CN100366275C - Method for controlling quality of Yunnan red tablet - Google Patents
Method for controlling quality of Yunnan red tablet Download PDFInfo
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Abstract
The present invention relates to a quality control method of a compound traditional Chinese medicine-Yunnan red drug tablet, which comprises the following steps: 1, effective constituents in samples are qualitatively identified and extracted by an ultrasonic treatment method, the index constituents of paris root saponin A, B in paris root are identified by using paris root saponin A, B as comparison articles, the index constituents of ginsenoside Rb1, Rg1 and notoginseng saponin R1 in notoginseng are identified by using ginsenoside Rb1, Rg1 and notoginseng R1 as comparison articles, and the medicinal material of dactylicapnos scandens in the prescription of Yunnan red drug is identified by using dactylicapnos scandens as comparison medicinal materials; 2, the limiting quantity of relevant alkaloids in aconitum in prepared aconitum vilnaorinianum kom is controlled by using aconitine for limiting toxic constituents as comparison articles; and 3, the content of the index constituent of ginsenoside Rg1 is quantitatively identified by a high performance liquid chromatography method.
Description
Technical field
The present invention relates to a kind of method of quality control of herbal mixture.
Background technology
Chinese patent CN03117723.9 discloses a kind of " Yunnan red tablet and its production and application ", described Yunnan red tablet is made by following materials based on weight: YUNNAN HONGYAO 210～283, microcrystalline Cellulose 5～8, cane sugar powder 18～30, dextrin 12～15, magnesium stearate 3, Pulvis Talci 3, carboxylic propyl methocel 60% ethanol liquid is an amount of; The parts by weight that wherein said YUNNAN HONGYAO raw material is formed are: Radix Notoginseng 20～30, Rhizoma Paridis 55～65, Radix dactylicapni (Radix Dactylicapnotis) 10～15, Radix seu Cortex Heteropanacis 15～20, Radix Psammosilenes 10～15, Pseudostreblus indica Bur. 20～28, Delavay ampelopsis root 20～30, manufacture-yellow Radix Aconiti Kusnezoffii 35～40, Rhizoma Acori Graminei 5～10, Radix Scutellariae 20～30.The pharmacological action of Yunnan red tablet safety non-toxic is strong, and indication is wide, can or prevent extensive use in stomach, duodenal ulcer and upper gastrointestinal hemorrhage, dysfunctional uterine hemorrhage, analgesia, the antiheumatic medicine in the preparation treatment.But document does not disclose the method for quality control of above-mentioned mercurochrome tablet.
Summary of the invention
The method of quality control that the purpose of this invention is to provide a kind of Yunnan red tablet.
Yunnan red tablet of the present invention is made by the crude drug of following parts by weight: Radix Notoginseng 20～30, Rhizoma Paridis 55～65, Radix dactylicapni (Radix Dactylicapnotis) 10～15, Radix seu Cortex Heteropanacis 15～20, Radix Psammosilenes 10～15, Pseudostreblus indica Bur. 20～28, Delavay ampelopsis root 20～30, manufacture-yellow Radix Aconiti Kusnezoffii 35～40, Rhizoma Acori Graminei 5～10, Radix Scutellariae 20～30.
Its method for making is: above ten flavors, be ground into fine powder, and sieve, mixing adds right amount of auxiliary materials, makes granule, tabletting, promptly get (or tabletting, the bag film-coat, promptly).
The method of quality control of Yunnan red tablet of the present invention, form by following steps:
One, qualitative identification extracts effective ingredient in the test sample with the method for supersound process, with Rhizoma Paridis saponin A, B in contrast product differentiate index components Rhizoma Paridis saponin A, B in the Rhizoma Paridis, with ginsenoside Rb1, Rg1, arasaponin R1 in contrast product differentiate index components ginsenoside Rb1, Rg1, arasaponin R1 in the Radix Notoginseng; With Radix dactylicapni (Radix Dactylicapnotis) is control medicinal material, differentiates the medical material Radix dactylicapni (Radix Dactylicapnotis) in the YUNNAN HONGYAO prescription;
Two, toxic component is limited the quantity of with aconitine product in contrast, in order to limiting the quantity of of relevant alkaloid (ester alkaloid) in the Aconitum in the control manufacture-yellow Radix Aconiti Kusnezoffii;
Three, quantitative identification is measured its index components ginsenoside Rg1's content with high performance liquid chromatography.
