CN101057895A - 'Fuyanshu' preparation for treating gynopathy and its preparation method and quality controlling method and application - Google Patents
'Fuyanshu' preparation for treating gynopathy and its preparation method and quality controlling method and application Download PDFInfo
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Abstract
The invention relates to a medicinal preparation for treating gynaecologic diseases, process for preparation, quality control method and use thereof, wherein the preparation is prepared from eleven kinds of Chinese herbs including honey-suckle stem, sargentgloryvine and licorice root through extracting, refining, and charging auxiliary material, thereby obtaining pharmacologically allowable dose forms such as micro-pellets. The preparation of the invention has better stability, high biological availability, easy administering, fine external appearance and thus is easily acceptable for the suffers.
Description
Technical field
The present invention is a kind of woman scorching relax preparation and method for making and quality control method and application for the treatment of gynaecopathia, belongs to technical field of Chinese medicine.
Technical background
Gynaecopathia pelvic inflammatory disease, endometritis, cervicitis etc. all are common disease and the frequently-occurring diseases that threaten WomanHealth now.Such disease or morbidity are rapid, or delay is difficult for a long time, and convenient, the evident in efficacy Therapeutic Method of present shortage has still brought great misery for numerous women.Methods such as that modern medicine is mainly used is antibiotic, operation are treated this disease, and these methods not only can make the patient that drug resistance takes place and easily cause double infection, and cause most of patients can not adhere to therapy discontinued for a long time because of treatment acceptance difference.Chinese medicine should disease accumulate rich experience, determined curative effect, and toxic and side effects is little, but often because of the preparation reason, takes inconvenience, has influenced clinical extensive use.The scorching relieving capsule of woman for example, reasonable recipe, curative effect is preferable.But this dosage form is suitable for crowd's narrow range, and bioavailability, medicine stability are undesirable, and the problem that especially bioavailability of effective ingredient is not high is badly in need of solving.In view of such circumstances, optimize technology, improve dosage form, the control method that improves the quality becomes the scorching relieving capsule of woman and is badly in need of the thing that solves.
Summary of the invention
Technical problem to be solved by this invention provides a kind of woman scorching relax preparation and method for making and quality control method and application for the treatment of gynaecopathia; The present invention is directed to prior art, the micropill that provides, disintegrative is good, and the bioavailability height is particularly suitable for the old people and swallow tablet or the inconvenient patient of capsule take; The preparation method that is provided can effectively prepare needs preparation, guarantee that the preparation production technique obtain is scientific and reasonable; The method of quality control that is provided, the means, technical method of the index that detects, detection etc. can be provided to relevant production, testing agency, so that better control the quality of said preparation, guarantee the safety of medication, can better instruct production, make controlling of production process rationally strict more, make consumer's energy full appreciation product quality.
Technical scheme of the present invention is to constitute like this: calculate according to weight, it is with Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g all the acceptable dosage forms on the pharmaceutics that are made comprise: injection, the powder pin, freeze-dried powder, tablet, dispersible tablet, capsule, soft capsule, microcapsule, granule, pill, micropill, powder, drop pill, slow releasing preparation, controlled release preparation, gel, oral liquid, soft extract, extractum and membrane.Say accurately: described preparation is dispersible tablet, soft capsule, granule, micropill, gel, oral liquid, drop pill, sustained-release preparation, tablet, effervescent tablet or capsule.
The method for making of the scorching preparation that relaxes of the woman of described treatment gynaecopathia: get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby; Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and makes different preparations then respectively.Pellet in the described preparation prepares like this: gets Rhizoma Corydalis (processed), is ground into fine powder, cross 100 mesh sieves, and standby; Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, general method for making or mould the method for making pill, promptly.Described general method for making pill is such: adding consumption is the starch of 30%-70%, with the medicine mixing, crosses 100 mesh sieves, and to be that the ethanol of 60-95% is general be micropill to concentration, and coating is after the drying, promptly.Say accurately: described general method for making pill is such: add consumption and be 50% starch, with the medicine mixing, cross 100 mesh sieves, to be that 95% ethanol is general be micropill to concentration, and coating is after the drying, promptly.The described method for making pill of moulding is such: do diluent, 90% ethanol and soybean oil system soft material with starch, with the soft material that makes micropill mechanism ball, wet feed pushed the 0.8mm sieve aperture, the wet grain of strip cuts off round as a ball, at 50～60 ℃ of drying and mouldings, cross 10 mesh sieves and select ball, promptly get described preparation and can be used to prepare the medicine for the treatment of pelvic inflammatory disease.
Content assaying method comprises following all or part of content:
A. the high performance liquid chromatography assay of peoniflorin in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and accurate adding Diluted Alcohol or methanol are an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol or methanol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol or alcoholic solution is made reference substance solution; With the alkyl silane bonded silica gel is filler, acetonitrile-0.01mol/L～0.08mol/L sodium dihydrogen phosphate or 0.01mol/L～0.08mol/L potassium dihydrogen phosphate=10～14: 90～86 is mobile phase, and the detection wavelength is among 220～240nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method or standard curve method, this product contains peoniflorin with dosage and must not be less than 3.0mg every day;
B. the high performance liquid chromatography assay of tanshinone in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, accurate add methanol or ethanol an amount of, claim decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol or ethanol, shakes up, filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol or ethanol is made reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-water=75～80: 25～20 is mobile phase; The detection wavelength is among 260～280nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 0.40mg every day;
C. the high performance liquid chromatography assay of salvianolic acid B in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and accurate 50～100% methanol that add are an amount of, claim to decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 50～100% methanol, shake up, filter, get subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 50～100% methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=25～30: 7～10: 1～3: 59～66 is mobile phase; The detection wavelength is among 283～289nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains salvianolic acid B with dosage and must not be less than 5.5mg every day.
Say accurately: content assaying method comprises following all or part of content:
A. the high performance liquid chromatography assay of peoniflorin in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and the accurate Diluted Alcohol that adds is an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol and make reference substance solution; With the octadecylsilane chemically bonded silica is filler, and acetonitrile-0.02mol/L sodium dihydrogen phosphate=12: 88 is a mobile phase, and the detection wavelength is 230nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains peoniflorin with dosage and must not be less than 6.7mg every day;
B. the high performance liquid chromatography assay of tanshinone in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, and the accurate methanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, and methanol-water=78: 22 is a mobile phase; The detection wavelength is 270nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 0.75mg every day;
C. the high performance liquid chromatography assay of salvianolic acid B in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, and accurate 75% methanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 75% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 75% methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=27: 8: 2: 63 is mobile phase; The detection wavelength is 286nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 11mg every day.
Discrimination method comprises following all or part of content:
A. one or both thin layer chromatography discriminating in rhubarb medicinal material, the chrysophanic acid in the preparation
It is an amount of to get this product powder, adds methanol or ethanol supersound process, filters, and filtrate evaporate to dryness, residue add methanol or ethanol makes dissolving, as need testing solution; It is an amount of that other gets the Radix Et Rhizoma Rhei control medicinal material, shines medical material solution in pairs with legal system; Get the chrysophanic acid reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively in same be on the silica gel thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with petroleum ether-Ethyl formate or ethyl acetate-formic acid or glacial acetic acid=12～18: 3～7: 0.5～1.5 solution or upper solution are developing solvent, launch, take out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
B. one or both thin layer chromatography discriminating in red rooted salvia, the tanshinone in the preparation
It is an amount of to get this product powder, and the supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate makes dissolving, as need testing solution; Other gets the Radix Salviae Miltiorrhizae control medicinal material, shines medical material solution in pairs with legal system; Get the tanshinone reference substance, add ethyl acetate and make reference substance solution; According to thin layer chromatography test, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with toluene or benzene-ethyl acetate=9.2～9.8: 0.8～0.2 is developing solvent, launch, take out, dry; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
C. one or both thin layer chromatography discriminating in corydalis tuber medicinal material, the tetrahydropalmatine in the preparation
It is an amount of to get this product powder, adds methanol or ethanol supersound process, filters, and filtrate evaporate to dryness, residue add ammonia solution makes dissolving, extracts with the ether jolting, and ether solution volatilizes, and residue adds methanol or ethanol makes dissolving, as need testing solution; Other gets the Rhizoma Corydalis control medicinal material, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with toluene or benzene-acetone=7～11: 1～3 is developing solvent, launch, take out, dry, put in the iodine steam and smoke, put under the uviol lamp and inspect to clear spot; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
D. one or both thin layer chromatography discriminating in licorice medicinal materials, the glycyrrhizic acid in the preparation
It is an amount of to get this product powder, add methanol or ethanol supersound process, the extracting solution evaporate to dryness, residue is dissolved in water, and water liquid extracts with water saturated n-butyl alcohol jolting, the saturated water washing of n-butyl alcohol of n-butyl alcohol liquid, discard water lotion, n-butyl alcohol liquid evaporate to dryness, residue adds methanol or ethanol makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds the water supersound process in addition, and extracting solution shines medical material solution in pairs with legal system; A kind of in extracting liquorice acid ammonium or the monoammonium glycyrrhizinate reference substance adds methanol and makes reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with ethyl acetate-formic acid-glacial acetic acid-water=12～18: be developing solvent at 0.5～1.5: 0.5～1.5: 1～3, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
E. one or both thin layer chromatography discriminating in Radix Paeoniae Rubra medical material, the peoniflorin in the preparation
It is an amount of to get this product powder, adds ethanol or methanol supersound process, and extracting solution evaporate to dryness, residue add ethanol or methanol makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol or methanol is made reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with chloroform or dichloromethane-ethyl acetate-methanol or ethanol-formic acid or glacial acetic acid=35～45: be developing solvent at 3～7: 12～18: 0.2～0.8, launch, take out, dry, spray is with the vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color.
Say accurately: discrimination method comprises following all or part of content:
A. one or both thin layer chromatography discriminating in rhubarb medicinal material, the chrysophanic acid in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution; It is an amount of that other gets the Radix Et Rhizoma Rhei control medicinal material, shines medical material solution in pairs with legal system; Get the chrysophanic acid reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with 30～60 ℃ of petroleum ether-Ethyl formate-formic acid=15: 5: 1 is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
B. one or both thin layer chromatography discriminating in red rooted salvia, the tanshinone in the preparation
It is an amount of to get this product powder, and the supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate makes dissolving, as need testing solution; Other gets the Radix Salviae Miltiorrhizae control medicinal material, shines medical material solution in pairs with legal system; Get the tanshinone reference substance, add ethyl acetate and make reference substance solution; According to thin layer chromatography test, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel g thin-layer plate, and be developing solvent with toluene-ethyl acetate=9.5: 0.5, launch, taking-up is dried; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
C. one or both thin layer chromatography discriminating in corydalis tuber medicinal material, the tetrahydropalmatine in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add ammonia solution makes dissolving, extracts with the ether jolting, and ether solution volatilizes, and residue adds methanol makes dissolving, as need testing solution; Other gets the Rhizoma Corydalis control medicinal material, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with toluene-acetone=9: 2 was developing solvent, launched, and took out, dry, put in the iodine cylinder after about 1 minute and take out, wave the iodine that adsorbs on the most plate after, put under the uviol lamp and inspect under the 365nm; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
D. one or both thin layer chromatography discriminating in licorice medicinal materials, the glycyrrhizic acid in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, the extracting solution evaporate to dryness, and residue is dissolved in water, water liquid extracts with water saturated n-butyl alcohol jolting, and n-butyl alcohol liquid discards water lotion with the saturated water washing of n-butyl alcohol, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds the water supersound process in addition, and aqueous extract extracts with water saturated n-butyl alcohol jolting, and n-butyl alcohol liquid discards water lotion with the saturated water washing of n-butyl alcohol, n-butyl alcohol liquid evaporate to dryness, and residue adds methanol makes dissolving, in contrast medical material solution; A kind of in extracting liquorice acid ammonium or the monoammonium glycyrrhizinate reference substance adds methanol and makes reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with ethyl acetate-formic acid-glacial acetic acid-water=14: 1: 1: 2 was developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
E. one or both thin layer chromatography discriminating in Radix Paeoniae Rubra medical material, the peoniflorin in the preparation
It is an amount of to get this product powder, adds the ethanol supersound process, and extracting solution evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol or methanol is made reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-ethanol-glacial acetic acid=40: 5: 15: 0.5 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color.
