CN102353744A - Method for detecting quality of Tong Jingning capsule - Google Patents
Method for detecting quality of Tong Jingning capsule Download PDFInfo
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Abstract
The invention discloses a method detecting the quality of a Tong Jingning capsule, which enables determination to radix paeoniae lactiflorae, tuber corydalidis and licorice; a high performance liquid chromatography is used, according to the test results of chromatography condition and system applicability, the effective ingredient of radix paeoniae lactiflorae-paeoniflorin is carried out a content determination, and the subliminal limitation is prescribed that the content of radix paeoniae lactiflorae in each capsule can not be less than 1.5mg by measuring with paeoniflorin (C23H28O11). The method of the invention achieves the purpose of controlling the quality of the Tong Jingning capsule and ensures the safety and effectiveness of drug administration.
Description
Technical field
The invention belongs to the Chinese traditional patent formulation formulation art, relate in particular to a kind of quality determining method of dysmenorrhoea Yiganning capsule.
Background technology
The dysmenorrhoea Yiganning capsule is by the peaceful syrup dosage changing form of dysmenorrhoea, and the peaceful syrup of dysmenorrhoea derives from the 11 in the Sanitation Ministry medicine standard Chinese traditional patent formulation preparation (standard numbering: WS3-B-2244-96).Prescription is: Radix Angelicae Sinensis (stir-fry) 160g rhizoma cyperi (system) the 160g root of herbaceous peony (stir-fry) 160g corydalis tuber (stir-fry) 144g Ligusticum wallichii (stir-fry) 96g Radix Glycyrrhizae (processing) 64g red sage root 160g Fructus meliae toosendan (stir-fry) 144g safflower 80g; Method for making is following: above nine flavors; Except that safflower; Eight flavor boiling secondaries such as all the other Radix Angelicae Sinensis add safflower after 2 hours for the first time, continue to decoct 0.5 hour; 1.5 hours for the second time; Collecting decoction filters, the filtrating standing over night; Get supernatant; Being concentrated into relative density is 1.10～1.12 (85 ℃), puts coldly, adds the ethanol of 2 times of amounts; Stir; Left standstill 24 hours, and got supernatant and reclaim ethanol to there not being the alcohol flavor, subsequent use; Other gets sucrose 300g and adds water boil; Filter, being concentrated into relative density is 1.08 (85 ℃), with above-mentioned soup mixing; Continuing to be concentrated into relative density is 1.07～1.08 (85 ℃); Put coldly, filter, filtrating adds benzoic acid 2.5g; Ethyl-para-hydroxybenzoate 0.3g; Add water to 1000ml again, stir, promptly get.It differentiates under the item to have only a chromogenic reaction, exclusive very strong discrimination test and assays such as no thin-layer chromatography discriminating.
Dysmenorrhoea is meant before and after the gynecologic menstrual and what occur between menstrual period is one type of illness of principal character with waist sacrum, underbelly pain and general malaise.Dysmenorrhoea is common disease, frequently-occurring disease, and weight person influences normal study and work.The traditional Chinese medical science thinks, dysmenorrhoea is mainly smooth or experience perverse trend because of feelings will, causes that qi and blood is not smooth in the body, invasion and attack women uterus, qi depression to blood stasis, the damage of cloudy blood, stagnation of QI and blood may bring about pain.With the passing of time do not heal, cause that very then the Secondary cases structural disease becomes.
The dysmenorrhoea Yiganning capsule is processed by Radix Angelicae Sinensis (stir-fry), rhizoma cyperi (system), the root of herbaceous peony (stir-fry), corydalis tuber (stir-fry), Ligusticum wallichii (stir-fry), Radix Glycyrrhizae (processing), the red sage root, Fructus meliae toosendan (stir-fry), safflower; Effect with menstruction regulating and pain relieving; Be used for irregular menstruation, suffer from abdominal pain through preceding, menstrual period.We are derived from the effective proved recipe of clinical menstruction regulating and pain relieving, and collection menstruction regulating and pain relieving, vital energy regualting and blood circulation-promoting, comprehensive therapies such as protecting this nourishes blood.Radix Angelicae Sinensis blood circulation promoting nourishes blood in the side is " the menstruation regulating panacea " of successive dynasties traditional Chinese medical science high praise; The kind especially regulating functional activities of vital QI of rhizoma cyperi and the pain relieving warp has the effect of separating Yu Chang feelings will simultaneously has " the women chief commander of section " good reputation.Two herbal medicines cooperate, embody vital energy regualting and blood circulation-promoting with pain relieving, nourish blood menstruction regulating and pain relieving heart method basic admittedly.Root of herbaceous peony preserving yin to nourish blood helps Radix Angelicae Sinensis to protect the cloudy red root of women admittedly originally with capsule, is good at gentle liver relieving spasm to stop pain again; Corydalis tuber, Fructus meliae toosendan, Ligusticum wallichii, safflower, the red sage root or be longer than promoting blood circulation and stopping pain or be longer than promoting qi circulation and relieving pain in the other drug are and strengthen the active drug of analgesic effect with thorough releasing dysmenorrhoea; The collaborative onset of all medicines in the honey-fried licorice root conditioning side also can relieving spasm to stop pain.All medicines share that then menstruction regulating and pain relieving power is strong, relieving dysmenorrhea symptom fast, can eliminate again dysmenorrhoea disease because of, the merit of playing menstruction regulating and pain relieving altogether reaches the purpose of treating both principal and secondary aspect of disease.
Summary of the invention
The object of the present invention is to provide a kind of control dysmenorrhoea Yiganning capsule product quality, guarantee the quality determining method of dysmenorrhoea Yiganning capsule safe and effective for medication.