In Yunnan red tablet of the present invention, Radix Notoginseng and Rhizoma Paridis are the monarch drug in the prescription, this method of quality control is to have adopted the method for supersound process to extract effective ingredient in the test sample, compares with existing hot reflux extracting method, and is more easy, advanced and practical.And index components Rhizoma Paridis saponin A, B in index components ginsenoside Rb1, Rg1, arasaponin R1 and the Rhizoma Paridis in the emphasis discriminating Radix Notoginseng.Owing to contain the manufacture-yellow Radix Aconiti Kusnezoffii in the Yunnan red tablet of the present invention, relevant alkaloid (ester alkaloid) is a therapeutic component in the Aconitum in the manufacture-yellow Radix Aconiti Kusnezoffii, it is again toxic component, the Dendrobium denneanum Kerr. crow makes this Alkaloid di esters hydrolysis can generate the little monoesters class alkali of toxicity after concocting, as continue to concoct, then become the littler amine alcohols alkali of toxicity, and curative effect is constant.With aconitine (severe toxicity is arranged, and oral 0.2mg can make the people poison, and 3-5mg can cause death) is reference substance, to limiting the quantity of of the xicity related composition of a class in the manufacture-yellow Radix Aconiti Kusnezoffii, is in order to make clinical drug safety effective.
Method of quality control of the present invention is to control drug quality simultaneously from qualitative and quantitative two aspects, also increased limiting the quantity of and ginsenoside Rg1's assay of aconitine simultaneously, make its method of quality control more comprehensively, science more, more can guarantee the safe and effective of medication.
The pharmacodynamics test of Yunnan red tablet of the present invention is as follows:
One. anastalsis
Yunnan red tablet is the Blood clotting time that dose dependent significantly shortens mice and tame rabbit whole blood, and shortens the mouse tail point bleeding time, and the prompting Yunnan red tablet has tangible anastalsis.Yunnan red tablet significantly improves isolated rat uterus tension force, shows to strengthen the uterus contraction, helps preventing and treating dysfunctional uterine hemorrhage.
Two. analgesic activity
In multiple pain reaction models such as electricity irritation Mus tail, mice hot plate method and acetic acid twisting method, Yunnan red tablet all shows significant analgesia role, and has dose-effect relationship.
Three, detumescence effect
The Yunnan red tablet of high, middle dosage (be equivalent to clinical consumption 20 and 10 times) obviously suppresses carrageenin and causes the rat paw edema effect, reduces the effect of mice caused by dimethylbenzene xylene auricle edema simultaneously, and certain amount one effect relationship of tool; Yunnan red tablet also is dosage correlation and significantly alleviates granulomatous dry weight of rat air bag and weight in wet base, points out Yunnan red tablet to have tangible resist inflammation on repercussive function thus.
Hemorrhagic stomach, duodenal ulcer and dysfunctional uterine hemorrhage Chang Bingfa stomachache and inflammation in various degree, Yunnan red tablet has stronger analgesia and antiinflammatory action, and this is very favourable to alleviating stomachache and preventing and treating mucosal inflammation.
Function of the present invention is hemostatic analgesia, promoting blood circulation to remove blood stasis, expelling wind and removing dampness with curing mainly.Be used for gastric hemorrhage, hemoptysis caused by bronchiectasis, dysfunctional uterine hemorrhage, menorrhagia, retinal hemorrhage, conjunctival hemorrhage, epistaxis, bleeding hemorrhoids, soft tissue contusion, rheumatic arthritis, rheumatic lumbago and scelalgia etc.Oral, one time 1～5,2-3 time on the one.Every of specification contains crude drug 0.1-0.5g.
The specific embodiment
Embodiment:
This product powder is got in [discriminating] (1), puts microscopically and observes: needle-like calcium oxalate crystal bunchy or be dispersed in long 40～80～240 μ m.Fiber is faint yellow, prismatic, and wall thickness, the hole ditch is thin.Fiber bunchy or loose from, wall thickness, there is longitudinal crack on the surface, and primary wall often separates with secondary wall, and the two ends normal off is cleaved into must shape, or truncate.Stone cell oval or class are square, wall thickness, and rill is obvious.
