CN100569251C - The method of quality control of KEKE JIAONANG - Google Patents
The method of quality control of KEKE JIAONANG Download PDFInfo
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- CN100569251C CN100569251C CNB2006102012333A CN200610201233A CN100569251C CN 100569251 C CN100569251 C CN 100569251C CN B2006102012333 A CNB2006102012333 A CN B2006102012333A CN 200610201233 A CN200610201233 A CN 200610201233A CN 100569251 C CN100569251 C CN 100569251C
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Abstract
The invention provides a kind of method of quality control of KEKE JIAONANG, described method of quality control mainly comprise in character, discriminating, inspection and the assay project partly or entirely; Wherein differentiate the thin layer chromatography discriminating that comprises microexamination, Herba Ephedrae and Radix Glycyrrhizae; Assay is the assay to the contained morphine of Pericarpium Papaveris in the preparation.Compared with prior art, method of quality control precision height of the present invention, favorable reproducibility, response rate height, measurement result is accurate, has improved the quality control standard of KEKE JIAONANG, can control product quality effectively, thereby definitely guarantee the healthy of its clinical efficacy and extensive patients.
Description
Technical field:
The present invention relates to a kind of method of quality control of KEKE JIAONANG, belong to the technical field of medicine being carried out quality control.
Background technology:
KEKE JIAONANG as far back as 1998 just publication in " Chinese traditional patent formulation preparation " 17.KEKE JIAONANG mainly is made up of Herba Ephedrae, Pericarpium Papaveris, Radix Glycyrrhizae, Semen Armeniacae Amarum, Semen Raphani, Radix Platycodonis, Gypsum Fibrosum, is used for cough, and dyspnea with rapid respiration is breathed hard etc.Because its determined curative effect is subjected to the welcome of extensive patients deeply, is the standing medicine in people's daily life.But the thin layer chromatography that has only recorded Herba Ephedrae and Radix Glycyrrhizae in the KEKE JIAONANG initial quality standard is differentiated, no assay index, and it is simple to exist quality control standard, can not control the shortcoming of end product quality well.Pericarpium Papaveris is the principal agent among the we, astringing lung-QI and relieving cough; Be mainly used in chronic cough of deficiency lung, pure nihility heresy person.Because of its effective ingredient morphine Alkaloid has addiction, should not be excessive and continue to take.Therefore for controlling the inherent quality of KEKE JIAONANG better, the content detection of morphine in the preparation is seemed very necessary.
Summary of the invention:
The objective of the invention is to: the method for quality control that a kind of KEKE JIAONANG is provided.The present invention is directed to the deficiency of existing quality control standard, method of quality control to KEKE JIAONANG is studied, replenish the content assaying method set up morphine, improved the quality control standard of KEKE JIAONANG, thereby guaranteed the healthy of the clinical efficacy of this medicine and extensive patients conscientiously.
KEKE JIAONANG of the present invention is to constitute like this: calculate according to composition by weight: it mainly is prepared from by Herba Ephedrae 120-240, Pericarpium Papaveris 120-240, Radix Glycyrrhizae 120-240, Semen Armeniacae Amarum 120-240, Semen Raphani 30-80, Radix Platycodonis 30-80 and Gypsum Fibrosum 30-80.Its preparation method is: above seven flavors, Herba Ephedrae and Pericarpium Papaveris are ground into fine powder, sieve, respectively stay fine powder 83g standby, the residue coarse powder decocts secondary with acidic aqueous solution (transferring about pH value to 5 with 10% hydrochloric acid solution), each 2 hours, filters, filtrate is transferred pH value to 7 with 10% sodium hydroxide solution, and medicinal liquid is standby; The five tastes such as all the other Radix Glycyrrhizaes decoct with water secondary, each 1 hour, filter, filtrate and above-mentioned solution merge, and are concentrated into the thick paste shape, add Herba Ephedrae and Pericarpium Papaveris fine powder, add an amount of calcium carbonate or microcrystalline Cellulose, mixing 60～70 ℃ of dryings, is ground into fine powder or makes granule, dry, incapsulate, make 1000, promptly.