The quality determining method of a kind of dysmenorrhoea Yiganning capsule of the present invention comprises the steps:
Assay: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2010);
(1) chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-0.1% phosphoric acid (volume ratio is 15:85) is moving phase; The detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 5000;
(2) preparation of reference substance solution: it is an amount of to get the Paeoniflorin reference substance, and accurate the title decides, and adds Diluted Alcohol and processes the solution that every 1ml contains 50 μ g, promptly gets;
(3) preparation of need testing solution: get these article content powder 0.25g, the accurate title, decide, and the accurate Diluted Alcohol 25ml that adds claims to decide weight; Ultrasonic Extraction 40 minutes is taken out, and puts coldly, claims decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and with the miillpore filter filtration of 0.45 μ m, gets subsequent filtrate, promptly gets;
(4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject high performance liquid chromatograph, measure, promptly get.
The quality determining method of above-mentioned dysmenorrhoea Yiganning capsule, wherein: every of these article contain the root of herbaceous peony with Paeoniflorin (C
23H
28O
11) meter, must not be less than 1.5mg.
The quality determining method of above-mentioned dysmenorrhoea Yiganning capsule, wherein:
Differentiate
A. get these article content 10g, the 50ml that adds diethyl ether, water-bath refluxed 30 minutes; Discard ether solution, the dregs of a decoction are flung to ether, add ethanol 50ml; Water-bath refluxed 30 minutes, took out, and put cold; Filter; The filtrating evaporate to dryness, residue adds water 20ml dissolving, with water saturation extracting n-butyl alcohol 2 times; Each 20ml merges normal butyl alcohol liquid; With the saturated water 20ml washing of normal butyl alcohol, discard water liquid again, normal butyl alcohol liquid evaporate to dryness, residue add ethanol 0.5ml dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and processes the solution that every 1ml contains 2mg, as reference substance solution; Test according to thin-layered chromatography (2010 editions appendix VIB of Chinese Pharmacopoeia); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; (volume ratio is 8: 1:4: 1) be developping agent with methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia; Launch, take out, dry; Spray is with 5% vanillic aldehyde sulfuric acid solution, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
B. get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes; Take out, put coldly, filter, filtrating is steamed near and is done, and residue adds water 15ml dissolving, with extracted with diethyl ether 3 times, and each 15ml, the merging ether solution volatilizes, and residue adds the 0.5ml dissolve with methanol, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2010); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with 1% sodium hydroxide solution, be developping agent with normal hexane-methenyl choloride-methyl alcohol (volume ratio is 10: 4: 1), presaturation 20 minutes; Launch; Take out, dry, put in the iodine vapor smoked after several minutes; In air, wave the iodine of most absorption, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
C. get these article content 5g, the 40ml that adds diethyl ether, water-bath refluxed 1 hour, discarded ether solution; The dregs of a decoction are flung to ether, add methyl alcohol 40ml, and water-bath refluxed 1 hour; Filter, filtrating evaporate to dryness, residue add water 40ml dissolving; With extracting n-butyl alcohol three times, each 20ml merges normal butyl alcohol liquid; With water washing 3 times, each 20ml discards water liquid; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution; Extracting liquorice control medicinal material powder 1g shines medicinal material solution in pairs with legal system in addition; Test according to thin-layered chromatography (2010 editions appendix VIB of Chinese Pharmacopoeia); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With methenyl choloride-methyl alcohol-water (volume ratio is 13: 7: 2) subnatant is developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
The quality determining method of above-mentioned dysmenorrhoea Yiganning capsule, wherein inspection: should meet each item regulation (an appendix I of Chinese Pharmacopoeia version in 2010 L) relevant under the capsule item.
The quality determining method of above-mentioned dysmenorrhoea Yiganning capsule; Wherein: the method for making of these article is Radix Angelicae Sinensis (stir-fry) 800g; Rhizoma cyperi (system) 800g; The root of herbaceous peony (stir-fry) 800g; Corydalis tuber (stir-fry) 720g; Ligusticum wallichii (stir-fry) 480g; Radix Glycyrrhizae (processing) 320g; Red sage root 800g; Fructus meliae toosendan (stir-fry) 720g; Safflower 400 g; More than nine the flavor; Except that safflower; Eight flavor boiling secondaries such as all the other Radix Angelicae Sinensis; Add for the first time 12 times of water gagings decoctions and add safflower after 2 hours; Continue to decoct 0.5 hour; Adding for the second time 10 times of water gagings decocted 1.5 hours; Collecting decoction; Filter the filtrating standing over night; Get supernatant, being concentrated into relative density is 1.10-1.12 (85 ℃), puts coldly, adds the ethanol of 2 times of amounts, stirs, and leaves standstill 24 hours; Get supernatant and reclaim ethanol, be concentrated into thick medicinal extract, drying under reduced pressure is pulverized, and adds appropriate amount of auxiliary materials, granulates, and drying incapsulates, and processes 1000, promptly gets.
The present invention compared with prior art; Has tangible beneficial effect; Can know from above technical scheme: through experimental study repeatedly in a large number; Employing is carried out assay to the effective ingredient Paeoniflorin of the main flavour of a drug root of herbaceous peony; The root of herbaceous peony, corydalis tuber and Radix Glycyrrhizae are differentiated; Reached control dysmenorrhoea Yiganning capsule product quality, guaranteed safe and effective for medication.
Embodiment
Below further specify beneficial effect of the present invention through Test Example:
" dysmenorrhoea Yiganning capsule " pressed " the peaceful syrup of dysmenorrhoea " former prescription, is made up of nine flavor medicinal materials such as Radix Angelicae Sinensis (stir-fry), rhizoma cyperi (system), the root of herbaceous peony (stir-fry), corydalis tuber (stir-fry), Ligusticum wallichii (stir-fry), Radix Glycyrrhizae (moxibustion), the red sage root." the peaceful syrup of dysmenorrhoea " (WS3-B-2244-96) differentiates under the item to have only a chromogenic reaction, exclusive very strong discrimination tests such as no thin-layer chromatography discriminating.During experimental study among the other side Radix Angelicae Sinensis, rhizoma cyperi etc. nine flavor medicinal materials carried out exclusive very strong thin-layer chromatography discrimination test, the wherein root of herbaceous peony, corydalis tuber and noiseless, the system's favorable reproducibility of Radix Glycyrrhizae three flavor medicinal material feminine genders are so list it in text; Other several flavors can not be listed text in owing to poor reproducibility or negative is disturbed reasons such as bigger.The result is following:
(1) discriminating of Radix Angelicae Sinensis, Ligusticum wallichii:
Radix Angelicae Sinensis, Ligusticum wallichii ingredient are close, influence each other greatly, are difficult to separately differentiate, with reference to " one one of Chinese pharmacopoeia " four thing mixture " is contrast with the forulic acid reference substance simultaneously, differentiates.