Compare with technology with existing dosage form, the invention solves dosage form and be suitable for crowd's narrow range, bioavailability, the unfavorable problem of medicine stability.Its preparation formulation bioavailability height, good stability, easy to carry, good mouthfeel, absorption is fast, anti humility performance good; The preparation method that is provided can effectively prepare needs preparation, guarantee that the preparation variety effect obtain is remarkable, production technology is scientific and reasonable, has overcome the problem that existing product exists; Institute provides throws away method of quality control, can more fully control the quality of said preparation; Reached purpose of the present invention.
Caulis Lonicerae among the we: heat-clearing and toxic substances removing, dispelling wind collateral dredging.Existing chemical medicine pharmacological research proof has effect antibacterial and the releasing smooth muscle spasm.Caulis Sargentodoxae: heat-clearing and toxic substances removing, invigorate blood circulation, dispel the wind.Has spasmolytic, the merit of analgesia.Rhizoma Polygoni Cuspidati: promoting the function of the gallbladder to alleviate jaundice, heat-clearing and toxic substances removing, blood circulation promoting and blood stasis dispelling.Compositions such as contained Polydatin, flavonoid all have inhibitory action to staphylococcus aureus, Hemolytic streptococcus, Bacillus typhi etc., and some virus is also had inhibitory action.Radix Paeoniae Rubra: promoting blood circulation to remove blood stasis, clearing away heat and cooling blood.Zoopery proves that wherein contained paeoniflorin has spasmolytic, antiinflammatory, antiulcer and refrigeration function.Effect characteristics: 1, bacteriostasis: various bacteria such as staphylococcus aureus, Hemolytic streptococcus, Bacillus typhi, bacillus pyocyaneus are all had powerful inhibitory action.2, antiinflammatory action: can effectively resist neutrophil accumulation, can resist the activity of inflammatory cell chemotactic factor, and accelerate the absorption of inflammatory exudate.3, analgesic activity: pharmacological testing shows that the ICR mice taking the back to the reduction of electricity irritation sensitivity, shows the threshold value rising, has analgesic activity.Long term toxicity test: irritate stomach to rat and carried out three trimestral long term toxicity tests of dosage, the result show large, medium and small dosage treated animal generally in order, hematological indices, blood biochemistry index and each organ-tissue pathological examination there is no unusually, shows that it belongs to nontoxic level medicine.
The applicant finds in development process, is the assurance product quality, the screening of adjuvant, process conditions, and the screening of all conditions of method of quality control is most important.The applicant has carried out a series of experiments, with method and parameter of the supplementary product kind of the preparation technology that selects pharmaceutical preparation provided by the invention, use and consumption and ratio, quality control etc.; To guarantee science, reasonability, the feasibility of invention.
Experimental example 1 Study on extraction
(1) factor is selected: the Chinese medicine extraction effect is subjected to the influence of factors such as solvent load, extraction time, extraction time.In the prior art, Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati extract with alcohol heating reflux, and this key factor of amount of water is not done any research, and the applicant's high spot reviews amount of water is to the influence of extraction effect.
(2) index is determined: select extractum recovery rate and paeoniflorin content as evaluation index, its reason and assay method are as follows:
1. extractum recovery rate: extractum is the material base of solid preparation performance curative effect, and its yield height directly influence preparation process, is reasonable, effective control device so be chosen as the extraction index.
2. assay: extractum recovery rate height can not reflect fully that active ingredient extracts situation, so measure the contained paeoniflorin content of extractum simultaneously as Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati alcohol reflux effect screening index.
(3) test:
1. the alcohol reflux solvent load is investigated: take by weighing Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, each 135g of Rhizoma Polygoni Cuspidati, nominal is got six parts, adds alcohol reflux twice respectively, each 2 hours, merge extractive liquid,, filter, filtrate decompression concentrates, vacuum drying, claim fixed dried cream weight, calculate paste-forming rate, and measure content of paeoniflorin, result of the test sees the following form.
The ethanol consumption is investigated table as a result
| Tested number | Amount of water (doubly) | Extractum recovery rate (%) | Paeoniflorin content (%) | |
| For the first time | For the second time | |||
| 1 2 3 4 5 6 | 6 8 10 10 10 8 | 6 8 10 8 6 6 | 9.25 12.68 16.19 16.16 14.27 10.83 | 0.51 0.68 0.87 0.86 0.74 0.60 |
As seen from the above table: extractum recovery rate and paeoniflorin content were higher when alcohol adding amount was 10,10 times and 10,8 times amount, and not significantly difference between the two, guaranteeing under the sufficient prerequisite of extracts active ingredients, in order to save cost and to shorten man-hour, determine to extract amount of water 10 times of amounts for the first time, for the second time 8 times of amounts.
2. the water boiling and extraction amount of water is investigated
Seven flavors such as the medicinal residues behind the alcohol reflux and all the other Caulis Loniceraes decoct with water, and heavily serve as to investigate index with extract cream still, and the amount of water that decocting is boiled carries out preferably, and experimental technique and result are as follows:
Taking by weighing Radix Ophiopogonis rises 150g, Caulis Sargentodoxae 150g, Radix Glycyrrhizae 15g, Folium Isatidis 15g, Herba Taraxaci 45g, Radix Et Rhizoma Rhei (system) 30g, Fructus Toosendan 45g is mixed in one, totally 5 parts, with " investigation of alcohol reflux solvent load " test 3,4 boil the last medicinal residues in back is mixed in one, be divided into 6 parts, wherein 5 parts are added respectively in above-mentioned 5 parts of medical materials, decoct with water three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, filtrate decompression is concentrated into the thick paste that relative density is 1.36 (60 ℃), and drying claims fixed dried cream weight, calculate paste-forming rate, experimental design and the results are shown in following table.
Amount of water is investigated
| Tested number | Amount of water (doubly) | Cream heavy (g) | Paste-forming rate (%) | ||
| For the first time | For the second time | For the third time | |||
| 1 2 3 4 5 | 6 6 8 8 8 | 6 6 6 6 8 | 4 6 4 6 6 | 44.64 48.96 55.52 66.40 66.57 | 7.63 8.37 9.49 11.35 11.38 |
As seen from the above table, it is that 8,8,6 times paste-forming rate difference is little with amount of water that amount of water is 8,6,6 times, from energy savings with become original and consider that the optimised process of amount of water is extracting in water three times, amount of water is 8,6,6 times.
(4) demonstration test: in order to verify determined preparation technology's feasibility, we have carried out confirmatory experiment three times to these process conditions.
Test method: taking by weighing Radix Ophiopogonis rises 900g, Caulis Sargentodoxae 900g, Radix Glycyrrhizae 90g, Folium Isatidis 90g, Herba Taraxaci 270g, Radix Et Rhizoma Rhei (system) 180g, Fructus Toosendan 270g, Rhizoma Polygoni Cuspidati 270g, Radix Salviae Miltiorrhizae 270g, Radix Paeoniae Rubra 270g, Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, the heating and refluxing extraction secondary, the ethanol that adds for the first time 10 times of amounts adds the ethanol of 8 times of amounts, each 2 hours the second time, merge extractive liquid, filters, and filtrate concentrates, drying, it is heavy to claim to decide cream, and measures content of paeoniflorin in the dried cream.Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted respectively 1.5 hours, 1 hour, collecting decoction filters, and filtrate concentrates, drying, it is heavy to claim to decide cream, calculates paste-forming rate, result such as following table.
Alcohol reflux condition demonstration test
| Medical material amount (g) | Cream heavy (g) | Paste-forming rate (%) | Average paste-forming rate (%) | Paeoniflorin content (%) |
| 810 810 810 | 131.14 130.73 131.06 | 16.19 16.14 16.18 | 16.17 | 0.87 0.86 0.85 |
From last table as seen, alcohol reflux condition more stable (average paste-forming rate is 16.17%), the content of gained sample is also more stable, illustrates that the experimental condition through screening is feasible.
The water boiling and extraction demonstration test
| Medical material amount (g) | Cream heavy (g) | Paste-forming rate (%) | Average paste-forming rate (%) |
| 3510 3510 3510 | 400.49 397.33 399.44 | 11.41 11.32 11.38 | 11.37 |
From last table as seen, the water boiling and extraction condition is more stable, illustrates to stablize feasible through the experimental condition of screening.
The investigation of experimental example 2 disintegrating process
Spice is mobile relevant with degree of grinding former, adjuvant.And flowability must form certain influence to micropill.But it is meticulous to pulverize, and has both increased pulverizing difficulty and loss, causes dust pollution again, according to the trial test result, pulverizes 100 orders and gets final product, and therefore test is measured the pulverizing flour extraction of this raw materials technology.
Test method: take by weighing corydalis tuber medicinal material 200g, be ground into fine powder, cross 100 mesh sieves, measure powder outlet quantity, flour extraction.Repeat three tests, calculate average flour extraction.The results are shown in following table.
The investigation of flour extraction
| Tested number | Medical material amount (g) | Powder outlet quantity (g) | Flour extraction (%) | Average flour extraction (%) |
| 1 2 3 | 200 200 200 | 193.70 195.06 196.24 | 96.85 97.53 98.12 | 97.50 |
The result is as seen: the flour extraction of three duplicate samples shows that all more than 95% breaking method is stable, feasible.
Experimental example 3 separates, concentration technology research
Separate and select: adopt 200 order filter clothes to filter.
The concentration technology condition: take all factors into consideration present plant produced and generally adopt the equipment situation, determine that tentatively filtrate employing triple effect concentration tank concentrates, be concentrated into the thick paste that relative density is about 1.20 (50 ℃), standby.Concentrated condition is: temperature is 84 ℃ of effects, two 80 ℃ of effects, 69 ℃ of triple effects, and vacuum is that an effect 0.025Mpa, two is imitated 0.046Mpa, triple effect 0.068Mpa.
Experimental example 4: pellet Study on Forming
The micropill diameter is less than 2.5mm, and class is in particle properties, the bioavailability height, and the applicant is when development product micropill of the present invention, and maximum difficulty is exactly that hygroscopicity is strong and mobile poor, and poor plasticity is difficult to molding.The micropill manufacturing technology and the adjuvant that adopt the applicant's screening to obtain make product be easy to disintegrate, and the bioavailability height is well-behaved.
The design of the prescription of micropill depends primarily on the physicochemical property, clinical medicine dose of crude drug etc.The character of its crude drug, hygroscopic investigation and clinical dosage are the bases of prescription design.Hygroscopicity is bigger to the influence of medicine, therefore needs its hygroscopicity is investigated when writing out a prescription design, writes out a prescription according to the investigation result then and designs and definite moulding process.
4.1 Study on Forming (general method for making)
4.1.1 the mensuration of hydroscopicity:
Test method: get the about 2g of dry powder, put respectively in the weighing botle of dry constant weight, thickness is no more than 5mm, places the close drying device of different relative humiditys then respectively.After 25 ℃ of water isolation type electrothermostats are placed 96 hours, accurately weigh, according to formula:
Calculate hydroscopicity, and observe the variation of appearance character.Experimental result sees the following form.
The hygroscopicity measurement result
| The solution kind | Relative humidity (%) | Hydroscopicity (%) | Average hydroscopicity (%) | ||
| 1 | 2 | 3 | |||
| NaCl saturated solution NaBr saturated solution 44%H 2SO 4 54％H 2SO 4 | 75.28 57.70 48.52 29.50 | 20.27 13.31 8.98 5.52 | 20.48 13.23 8.93 5.41 | 20.30 13.33 8.97 5.51 | 20.35 13.29 8.96 5.48 |
As seen from table, the hygroscopicity of material is less.