Method of quality control of the present invention mainly comprise in character, discriminating, inspection and the assay project partly or entirely; Wherein differentiate the thin layer chromatography discriminating that comprises microexamination, Herba Ephedrae and Radix Glycyrrhizae; Assay is the assay to the contained morphine of Pericarpium Papaveris in the preparation.
The discrimination method of its epheday intermedia is to be contrast with the ephedrine hydrochloride reference substance, and with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is the thin layer chromatography of developing solvent.The discrimination method of Radix Glycyrrhizae is to be contrast with the enoxolone reference substance, and with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is the thin layer chromatography of developing solvent.
Discrimination method comprises the part or all of of following project:
(1) get this preparation, put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped.
(2) get this preparation content, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes solution, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this preparation content, porphyrize adds hydrochloric acid and chloroform, puts reflux in the water-bath, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes solution, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Concrete discrimination method comprises the part or all of of following project:
(1) get this preparation, put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped.
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The content assaying method of morphine is to be contrast with the morphine reference substance in the Pericarpium Papaveris, and with methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is the high performance liquid chromatography of mobile phase.
The concrete content assaying method of morphine is:
According to Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak.
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly.
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is put in the tool plug conical flask, adds dichloromethane, ammonia, supersound process; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane.
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly.
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
The morphine content assay method is more specifically:
According to Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak.
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly.
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane.
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly.
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Method of quality control of the present invention comprises:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes solution, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content, porphyrize adds hydrochloric acid and chloroform, puts reflux in the water-bath, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes solution, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is put in the tool plug conical flask, adds dichloromethane, ammonia, supersound process; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane;
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
The applicant finds after deliberation, adopts the quality of following method of quality control with easier control KEKE JIAONANG, is more conducive to guarantee the clinical efficacy of preparation.So described method of quality control also can comprise:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane;
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Find that after deliberation following method of quality control is an optimal choice, the quality of easier control KEKE JIAONANG guarantees its clinical efficacy.So described method of quality control can also comprise:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=10: 2: 0.1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=19: 1 mixed solutions, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=10: 20: 7: 0.5 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=25: 74: 1 is a mobile phase; The detection wavelength is 215nm; Flow velocity is: 1.0ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane;
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Method of quality control of the present invention is the preferred plan that obtains through a large amount of screening tests, and following experimentation is a preferred process of the present invention:
The morphine content study on determination method:
1, need testing solution preparation method research
Method 1: get the content of KEKE JIAONANG, porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes.Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with microporous filter membrane (0.45um).
Method 2: get the KEKE JIAONANG content, porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds 1ml ammonia, 25ml chloroform, supersound process 1h.Put coldly, filter, residue filters with minimum of chloroform washing 3 times, combined chloroform liquid, water-bath volatilizes chloroform, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, be diluted to scale with methanol, shake up, filter, get subsequent filtrate as need testing solution with microporous filter membrane (0.45um).
| Sample | Method 1 (mg/g) | Method 2 (mg/g) |
| 1 | 0.0658 | 0.0452 |
| 2 | 0.0629 | 0.0495 |
| 3 | 0.0680 | 0.0510 |
According to result of the test as can be known, employing method 1 preparation need testing solution, morphine extracts more complete.
2, the selection of mobile phase:
Mobile phase 1: with 0.012molL
-1Potassium dihydrogen phosphate-acetonitrile (90: 10, contain 0.05mmolL
-1Triethylamine) is mobile phase
Mobile phase 2: with methanol-0.5% ammonium acetate aqueous solution-1% triethylamine (25: 74: 1) is mobile phase
Mobile phase 3: with 0.1molL
-1Sodium dihydrogen phosphate-methanol (78: 22) is mobile phase
Mobile phase 4: with A is the heptanesulfonic acid sodium solution that contains the 0.01mol/L of 0.15% acetic acid, and B is the mixed solution of 10% acetonitrile and 90% methanol, A: B=40: 60 is mobile phase
The result: with methanol-0.5% ammonium acetate aqueous solution-1% triethylamine (25: 74: 1) is mobile phase; The negative sample chromatogram is at noiseless peak, morphine position, and morphine separates fully (separating degree＞1.5) with close impurity peaks, and promptly morphine separates with other components fully under this condition.Therefore optimal flow is mutually: methanol-0.5% ammonium acetate aqueous solution-1% triethylamine (25: 74: 1).