(2) get this product 5-12 sheet, porphyrize adds ethanol 5-20ml, and supersound process 5-20 minute, filter, get filtrate 1-5ml, the sodium chloride gelatin test solution 1-4 that adds new system drips, and produces white casse; Other gets filtrate 1-10ml, adds 1% ferric chloride alcoholic solution 1-6 and drips, and shows blue.
(3) get this product 5-80 sheet, porphyrize, 20-120ml adds diethyl ether, supersound process 1-20 minute, filter, discard ether solution, medicinal residues volatilize solvent, add n-butyl alcohol 10-80ml, and placement is spent the night, supersound process 5-50 minute, filter, filtrate is put in the separatory funnel, wash 1-6 time with ammonia solution, each 5-50ml divides and gets n-butyl alcohol liquid evaporate to dryness, residue adds methanol 0.5-10ml makes dissolving, as need testing solution.Other gets Rhizoma Paridis saponin A, B reference substance, adds methanol and makes the mixed solution that every 1ml contains 1-6mg, in contrast product solution.According to thin layer chromatography (" Chinese pharmacopoeia appendix) test, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid (1 → 10), and it is clear to dry by the fire to the speckle colour developing at 80-120 ℃, puts respectively under daylight and the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle or the fluorescence speckle of same color.
(4) get the need testing solution of [discriminating] (3) item.Other gets ginsenoside Rb1, Rg1 and arasaponin R1 reference substance, adds methanol and makes the mixed solution that every 1ml contains 1-5mg, in contrast product solution.According to thin layer chromatography (" Chinese pharmacopoeia appendix) test, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water) be developing solvent, launch, take out, dry, spray is with ethanol solution of sulfuric acid (1 → 10), and it is clear to dry by the fire to the speckle colour developing at 80-120 ℃, puts respectively under daylight and the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle or the fluorescence speckle of same color.
(5) get this product 5-60 sheet, porphyrize adds ammonia solution 1-10ml, chloroform 20-100ml, and jolting 0.5-2 hour, filter, filtrate evaporate to dryness, residue add methanol 0.5-5ml makes dissolving, as need testing solution.Other gets Radix dactylicapni (Radix Dactylicapnotis) control medicinal material 0.1-2g, is equipped with control medicinal material solution with legal system.According to thin layer chromatography (" Chinese pharmacopoeia appendix) test, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, lower floor's solution with placement below chloroform-ethyl acetate-methanol-water 5-20 ℃ is developing solvent, launch, take out, dry, spray is with rare bismuth potassium iodide test solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical orange red principal spot.
[inspection] aconitine limit is got this product 50-200 sheet, and porphyrize is put in the tool plug conical flask, and 80-300ml adds diethyl ether, jolting 5-30 minute, add ammonia solution 5-30ml, jolting 10-60 minute, placed 0.5-3 hour, divide and get the ether layer, evaporate to dryness adds dehydrated alcohol 1-10ml and makes dissolving, as need testing solution.Other gets the aconitine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1-5mg, in contrast product solution.According to thin layer chromatography (" appendix of Chinese pharmacopoeia) test, draw need testing solution 5-20 μ l, reference substance solution 1-15 μ l, put respectively on the alumina G lamellae of same 0.1-0.5% sodium hydroxide preparation, with normal hexane-ethyl acetate is developing solvent, launches, and takes out, dry, spray is with rare bismuth potassium iodide solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on the speckle of speckle or appearance or not and should be less than the speckle of reference substance.
Other (1) microbial limits detect
Antibacterial: get 1: 10 test liquid 2ml, every 1ml injects in 5 flat boards, each flat board adds 0.2ml, pour into and melt and be cooled to 45 ℃ TTC nutrient agar 15ml, abundant mixing, coagulate the back and place 30-35 ℃ of incubator cultivation 48 hours, the counting clump count multiply by extension rate report (10 dull and stereotyped sum/2 * extension rates)
Mycete, yeast: get 1: 10 test liquid 2ml, every 1ml injects in 10 flat boards, each flat board adds 0.1ml, pour into and melt and be cooled to 45 ℃ the about 15ml of Rose Bengal Sodium agar culture medium, abundant mixing, coagulate the back and place 25-28 ℃ of incubator cultivation 72 hours, the counting clump count multiply by extension rate report (20 dull and stereotyped sum/2 * extension rates)
Control bacterium: by " an appendix XIII of Chinese pharmacopoeia version in 2000 microbial limit test detects.
(2) except that the microbial limit item, should meet every regulation relevant under the tablet item (" appendix of Chinese pharmacopoeia).