Radix Angelicae Sinensis contains volatile oil and fixed oil composition.Wherein volatile oil content accounts for 0.4%, be divided into neutrality, phenol property, acid three kinds.Neutral volatile oil mainly contains Ligustilide, accounts for multiple composition such as 49% of volatile oil total amount; Phenol property volatile oil contains carvacrol, phenol, p-cresol etc.; Acid volatile oil mainly contains repefral, dimethyl azelate etc.Involatile constituent contains forulic acid, vanillic acid, new angelica lactone, nicotinic acid or the like compound.
The chemical constitution study of Ligusticum wallichii is more, contains volatile oil 1%; Alkaloid has acyl ammonia, perlolyrine etc. in ligustrazine, the isoleucyl-valine; Phenols component has forulic acid, chuanxingol, archen, caffeic acid etc.; The lactone composition has Ligustilide, new frutus cnidii lactone; Terpenoid has compositions such as spainulenol, cupreol.
Method:
The preparation of need testing solution: get these article content 4g, add 80% ethanol 30ml sonicated 30 minutes, put cold; Filter, filtrating is steamed near and is done, and residue adds water 20 ml heating for dissolving; Put cold, with ethyl acetate extraction 3 (20ml, 15ml; 15ml); Combined ethyl acetate liquid extracts 3 times with 2% sodium carbonate liquor, each 15ml; Merge alkali lye; Add watery hydrochloric acid and transfer PH=2～3, add benzene 15ml washing, abandon benzene liquid; Use ethyl acetate extraction again 3 times; Each 15ml, combined ethyl acetate liquid volatilizes; Residue adds methyl alcohol 0.5ml dissolving, as need testing solution.
The preparation of reference substance solution: it is an amount of that other gets the forulic acid reference substance, adds methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution.
The preparation of negative control solution: get the negative sample of scarce Radix Angelicae Sinensis, Ligusticum wallichii, prepare negative control article solution by the need testing solution preparation method.
Draw each 5～15 μ l of above need testing solution, reference substance solution, negative control solution respectively, put in same be on the silica gel g thin-layer plate of bonding agent with the sodium carboxymethyl cellulose, launch with following developping agent, take out, dry, the result is following:
The result shows: reasons such as negative interference, system's poor reproducibility, can't list text in.
(2) discriminating of rhizoma cyperi
The rhizoma cyperi rhizome contains volatile oil about 1%; Contain multiple sequiterpene and oxide thereof in the oil: rhizome of nutgrass flatsedge alkene, selinatriene, β-selinene, α-cyperolone, β-cyperolone, Pogostone, cyperolone, cyperol etc., also contain a small amount of monoterpene: nopinene, amphene, citrene, to umbelliprenin, lauro lene.
Method:
The preparation of need testing solution: get these article content 3g, the 25ml sonicated that adds diethyl ether 30 minutes is put coldly, filters, and filtrating evaporate to dryness, residue add ethyl acetate 0.5ml dissolving, as need testing solution.
The preparation of control medicinal material solution: get rhizoma cyperi control medicinal material powder 1g, shine medicinal material solution in pairs with legal system.
Negative control solution: get the negative sample that lacks rhizoma cyperi, the preparation method prepares negative control solution by need testing solution.
Draw each 5～15 μ l of above-mentioned need testing solution, control medicinal material solution, negative control solution respectively, put in same be on the thin layer plate of bonding agent with the sodium carboxymethyl cellulose, launch with different developping agents respectively, take out, dry, the result is following:
The result shows: need testing solution chromatogram and control medicinal material solution chromatogram do not have corresponding spot, so do not list it in text on identical position.
(3) discriminating of the root of herbaceous peony:
Root of herbaceous peony principal ingredient has: compositions such as Paeoniflorin, peonin, paeonol, albiflorin, benzoylpaeoniflorin, lacdtlorin.
Method A:
The preparation of need testing solution: get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes; Put cold; Filter, filtrating evaporate to dryness, residue add water 15ml dissolving; With water saturation extracting n-butyl alcohol 3 times; Each 15ml merges normal butyl alcohol liquid, evaporate to dryness; Residue adds ethanol 1ml dissolving; Add the 1g neutral alumina, in water-bath, mix drying thoroughly; Neutral alumina post (200 orders of packing into; 4g), collect cleansing solution, evaporate to dryness with ethyl acetate-methyl alcohol (1: 1) 30ml washing; Residue adds ethanol 1ml dissolving, as need testing solution.
The preparation of reference substance solution: it is an amount of to get the Paeoniflorin reference substance, adds ethanol and processes every 1ml and contain 2mg solution, as reference substance solution.
The preparation of negative control solution: get scarce root of herbaceous peony negative control sample, the preparation method prepares negative control solution by need testing solution.