4.1.2 preparation prescription screening study
4.1.2.1 binding agent is preferred:
Owing in the material ointment powder is arranged, certain viscosity is arranged, adopt water to prepare micropill as binding agent, micropill is adhesion very easily, therefore we consider to select for use alcoholic solution as binding agent, we prepare micropill with Different concentrations of alcohol solution as binding agent for this reason, serve as to investigate index with the rounding property of micropill.Experimental result sees the following form.
Binding agent preferred
| The kind of binding agent | The micropill outward appearance |
| 60% ethanol, 80% alcohol 95 % ethanol | Ball shape is not round, and easily adhesion ball shape has change slightly, and part adhesion ball shape is round slightly, indivedual adhesions |
By last table result as seen, the viscosity of powder is very strong, and to adopt 95% ethanol be binding agent than other two alcoholic degrees is that the micropill outward appearance for preparing of binding agent is quite a lot of, thus we to consider to adopt 95% ethanol be binding agent.
4.1.2.2 the selection of adjuvant:
4.1.2.2.1 the screening of supplementary product kind
Because the viscosity of crude drug is big, even adopt 95% ethanol also to influence the mouldability of micropill as binding agent, better for the mouldability that makes micropill, need in prescription, to add the suitable dilution agent, so we screen adjuvant medical starch, dextrin commonly used in the test.
Get the medicated powder of two parts of identical weight, a medical starch that adds, a dextrin that adds, mix homogeneously is put into sugar coating machine respectively, sprays into 95% ethanol, makes micropill, and result of the test sees Table.
Adjuvant optimization experiment result
| Supplementary product kind and consumption | Medical starch (70%) | Dextrin (70%) |
| The micropill outward appearance | Rounding is even | Ball shape is not round |
As seen from the above table, the pill effect of starch is better than dextrin, and therefore selecting medical starch for use is adjuvant.
4.1.2.2.2 determining of supplementary product consumption
The rounding property of micropill is not only relevant with the kind of adjuvant, and is also relevant with the consumption of adjuvant, therefore the consumption of adjuvant investigated.Get the medicated powder of three parts of identical weight, the ratio that adds starch is respectively 30%, 50%, 70%, drops in the sugar coating machine, sprays into 95% ethanol, makes micropill, observes the mouldability of micropill.
Determining of supplementary product consumption
| The starch consumption | The micropill outward appearance |
| 30％ 50％ 70％ | Not round, adhesion ball shape rounding ball shape rounding |
By experimental result as can be known, the starch consumption is 50% and 70% o'clock, and gained micropill ball shape is better, because of both pill effects are the same, for saving supplementary product consumption, determines that supplementary product consumption is 50%.
4.1.3 pill:
Taking by weighing Radix Ophiopogonis rises 450g, Caulis Sargentodoxae 450g, Radix Glycyrrhizae 45g, Folium Isatidis 45g, Herba Taraxaci 135g, Radix Et Rhizoma Rhei (system) 135g, Fructus Toosendan 135g, Rhizoma Polygoni Cuspidati 135g, Radix Salviae Miltiorrhizae 135g, Radix Paeoniae Rubra 135g, Rhizoma Corydalis 135g, more than ten simply, Rhizoma Corydalis (system) is ground into fine powder, crosses 100 mesh sieves, and is standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, adds the ethanol of 8 times of amounts for the second time, each 2 hours, merge extractive liquid, filtered, filtrate recycling ethanol also is concentrated into the thick paste that relative density is 1.36 (60 ℃), and drying is pulverized.Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted respectively 1.5 hours, 1 hour, collecting decoction filters, filtrate decompression is concentrated into the thick paste that relative density is 1.36 (60 ℃), and drying is pulverized.Two kinds of extract powders and corydalis tuber medicinal material powder are mixed in one, add appropriate amount of starch, mixing is crossed 100 mesh sieves, and ethanol is general to be micropill, after the drying, promptly.Gained micropill rounding is even, illustrates that the prescription through screening is feasible.
4.2 Study on Forming (moulding method for making)
4.2.1 system soft material
Get extractum fine powder, crude drug powder and starch, soybean oil and ethanol and make soft material with wet granulation process in right amount, make it to reach and hold agglomeratingly, that pinches can loose, standby.Primary study concentration of alcohol and soybean oil consumption influence pill, and experimental result sees the following form.
| Tested number | Concentration of alcohol | System soft material situation |
| 1 2 3 | 95% ethanol, 90% ethanol, 85% ethanol | The moderate soft material of the not enough soft material of soft material viscosity easily bonds |
The soybean oil consumption is investigated
| Tested number | The soybean oil consumption | The pill situation |
| 1 2 3 | 90% ethanol, 1% soybean oil, 90% ethanol, 2% soybean oil, 90% ethanol, 3% soybean oil | Lubricity is not enough, and it is moderate to push difficult lubricity, and suitable pill lubricity is excessive, can't become ball |
4.2.2 pill
The soft material that makes is with micropill mechanism ball, and wet feed pushed the 0.8mm sieve aperture, and the wet grain of strip cuts off round as a ball, at 50～60 ℃ of drying and mouldings, crosses 10 mesh sieves and selects ball.
The pharmacological research of experimental example 5 antiinflammatory actions
5.1 the influence of xylol induced mice auricle edema
Experimental technique faces with preceding that micropill of the present invention (general method for making) is mixed with the 0.10g/ml suspension with 0.5% Carboxymethyl cellulose sodium (CMC-Na) is standby, and animal is used healthy Kunming mouse, body weight 20 grams.Mice is divided into 4 groups (matched group normal saline) at random, irritate the long-pending 20ml/kg of being of body of stomach, two weeks of continuous irrigation stomach, once a day, the last administration after 30 minutes with microsyringe only with 0.05ml/, dimethylbenzene is applied to mouse right ear, put to death mice after 15 minutes, cut two ears, dash with the 8mm diameter steel and lay round auricle in left and right sides auricle same area respectively along the auricle baseline, torsion balance claims two auricle weight in wet bases, with two auricle weight differences as the swelling level index.Inhibitory rate of intumesce equals the difference of average swelling degree of matched group and the average swelling degree of administration group and takes advantage of 100% again divided by the average swelling degree of matched group.
| Group | Dosage (g/kg) | Animal (only) | Average swelling degree (mg) | Suppression ratio (%) |
| The scorching relieving capsule group of matched group hydrocortisone group woman micropill group of the present invention | 20ml/kg 0.04 2.0 2.0 | 8 8 8 8 | 20.15±3.21 7.11±1.36 15.24±3.30 14.02±4.32 | - 65.75 30.85 31.12 |
The result shows that preparation of the present invention has good antiinflammatory action, and effect is better than the scorching relieving capsule of commercially available woman.
5.2 influence to the rat uterus inflammation
Experimental technique: animal is selected the female rat of SD kind for use, about 200 grams of body weight.Animal is divided 4 groups, each treated animal under etherization cuts off the hypogastric region hair, and long mouthful of 2cm is cut in the sterilization back in the abdomen center, expose the uterus, make a kerf in the place along 1cm on the left hand corner of uterus, with a plastic hoop (caliber 2cm, long 0.5cm, heavy 2mg, alcohol disinfecting) be positioned over intrauterine, with the uterine incision sutured, postoperative beginning in 2 hours administration, once a day, the administration volume is the 20ml/kg body weight, puts to death animal after 7 days, takes out the uterus, both sides, remove fat, analytical balance is weighed, and left side, every Mus uterus is inflammation swelling degree with the difference on right side, calculates the swelling rate and the suppression ratio of administration group.The swelling rate equals to cause scorching uterus average weight and not multiply by 100% with the difference that does not cause scorching uterus average weight divided by causing scorching uterus average weight, and the difference that suppression ratio equals average swelling rate in matched group uterus and the average swelling rate in administration group uterus multiply by 100% divided by the average swelling rate in matched group uterus.
| Group | Dosage (g/kg) | Animal (only) | Swelling rate (%) | Suppression ratio (%) |
| The scorching relieving capsule group micropill group of the present invention (system of moulding) of matched group hydrocortisone group woman | 20ml/kg 0.04 2.0 2.0 | 10 10 10 10 | 208.12 6.72 17.08 16.27 | 96.17 90.37 91.94 |
The result shows that preparation of the present invention has good anti-inflammation of uterus effect, and effect is better than the scorching relieving capsule of woman.The thin layer chromatography discrimination method of rhubarb medicinal material, chrysophanic acid research in experimental example 6 dispersible tablets:
Feature for outstanding Radix Et Rhizoma Rhei, selected with chrysophanic acid as the feature speckle, but owing to have composition, for example chlorogenic acid and the caffeic acid in Caulis Lonicerae, the Herba Taraxaci like the more or polar phase close with the chrysophanic acid structure in the preparation, glycyrrhizic acid in the Radix Glycyrrhizae, compositions such as the peoniflorin in the Radix Paeoniae Rubra.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:
The thin layer chromatography discrimination method of rhubarb medicinal material research in the dispersible tablet
| Condition | The result |
| The upper solution silica gel g thin-layer plate of the upper solution silica gel g thin-layer plate benzinum (30～60 ℃) of the upper solution silica gel g thin-layer plate benzinum (30～60 ℃) of benzinum (30～60 ℃)-Ethyl formate=15: 4 silica gel H lamellae benzinum (30～60 ℃)-isopropanol-water=15: 3: 2 silica gel H lamellae chloroform-acetone=10: 7 silica gel g thin-layer plate ethyl acetate-ethanol=13: 7 silica gel H lamellae benzinum (60～90 ℃)-Ethyl formate-formic acid=20: 5: 1-Ethyl formate-formic acid=18: 3: 0.5 silica gel g thin-layer plate benzinum (60～90 ℃)-Ethyl formate-formic acid=12: 7: 1.5-Ethyl formate-formic acid=15: 5: 1 | Feminine gender has the unintelligible feminine gender of interference separation to have the feminine gender of interference to have the interference separation poor separation more clear, Rf value is moderate, negative noiseless separation is more clear, Rf value is moderate, negative noiseless separation is the most clear, Rf value is moderate, and is negative noiseless |
Through screening, determined optimum condition: being immobile phase with the silica gel g thin-layer plate, is developing solvent with the upper solution of petroleum ether (30～60 ℃)-Ethyl formate-formic acid=15: 5: 1, with this understanding, the Rf value of chrysophanic acid feature speckle is moderate, and it is clear to separate with other speckle, and is negative noiseless.
The thin layer chromatography discrimination method of red rooted salvia, tanshinone research in experimental example 7 pellets:
Feature for outstanding Radix Salviae Miltiorrhizae, selected to contrast speckle as it with tanshinone, but owing to there is composition like the more or polar phase close in the preparation with the tanshinone structure, the caffeic acid in Caulis Lonicerae, the Herba Taraxaci for example, the indirubin in the Folium Isatidis, composition such as indigo.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:
The thin layer chromatography discrimination method of red rooted salvia, tanshinone research in the pellet
| Condition | The result |
| Benzinum, (30～60 ℃)-ethyl acetate=15: 4 silica gel H lamellae benzinums, (30～60 ℃)-ethanol=15: 2 silica gel H lamellae chloroform-acetone=10: 1 silica gel g thin-layer plate benzene-ethyl acetate=10: 0.1 silica GF254 lamellae ethyl acetate-ethanol=13: 7 silica gel H lamellae benzene-ethyl acetate=9: 1 silica gel g thin-layer plate benzene-ethyl acetate=9.2: 0.8 silica gel g thin-layer plate toluene-ethyl acetate=9.8: 0.2 silica gel g thin-layer plate toluene-ethyl acetate=9.5: 0.5 silica gel g thin-layer plates | It is unintelligible to separate poor separation, feminine gender has the feminine gender of interference to have the higher separation poor separation of the Rf value Rf value on the low side of interference more clear, Rf value is moderate, negative noiseless separation is more clear, Rf value is moderate, negative noiseless separation is the most clear, Rf value is moderate, and is negative noiseless |
Through screening, determined optimum condition: be immobile phase with the silica gel g thin-layer plate, with toluene-ethyl acetate=developing solvent in 9.5: 0.5, with this understanding, the Rf value of tanshinone feature speckle is moderate, and it is clear to separate with other speckle, and is negative noiseless.
