, filter, the filtrate evaporate to dryness. residue adds water 30ml slight fever makes dissolving, extracts 2 times with water saturated n-butyl alcohol jolting, each 30ml, merge n-butanol extracting liquid, with ammonia solution washing 2 times, each 30ml discards ammonia solution, n-butyl alcohol liquid evaporate to dryness, residue adds water 5ml makes dissolving, by D101 type macroporous adsorptive resins (internal diameter 1cm, the post height is 15cm), water 50ml eluting, discard water lotion, reuse 40% ethanol 30ml eluting discards 40% ethanol liquid; Continue with 70% ethanol 80ml eluting, collect eluent, evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution. other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution. according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw above-mentioned need testing solution 10 μ l, reference substance solution 5 μ l put respectively on same silica gel g thin-layer plate, are developing solvent with lower floor's solution of chloroform-methanol-water (13: 7: 2), launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to speckle colour developing at 105 ℃, puts respectively to reach under the ultra-violet lamp (365nm) daylight under and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color under the sight; Ultra-violet lamp (365nm) shows identical orange-yellow fluorescence speckle down.
2, Hesperidin is differentiated: get this product content 3g, porphyrize adds ethyl acetate 15ml, reflux 1 hour filters polyamide column (14～30 orders of filtrate by having handled well, 4g, internal diameter 1.0cm) on, water 80ml eluting is collected eluent, evaporate to dryness, residue adds methanol 1ml makes dissolving, leaves standstill, and gets supernatant as need testing solution.Other gets the Hesperidin reference substance, adds methanol and makes saturated solution, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-methanol-water (20: 3: 2) is developing solvent, launches, and takes out, dry, spray dries up with the aluminum chloride test solution, puts under the ultra-violet lamp (365nm) and inspects.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
3, Radix Codonopsis is differentiated: get this product content 4g, porphyrize adds water 10ml and makes dissolving, slowly add ethanol 80ml again, left standstill 30 minutes, filter, filtrate evaporate to dryness, residue add dilute hydrochloric acid 20ml, ether 30ml, reflux 1 hour, put coldly, divide and to get ether layer, acid solution is extracted 1 time with ether 20ml jolting, merge ether solution, volatilize, residue adds ethanol 1ml makes dissolving, as need testing solution.Other gets Radix Codonopsis control medicinal material 2g, adds water 30ml, reflux 30 minutes, filters, and filtrate is steamed to about 10ml, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (10: 7: 0.5) is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, inspects under daylight.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
4, psoralen and isopsoralen are differentiated: get this product content 2g, and porphyrize, the 30ml that adds diethyl ether, reflux 30 minutes is put coldly, filters, and filtrate volatilizes, and residue adds ethanol 1ml makes dissolving, as need testing solution.Other gets psoralen reference substance, isopsoralen reference substance, adds ethanol and makes the mixed solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw above-mentioned need testing solution 5 μ l, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with normal hexane one ethyl acetate (4: 1) is developing solvent, launches, and takes out, dry, spray is dried with 10% sodium hydrate methanol solution, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
The aconitine limit test:
It is tolerant to get this product 24 intragranulars, and porphyrize is put in the tool plug conical flask, the 150ml that adds diethyl ether, and jolting 10 minutes adds ammonia solution 8ml, and jolting was extracted 30 minutes, placed 2 hours, divided and got the ether layer, and evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution.Other gets the aconitine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw above-mentioned need testing solution 15 μ l, reference substance solution 5 μ l, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane-ethyl acetate-dehydrated alcohol (7: 3: 1) is developing solvent, launch, take out, dry, spray is with rare bismuth potassium iodide test solution, under daylight, inspect. in the test sample chromatograph, with the corresponding position of reference substance chromatograph on the speckle that occurs should be less than the speckle of reference substance, or speckle does not appear.
The WENWEISHU JIAONANG assay:
Get this product content, porphyrize, powder are crossed sieve No. 4, mixing is got 0.6g, and accurate the title decides, put in the apparatus,Soxhlet's, it is an amount of to add 1% hydrochloric acid methanol, is extracted into colourless, extracting solution is concentrated into about 10ml, is transferred in the 25ml measuring bottle, adds 1% hydrochloric acid methanol again to scale, shake up, filter, get subsequent filtrate, as need testing solution; Other gets the psoralen reference substance, each is an amount of for the isopsoralen reference substance, accurate claims surely, adds methanol and makes every 1ml and contain 10 μ g mixed solution, product solution in contrast; According to high effective liquid chromatography for measuring, be filler with the octadecylsilane chemically bonded silica, methanol-0.4% phosphoric acid solution (45: 55) is a mobile phase, the detection wavelength is 246nm.Theoretical cam curve is calculated by the psoralen peak should be not less than 3000, and accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, and every of WENWEISHU JIAONANG contains Fructus Psoraleae to mend ossein (C
11H
6O
3) and isopsoralen (C
11H
6O
3) the total amount meter, must not be less than 0.30mg.
Claims (8)
1. the quality determining method of a WENWEISHU JIAONANG is characterized in that this method comprises:
A, astragaloside are differentiated: get this product content, add the methanol supersound extraction, the extracting solution evaporate to dryness, residue adds water makes dissolving, extracts with water saturated n-butyl alcohol jolting, washs with ammonia solution, get n-butyl alcohol liquid evaporate to dryness, residue is dissolved in water, by D101 type macroporous adsorptive resins, and difference water, different concentration ethanol eluant solution, collect high concentration ethanol eluant solution liquid, evaporate to dryness, residue adds dissolve with methanol, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes solution, in contrast product solution.Test according to thin layer chromatography, draw above-mentioned need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts respectively to reach under the ultra-violet lamp (365nm) daylight under and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, the speckle of apparent same color under the daylight; Ultra-violet lamp (365nm) shows identical fluorescence speckle down.