3, repeated experiment:
Get KEKE JIAONANG,, measure by above-mentioned chromatographic condition by 5 parts of test liquids of preparation method preparation of test liquid under the method for quality control assay item of the present invention, morphine average content 0.0669mg/g, RSD%=0.88% wherein, the method repeatability is good.
The repeatability of morphine test in the preparation test sample
According to the result as seen, this method repeatability is good.
4, response rate experiment
Get 5 parts in the sample of known content, every part of about 0.3g accurately claims surely, puts in the 50ml tool plug triangular flask, and accurate respectively morphine reference substance solution (0.0104mg/ml) 2ml that adds according to the operation of need testing solution preparation method, makes application of sample and reclaims test sample liquid.Measure morphine content, recording average average recovery is 97.21%, RSD%=1.55%.
| Experiment number | Sample size (g) | Contain morphine amount (mg) | Add morphine amount (mg) | The amount of recording (mg) | The response rate (%) | Average recovery rate (%) | RSD(％) |
| 1 2 3 4 5 | 0.3012 0.2988 0.3010 0.2985 0.2978 | 0.0201 0.0199 0.0201 0.0199 0.0199 | 0.0208 | 0.0399 0.0397 0.0394 0.0398 0.0393 | 97.55 97.54 96.33 97.78 96.56 | 97.15 | 1.55 |
The applicant has also carried out following experiment:
The morphine addiction Journal of Sex Research
1 materials and methods
(1) animal grouping: rat Wistar kind, 70, ♀ ♂ half and half, body weight 180～220g is divided into 7 groups, 10 every group.One group is the blank group, and all the other are experimental group, freely ingest, drink water.
(2) medication: irritate the morphine hydrochloride of stomach experimental group rat 40%, irritate stomach 5ul, 10ul, 25ul, 50ul, 100ul, 125ul respectively for 1～6 group every group at every turn, every day 2 times, 4d continuously.Matched group is irritated stomach equal-volume normal saline.
(3) index of judgement rat morphine addiction: every rat is observed withdrawal symptom 20min.Direct record withdrawal symptom: the number of times of the battle array number of gritting one's teeth, upright, reason hair, wet Canis familiaris L. sample shake, stretching, extension, cunnilingus etc.
2 results
Observe the result of morphine-addicted rats and matched group withdrawal symptom, see the following form.
Morphine-addicted rats and matched group withdrawal symptom (x ± s)
* P＜0.05 of comparing with matched group, * * P＜0.01.
By the result as seen, the 6th group of rat naloxone bring out withdrawal symptom result and matched group as a result statistical procedures show the very remarkable P of group difference＜0.01; 2,3,4,5 groups of rat naloxones bring out withdrawal symptom result and matched group statistical procedures demonstration as a result P＜0.05; The 1st group with matched group do not have significant difference.
The KEKE JIAONANG pharmacodynamic study
1 test
Be subjected to the KEKE JIAONANG of reagent thing the present invention control
The experimental control medicine
(1) blank: equal-volume normal saline.
(2) known positive control drug: the KEKE JIAONANG of proper mass standard control
Modeling medicine: 1% Pilocarpus jaborandi aqueous slkali.
2 animal subjects
Kunming mouse, ♀ ♂ half and half, body weight is 18～22g; Rat Wistar kind, body weight 150～200g, ♀ ♂ half and half.