Prolong the thin layer chromatography discrimination method research of corydalis tuber medicinal material, Rhizoma Corydalis second element in experimental example 8 soft capsules:
For the feature of outstanding Rhizoma Corydalis, selected to contrast speckle as it, but owing to there is composition like the more or polar phase close in the preparation, for example the indirubin in the Folium Isatidis, composition such as indigo with the tetrahydropalmatine structure with tetrahydropalmatine.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:
The thin layer chromatography discrimination method of corydalis tuber medicinal material, tetrahydropalmatine research in the soft capsule
| Condition | The result |
| Benzinum (60～90 ℃)-methyl alcohol=7: 4 silica gel g thin-layer plate benzinum (30～60 ℃)-acetone=7: 2 silica gel g thin-layer plate chloroform-acetone=10: 3 silica G F254 lamellae benzene-chloroform=10: 1 silica gel g thin-layer plate toluene-acetone=12: 1 silica gel H lamellae benzene-ethyl acetate=9: 1 silica gel g thin-layer plate toluene-acetone=7: 1 silica gel g thin-layer plate benzene-acetone=11: 3 silica gel g thin-layer plate toluene-acetone=9: 2 silica gel g thin-layer plates | Separating the unintelligible feminine gender of poor separation has the interference separation poor separation unintelligible, the higher separation poor separation of Rf value is more clear, Rf value is moderate, negative noiseless separation is more clear, Rf value is moderate, negative noiseless separation is the most clear, Rf value is moderate, and is negative noiseless |
Through screening, determined optimum condition: be immobile phase with the silica gel g thin-layer plate, with toluene-acetone=developing solvent in 9: 2, with this understanding, the Rf value of tetrahydropalmatine feature speckle is moderate, and it is clear to separate with other speckle, and is negative noiseless.
The thin layer chromatography discrimination method of Radix Paeoniae Rubra medical material, peoniflorin research in experimental example 9 pellets:
For the feature of outstanding Radix Paeoniae Rubra, selected peoniflorin as the characteristic component speckle, but owing to there is composition like the more or polar phase close in the preparation, the glycyrrhizic acid in the Radix Glycyrrhizae for example, compositions such as the anthraquinone glycoside in the Radix Et Rhizoma Rhei with the peoniflorin structure.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:
The thin layer chromatography discrimination method of Radix Paeoniae Rubra medical material, peoniflorin research in the pellet
| Condition | The result |
| Chloroform-methyl alcohol=7: 1 silica gel H lamellae chloroform-ethyl acetate-ethanol=35: 3: 12 silica gel H lamellae chloroform-acetone=10: 7 silica gel g thin-layer plate chloroform-ethyl acetate-ethanol-formic acid=35: 3: 12: 0.2 silica gel H lamellae chloroform-ethyl acetate-methyl alcohol-glacial acetic acid=45: 7: 18: 0.8 silica gel g thin-layer plate chloroform-ethyl acetate-ethanol-glacial acetic acid=40: 5: 15: 0.5 silica gel g thin-layer plate | Feminine gender has the peak shape of interference hangover feminine gender to have interference separation more clear, Rf value is moderate, negative noiseless separation is more clear, Rf value is moderate, negative noiseless separation is the most clear, Rf value is moderate, and is negative noiseless |
Through screening, determined optimum condition: be immobile phase with the silica gel g thin-layer plate, chloroform-ethyl acetate ethanol glacial acetic acid=40: 5: 15: 0.5 is developing solvent, with this understanding, the Rf value of peoniflorin feature speckle is moderate, and it is the most clear to separate with other speckle, and is negative noiseless.
The thin layer chromatography discrimination method of licorice medicinal materials, glycyrrhizic acid research in experimental example 10 dispersible tablets:
For the feature of outstanding Radix Glycyrrhizae, the mattress quinone glycoside class in the Radix Et Rhizoma Rhei for example, compositions such as the peoniflorin in the Radix Paeoniae Rubra have been selected with glycyrrhizic acid as its feature speckle, but owing to there is composition like the more or polar phase close in the preparation with the glycyrrhizic acid structure.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of thin layer chromatography effect quality is the composition of unfolding condition, particularly developing solvent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:
The thin layer chromatography discrimination method of licorice medicinal materials, glycyrrhizic acid research in the dispersible tablet
| Condition | The result |
| Methyl alcohol-formic acid=15: 2 silica gel H lamellae chloroform-ethanol=10: 3 silica gel g thin-layer plate ethyl acetate-methyl alcohol=13: 7 silica gel H lamellae ethyl acetate-methyl alcohol-glacial acetic acid=12: 2: 0.5 silica gel g thin-layer plate ethyl acetate-methyl alcohol-glacial acetic acid-water=18: 0.5: 1.5: the silica gel g thin-layer plate ethyl acetate-methyl alcohol of 1 usefulness, 1% sodium hydroxide solution preparation-glacial acetic acid-water=12: 1.5: 0.5: 3 silica gel g thin-layer plate ethyl acetate-methyl alcohol-glacial acetic acid-water=14: 1: 1: the silica gel g thin-layer plate that 2 usefulness, 1% sodium hydroxide solution prepares | Separate unintelligible, feminine gender has the feminine gender of interference to have interference separation unintelligible, feminine gender has the interference separation poor separation more clear, Rf value is moderate, negative noiseless separation is more clear, Rf value is moderate, negative noiseless separation is the most clear, Rf value is moderate, and is negative noiseless |
Through screening, determined optimum condition: with the silica gel g thin-layer plate with the preparation of 1% sodium hydroxide solution is immobile phase, ethyl acetate-methanol-glacial acetic acid-water=14: 1: 1: 2 are developing solvent, with this understanding, the Rf value of glycyrrhizic acid characteristic component speckle is moderate, it is the most clear to separate with other speckle, negative noiseless.
The high performance liquid chromatography content assaying method of peoniflorin research in experimental example 11 pellets:
1 instrument and reagent
1.1 key instrument
High performance liquid chromatograph 2010Aht SHIMADZU
The general all purpose instrument company limited of analysing in ultraviolet spectrophotometer TU-1800SPC Beijing
Electronic analytical balance BP211D SARTORIUS
Ultrasonic washing unit KQ250B Kunshan Ultrasonic Instruments Co., Ltd.
1.2 reagent
Methanol analytical pure Beijing Chemical Plant
Acetonitrile chromatographically pure CALEDON
The pure Beijing of phosphoric acid top grade chemical reagents corporation
The pure water WAHAHA
It is an amount of that the 2 selection precisions that detect wavelength take by weighing the peoniflorin reference substance, adds methanol and make the solution that every 1ml contains 15 μ g, in the interscan of 200～400nm wave-length coverage.The result shows that peoniflorin has absorption maximum at the 230nm place, therefore selects 230nm as the detection wavelength of measuring paeoniflorin content in the scorching micropill that relaxes of woman.
3 chromatographic conditions
Chromatographic column: Diamonsil ODS 250mm * 4.6mm 5 μ m
Mobile phase: acetonitrile-0.02mol/L sodium dihydrogen phosphate (12: 88)
Flow velocity: 1.0ml/min
Column temperature: 30 ℃
Sample size: 10 μ l
Detect wavelength: 220nm
Test and Selection peoniflorin as its index components, but owing to have composition, for example glycyrrhizic acid in the Radix Glycyrrhizae, the compositions such as anthraquinone glycoside in the Radix Et Rhizoma Rhei like the more or polar phase close in the preparation with the peoniflorin structure.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of high performance liquid chromatography effect quality is the composition of elution requirement, particularly mobile phase.Therefore, experiment sieving multiple elution requirement, part elution requirement and result are as follows:
The investigation of chromatographic condition
| Condition | The result |
| Methanol-water=80: 20 octadecylsilane chemically bonded silica methyl alcohol-0.05mol/L sodium dihydrogen phosphate=15: 85 eight alkyl silane bonded silica gel methanol-waters=20: 80 dialkyl silane bonded silica gel acetonitrile-0.05mol/L potassium dihydrogen phosphate buffer solution=30: 70 octadecylsilane chemically bonded silica acetonitrile-water=15: 85 octadecylsilane chemically bonded silica acetonitrile-0.02mol/L sodium dihydrogen phosphate=10: 90 octadecylsilane chemically bonded silica acetonitrile-0.02mol/L sodium dihydrogen phosphate=14: 86 octadecylsilane chemically bonded silica acetonitrile-0.02mol/L sodium dihydrogen phosphate=12: 88 octadecylsilane chemically bonded silicas | It is long partially to separate the incomplete longer retention time of the incomplete appearance time of the long separation of incomplete disengaging time of separating, it is more clear to separate, negative noiseless retention time is moderate, it is more clear to separate, negative noiseless retention time is moderate, it is the most clear to separate, negative noiseless |
Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, acetonitrile-0.02mol/L sodium dihydrogen phosphate=12: 88 is a mobile phase, with this understanding, the peoniflorin retention time is moderate, and the peak is capable sharp-pointed, symmetry is separated fully with adjacent peak, and is negative noiseless.
This product under the content uniformity item is got in the preparation of 4 need testing solutions, and porphyrize is got about 1.5g, the accurate title, decide, and puts in the tool plug conical flask, the accurate Diluted Alcohol 20ml that adds, close plug claims to decide weight, supersound process (power 250W, frequency 33KHz) 30 minutes, take out, put to room temperature, claim to decide weight, supply the weight that subtracts mistake, shake up with Diluted Alcohol, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.
The investigation precision of 5 linear relationships is measured peoniflorin reference substance solution (1.134mg/ml) 0.4ml, 0.8ml, 1.2ml, 1.6ml, 2.0ml, split in the 10ml measuring bottle, add mobile phase and be diluted to scale, shake up, be mixed with the reference substance solution of 0.04536mg/ml, 0.09072mg/ml, 0.13608mg/ml, 0.18144mg/ml, 0.22680mg/ml, the therefrom accurate respectively 10 μ l that draw, inject chromatograph of liquid, measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).With the peak area is abscissa, and peoniflorin sample size (μ g) is figure for vertical coordinate, the drawing standard curve.The result is as follows:
The peoniflorin linear relationship
| Numbering | Peak area | Peoniflorin sample size (μ g) |
| 1 2 3 4 5 | 422713 862241 1292242 1725801 2139741 | 0.4536 0.9072 1.3608 1.8144 2.2680 |
Regression equation: Y=0.000001X+0.000919
Correlation coefficient: γ=0.9999
The result shows that peoniflorin linear relationship between 0.4536 μ g～2.2680 μ g is good.
Through calculating, the peoniflorin standard curve is one to cross the straight line of initial point, therefore selects one point external standard method to measure content of paeoniflorin in the scorching micropill that relaxes of woman.
The test of 6 precision is accurate draws with a peoniflorin reference substance solution 10 μ l, injects chromatograph of liquid, and replication 5 times is investigated reference substance solution precision, and the result is as follows:
The precision test
| Test number (TN) | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD(％) |
| Peak area | 1082259 | 1075463 | 1080794 | 1090023 | 1079536 | 1081615 | 0.49 |
The result shows that reference substance solution precision is good.