B, Hesperidin are differentiated: get this product content, with the ethyl acetate heating and refluxing extraction, extracting solution is by polyamide column, and the water eluting is collected eluent, evaporate to dryness, and residue adds dissolve with methanol, leaves standstill, and gets supernatant as need testing solution.Other gets the Hesperidin reference substance, adds methanol and makes saturated solution, in contrast product solution.According to the thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-methanol-water, launch, take out, to dry, spray dries up with the aluminum chloride test solution, puts under the ultra-violet lamp (365nm) and inspects.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
C, Radix Codonopsis are differentiated: get this product content, add water and make dissolving, add ethanol again, leave standstill, filter, filtrate evaporate to dryness, residue add dilute hydrochloric acid, ether reflux, put coldly, divide and to get ether layer, acid solution ether extraction, merge ether solution, volatilize, residue adds dissolve with ethanol, as need testing solution.Other gets the Radix Codonopsis control medicinal material, adds the water reflux, filters, and filtrate is shone medical material solution in pairs with legal system.According to the thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with chloroform-ethyl acetate-formic acid, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, inspects under daylight.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
D, psoralen and isopsoralen are differentiated: get this product content, the reflux that adds diethyl ether is put coldly, filters, and filtrate volatilizes, and residue adds dissolve with ethanol, as need testing solution.Other gets psoralen reference substance, isopsoralen reference substance, adds ethanol and makes mixed solution, in contrast product solution.According to the thin layer chromatography test, draw above-mentioned need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with normal hexane one ethyl acetate, launch, take out, dry, spray is dried with 10% sodium hydrate methanol solution, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
E, aconitine limit test: it is a certain amount of to get this product content, and porphyrize is put in the tool plug conical flask, and the jolting that adds diethyl ether is extracted, and adds the ammonia solution jolting again and extracts washing, places, and divide again and get the ether layer, evaporate to dryness, residue adds anhydrous alcohol solution, as need testing solution.Other gets the aconitine reference substance, adds dehydrated alcohol and makes certain density solution, in contrast product solution.Test according to thin layer chromatography, draw a certain amount of need testing solution and reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane-ethyl acetate-dehydrated alcohol is developing solvent, launch, take out, dry, spray is inspected under daylight with rare bismuth potassium iodide test solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on the speckle that occurs should be less than the speckle of reference substance, or speckle does not appear.
F, psoralen and isopsoralen assay: get a certain amount of this product content, with 1% hydrochloric acid methanol Soxhlet reflux, extract,, the extracting solution standardize solution filters, and gets subsequent filtrate as need testing solution; Other gets the psoralen reference substance, each is an amount of for the isopsoralen reference substance, with dissolve with methanol, makes mixed solution, in contrast product solution; According to high effective liquid chromatography for measuring, with the octadecylsilane chemically bonded silica is filler, methanol-0.4% phosphoric acid solution is a mobile phase, detecting wavelength is that the 246nm. theoretical cam curve should be not less than 3000 by the calculating of psoralen peak, accurate respectively reference substance solution and the need testing solution drawn, inject chromatograph of liquid, measure, the result should be: every of WENWEISHU JIAONANG contains Fructus Psoraleae with psoralen (C
11H
6O
3) and isopsoralen (C
11H
6O
3) the total amount meter, must not be less than 0.30mg.
2. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: described low-concentration ethanol solution concentration scope is 30-40%, and high concentration ethanol solution concentration scope is 70-80%.The ratio of its described " chloroform-methanol-water " developing solvent is 13: 7: 2.
3. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: the model of polyamide column is: 14～30 orders, 4g, internal diameter 1.0cm; The ratio of its described " ethyl acetate-methanol-water " developing solvent is 20: 3: 2.
4. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: the ratio of its described " chloroform-ethyl acetate-formic acid " developing solvent is 10: 7: 0.5.
5. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: the ratio of its described " normal hexane-ethyl acetate " developing solvent is 4: 1.
6. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: the ratio of its described " normal hexane-ethyl acetate-dehydrated alcohol " developing solvent is 7: 3: 1.
7. the quality determining method of WENWEISHU JIAONANG according to claim 1 is characterized in that: sample extraction with 1% hydrochloric acid methanol put be extracted in the apparatus,Soxhlet's colourless.
8. the quality determining method of WENWEISHU JIAONANG according to claim 1, it is characterized in that: described, the volume proportioning of methanol-0.4% phosphoric acid solution is 45: 55.
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| CN109959751A (en) * | 2019-04-09 | 2019-07-02 | 浙江大学 | Water-soluble natural product thin layer quantitative image recognition detection method |
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| CN108760928A (en) * | 2018-06-11 | 2018-11-06 | 合肥华润神鹿药业有限公司 | A kind of construction method of temperature stomach relaxation grain finger-print |
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| CN109959751A (en) * | 2019-04-09 | 2019-07-02 | 浙江大学 | Water-soluble natural product thin layer quantitative image recognition detection method |
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| CP03 | Change of name, title or address |
Address after: Peach Industrial Park, economic and Technological Development Zone 230051 Anhui city in Hefei Province, Hefei SHANGPAI Highway No. 12 Patentee after: Hefei shenlu Huarun Pharmaceutical Co. Ltd. Address before: 230051 Anhui city in Hefei Province, Hefei SHANGPAI Highway No. 12 Patentee before: Hefei Shenlu Shuanghe Pharmaceutical Industry Co., Ltd. |