3 test methods
(1) mice ammonia draws the method for coughing experiment mice is divided into 5 groups at random by body weight, and 10 every group, ♀ ♂ half and half is by following method grouping.Normal saline group: use and the normal saline gastric infusion 0.2ml/10g of administration with volume, continuous 14 days, after the last administration, tested in 1 hour, the KEKE JIAONANG of the KEKE JIAONANG of the present invention's control and the control of proper mass standard respectively is one group, gastric infusion is 14 days continuously, after the last administration, tested in 1 hour, each group mice is put into successively respectively to be equipped with and uses 0.5ml, the 500ml inverted cup of the 0.25g cotton balls that 25%～28% ammonium hydroxide injects, should be rapid when being inverted beaker, in order to avoid gas is omitted.Observe mice immediately, shrink (breast contracts) with its abdominal muscle and magnify mouth simultaneously and be as the criterion, have sometimes cough sound show as the cough observation index.Begin until required second number of cough taking place as cough latent period first to put into the ammonia cotton balls.Examine with itemized record white mice 3 minutes in cough number of times and cough latent period, each group of statistics is calculated its antitussive percentage rate and prolongation of latency rate with the difference between the physiology saline group according to following formula.
Antitussive percentage rate (%)=(N-N
0)/N
0* 100%
(N
0: normal saline group cough number of times average, N: administration group cough number of times average)
Prolongation of latency rate (%)=(S-S
0)/S
0* 100%
(S
0: normal saline group average incubation period, S: administration group average incubation period)
(2) the phenol red expelling phlegm method test mice of mice is divided into 5 groups, every group 10, ♀ ♂ half and half, medication is tested with antitussive, and the KEKE JIAONANG group of normal saline matched group, the present invention's control and the KEKE JIAONANG group of proper mass standard control 30 minutes pneumoretroperitoneums after the last administration are only injected 0.5% phenol red solution 0.5ml/, inject and take off cervical vertebra execution mice in back 30 minutes, facing upward the position is fixed on the operation plate, cut off neck center skin, separate trachea, draw 5% NaHC with 1ml syringe, No. 7 indenting balls
3C solution 0.5ml lavation respiratory tract, 3 times, whenever recharge and wash respiratory tract 3 times, irrigating solution is injected a color comparison tube, shared 1.5mlNaHC like this
3C takes out and washes 9 times, and 3 times every time irrigating solutions are merged, and puts in the color comparison tube, carries out colorimetric with the phenol red color comparison tube of standard.
4 result of the tests
(1) mice ammonia draws the method result of the test of coughing and shows that the KEKE JIAONANG of the present invention's control compares with the KEKE JIAONANG of proper mass standard control, and through the biometrics analysis, the result shows that its difference all has the meaning of highly significant.Cough-relieving (P＜0.01) and prolongation cough latent period effect (P＜0.01) the results are shown in following table.
The KEKE JIAONANG antitussive action (n=10, X ± SD)
| Medicine | Dosage (g/kgd) | Cough number of times (inferior/3 minute) | Antitussive percentage rate (%) |
| Normal saline | Isometric(al) | 106.00±15.30 | |
| Former KEKE JIAONANG | 0.004 | 47.80±24.27 * | 56.84 * |
| KEKE JIAONANG of the present invention | 5.00 | 37.90±16.84 * | 64.58 * |
| KEKE JIAONANG of the present invention | 2.50 | 48.90±17.43 * | 55.62 * |
| KEKE JIAONANG of the present invention | 1.25 | 52.41±22.95 * | 50.26 * |
* compare P＜0.01 with former KEKE JIAONANG group.
The effect of KEKE JIAONANG prolongation cough latent period (n=10, X ± SD)
| Medicine | Dosage (g/kgd) | Cough latent period (s) | Percentage rate incubation period (%) |
| Normal saline | Isometric(al) | 6.80±3.26 | |
| Former KEKE JIAONANG | 0.004 | 46.80±41.05 * | 625.26 * |
| KEKE JIAONANG of the present invention | 5.00 | 53.60±9.10 * | 702.12 * |
| KEKE JIAONANG of the present invention | 2.50 | 49.50±13.25 | 638.89 * |
| KEKE JIAONANG of the present invention | 1.25 | 29.20±10.22 * | 325.88 * |
* compare P＜0.01 with former KEKE JIAONANG group.
(2) the phenol red expelling phlegm method test of mice result of the test shows, the KEKE JIAONANG of the present invention's control compares with the KEKE JIAONANG of proper mass standard control, and through the biometrics analysis, the result shows that its difference all has the meaning (P＜0.01) of highly significant.See the following form.