[assay] measures ginsenoside Rg1's content according to high performance liquid chromatography (" Chinese pharmacopoeia appendix).
Chromatographic condition and systemic suitability experiment are filler with the octyl bonded silica gel; Acetonitrile-water is a mobile phase, and flow velocity 0.5-2.0ml/min, detects wavelength 203nm by column temperature 30-45 ℃.Number of theoretical plate calculates by the ginsenoside Rg1 peak should be not less than 4000.
It is an amount of that the preparation precision of reference substance solution takes by weighing ginsenoside Rg1's reference substance, makes the solution that every 1ml contains the 0.1-0.5mg ginsenoside Rg1 with methanol, promptly.
This product 10-40 sheet is got in the preparation of need testing solution, and accurate the title decided porphyrize, get fine powder 0.5-5g, the accurate title, decide, and 20-80ml adds diethyl ether, flooded 0.5-3 hour, and supersound process 1-30 minute, filtered, discard ether solution, medicinal residues volatilize ether, the accurate n-butyl alcohol 20-80ml that adds, weigh, placement is spent the night, supersound process 5-30 minute,, put coldly, supply weight with n-butyl alcohol, mixing filters, and precision is measured subsequent filtrate 10-50ml, put in the separatory funnel, wash 1-5 time with ammonia solution, each 10-50ml places layering, branch is got n-butyl alcohol liquid and is put evaporate to dryness in the water-bath, and residue also quantitatively is transferred in the 2-10ml measuring bottle with dissolve with methanol.Add methanol to scale, shake up, filter, get subsequent filtrate as need testing solution.
Algoscopy is got reference substance solution and need testing solution 1-40 μ l respectively, injects chromatograph of liquid, calculates with external standard method, promptly.
This product contains Radix Notoginseng with ginsenoside Rg1 (C
24H
72O
14) calculate, every must not be less than 0.1-1mg.Every must not be less than 0.2mg in the present embodiment.
Claims (1)
1. the method for quality control of a Yunnan red tablet is characterized in that being made up of following steps:
One, qualitative identification
(1) gets this product powder, put microscopically and observe: needle-like calcium oxalate crystal bunchy or be dispersed in long 40～80～240 μ m; Fiber is faint yellow, prismatic, and wall thickness, the hole ditch is thin; Fiber bunchy or loose from, wall thickness, there is longitudinal crack on the surface, and primary wall often separates with secondary wall, and the two ends normal off is cleaved into must shape, or truncate; Stone cell oval or class are square, wall thickness, and rill is obvious;
(2) get this product 5-12 sheet, porphyrize adds ethanol 5-20ml, and supersound process 5-20 minute, filter, get filtrate 1-5ml, the sodium chloride gelatin test solution 1-4 that adds new system drips, and produces white casse; Other gets filtrate 1-10ml, adds 1% ferric chloride alcoholic solution 1-6 and drips, and shows blue;
(3) get this product 5-80 sheet, porphyrize, 20-120ml adds diethyl ether, supersound process 1-20 minute, filter, discard ether solution, medicinal residues volatilize solvent, add n-butyl alcohol 10-80ml, and placement is spent the night, supersound process 5-50 minute, filter, filtrate is put in the separatory funnel, wash 1-6 time with ammonia solution, each 5-50ml divides and gets n-butyl alcohol liquid evaporate to dryness, residue adds methanol 0.5-10ml makes dissolving, as need testing solution; Other gets Rhizoma Paridis saponin A, B reference substance, adds methanol and makes the mixed solution that every 1ml contains 1-6mg, in contrast product solution; Test according to thin layer chromatography, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid 1 → 10, and it is clear to dry by the fire to the speckle colour developing at 80-120 ℃, puts respectively under daylight and the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle or the fluorescence speckle of same color;
(4) get the need testing solution of (3) item; Other gets ginsenoside Rb1, Rg1 and arasaponin R1 reference substance, adds methanol and makes the mixed solution that every 1ml contains 1-5mg, in contrast product solution; Test according to thin layer chromatography, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid 1 → 10, and it is clear to dry by the fire to the speckle colour developing at 80-120 ℃, puts respectively under daylight and the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle or the fluorescence speckle of same color;