7 need testing solution stability tests are accurate to be drawn with a need testing solution 10 μ l, injects chromatograph of liquid, measures at 0,2,4,8,24 hour sample introduction respectively, and the result is as follows:
The need testing solution stability test
| Testing time (h) | 0 | 2 | 4 | 8 | 24 | Meansigma methods | RSD(％) |
| Content (mg/ bag) | 2.805 | 2.751 | 2.803 | 2.783 | 2.832 | 2.795 | 1.09 |
The result shows that need testing solution is good at 24 hours internal stabilities.
8 replica tests are got this product, and porphyrize is got about 1.5g (totally 5 parts), and accurate the title decides, and press the preparation and the operation down of algoscopy item of text need testing solution, and the result is as follows:
Replica test
| Numbering | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD(％) |
| Content (mg/ bag) | 2.754 | 2.762 | 2.811 | 2.796 | 2.783 | 2.781 | 0.85 |
The result shows that repeatability is good.
This product under the content uniformity item is got in the test of 9 average recoveries, and porphyrize is got about 0.75g (totally 6 parts), and accurate the title decides, and splits in the tool plug conical flask; Precision is measured peoniflorin reference substance solution (0.5144mg/ml) 1.0ml, 1.2ml, 1.4ml (each 2 parts), splits in the above-mentioned tool plug conical flask the accurate Diluted Alcohol 20ml that adds, close plug claims to decide weight, supersound process (power 250W, frequency 33KHz) 30 minutes, take out, put to room temperature, claim to decide weight, supply the weight that subtracts mistake, shake up with Diluted Alcohol, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, press the algoscopy item operation down of text need testing solution, measure, promptly.Measurement result is as follows:
The test of peoniflorin average recovery
| Numbering | Weighing (g) | Peoniflorin amount (mg) in the test sample | Peoniflorin addition (mg) | Measured value (mg) | The response rate (%) |
| 1 2 3 4 5 6 | 0.74809 0.75532 0.74334 0.73917 0.75508 0.74096 | 0.6876 0.6943 0.6833 0.6794 0.6941 0.6811 | 0.51440 0.51440 0.61728 0.61728 0.72016 0.72016 | 1.1890 1.1990 1.2933 1.2818 1.4008 1.3958 | 97.47 98.11 98.82 97.59 98.13 99.24 |
Average recovery rate=98.23%, RSD=0.70%.
The result shows that the response rate is good.
10 sample sizes are measured the preparation and the operation down of chromatographic condition item of pressing need testing solution, measure ten batch samples, and the result is as follows:
Content of paeoniflorin in ten batches of scorching micropills that relax of woman
| Lot number | Peoniflorin average content (mg/ bag) |
| 1 2 3 4 5 6 7 8 9 10 | 3.021 2.855 3.244 2.907 2.531 3.009 2.485 2.733 2.814 2.832 |
The high performance liquid chromatography content assaying method of tanshinone research in experimental example 12 dispersible tablets:
1 instrument and reagent
1.1 key instrument
High performance liquid chromatograph 2010Aht SHIMADZU
The general all purpose instrument company limited of analysing in ultraviolet spectrophotometer TU-1800SPC Beijing
Electronic analytical balance BP211D SARTORIUS
Ultrasonic washing unit KQ250B Kunshan Ultrasonic Instruments Co., Ltd.
1.2 reagent
Methanol analytical pure Beijing Chemical Plant
Acetonitrile chromatographically pure CALEDON
The pure Beijing of phosphoric acid top grade chemical reagents corporation
The pure water WAHAHA
It is an amount of that the 2 selection precisions that detect wavelength take by weighing the tanshinone reference substance, adds methanol and make the solution that every 1ml contains 15 μ g, in the interscan of 200～400nm wave-length coverage.The result shows that tanshinone has absorption maximum at the 270nm place, therefore selects 270nm as the detection wavelength of measuring tanshinone glycosides content in the scorching dispersible tablet that relaxes of woman.
3 chromatographic conditions
Chromatographic column: Diamonsil ODS 250mm * 4.6mm 5 μ m
Mobile phase: methanol-water=78: 22
Flow velocity: 1.0ml/min
Sample size: 10 μ l
Detect wavelength: 270nm
Test and Selection tanshinone as its index components, but owing to there is composition like the more or polar phase close in the preparation with the tanshinone structure, the caffeic acid in Caulis Lonicerae, the Herba Taraxaci for example, indigo composition such as grade in the Folium Isatidis.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of high performance liquid chromatography effect quality is the composition of elution requirement, particularly mobile phase.Therefore, experiment sieving multiple elution requirement, part elution requirement and result are as follows:
The investigation of chromatographic condition
| Condition | The result |
| Methanol-water=60: 20 octadecylsilane chemically bonded silica methyl alcohol-0.05mol/L sodium dihydrogen phosphate=15: 85 eight alkyl silane bonded silica gel acetonitrile-waters=50: 50 dialkyl silane bonded silica gel methanol-water=80: 20 octadecylsilane chemically bonded silica methanol-water=75: 25 octadecylsilane chemically bonded silica methanol-water=78: 22 octadecylsilane chemically bonded silicas | Separating incomplete appearance time, to separate incomplete retention time more slowly moderate, it is more clear to separate, negative noiseless retention time is moderate, it is more clear to separate, negative noiseless retention time is moderate, it is the most clear to separate, negative noiseless |
Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, methanol-water=78: 22 be a mobile phase, and with this understanding, the tanshinone retention time is moderate, and the peak is capable sharp-pointed, and symmetry is separated fully with adjacent peak, and feminine gender is noiseless.
This product powder 1g is got in the preparation of 4 need testing solutions, and accurate the title decides, and puts in the tool plug conical flask, and the accurate methanol 20ml that adds claims to decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, as need testing solution.
The investigation precision of 5 linear relationships is measured the tanshinone reference substance solution of variable concentrations, and the therefrom accurate respectively 10 μ l that draw inject chromatograph of liquid, according to high effective liquid chromatography for measuring.With the peak area is vertical coordinate, and tanshinone sample size (ng) is figure for abscissa, the drawing standard curve.The result is as follows:
The tanshinone linear relationship
| Numbering | Sample size (ng) | Peak area |
| 1 2 3 4 5 | 115.92 173.88 231.84 289.80 347.76 | 679.891 1017.30 1369.29 1709.56 2047.02 |
Regression equation: Y=5.9119x-5.9966
Correlation coefficient: γ=0.9999
The result shows that tanshinone linear relationship between 115.92ng～347.76ng is good.
Through calculating, the tanshinone standard curve is one to cross the straight line of initial point, therefore selects one point external standard method to measure the content of tanshinone in the scorching dispersible tablet that relaxes of woman.
The test of 6 precision is accurate draws with a tanshinone reference substance solution 10 μ l, injects chromatograph of liquid, and replication 5 times is investigated reference substance solution precision, and the result is as follows:
The precision test
| Test number (TN) | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD(％) |
| Peak area | 1320.2 | 1352.6 | 1347 | 1348.7 | 1329.7 | 1339.6 | 0.93 |
The result shows that reference substance solution precision is good.
7 need testing solution stability tests are accurate to be drawn with a need testing solution 10 μ l, injects chromatograph of liquid, measures at 0,2,4,8,24 hour sample introduction respectively, and the result is as follows:
The need testing solution stability test
| Testing time (h) | 0 | 2 | 4 | 8 | 24 | Meansigma methods | RSD(％) |
| Peak area | 1235.1 | 1225.4 | 1238.7 | 1220.9 | 1236.7 | 1231.36 | 0.56 |
The result shows that need testing solution is good at 24 hours internal stabilities.
8 replica tests are got this product of same lot number, and porphyrize is got about 1g (totally 5 parts), and accurate the title decides, and press the preparation and the operation down of algoscopy item of text need testing solution, and the result is as follows:
Replica test
| Numbering | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD(％) |
| Content (mg/ sheet) | 0.095 | 0.103 | 0.098 | 0.097 | 0.101 | 0.099 | 2.89 |
The result shows that repeatability is good.
This product that same lot number is got in the test of 9 average recoveries, porphyrize is got about 0.5g (totally 6 parts), and accurate the title, decide, and splits in the tool plug conical flask; The respectively accurate tanshinone reference substance solution that adds is an amount of, makes the tanshinone of adding suitable with the amount of the contained tanshinone of sample reality.The accurate methanol 20ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.Measure tanshinone average recovery rate=98.57% as a result, RSD=1.87% in accordance with the law.
The result shows that the response rate is good.
10 sample sizes are measured by operating under chromatographic condition and the need testing solution preparation, measure ten batch samples, and the result is as follows:
The content of tanshinone in the scorching dispersible tablet that relaxes of the ten crowdes of woman
| Lot number | Tanshinone average content (mg/ sheet) |
| 1 2 3 4 5 6 7 8 9 10 | 0.098 0.085 0.121 0.115 0.089 0.096 0.092 0.105 0.097 0.083 |
The high performance liquid chromatography content assaying method of salvianolic acid B research in experimental example 13 drop pills:
1 instrument and reagent
1.1 key instrument
High performance liquid chromatograph 2010Aht SHIMADZU
The general all purpose instrument company limited of analysing in ultraviolet spectrophotometer TU-1800SPC Beijing
Electronic analytical balance BP211D SARTORIUS
Ultrasonic washing unit KQ250B Kunshan Ultrasonic Instruments Co., Ltd.
1.2 reagent
Methanol analytical pure Beijing Chemical Plant
Acetonitrile chromatographically pure CALEDON
The pure Beijing of formic acid top grade chemical reagents corporation
The pure water WAHAHA
It is an amount of that the 2 selection precisions that detect wavelength take by weighing the salvianolic acid B reference substance, adds methanol and make the solution that every 1ml contains 100 μ g, in the interscan of 200～400nm wave-length coverage.The result shows that salvianolic acid B has absorption maximum at the 286nm place, therefore selects 286nm as the detection wavelength of measuring content of danshinolic acid B in the scorching drop pills of woman.
3 chromatographic conditions
Chromatographic column: Diamonsil ODS 250mm * 4.6mm 5 μ m
Mobile phase: methanol-acetonitrile-formic acid-water=27: 8: 2: 63
Flow velocity: 1.0ml/min
Sample size: 10 μ l
Detect wavelength: 286nm
Test and Selection salvianolic acid B as its index components, but owing to there is composition like the more or polar phase close in the preparation with the salvianolic acid B structure, the chlorogenic acid in Caulis Lonicerae, the Herba Taraxaci for example, compositions such as the glycyrrhizic acid in the Radix Glycyrrhizae.Have only the interference of getting rid of these compositions, could obtain ideal chromatograph effect.The key factor of high performance liquid chromatography effect quality is the composition of elution requirement, particularly mobile phase.Therefore, experiment sieving multiple elution requirement, part elution requirement and result are as follows:
The investigation of chromatographic condition
| Condition | The result |
| Acetonitrile-formic acid-water=30: 10: 60 octadecylsilane chemically bonded silica methanol-water=60: 20 octadecylsilane chemically bonded silica methyl alcohol-0.05mol/L sodium dihydrogen phosphate=15: 85 eight alkyl silane bonded silica gel methyl alcohol-acetonitrile-formic acid-water=35: 8: 2: 55 dialkyl silane bonded silica gel methyl alcohol-acetonitrile-formic acid-water=26: 8: 3: 66 octadecylsilane chemically bonded silica methyl alcohol-acetonitrile-formic acid-water=30: 10: 2: 60 octadecylsilane chemically bonded silica methyl alcohol-acetonitrile-formic acid-water=25: 7: 1: 59 octadecylsilane chemically bonded silica methyl alcohol-acetonitrile-formic acid-water=27: 8: 2: 63 octadecylsilane chemically bonded silicas | Not exclusively the incomplete separation of separation is more clear to separate the incomplete separation of incomplete separation, negative noiseless separation is more clear, negative noiseless separation is more clear, negative noiseless retention time is moderate, it is the most clear to separate, negative noiseless |
Through screening, determined with the octadecylsilane chemically bonded silica to be immobile phase, methanol-acetonitrile-formic acid-water=27: 8: 2: 63 is mobile phase, with this understanding, the salvianolic acid B retention time is moderate, and the peak is capable sharp-pointed, symmetry is separated fully with adjacent peak, and is negative noiseless.