The KEKE JIAONANG phlegm-dispelling functions (n=10, X ± SD)
| Medicine | Dosage (g/kgd) | The phenol red excretion amount of trachea (ug/ml) | The P value |
| Normal saline | Isometric(al) | 1.200±0.953 | P＜0.01 |
| Former KEKE JIAONANG | 0.004 | 6.460±2.783 | P＜0.01 |
| KEKE JIAONANG of the present invention | 5.00 | 7.210±1.286 | P＜0.01 |
| KEKE JIAONANG of the present invention | 2.50 | 6.920±1.055 | P＜0.01 |
| KEKE JIAONANG of the present invention | 1.25 | 5.560±2.369 | P＜0.01 |
Experimental results show that by above-mentioned: the present invention can reach very effective effect for the content detection and the control of morphine in the KEKE JIAONANG; The KEKE JIAONANG of the present invention's control compares with the KEKE JIAONANG of proper mass standard control, and through experimental analysis, the result shows that its difference all has the meaning of highly significant.Can control product from links such as the requirement of the buying of raw materials for production, production technology, quality controls by the present invention, thereby guarantee the quality of KEKE JIAONANG.
Compared with prior art, method of quality control precision height of the present invention, favorable reproducibility, response rate height, measurement result is accurate, has improved the quality control standard of KEKE JIAONANG, can control product quality effectively, thereby definitely guarantee the healthy of its clinical efficacy and extensive patients.
The specific embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Embodiments of the invention 1: the method for quality control of KEKE JIAONANG comprises:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped.
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=10: 2: 0.1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=19: 1 mixed solutions, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=10: 20: 7: 0.5 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=25: 74: 1 is a mobile phase; The detection wavelength is 215nm; Flow velocity is: 1.0ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak.
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly.
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane.
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly.
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Embodiments of the invention 2: the method for quality control of KEKE JIAONANG can comprise:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped.
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5: 4: 0.05 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15: 2 mixed solutions, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20: 80: 0.5 is a mobile phase; The detection wavelength is 150nm; Flow velocity is: 0.5ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak.
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly.
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane.
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly.
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Embodiments of the invention 3: the method for quality control of KEKE JIAONANG can comprise:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, put cold, filtration, filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, respectively on the same silica gel g thin-layer plate of idea, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5: 30: 3: 0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=30: 60: 2 is a mobile phase; The detection wavelength is 280nm; Flow velocity is: 3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak.
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40ug, promptly.
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with the 0.45um microporous filter membrane.
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, promptly.
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
Embodiments of the invention 4: the method for quality control of KEKE JIAONANG can comprise:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor.
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped.
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=15: 0.5: 1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=25: 0.5 mixed solutions, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=15: 10: 10: 0.2 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix IL capsule item.
Claims (3)
1. the detection method of a KEKE JIAONANG, described KEKE JIAONANG are to constitute like this: calculate according to composition by weight: it mainly is prepared from by Herba Ephedrae 120-240, Pericarpium Papaveris 120-240, Radix Glycyrrhizae 120-240, Semen Armeniacae Amarum 120-240, Semen Raphani 30-80, Radix Platycodonis 30-80 and Gypsum Fibrosum 30-80; Its preparation method is: above seven flavors, Herba Ephedrae and Pericarpium Papaveris are ground into fine powder, sieve, respectively stay fine powder 83g standby, the acidic aqueous solution of residue coarse powder about with 10% hydrochloric acid solution adjust pH to 5 decocts secondary, and each 2 hours, filtration, filtrate is with 10% sodium hydroxide solution adjust pH to 7, and medicinal liquid is standby; The five tastes such as all the other Radix Glycyrrhizaes decoct with water secondary, each 1 hour, filter, filtrate and above-mentioned solution merge, and are concentrated into the thick paste shape, add Herba Ephedrae and Pericarpium Papaveris fine powder, add an amount of calcium carbonate or microcrystalline Cellulose, mixing 60～70 ℃ of dryings, is ground into fine powder or makes granule, dry, incapsulate, make 1000, promptly; It is characterized in that: described detection method is:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor;