(5) get this product 5-60 sheet, porphyrize adds ammonia solution 1-10ml, chloroform 20-100ml, and jolting 0.5-2 hour, filter, filtrate evaporate to dryness, residue add methanol 0.5-5ml makes dissolving, as need testing solution; Other gets Radix dactylicapni (Radix Dactylicapnotis) control medicinal material 0.1-2g, is equipped with control medicinal material solution with legal system; Test according to thin layer chromatography, draw each 1～10 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, lower floor's solution with placement below chloroform-ethyl acetate-methanol-water 5-20 ℃ is developing solvent, launch, take out, dry, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical orange red principal spot;
Two, toxic component is limited the quantity of
Aconitine limit is got this product 50-200 sheet, and porphyrize is put in the tool plug conical flask, and 80-300ml adds diethyl ether, jolting 5-30 minute, add ammonia solution 5-30ml, jolting 10-60 minute, placed 0.5-3 hour, divide and get the ether layer, evaporate to dryness adds dehydrated alcohol 1-10ml and makes dissolving, as need testing solution; Other gets the aconitine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1-5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 5-20 μ l, reference substance solution 1-15 μ l, putting respectively on the alumina G lamellae of same 0.1-0.5% sodium hydroxide preparation, is developing solvent with normal hexane-ethyl acetate, launches, take out, dry, spray is with rare bismuth potassium iodide solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on the speckle of speckle or appearance or not and should be less than the speckle of reference substance;
Three, quantitative identification is according to high effective liquid chromatography for measuring ginsenoside Rg1's content;
Chromatographic condition and systemic suitability experiment are filler with the octyl bonded silica gel; Acetonitrile-water is a mobile phase, and flow velocity 0.5-2.0ml/min, detects wavelength 203nm by column temperature 30-45 ℃; Number of theoretical plate calculates by the ginsenoside Rg1 peak should be not less than 4000;
It is an amount of that the preparation precision of reference substance solution takes by weighing ginsenoside Rg1's reference substance, makes the solution that every 1ml contains the 0.1-0.5mg ginsenoside Rg1 with methanol, promptly;
This product 10-40 sheet is got in the preparation of need testing solution, and accurate the title decided porphyrize, get fine powder 0.5-5g, the accurate title, decide, and 20-80ml adds diethyl ether, flooded 0.5-3 hour, and supersound process 1-30 minute, filtered, discard ether solution, medicinal residues volatilize ether, the accurate n-butyl alcohol 20-80ml that adds, weigh, placement is spent the night, supersound process 5-30 minute,, put coldly, supply weight with n-butyl alcohol, mixing filters, and precision is measured subsequent filtrate 10-50ml, put in the separatory funnel, wash 1-5 time with ammonia solution, each 10-50ml places layering, branch is got n-butyl alcohol liquid and is put evaporate to dryness in the water-bath, and residue also quantitatively is transferred in the 2-10ml measuring bottle with dissolve with methanol; Add methanol to scale, shake up, filter, get subsequent filtrate as need testing solution;
Algoscopy is got reference substance solution and need testing solution 1-40 μ l respectively, injects chromatograph of liquid, calculates with external standard method, promptly;
This product contains Radix Notoginseng with ginsenoside Rg1 (C
24H
72O
14) calculate, every must not be less than 0.1-1mg.
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| CN101011474B (en) * | 2007-02-08 | 2011-01-05 | 南昌弘益科技有限公司 | Limitary determination of traditional medicine aconitine containing radix aconiti kusnezoffii praeparata |
| CN101590171B (en) * | 2009-07-15 | 2011-03-02 | 李志光 | Chinese medicinal composition for treating acute ankle sprain |
| CN102172409B (en) * | 2011-01-14 | 2012-07-18 | 中国人民解放军第三〇二医院 | Biological assay method and device for detecting toxicity potency of aconite traditional Chinese medicines |
| CN105974025A (en) * | 2016-06-08 | 2016-09-28 | 健民药业集团股份有限公司 | Detection method of traditional Chinese preparation for treating stomach illness |
| CN106353417A (en) * | 2016-08-14 | 2017-01-25 | 西南民族大学 | UPLC (ultra-performance liquid chromatography) detection method of multiple steroid saponins in Yunnan rhizoma paridis or its polygerm strain |
| CN111610271A (en) * | 2020-06-04 | 2020-09-01 | 成都中医药大学 | Identification method of authentic rhizoma paridis |
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| 中药成方制剂第15册. 药典委员会，26，药典委员会. 1998 * |
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