This product powder 0.3g is got in the preparation of 4 need testing solutions, and accurate the title decides, and puts in the tool plug conical flask, and the accurate 75% methanol 20ml that adds claims to decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 75% methanol, shake up, filter, get subsequent filtrate, as need testing solution.
The investigation precision of 5 linear relationships is measured the salvianolic acid B reference substance solution of variable concentrations, and the therefrom accurate respectively 10 μ l that draw inject chromatograph of liquid, according to high effective liquid chromatography for measuring.With the peak area is vertical coordinate, and salvianolic acid B sample size (μ g) is figure for abscissa, the drawing standard curve.The result is as follows:
The salvianolic acid B linear relationship
| Numbering | Sample size (μ g) | Peak area |
| 1 2 3 4 5 | 0.5057 0.7586 1.0115 1.2643 1.5172 | 1342.49 2043.64 2699.37 3367.11 4036.29 |
Regression equation: y=2654x+13.349
Correlation coefficient: γ=0.9999
The result shows that salvianolic acid B linear relationship between 0.5057 μ g～1.5172 μ g is good.
Through calculating, the salvianolic acid B standard curve is one to cross the straight line of initial point, therefore selects one point external standard method to measure the content of salvianolic acid B in the scorching drop pills of woman.
The test of 6 precision is accurate draws with a salvianolic acid B reference substance solution 10 μ l, injects chromatograph of liquid, and replication 5 times is investigated reference substance solution precision, and the result is as follows:
The precision test
| Test number (TN) | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD (％) |
| Peak area | 2803.9 | 2873.4 | 2845.1 | 2812.5 | 2804.6 | 2827.9 | 0.96 |
The result shows that reference substance solution precision is good.
7 need testing solution stability tests are accurate to be drawn with a need testing solution 10 μ l, injects chromatograph of liquid, measures at 0,2,4,8,24 hour sample introduction respectively, and the result is as follows:
The need testing solution stability test
| Testing time (h) | 0 | 2 | 4 | 8 | 24 | Meansigma methods | RSD(％) |
| Peak area | 2954.1 | 2905.7 | 2941.2 | 2948.6 | 2926.5 | 2935.2 | 0.59 |
The result shows that need testing solution is good at 24 hours internal stabilities.
8 replica tests are got this product of same lot number, and porphyrize is got about 0.3g (totally 5 parts), and accurate the title decides, and press the preparation and the operation down of algoscopy item of text need testing solution, and the result is as follows:
Replica test
| Numbering | 1 | 2 | 3 | 4 | 5 | Meansigma methods | RSD(％) |
| Content (mg/ bag) | 4.62 | 4.57 | 4.76 | 4.67 | 4.74 | 4.67 | 1.53 |
The result shows that repeatability is good.
This product that same lot number is got in the test of 9 average recoveries, porphyrize is got about 0.15g (totally 6 parts), and accurate the title, decide, and splits in the tool plug conical flask; The respectively accurate salvianolic acid B reference substance solution that adds is an amount of, makes the salvianolic acid B of adding suitable with the amount of the contained salvianolic acid B of sample reality.The accurate 75% methanol 20ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 75% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.Measure salvianolic acid B average recovery rate=98.7% as a result, RSD=2.07% in accordance with the law.
The result shows that the method response rate is good.
10 sample sizes are measured the preparation and the operation down of chromatographic condition item of pressing need testing solution, measure ten batch samples, and the result is as follows:
The content of salvianolic acid B in the scorching drop pills of the ten crowdes of woman
| Lot number | Salvianolic acid B average content (mg/ bag) |
| 1 2 3 4 5 6 7 8 9 10 | 4.75 4.62 4.59 4.87 4.10 4.57 4.29 4.28 4.06 4.71 |
Concrete embodiment
Embodiments of the invention 1: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and adds consumption and be 50% starch, and mixing is crossed 100 mesh sieves, and to be that 95% ethanol is general be micropill to concentration, uses coating, after the drying, promptly gets micropill, oral, and three times on the one, 3g/ time.
Embodiments of the invention 2: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and adds consumption and be 30%% starch, and mixing is crossed 100 mesh sieves, and to be that 60% ethanol is general be micropill to concentration, uses coating, after the drying, promptly gets micropill.
Embodiments of the invention 3: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and adds consumption and be 70% starch, and mixing is crossed 100 mesh sieves, and to be that 80% ethanol is general be micropill to concentration, uses coating, after the drying, promptly gets micropill.
Embodiments of the invention 4: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized; Get CMC-Na: PVPP: mannitol=add an amount of pigment mixing as pharmaceutical adjunct at 1: 2: 3, get 3/5 pharmaceutical adjunct and the medicated powder mix homogeneously that is equivalent to 10 times of amounts of CMC-Na approximately, PVP-K30 anhydrous alcohol solution with 2% is made binding agent, 40 order system material, granulate, remain 2/5 pharmaceutical adjunct and an amount of caramel color mixing, be added on outward in the particle that makes, tabletting promptly gets dispersible tablet.
Embodiments of the invention 5: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and crosses 80 mesh sieves, presses medication amount: substrate amount=1: 1.2 adding soybean oil, mixing; The prescription of rubber is a gelatin: glycerol: water: titanium dioxide=100g: 40g: 100g: 1g, batchingization adhesive tape part is: weigh batching, in the inputization glue jar, merceration is warming up to 65 ± 5 ℃ gradually after 30 minutes, stirred 5 hours and simultaneously evacuation remove bubble, treat evenly back blowing of sizing material, incapsulate after the filtration in the sizing material bucket of machine; The debugging pellet press, 65 ℃ of gelatin box temperature controls, mould rotating speed 2.0 is rolled in 45 ℃ of sprinkler body temperature controls, rubber thickness 0.8mm, 18～25 ℃ of indoor temperatures, relative humidity＜40%, pelleting; The dry typing drying of rolling that adopts combined with two steps of tray dried, dry 2 hours of the typing of rolling, and 22 ℃ of baking temperatures, dry relative humidity should be lower than 40%, and promptly got soft capsule at 24～48 hours drying time.
Embodiments of the invention 6: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, the powder portion of getting it filled, PEG4000 portion, mix homogeneously, fuse in the water-bath, stir evenly, drip and in dimethicone, to become ball, drip apart from 5cm, water dropper bore 2.5mm/2mm, mix 70 ℃ of ointment temperature, liquid coolant height 60cm promptly gets drop pill.
Embodiments of the invention 7: Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g
Get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby.Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, vacuum drying, pulverize, do diluent, 90% ethanol and soybean oil system soft material with starch, with the soft material that makes micropill mechanism ball, wet feed pushed the 0.8mm sieve aperture, the wet grain of strip cuts off round as a ball, at 50～60 ℃ of drying and mouldings, cross 10 mesh sieves and select ball, promptly
The thin layer chromatography of rhubarb medicinal material, chrysophanic acid is differentiated in embodiment 8 pellets
Get this product powder 4g, add methanol 30ml supersound process, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the chrysophanic acid reference substance, add methanol and make the reference substance solution that every ml contains 1mg; Test according to thin layer chromatography, draw above-mentioned need testing solution 20 μ l, control medicinal material solution 5 μ l respectively, contrast drug solution 5 μ l, the point in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid=15: 5: 1 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color.
The thin layer chromatography of rhubarb medicinal material is differentiated in embodiment 9 dispersible tablets
Get this product powder 1g, add ethanol 20ml supersound process, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 1g, shines medical material solution in pairs with legal system; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, upper solution with petroleum ether (60～90 ℃)-Ethyl formate-formic acid=12: 7: 1.5 is developing solvent, launches, and takes out, dry, put under the 254nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
The thin layer chromatography of rhubarb medicinal material is differentiated in embodiment 10 dispersible tablets
Get this product powder 2g, add ethanol 30ml supersound process, filter, filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.5g, shines medical material solution in pairs with legal system; According to thin layer chromatography test, draw each 3 μ l of above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, be developing solvent with petroleum ether (60～90 ℃)-Ethyl formate-formic acid=18: 3: 0.5, launch, taking-up is dried, and puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
The thin layer chromatography of tanshinone is differentiated in the embodiment 11 micropill tablets
Get this product powder 8g, the 30ml supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets the tanshinone reference substance, adds ethyl acetate and makes the reference substance solution that every ml contains 0.5mg; According to thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate=9.5: 0.5, launch, taking-up is dried; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of red rooted salvia, tanshinone is differentiated in embodiment 12 dispersible tablets
Get this product powder 3g, the 20ml supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets red rooted salvia 0.5g, shines medical material solution in pairs with legal system; Get the tanshinone reference substance, add ethyl acetate and make the reference substance solution that every ml contains 0.5mg; According to thin layer chromatography test, draw above-mentioned reference substance solution, control medicinal material solution, each 3 μ l of need testing solution respectively, put on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate=9.2: 0.8, launch, taking-up is dried; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color.