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes solution, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content, porphyrize adds hydrochloric acid and chloroform, puts reflux in the water-bath, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes solution, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix I L capsule item;
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40 μ g, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is put in the tool plug conical flask, adds dichloromethane, ammonia, supersound process; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with 0.45 μ m microporous filter membrane;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, calculate, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
2. according to the detection method of the described KEKE JIAONANG of claim 1, it is characterized in that: described detection method is:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor;
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=5-15: 0.5-4: 0.05-1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=15-25: the 0.5-2 mixed solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=5-15: 10-30: 3-10: 0.2-0.8 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix I L capsule item;
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=20-30: 60-80: 0.5-2 is a mobile phase; The detection wavelength is 150-280nm; Flow velocity is: 0.5-3ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40 μ g, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with 0.45 μ m microporous filter membrane;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, calculate, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
3. according to the detection method of the described KEKE JIAONANG of claim 1, it is characterized in that: described detection method is:
Character: content is that pale brown color is to tan powder or granule; Mildly bitter flavor;
Differentiate: (1) gets this preparation, and put microscopically and observe: the outer peel cell is pentagon or rectangle like, and wall is the beads shape and thickens, and the guard cell side is seen and is dumbbell shaped;
(2) get this preparation content 3g, porphyrize adds ammonia solution number droplet, mixes thoroughly, adds chloroform 20ml, and merceration spends the night, and filters, and filtrate adds dilute hydrochloric acid 5ml, and jolting divides and gets the sour water layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the ephedrine hydrochloride reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to Chinese Pharmacopoeia appendix VIB thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: strong ammonia solution=10: 2: 0.1 is developing solvent, launch, take out, dry, spray is with 0.3% 1,2,3-indantrione monohydrate butanol solution: acetic acid=19: 1 mixed solutions, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this preparation content 2g, porphyrize adds hydrochloric acid 1ml and chloroform 20ml, puts in the water-bath reflux 1 hour, puts coldly, filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Extracting liquorice subacid reference substance adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution in addition; Test according to Chinese Pharmacopoeia appendix VI B thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: benzene: ethyl acetate: glacial acetic acid=10: 20: 7: 0.5 is developing solvent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, 105 ℃ of heating 5 minutes; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Check: measure according to Chinese Pharmacopoeia appendix IX H aquametry first method, must not surpass 12.0%;
All other should meet relevant every regulation under the Chinese Pharmacopoeia appendix I L capsule item;
Assay: shine Chinese Pharmacopoeia appendix VI D high effective liquid chromatography for measuring:
Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With methanol: 0.5% ammonium acetate aqueous solution: 1% triethylamine=25: 74: 1 is a mobile phase; The detection wavelength is 215nm; Flow velocity is: 1.0ml/min; Number of theoretical plate should be not less than 3000 by the morphine peak;
It is an amount of that the morphine reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 40 μ g, promptly;
The content under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, and accurate the title decides, and puts in the 50ml tool plug conical flask, adds the 30ml dichloromethane, 1ml ammonia, supersound process 30 minutes; Put coldly, filter, residue is with a small amount of washed with dichloromethane three times, filtration, combined dichloromethane liquid, water-bath volatilizes dichloromethane, and residue adds methanol makes dissolving, and quantitatively is transferred in the 5ml measuring bottle, is diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly with 0.45 μ m microporous filter membrane;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, calculate, promptly;
Every of this capsule contains morphine C
17H
19O
3N is 0.03～0.30mg.
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| CN102914498A (en) * | 2012-10-15 | 2013-02-06 | 辽源誉隆亚东药业有限责任公司 | Yanling Changchun capsule detection method |
| CN108956798A (en) * | 2018-04-27 | 2018-12-07 | 青海省药品检验检测院 | A kind of morphine hydrochloride preparation content determining method |
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| 高效液相色谱法测定克咳胶囊中吗啡的含量. 李娜等.时珍国医国药，第16卷第4期. 2005 |
| 高效液相色谱法测定克咳胶囊中吗啡的含量. 李娜等.时珍国医国药，第16卷第4期. 2005 * |
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