Draw above-mentioned need testing solution, reference substance solution, each 5～15 μ l of negative control solution respectively; The point in same be on the silica gel g thin-layer plate of bonding agent with CMC-Na; Launch with different developping agents respectively; Take out; Dry, spray different developers, 105 ℃ to be heated to spot colour developing clear; Inspect under the daylight, the result is following:
| Numbering | Developping agent | Developer | The result |
| 1 | Methenyl choloride-ethyl acetate-methyl alcohol-formic acid (40: 5: 10: 0.3) | Spray 5% vanillic aldehyde sulfuric acid solution | There is not corresponding spot |
| 2 | Methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia (8: 1: 4: 1) | Spray 5% vanillic aldehyde sulfuric acid solution | There is not corresponding spot |
| 3 | The subnatant of methenyl choloride-methanol-water (13: 7: 2) | Spray 10% ethanol solution of sulfuric acid | There is not corresponding spot |
Method B:
The preparation of need testing solution: get these article content 10g, the 50ml that adds diethyl ether, water-bath refluxed 30 minutes, discarded ether solution; The dregs of a decoction are flung to ether, add ethanol 50ml, and water-bath refluxed 30 minutes, take out; Put coldly, filter the filtrating evaporate to dryness; Residue adds water 20ml dissolving, with water saturation extracting n-butyl alcohol twice, and each 20ml; Merge normal butyl alcohol liquid, with the saturated water 20ml washing of normal butyl alcohol, discard water liquid again; Normal butyl alcohol liquid evaporate to dryness, residue add ethanol 0.5ml dissolving, as need testing solution.
The preparation of reference substance solution: get the Paeoniflorin reference substance, add ethanol and process every 1ml and contain 2mg solution, as reference substance solution.
The preparation of negative control solution: get scarce root of herbaceous peony negative sample, the preparation method prepares negative control solution by need testing solution.
Draw above need testing solution, reference substance solution, each 5～15 μ l of negative control solution respectively; The point in same be on the silica gel g thin-layer plate of bonding agent with CMC-Na; Launch with different developping agents respectively; Take out; Dry, spray different developers, 105 ℃ to be heated to spot colour developing clear; Inspect under the daylight, the result is following:
| Numbering | Developping agent | Developer | The result |
| 1 | Methenyl choloride-ethyl acetate-methyl alcohol-formic acid (40: 5: 10: 0.3) | Spray 5% vanillic aldehyde sulfuric acid solution | Spot is unintelligible |
| 2 | Methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia (8: 1: 4: 1) | Spray 5% vanillic aldehyde sulfuric acid solution | Clear spot, system's favorable reproducibility |
| 3 | The subnatant of methenyl choloride-methanol-water (13: 7: 2) | Spray 10% ethanol solution of sulfuric acid | There is not corresponding spot |
The result shows that system 2 among the method B, clear spot, and favorable reproducibility, negative noiseless, so list it in text.That is: get these article content 10g, the 50ml that adds diethyl ether, water-bath refluxed 30 minutes, discarded ether solution; The dregs of a decoction are flung to ether, add ethanol 50ml, and water-bath refluxed 30 minutes, take out; Put coldly, filter the filtrating evaporate to dryness; Residue adds water 20ml dissolving, with water saturation extracting n-butyl alcohol twice, and each 20ml; Merge normal butyl alcohol liquid, with the saturated water 20ml washing of normal butyl alcohol, discard water liquid again; Normal butyl alcohol liquid evaporate to dryness, residue add ethanol 0.5ml dissolving, as need testing solution.Other gets the Paeoniflorin reference substance, adds methyl alcohol and processes every 1ml and contain 2mg solution, as reference substance solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B); Draw each 5～15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia (8: 1: 4: 1) be developping agent; Launch, take out, dry; Spray 5% vanillic aldehyde sulfuric acid solution, 105 ℃ to be heated to spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the same color spot.
(4) discriminating of corydalis tuber:
Get about more than 20 alkaloids in the corydalis tuber: d-corydaline, dl-Tetrahydropalmatine, protopine, l-tetrahydrochysene coptis root alkali, l-tetrahydrocolumbamine, corydalis tuber first element, tetrahydropalmatine, corydalis tuber element in the last of the ten Heavenly stems, the sub-element of corydalis tuber, corydalis etc.
Method:
The preparation of need testing solution: get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes; Put coldly, filter, filtrating is steamed near and is done; Residue adds water 15ml dissolving; Add ammonia solution and regulate PH to 9, with extracted with diethyl ether 3 times, 15ml at every turn; Merge ether solution; Volatilize, residue adds the 0.5ml dissolve with methanol, as need testing solution.
The preparation of reference substance solution: it is an amount of to get the tetrahydropalmatine reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.
The preparation of negative control: get scarce corydalis tuber negative sample, the preparation method prepares negative control solution by need testing solution.
Draw above need testing solution, reference substance solution, each 5～15 μ l of negative control solution respectively, put on same thin layer plate, launch with different developping agents respectively; Take out, dry, iodine vapor is smoked after several minutes; Put the iodine of waving most absorption in the air again, uviol lamp (365nm) is inspected, and the result is following:
| Numbering | Chromatoplate | Developping agent | Colour developing | The result |
| 1 | The CMC-Na-silica G | Cyclohexane-methenyl choloride-methyl alcohol (5: 3: 0.5) | Iodine vapor is smoked puts the iodine of waving most absorption in the air after several minutes again, and uviol lamp (365nm) is inspected | Spot separates bad |
| 2 | The silica G of 1%NaOH preparation | Cyclohexane-methenyl choloride-methyl alcohol (10: 4: 1) | Iodine vapor is smoked puts the iodine of waving most absorption in the air after several minutes again, and uviol lamp (365nm) is inspected | Clear spot, favorable reproducibility |
The result shows: system's 2 clear spots, and favorable reproducibility, negative noiseless, so list it in text.That is: get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes is taken out, and puts cold; Filter, filtrating is steamed near and is done, and residue adds water 15ml dissolving, with extracted with diethyl ether 3 times, and 15ml at every turn; Merge ether solution, volatilize, residue adds the 0.5ml dissolve with methanol, as need testing solution.Other gets the tetrahydropalmatine reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B); Draw each 5～15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with 1% sodium hydroxide solution, be developping agent with normal hexane-methenyl choloride-methyl alcohol (10: 4: 1), presaturation 20 minutes; Launch; Take out, dry, put in the iodine vapor smoked after several minutes; In air, wave the iodine of most absorption, put under the ultraviolet (365nm) and inspect.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the same color spot.