The thin layer chromatography of tanshinone is differentiated in embodiment 13 soft capsules
Get this product powder 5g, the 30ml supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets the tanshinone reference substance, adds ethyl acetate and makes the reference substance solution that every ml contains 1mg; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, be developing solvent with benzene-ethyl acetate=9.8: 0.2, launch, taking-up is dried; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of tetrahydropalmatine is differentiated in embodiment 14 pellets
Get this product powder 8g, add methanol 30ml supersound process, filter, filtrate evaporate to dryness, residue add ammonia solution 40ml makes dissolving, extracts 2 times with the ether jolting, and each 20ml merges ether solution, volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methanol and makes the reference substance solution that every ml contains 0.5mg; Test according to thin layer chromatography, draw above-mentioned need testing solution 5 μ l, control medicinal material solution 2 μ l respectively, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with toluene-acetone=9: 2 was developing solvent, launched, and took out, dry, put in the iodine cylinder after about 1 minute and take out, wave the iodine that adsorbs on the most plate after, put under the uviol lamp and inspect under the 365nm; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of corydalis tuber medicinal material, tetrahydropalmatine is differentiated in embodiment 15 dispersible tablets
Get this product powder 3g, add ethanol 20ml supersound process, filter, filtrate evaporate to dryness, residue add ammonia solution 40ml makes dissolving, extracts with ether 30ml jolting, and ether solution volatilizes, and residue adds methanol 2ml makes dissolving, as need testing solution; Other gets corydalis tuber medicinal material 1g, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make the reference substance solution that every ml contains 0.5mg; Test according to thin layer chromatography, draw above-mentioned control medicinal material solution, reference substance solution, each 5 μ l of need testing solution respectively, point is on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with toluene-acetone=11: 1 was developing solvent, launched, and took out, dry, put in the iodine cylinder after about 1 minute and take out, wave the iodine that adsorbs on the most plate after, put under the uviol lamp and inspect under the 254nm; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of corydalis tuber medicinal material, tetrahydropalmatine is differentiated in embodiment 16 soft capsules
Get this product content 2g, add ethanol 20ml supersound process, filter, the filtrate evaporate to dryness, residue ammonification 30ml test solution makes dissolving, extracts with ether 40ml jolting, and ether solution volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets corydalis tuber medicinal material 1g, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make the reference substance solution that every ml contains 0.5mg; According to the thin layer chromatography test, draw above-mentioned control medicinal material solution, reference substance solution, each 5 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, with benzene-acetone=7: 3 was developing solvent, launched, and took out, dry, put in the iodine steam and smoke, put under the uviol lamp and inspect under the 365nm to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of licorice medicinal materials, glycyrrhizic acid is differentiated in embodiment 17 granules
Get this product 2g, add methanol 30ml supersound process, extracting solution evaporate to dryness, residue add water 15ml dissolving, water liquid extracts 2 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol, discard water lotion, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds the water supersound process in addition, and extracting solution extracts 2 times with water saturated n-butyl alcohol jolting, each 20ml, merge n-butyl alcohol liquid,, discard water lotion with the saturated water washing of n-butyl alcohol, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Extracting liquorice acid ammonium reference substance adds methanol and makes the reference substance solution that every ml contains 0.5g; Test according to thin layer chromatography, draw above-mentioned control medicinal material solution, reference substance solution, each 5 μ l of need testing solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with ethyl acetate-formic acid-glacial acetic acid-water=14: 1: 1: 2 was developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
The thin layer chromatography of licorice medicinal materials, glycyrrhizic acid is differentiated in embodiment 18 capsules
Get this product content 3g, add ethanol 20ml supersound process, extracting solution evaporate to dryness, residue add water 15ml dissolving, water liquid extracts 2 times with water saturated n-butyl alcohol jolting, each 20ml merges n-butyl alcohol liquid, with the saturated water washing of n-butyl alcohol, discard water lotion, n-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds water 20ml supersound extraction in addition, and extracting solution extracts with water saturated n-butyl alcohol 30ml jolting, n-butyl alcohol liquid discards water lotion, n-butyl alcohol liquid evaporate to dryness with the saturated water 10ml washing of n-butyl alcohol, residue adds ethanol 1ml makes dissolving, in contrast medical material solution; Extracting liquorice acid mono-ammonium reference substance adds methanol and makes the reference substance solution that every ml contains 0.5g; Test according to thin layer chromatography, draw above-mentioned control medicinal material solution, reference substance solution, each 5 μ l of need testing solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with ethyl acetate-formic acid-glacial acetic acid-water=12: 1.5: 0.5: 3 was developing solvent, launches, and takes out, dry, spray is with 20% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts under the ultra-violet lamp 254nm and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
The thin layer chromatography of licorice medicinal materials is differentiated in embodiment 19 pellets
Get this product powder 2g, add ethanol 30ml supersound process, the extracting solution evaporate to dryness, residue adds water 20ml dissolving, and water liquid extracts with water saturated n-butyl alcohol 30ml jolting, and n-butyl alcohol liquid washs with the saturated water 10ml of n-butyl alcohol, discard water lotion, n-butyl alcohol liquid evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds water 20ml supersound extraction in addition, and extracting solution extracts with water saturated n-butyl alcohol 30ml jolting, n-butyl alcohol liquid discards water lotion, n-butyl alcohol liquid evaporate to dryness with the saturated water 10ml washing of n-butyl alcohol, residue adds ethanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution respectively, point is on same silica gel g thin-layer plate, with ethyl acetate-formic acid-glacial acetic acid-water=18: 0.5: 1.5: 1 was developing solvent, launches, and takes out, dry, spray is with 20% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts under the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
The thin layer chromatography of Radix Paeoniae Rubra medical material, peoniflorin is differentiated in embodiment 20 pellets
Get this product powder 1g, add ethanol 30ml supersound process, extracting solution evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol and make the reference substance solution that every ml contains 0.5mg; Test according to thin layer chromatography, draw above-mentioned control medicinal material solution, reference substance solution, each 3 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-ethanol-glacial acetic acid=40: 5: 15: 0.5 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of Radix Paeoniae Rubra medical material, peoniflorin is differentiated in embodiment 21 dispersible tablets
Get this product powder 2g, add methanol 25ml supersound process, extracting solution evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol and make the reference substance solution that every ml contains 1mg; Test according to thin layer chromatography, draw above-mentioned control medicinal material solution, reference substance solution, each 3 μ l of need testing solution, put respectively on same silica gel g thin-layer plate, with dichloromethane-ethyl acetate-methanol-formic acid=35: 7: 12: 0.8 was developing solvent, launch, take out, dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The thin layer chromatography of peoniflorin is differentiated in embodiment 22 microcapsules
Get this product powder 3g, add methanol 20ml supersound process, extracting solution evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds ethanol and makes the reference substance solution that every ml contains 1mg; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, with dichloromethane-ethyl acetate-methanol-formic acid=45: 3: 18: 0.2 was developing solvent, launches, and takes out, dry, spray is with 2% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The high performance liquid chromatography assay of peoniflorin in embodiment 23 pellets
Get this product powder 1.5g, the accurate title, decide, and puts in the tool plug conical flask, and the accurate Diluted Alcohol 20ml that adds is an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol and make the reference substance solution that every ml contains 80 μ g; With the octadecylsilane chemically bonded silica is filler, and acetonitrile-0.02mol/L sodium dihydrogen phosphate=12: 88 is a mobile phase, and the detection wavelength is 230nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains peoniflorin with dosage and must not be less than 6.7mg every day.
The high performance liquid chromatography assay of peoniflorin in embodiment 24 dispersible tablets
Get this product powder 2.0g, the accurate title, decide, and puts in the tool plug conical flask, and the accurate methanol 20ml that adds is an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol and make the reference substance solution that every ml contains 40 μ g; With the octadecylsilane chemically bonded silica is filler, and acetonitrile-0.01mol/L sodium dihydrogen phosphate=10: 90 is a mobile phase, and the detection wavelength is 220nm; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains peoniflorin with dosage and must not be less than 3.0mg every day.
The high performance liquid chromatography assay of peoniflorin in embodiment 25 granules
Get this product powder 1.0g, the accurate title, decide, and puts in the tool plug conical flask, and the accurate Diluted Alcohol 30ml that adds is an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol and make the reference substance solution that every ml contains 80 μ g; With the octadecylsilane chemically bonded silica is filler, and acetonitrile-0.08mol/L potassium dihydrogen phosphate=14: 86 is a mobile phase, and the detection wavelength is 240nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with standard curve method, this product contains peoniflorin with dosage and must not be less than 6.0mg every day.
The high performance liquid chromatography assay of tanshinone in embodiment 26 dispersible tablets
Get this product powder 0.1g, accurate claim surely, put in the tool plug conical flask, the accurate ethanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with ethanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol and make the reference substance solution that every ml contains 20 μ g; With octadecylsilane chemically bonded silica is filler, and methanol-water=75: 25 is a mobile phase; The detection wavelength is 260nm; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 5.0mg every day.
The high performance liquid chromatography assay of tanshinone in embodiment 27 pellets
Get this product powder 0.2g, accurate claim surely, put in the tool plug conical flask, the accurate methanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol and make the reference substance solution that every ml contains 30 μ g; With octadecylsilane chemically bonded silica is filler, and methanol-water=78: 22 is a mobile phase; The detection wavelength is 270nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 10mg every day.
The high performance liquid chromatography assay of tanshinone in embodiment 28 pellets
Get this product powder 0.1g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 30ml that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol and make the reference substance solution that every ml contains 20 μ g; With octadecylsilane chemically bonded silica is filler, and methanol-water=80: 20 is a mobile phase; The detection wavelength is 280nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with standard curve method, this product contains tanshinone with dosage and must not be less than 10mg every day.
The high performance liquid chromatography assay of salvianolic acid B in embodiment 29 drop pills
Get this product powder 0.3g, accurate claim surely, put in the tool plug conical flask, the accurate 75% methanol 20ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 75% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 75% methanol and make the reference substance solution that every ml contains 0.1mg; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=27: 8: 2: 63 is mobile phase; The detection wavelength is 286nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 11mg every day.
The high performance liquid chromatography assay of salvianolic acid B in embodiment 30 dispersible tablets
Get this product powder 0.5g, accurate claim surely, put in the tool plug conical flask, the accurate 50% methanol 25ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 50% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 50% methanol and make the reference substance solution that every ml contains 0.1mg; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=25: 7: 3: 66 is mobile phase; The detection wavelength is 283nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 11mg every day.
The high performance liquid chromatography assay of salvianolic acid B in embodiment 31 pellets
Get this product powder 0.5g, accurate claim surely, put in the tool plug conical flask, the accurate 50% methanol 25ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 50% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 50% methanol and make the reference substance solution that every ml contains 0.8mg; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=28: 10: 2: 59 is mobile phase; The detection wavelength is 283nm; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 5.5mg every day.
The high performance liquid chromatography assay of salvianolic acid B in embodiment 32 microcapsules
Get this product powder 0.3g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 25ml that adds claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution.It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds methanol and make the reference substance solution that every ml contains 0.8mg; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=30: 9: 1: 61 is mobile phase; The detection wavelength is 289nm; Accurate respectively reference substance solution and each 15 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 5.5mg every day.
Claims (12)
1, a kind of scorching preparation that relaxes of woman for the treatment of gynaecopathia, it is characterized in that: calculate according to weight, it is with Caulis Lonicerae 750g, Caulis Sargentodoxae 750g, Radix Glycyrrhizae 75g, Folium Isatidis 75g, Herba Taraxaci 225g, Radix Paeoniae Rubra 225g, Radix et Rhizoma Rhei (processed) 150g, Radix Salviae Miltiorrhizae 225g, Rhizoma Polygoni Cuspidati 225g, Fructus Toosendan 225g, Rhizoma Corydalis (processed) 225g all the acceptable dosage forms on the pharmaceutics that are made comprise: injection, the powder pin, freeze-dried powder, tablet, dispersible tablet, capsule, soft capsule, microcapsule, granule, pill, micropill, powder, drop pill, slow releasing preparation, controlled release preparation, gel, oral liquid, soft extract, extractum and membrane.
2, according to the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 1, it is characterized in that: described preparation is dispersible tablet, soft capsule, granule, micropill, gel, oral liquid, drop pill, sustained-release preparation, tablet, effervescent tablet or capsule.
3, according to the method for making of the scorching preparation that relaxes of the woman of claim 1 or 2 described treatment gynaecopathias, it is characterized in that: get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, standby; Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, and makes different preparations then respectively.
4, according to the method for making of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 3, it is characterized in that: the pellet in the described preparation prepares like this: get Rhizoma Corydalis (processed), be ground into fine powder, cross 100 mesh sieves, and standby; Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Rhizoma Polygoni Cuspidati add ethanol, and the heating and refluxing extraction secondary adds the ethanol of 10 times of amounts the first time, the ethanol that adds for the second time 8 times of amounts, each 2 hours, merge extractive liquid,, filter, filtrate recycling ethanol and when being concentrated into 60 ℃ relative density be 1.36 thick paste, standby; Seven flavors such as medicinal residues and all the other Caulis Loniceraes decoct with water three times, add 8 times of water gagings for the first time and decoct 2 hours, all add 6 times of water gagings for the second time, for the third time, decocted collecting decoction respectively 1.5 hours, 1 hour, filter, relative density was 1.36 thick paste when filtrate decompression was concentrated into 60 ℃; Two kinds of thick pastes and corydalis tuber medicinal material powder are mixed in one, and vacuum drying is pulverized, general method for making or mould the method for making pill, promptly.
5, according to the method for making of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 4, it is characterized in that: described general method for making pill is such: adding consumption is the starch of 30%-70%, with the medicine mixing, cross 100 mesh sieves, concentration is that the ethanol of 60-95% is general for micropill, coating is after the drying, promptly.
6, according to the method for making of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 5, it is characterized in that: described general method for making pill is such: the adding consumption is 50% starch, with the medicine mixing, cross 100 mesh sieves, to be that 95% ethanol is general be micropill, coating to concentration, after the drying, promptly.
7, according to the method for making of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 4, it is characterized in that: the described method for making pill of moulding is such: do diluent, 90% ethanol and soybean oil system soft material with starch, with the soft material that makes micropill mechanism ball, wet feed pushed the 0.8mm sieve aperture, the wet grain of strip cuts off round as a ball, at 50～60 ℃ of drying and mouldings, cross 10 mesh sieves and select ball, promptly.
8, according to the scorching preparation that relaxes of the woman of any described treatment gynaecopathia in the claim 1～7, it is characterized in that: described preparation can be used to prepare the medicine for the treatment of pelvic inflammatory disease.