(5) discriminating of Radix Glycyrrhizae:
The chemical constitution of Radix Glycyrrhizae is very complicated, and up till now, Beijing Medical University has separated 160 number of chemical compositions from Radix Glycyrrhizae.Its main type has: 1. triterpene compound: the potassium of glycyrrhizic acid, calcium salt are the sweet ingredient of Radix Glycyrrhizae.Also contain glycyrrhizic acid A
3, B
2, C
2, D
3, F
3, G
2, H
2, J
2And K
2And multiple triterpenes components such as enoxolone, different enoxolone, glycyrrhetol, licorice root lactone.2. flavone compound, the principal ingredient of Radix Glycyrrhizae spasmolysis, content is more has liquiritin, liquiritigenin etc.
Method:
The preparation of need testing solution: get these article content 5g, the 40ml that adds diethyl ether, refluxing extraction 1 hour is removed ether solution; The medicine residue is flung to ether, adds 40ml methyl alcohol, refluxing extraction 1 hour; Filter, filtrating evaporate to dryness, residue add water 40ml dissolving; With extracting n-butyl alcohol three times, each 20ml merges normal butyl alcohol liquid; With water washing 3 times, each 20ml, liquid anhydrates; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution.
The preparation of control medicinal material solution: extracting liquorice control medicinal material 1g in addition, shine medicinal material solution in pairs with legal system.
The preparation of negative sample solution: get scarce Radix Glycyrrhizae negative sample, prepare negative control solution according to the need testing solution preparation method.
Draw above need testing solution, control medicinal material solution, each 5～15 μ l of negative control solution respectively; The point in same be on the silica gel g thin-layer plate of bonding agent with CMC-Na; Launch with different developping agents respectively; Take out; Dry, spray different developers, 105 ℃ are heated to clear spot; Inspect under the daylight, the result is following:
The result shows: system's 5 clear spots, and favorable reproducibility, negative noiseless, so list it in text.That is: get these article content 5g, the 40ml that adds diethyl ether, water-bath refluxed 1 hour, discarded ether solution; The dregs of a decoction are flung to ether, add methyl alcohol 40ml, and water-bath refluxed 1 hour; Filter, filtrating evaporate to dryness, residue add water 40ml dissolving; With extracting n-butyl alcohol three times, each 20ml merges normal butyl alcohol liquid; With water washing 3 times, each 20ml discards water liquid; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution.Extracting liquorice control medicinal material 1g shines medicinal material solution in pairs with legal system in addition.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B); Draw each 5～15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With methenyl choloride-methanol-water (13: 7: 2) subnatant is developping agent; Launch, take out, dry; Spray 10% ethanol solution of sulfuric acid, 105 ℃ to be heated to spot colour developing clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the same color spot.
(6) discriminating of the red sage root:
The red sage root mainly contains fat-soluble diterpene phenanthrenequione pigment tanshinone component and water miscible liposoluble ingredient.Also contain flavonoids, triterpenes and sterols composition in addition.
Method:
The preparation of need testing solution: get these article content 5g, add 80% ethanol 40ml, sonicated 30 minutes; Put coldly, filter the filtrating evaporate to dryness; Residue adds 0.2mol/L hydrochloric acid 20ml stirring makes dissolving; Water-bath heating 10 minutes, with ethyl acetate extraction 3 times, 15ml at every turn; Combined ethyl acetate liquid; Evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution.
The preparation of reference substance, control medicinal material solution: other gets former tea acid reference substance, adds methyl alcohol and processes every 1ml and contain 1mg solution, as reference substance solution.Get red sage root control medicinal material 1g again, boiling 30 minutes, filtrating is concentrated into 10ml, and with ethyl acetate extraction 2 times, 10ml/ time, combined ethyl acetate liquid, evaporate to dryness, residue add methyl alcohol 1ml and dissolve, as control medicinal material liquid.
The preparation of negative control solution: get scarce red sage root negative sample, the preparation method prepares negative control solution by need testing solution.
Draw each 5～15 μ l of above need testing solution, reference substance solution, control medicinal material solution, negative control solution respectively, put in same be on the silica gel g thin-layer plate of bonding agent with CMC-Na, launch with different developping agents respectively, take out, dry, the result is following:
The result shows that because negative the interference, spot separates bad, and reasons such as poor reproducibility can't be listed it in text.
(7) discriminating of Fructus meliae toosendan:
Principal ingredient is a triterpene compound in the Fructus meliae toosendan, secondly is the glycoside compounds of the sequiterpene of ionone type.
Tetracyclic triterpene has toosendanin, isochuanliansu, and latter's toxicity is bigger.
Method:
The preparation of need testing solution: get these article content 3g, add ethanol 30ml, sonicated 30 minutes is put coldly, filters, and the filtrating evaporate to dryness, residue adds the 0.5ml dissolve with ethanol, as need testing solution.
The preparation of control medicinal material solution: get Fructus meliae toosendan control medicinal material 1g, shine medicinal material solution in pairs with legal system.
The preparation of negative control: get scarce Fructus meliae toosendan negative sample, the preparation method prepares negative control solution by need testing solution.
Draw each 5～15 μ l of above-mentioned need testing solution, control medicinal material solution, negative control solution respectively, put in same be on the silica gel g thin-layer plate of bonding agent with methylol fiber sodium, launch with different developping agents respectively, take out, dry, the result is following:
| Numbering | Developping agent | Colour developing | The result |
| 1 | Toluene-acetone (9: 1) | Iodine vapor is smoked clear to the spot colour developing, inspects under the daylight | Spot separates bad |
| 2 | Benzene-acetone (9: 1) | Iodine vapor is smoked clear to the spot colour developing, inspects under the daylight | Spot separates bad |
| 3 | Benzene-ethyl acetate (17: 3) | Inspect under the uviol lamp (365nm) | The negative interference |
The result shows: because negative the interference, spot separates bad, and reasons such as poor reproducibility can't be listed it in text.
(8) discriminating of safflower:
Safflower mainly contains carthamin yellow, carthamin, neocarthamin, kaempferol-3-rhamnoside.Its bitter principle is podocarpus resinol-list-β-D-glycoside.Fat oil is still arranged in addition, nonacosane-cupreol, palmitic acid, myristic acid and lauric acid etc.
Method:
The preparation of need testing solution: get these article content 4g, add 80% acetone soln 30ml, sonicated 15 minutes filters, and filtrating evaporate to dryness, residue add acetone 0.5ml dissolving, as need testing solution.
The preparation of control medicinal material solution: get safflower control medicinal material 1g, shine medicinal material solution in pairs with legal system.
The preparation of negative sample solution: get scarce safflower negative sample, prepare negative control solution according to the need testing solution preparation method.
Draw each 5～15 μ l of above-mentioned need testing solution, control medicinal material solution, negative control solution respectively, put in same be on the silica gel thin-layer plate of bonding agent with CMC-Na, launch with different developping agents respectively, take out, dry, the result is following:
The result shows: because the spot separation is bad, reasons such as poor reproducibility can't be listed it in text.
[assay] is that the effective constituent content of paeoniflorin of the root of herbaceous peony in these article is measured.
(1) instrument and reagent:
High performance liquid chromatograph: LC-10ATVP infusion pump (double pump), the SPD-10AVP UV-detector, Tianjin, island CS-Light Chinese edition chromatographic work station (version: 200.00 Sub), HT-230A chromatographic column constant temperature oven (homemade)
Chromatographic column: Hypersil C
18Post (ODS2), 5 μ m, 250 * 4.6mm, post number: E1612780 (Dalian Yi Lite scientific instrument company)
UV-1700 type ultraviolet-visible pectrophotometer (day island proper Tianjin)
AUW 220D type (scale division value 0.01mg) electronic balance (day island proper Tianjin)
CH-250 ultrasonic cleaner (Beijing innovation moral ultrasonic electronic research institute)
YY 9105 type electric-heated thermostatic water baths (Tianjin Tai Site Instr Ltd.)
Paeoniflorin reference substance (supply assay with): purchase in Nat'l Pharmaceutical & Biological Products Control Institute
Methyl alcohol, acetonitrile are chromatographically pure, and water is double distilled water.All the other reagent are pure for analyzing
(2) preparation of reference substance solution
Precision takes by weighing 36 hours Paeoniflorin reference substance 12.52mg of drying in the phosphorus pentoxide vacuum drying apparatus, puts in the 25ml measuring bottle, adds the Diluted Alcohol dissolving and is diluted to scale, shakes up, and promptly gets Paeoniflorin reference substance storing solution (every 1ml contains 0.5008mg).
The accurate storing solution 5ml that draws places the 50ml measuring bottle, adds dilute methanol and is diluted to scale, shakes up, and promptly gets Paeoniflorin reference substance solution (every 1ml contains 50.08 μ g).
(3) selection of chromatographic condition
1) chromatographic column: Hypersil C
18Post (ODS2), 5 μ m, 250 * 4.6mm, post number: E1612780 (Dalian Yi Lite scientific instrument company)
2) detect wavelength: according to the UV scanning figure of Paeoniflorin, Paeoniflorin has absorption maximum at 229.5nm, and with reference to interrelated data, selecting to detect wavelength is 230nm.
3) column temperature: 30 ℃
4) flow velocity: 1.0ml/min
5) sample size: 10 μ l
6) selection of moving phase, the result is following:
| Numbering | Moving phase | Retention time (min) | Peak area | The result |
| 1 | Methanol-water (25:75) | 15.086 | 194702 | The main peak peak shape is bad |
| 2 | Methyl alcohol-0.1% phosphoric acid (25:75) | 14.799 | 389968 | Main peak peak shape symmetry is bad |
| 3 | Acetonitrile-water (15:85) | 11.352 | 398796 | Do not separate fully with impurity peaks |
| 4 | Acetonitrile-0.1% phosphoric acid (15:85) | 11.355 | 338200 | Separate fully with impurity peaks |
| 5 | Acetonitrile-0.1% phosphoric acid-isopropyl alcohol (15:85:0.8) | 10.974 | 377980 | Do not separate fully with impurity peaks |
The result shows that moving phase is better with acetonitrile-0.1% phosphoric acid (15: 85), and the peak shape symmetry at Paeoniflorin peak is good, and number of theoretical plate is high, and separates fully with other impurity peaks.
7) system suitability test
1. theoretical cam curve: be calculated as follows theoretical cam curve
Retention time
2
Number of theoretical plates =? 5.54 ×
The half-peak height is wide
2
Measure the system suitability condition under the assay item in the related data with reference to paeoniflorin content, the theoretical cam curve of this test is decided to be by the Paeoniflorin peak should be not less than 5000.
2. degree of separation: be calculated as follows the Paeoniflorin peak becomes swarming with other degree of separation.
2×（t
R2－t
R1）
Separation =?
W
1+?W
2
The result shows that in this chromatographic condition, the Paeoniflorin peak becomes swarming all can separate fully with other, and degree of separation meets the requirements greater than 1.5.
(4) preparation of need testing solution
The related data of measuring with reference to paeoniflorin content is studied the preparation method of need testing solution, and the result is following:
Test findings shows that ultrasonic Extraction 40 minutes, 50 minutes and 60 minutes effects are approaching, and the completeness of considering to handle sample is decided to be 40 minutes with quick with the sample ultrasonic time.