9, according to the quality control method of the scorching preparation that relaxes of the woman of any described treatment gynaecopathia in the claim 1～7, it is characterized in that content assaying method comprises following all or part of content:
A. the high performance liquid chromatography assay of peoniflorin in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and accurate adding Diluted Alcohol or methanol are an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol or methanol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol or alcoholic solution is made reference substance solution; With the alkyl silane bonded silica gel is filler, acetonitrile-0.01mol/L～0.08mol/L sodium dihydrogen phosphate or 0.01mol/L～0.08mol/L potassium dihydrogen phosphate=10～14: 90～86 is mobile phase, and the detection wavelength is among 220～240nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method or standard curve method, this product contains peoniflorin with dosage and must not be less than 3.0mg every day;
B. the high performance liquid chromatography assay of tanshinone in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, accurate add methanol or ethanol an amount of, claim decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol or ethanol, shakes up, filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol or ethanol is made reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-water=75～80: 25～20 is mobile phase; The detection wavelength is among 260～280nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 0.40mg every day;
C. the high performance liquid chromatography assay of salvianolic acid B in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and accurate 50～100% methanol that add are an amount of, claim to decide weight, supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 50～100% methanol, shake up, filter, get subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 50～100% methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=25～30: 7～10: 1～3: 59～66 is mobile phase; The detection wavelength is among 283～289nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains salvianolic acid B with dosage and must not be less than 5.5mg every day.
10, according to the quality control method of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 9, it is characterized in that: content assaying method comprises following all or part of content:
A. the high performance liquid chromatography assay of peoniflorin in the preparation
It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, and the accurate Diluted Alcohol that adds is an amount of, close plug, claim decide weight, supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, as need testing solution with microporous filter membrane; Get peoniflorin, add methanol and make reference substance solution; With the octadecylsilane chemically bonded silica is filler, and acetonitrile-0.02mol/L sodium dihydrogen phosphate=12: 88 is a mobile phase, and the detection wavelength is 230nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains peoniflorin with dosage and must not be less than 6.7mg every day;
B. the high performance liquid chromatography assay of tanshinone in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, and the accurate methanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the tanshinone reference substance, adds methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, and methanol-water=78: 22 is a mobile phase; The detection wavelength is 270nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 0.75mg every day;
C. the high performance liquid chromatography assay of salvianolic acid B in the preparation
It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, and accurate 75% methanol that adds is an amount of, claims decide weight, and supersound process is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 75% methanol, shakes up, and filtration is got subsequent filtrate, as need testing solution; It is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds 75% methanol and make reference substance solution; With octadecylsilane chemically bonded silica is filler, methanol-acetonitrile-formic acid-water=27: 8: 2: 63 is mobile phase; The detection wavelength is 286nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject chromatograph of liquid, measure, promptly; Calculate with one point external standard method, this product contains tanshinone with dosage and must not be less than 11mg every day.
11, according to the quality control method of the scorching preparation that relaxes of the woman of any described treatment gynaecopathia in the claim 1～7, it is characterized in that discrimination method comprises following all or part of content:
A. one or both thin layer chromatography discriminating in rhubarb medicinal material, the chrysophanic acid in the preparation
It is an amount of to get this product powder, adds methanol or ethanol supersound process, filters, and filtrate evaporate to dryness, residue add methanol or ethanol makes dissolving, as need testing solution; It is an amount of that other gets the Radix Et Rhizoma Rhei control medicinal material, shines medical material solution in pairs with legal system; Get the chrysophanic acid reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively in same be on the silica gel thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with petroleum ether-Ethyl formate or ethyl acetate-formic acid or glacial acetic acid=12～18: 3～7: 0.5～1.5 solution or upper solution are developing solvent, launch, take out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
B. one or both thin layer chromatography discriminating in red rooted salvia, the tanshinone in the preparation
It is an amount of to get this product powder, and the supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate makes dissolving, as need testing solution; Other gets the Radix Salviae Miltiorrhizae control medicinal material, shines medical material solution in pairs with legal system; Get the tanshinone reference substance, add ethyl acetate and make reference substance solution; According to thin layer chromatography test, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with toluene or benzene-ethyl acetate=9.2～9.8: 0.8～0.2 is developing solvent, launch, take out, dry; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
C. one or both thin layer chromatography discriminating in corydalis tuber medicinal material, the tetrahydropalmatine in the preparation
It is an amount of to get this product powder, adds methanol or ethanol supersound process, filters, and filtrate evaporate to dryness, residue add ammonia solution makes dissolving, extracts with the ether jolting, and ether solution volatilizes, and residue adds methanol or ethanol makes dissolving, as need testing solution; Other gets the Rhizoma Corydalis control medicinal material, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with toluene or benzene-acetone=7～11: 1～3 is developing solvent, launch, take out, dry, put in the iodine steam and smoke, put under the uviol lamp and inspect to clear spot; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
D. one or both thin layer chromatography discriminating in licorice medicinal materials, the glycyrrhizic acid in the preparation
It is an amount of to get this product powder, add methanol or ethanol supersound process, the extracting solution evaporate to dryness, residue is dissolved in water, and water liquid extracts with water saturated n-butyl alcohol jolting, the saturated water washing of n-butyl alcohol of n-butyl alcohol liquid, discard water lotion, n-butyl alcohol liquid evaporate to dryness, residue adds methanol or ethanol makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds the water supersound process in addition, and extracting solution shines medical material solution in pairs with legal system; A kind of in extracting liquorice acid ammonium or the monoammonium glycyrrhizinate reference substance adds methanol and makes reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with ethyl acetate-formic acid-glacial acetic acid-water=12～18: be developing solvent at 0.5～1.5: 0.5～1.5: 1～3, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
E. one or both thin layer chromatography discriminating in Radix Paeoniae Rubra medical material, the peoniflorin in the preparation
It is an amount of to get this product powder, adds ethanol or methanol supersound process, and extracting solution evaporate to dryness, residue add ethanol or methanol makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol or methanol is made reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel thin-layer plate, with chloroform or dichloromethane-ethyl acetate-methanol or ethanol-formic acid or glacial acetic acid=35～45: be developing solvent at 3～7: 12～18: 0.2～0.8, launch, take out, dry, spray is with the vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color.
12, according to the quality control method of the scorching preparation that relaxes of the woman of the described treatment gynaecopathia of claim 11, it is characterized in that: discrimination method comprises following all or part of content:
A. one or both thin layer chromatography discriminating in rhubarb medicinal material, the chrysophanic acid in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution; It is an amount of that other gets the Radix Et Rhizoma Rhei control medicinal material, shines medical material solution in pairs with legal system; Get the chrysophanic acid reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with 30～60 ℃ of petroleum ether-Ethyl formate-formic acid=15: 5: 1 is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
B. one or both thin layer chromatography discriminating in red rooted salvia, the tanshinone in the preparation
It is an amount of to get this product powder, and the supersound process that adds diethyl ether filters, and filtrate volatilizes, and residue adds ethyl acetate makes dissolving, as need testing solution; Other gets the Radix Salviae Miltiorrhizae control medicinal material, shines medical material solution in pairs with legal system; Get the tanshinone reference substance, add ethyl acetate and make reference substance solution; According to thin layer chromatography test, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel g thin-layer plate, and be developing solvent with toluene-ethyl acetate=9.5: 0.5, launch, taking-up is dried; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
C. one or both thin layer chromatography discriminating in corydalis tuber medicinal material, the tetrahydropalmatine in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add ammonia solution makes dissolving, extracts with the ether jolting, and ether solution volatilizes, and residue adds methanol makes dissolving, as need testing solution; Other gets the Rhizoma Corydalis control medicinal material, shines medical material solution in pairs with legal system; Get the tetrahydropalmatine reference substance, add methanol and make reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with toluene-acetone=9: 2 was developing solvent, launched, and took out, dry, put in the iodine cylinder after about 1 minute and take out, wave the iodine that adsorbs on the most plate after, put under the uviol lamp and inspect under the 365nm; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color;
D. one or both thin layer chromatography discriminating in licorice medicinal materials, the glycyrrhizic acid in the preparation
It is an amount of to get this product powder, adds the methanol supersound process, the extracting solution evaporate to dryness, and residue is dissolved in water, water liquid extracts with water saturated n-butyl alcohol jolting, and n-butyl alcohol liquid discards water lotion with the saturated water washing of n-butyl alcohol, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g adds the water supersound process in addition, and aqueous extract extracts with water saturated n-butyl alcohol jolting, and n-butyl alcohol liquid discards water lotion with the saturated water washing of n-butyl alcohol, n-butyl alcohol liquid evaporate to dryness, and residue adds methanol makes dissolving, in contrast medical material solution; A kind of in extracting liquorice acid ammonium or the monoammonium glycyrrhizinate reference substance adds methanol and makes reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with ethyl acetate-formic acid-glacial acetic acid-water=14: 1: 1: 2 was developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp 365nm and inspects; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color;
E. one or both thin layer chromatography discriminating in Radix Paeoniae Rubra medical material, the peoniflorin in the preparation
It is an amount of to get this product powder, adds the ethanol supersound process, and extracting solution evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Other gets the Radix Paeoniae Rubra medical material, shines medical material solution in pairs with legal system; Get the peoniflorin reference substance, add ethanol or methanol is made reference substance solution; Test according to thin layer chromatography, one or both and the need testing solution drawn in above-mentioned reference substance solution, the control medicinal material solution are an amount of, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-ethanol-glacial acetic acid=40: 5: 15: 0.5 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing in 105 ℃; In the test sample chromatograph, with contrast chromatograph corresponding position on, show the speckle of same color.
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| CN2006101503111A CN101057895B (en) | 2005-10-26 | 2006-10-26 | 'Fuyanshu' preparation for treating gynopathy and its quality controlling method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104459005A (en) * | 2014-11-21 | 2015-03-25 | 四川中方制药有限公司 | Quality control method of brain-clearing tranquilization liquid |
| CN104965046A (en) * | 2015-07-17 | 2015-10-07 | 江苏天晟药业有限公司 | Quality control method for cosmetics containing traditional Chinese medicinal ingredients |
| CN107014945A (en) * | 2017-05-23 | 2017-08-04 | 四川逢春制药有限公司 | A kind of detection method for the Chinese medicine preparation for treating acute infectious hepatitis |
| CN108362690A (en) * | 2018-01-10 | 2018-08-03 | 广西大学 | A kind of method of quick discriminating red sandalwood and dyestuff red sandalwood |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1891262A (en) * | 2005-07-08 | 2007-01-10 | 北京润德康医药技术有限公司 | Chinese medicine compound preparation for treating gynaecologic inflammation and its preparing method |
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2006
- 2006-10-26 CN CN2006101503111A patent/CN101057895B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104459005A (en) * | 2014-11-21 | 2015-03-25 | 四川中方制药有限公司 | Quality control method of brain-clearing tranquilization liquid |
| CN104459005B (en) * | 2014-11-21 | 2016-02-03 | 四川中方制药有限公司 | A kind of method of quality control of Qingnaofushenye |
| CN104965046A (en) * | 2015-07-17 | 2015-10-07 | 江苏天晟药业有限公司 | Quality control method for cosmetics containing traditional Chinese medicinal ingredients |
| CN107014945A (en) * | 2017-05-23 | 2017-08-04 | 四川逢春制药有限公司 | A kind of detection method for the Chinese medicine preparation for treating acute infectious hepatitis |
| CN107014945B (en) * | 2017-05-23 | 2018-07-31 | 四川逢春制药有限公司 | A kind of detection method of Chinese medicine preparation that treating acute infectious hepatitis |
| CN108362690A (en) * | 2018-01-10 | 2018-08-03 | 广西大学 | A kind of method of quick discriminating red sandalwood and dyestuff red sandalwood |
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| CN101057895B (en) | 2010-05-19 |
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