That is: the preparation method of need testing solution is:
These article of getting capsule 's content powder 0.25g, the accurate title, decide, and adds Diluted Alcohol 25ml, claims to decide weight.Ultrasonic Extraction 40 minutes is taken out, and puts coldly, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and with the filtering with microporous membrane of 0.45 μ m, gets subsequent filtrate as need testing solution.
(5) mensuration of negative control and solvent peak
Prepare the negative sample solution that lacks the root of herbaceous peony by selected need testing solution preparation method.
Draw each 10 μ l of negative sample solution, reference substance solution, need testing solution and solvent (Diluted Alcohol) respectively, inject liquid chromatograph, make an experiment by above-mentioned chromatographic condition, the result is following:
The result shows; The negative control solution and the solvent (Diluted Alcohol) that lack the root of herbaceous peony are compared with reference substance solution and need testing solution chromatogram; The corresponding retention time at the Paeoniflorin peak (about 11.3min) locates not have absorption peak, shows that feminine gender and solvent are noiseless to the mensuration of test sample.
(6) linear relationship is investigated test
Use the reference substance solution of reference substance storing solution (every 1ml contains Paeoniflorin 0.5008mg) preparation variable concentrations respectively, accurate each solution 10 μ l that draw inject liquid chromatograph, make an experiment by above-mentioned chromatographic condition, and the result is following:
With the peak area is ordinate, and sample size (μ g) is a horizontal ordinate drawing standard curve, and the result shows that Paeoniflorin has good linear relationship (r=0.99999) in sample size is 125.20～1502.4ng scope.
(7) precision test
Get the Paeoniflorin reference substance solution (concentration: 50.08 μ g/ml) 10 μ l injecting chromatographs make an experiment, and repeat 5 times, and the result is following:
The result shows that the peak area relative standard deviation of Paeoniflorin illustrates that less than 2.0% this test precision is good.
(8) stability test
Take by weighing sample contents powder 0.2507g, prepare need testing solution by the text method, need testing solution was put the room temperature held 0,1,2,4,6,8,10 hour, draw 10 μ l sample introductions respectively and measure, the result is following:
The result shows, the relative standard deviation of Paeoniflorin peak area is less than 2.0%, illustrates that this solution has good stable property at room temperature 10 hours.
(9) replica test
Precision takes by weighing 5 parts in sample contents powder respectively, every part of 0.25g, and accurate the title, decide, and prepares need testing solution by the text method, and make an experiment, and the result is following:
The result shows that the relative standard deviation of paeoniflorin content is 0.49%, less than 2.0%, shows that repeatability is good.
(10) recovery test
Adopt the application of sample absorption method: precision take by weighing known content sample contents (paeoniflorin content: 0.5004%) 6 parts, every part of 0.125g.Add Paeoniflorin reference substance solution (concentration: 50.08 μ g/ml) an amount of respectively; It is an amount of to add Diluted Alcohol again; To liquor capacity be 25ml; From " claim decide weight; ultrasonic Extraction 40 minutes ... ", prepare need testing solution by the preparation method of text need testing solution; And press the text condition test, be calculated as follows the recovery:
The amount of reference substance total amount-sample reference substance of measuring
Recovery (%) =?
× 100%
Add the amount of reference substance
The result is following:
The result shows that the average recovery rate of Paeoniflorin is 99.49%, and the relative standard deviation of the recovery is 0.91%, less than 2.0%, illustrates that measuring Paeoniflorin with this law has the good recovery.
(11) sample determination
Test 10 lot sample article by text method and chromatographic condition, be calculated as follows content of paeoniflorin in the sample:
Sample peak area * reference substance content * 25
Content (%) =
? × 100%
Reference substance peak area * sample sample weighting amount * 1000 * 1000
The result is following:
Measure the result according to above 10 lot sample article, tentative these article contain the root of herbaceous peony with Paeoniflorin (C
23H
28O
11) meter, every must not be less than 1.5mg.
Embodiment:
A kind of quality determining method of dysmenorrhoea Yiganning capsule comprises the steps:
(1) differentiates
A. get these article content 10g, the 50ml that adds diethyl ether, water-bath refluxed 30 minutes; Discard ether solution, the dregs of a decoction are flung to ether, add ethanol 50ml; Water-bath refluxed 30 minutes, took out, and put cold; Filter; The filtrating evaporate to dryness, residue adds water 20ml dissolving, with water saturation extracting n-butyl alcohol 2 times; Each 20ml merges normal butyl alcohol liquid.With the saturated water 20ml washing of normal butyl alcohol, discard water liquid again, normal butyl alcohol liquid evaporate to dryness, residue add ethanol 0.5ml dissolving, as need testing solution.Other gets the Paeoniflorin reference substance, adds methyl alcohol and processes the solution that every 1ml contains 2mg, as reference substance solution.Test according to thin-layered chromatography (2010 editions appendix VIB of Chinese Pharmacopoeia); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia (8: 1:4: 1) be developping agent; Launch, take out, dry; Spray is with 5% vanillic aldehyde sulfuric acid solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
B. get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes.Take out, put coldly, filter, filtrating is steamed near and is done, and residue adds water 15ml dissolving, with extracted with diethyl ether 3 times, and each 15ml, the merging ether solution volatilizes, and residue adds the 0.5ml dissolve with methanol, as need testing solution.Other gets the tetrahydropalmatine reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2010); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with 1% sodium hydroxide solution, be developping agent with normal hexane-methenyl choloride-methyl alcohol (10: 4: 1), presaturation 20 minutes; Launch; Take out, dry, put in the iodine vapor smoked after several minutes; In air, wave the iodine of most absorption, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
C. get these article content 5g, the 40ml that adds diethyl ether, water-bath refluxed 1 hour, discarded ether solution; The dregs of a decoction are flung to ether, add methyl alcohol 40ml, and water-bath refluxed 1 hour; Filter, filtrating evaporate to dryness, residue add water 40ml dissolving; With extracting n-butyl alcohol three times, each 20ml merges normal butyl alcohol liquid; With water washing 3 times, each 20ml discards water liquid; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution.Extracting liquorice control medicinal material powder 1g shines medicinal material solution in pairs with legal system in addition.Test according to thin-layered chromatography (2010 editions appendix VIB of Chinese Pharmacopoeia); Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With methenyl choloride-methyl alcohol-water (13: 7: 2) subnatant is developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) inspection: should meet each item regulation (an appendix I of Chinese Pharmacopoeia version in 2010 L) relevant under the capsule item.
(3) assay: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2010).
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-0.1% phosphoric acid (15:85) is moving phase; The detection wavelength is 230nm.Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 5000.
The preparation of reference substance solution: it is an amount of to get the Paeoniflorin reference substance, and accurate the title decides, and adds Diluted Alcohol and processes the solution that every 1ml contains 50 μ g, promptly gets.
The preparation of need testing solution: get these article content powder 0.25g, the accurate title, decide, and the accurate Diluted Alcohol 25ml that adds claims to decide weight.Ultrasonic Extraction 40 minutes is taken out, and puts coldly, claims decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and with the miillpore filter filtration of 0.45 μ m, gets subsequent filtrate, promptly gets.
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject high performance liquid chromatograph, measure, promptly get.
Every of these article contain the root of herbaceous peony with Paeoniflorin (C
23H
28O
11) meter, must not be less than 1.5mg.
Wherein: the method for making of these article is Radix Angelicae Sinensis (stir-fry) 800g, rhizoma cyperi (system) 800g, the root of herbaceous peony (stir-fry) 800g, corydalis tuber (stir-fry) 720g, Ligusticum wallichii (stir-fry) 480g, Radix Glycyrrhizae (processing) 320g, red sage root 800g, Fructus meliae toosendan (stir-fry) 720g, safflower 400 g; More than nine the flavor; Except that safflower; Eight flavor boiling secondaries such as all the other Radix Angelicae Sinensis; Add for the first time 12 times of water gagings decoctions and add safflower after 2 hours; Continue to decoct 0.5 hour; Adding for the second time 10 times of water gagings decocted 1.5 hours; Collecting decoction; Filter the filtrating standing over night.Get supernatant, being concentrated into relative density is 1.10 ~ 1.12 (85 ℃), puts coldly, adds the ethanol of 2 times of amounts, stirs, and leaves standstill 24 hours.Get supernatant and reclaim ethanol, be concentrated into thick medicinal extract, drying under reduced pressure is pulverized, and adds appropriate amount of auxiliary materials, granulates, and drying incapsulates, and processes 1000, promptly gets.
Claims (5)
1. the quality determining method of a dysmenorrhoea Yiganning capsule comprises the steps:
(1) chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-0.1% phosphoric acid (15:85) is moving phase; The detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 5000;
(2) preparation of reference substance solution: it is an amount of to get the Paeoniflorin reference substance, and accurate the title decides, and adds Diluted Alcohol and processes the solution that every 1ml contains 50 μ g, promptly gets;
(3) preparation of need testing solution: get these article content powder 0.25g, the accurate title, decide, and the accurate Diluted Alcohol 25ml that adds claims to decide weight; Ultrasonic Extraction 40 minutes is taken out, and puts coldly, claims decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and with the miillpore filter filtration of 0.45 μ m, gets subsequent filtrate, promptly gets;
(4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject high performance liquid chromatograph, measure, promptly get.
2. the quality determining method of dysmenorrhoea Yiganning capsule as claimed in claim 1, wherein: every of these article contain the root of herbaceous peony in Paeoniflorin, must not be less than 1.5mg.
3. the quality determining method of dysmenorrhoea Yiganning capsule as claimed in claim 1 or 2, wherein:
Differentiate
A. get these article content 10g, the 50ml that adds diethyl ether, water-bath refluxed 30 minutes; Discard ether solution, the dregs of a decoction are flung to ether, add ethanol 50ml; Water-bath refluxed 30 minutes, took out, and put cold; Filter; The filtrating evaporate to dryness, residue adds water 20ml dissolving, with water saturation extracting n-butyl alcohol 2 times; Each 20ml merges normal butyl alcohol liquid; With the saturated water 20ml washing of normal butyl alcohol, discard water liquid again, normal butyl alcohol liquid evaporate to dryness, residue add ethanol 0.5ml dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and processes the solution that every 1ml contains 2mg, as reference substance solution; Test according to thin-layered chromatography; Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With volume ratio is 8: 1:4: 1 methenyl choloride-ethyl acetate-methyl alcohol-dense ammonia is developping agent; Launch, take out, dry; Spray is with 5% vanillic aldehyde sulfuric acid solution, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
B. get these article content 4g, add 80% ethanol 40ml, sonicated 30 minutes; Take out, put coldly, filter, filtrating is steamed near and is done, and residue adds water 15ml dissolving, with extracted with diethyl ether 3 times, and each 15ml, the merging ether solution volatilizes, and residue adds the 0.5ml dissolve with methanol, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; Test according to thin-layered chromatography; Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with 1% sodium hydroxide solution, be that 10: 4: 1 normal hexane-methenyl choloride-methyl alcohol is developping agent with volume ratio, presaturation 20 minutes; Launch; Take out, dry, put in the iodine vapor smoked after several minutes; In air, wave the iodine of most absorption, put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
C. get these article content 5g, the 40ml that adds diethyl ether, water-bath refluxed 1 hour, discarded ether solution; The dregs of a decoction are flung to ether, add methyl alcohol 40ml, and water-bath refluxed 1 hour; Filter, filtrating evaporate to dryness, residue add water 40ml dissolving; With extracting n-butyl alcohol three times, each 20ml merges normal butyl alcohol liquid; With water washing 3 times, each 20ml discards water liquid; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml dissolving, as need testing solution; Extracting liquorice control medicinal material powder 1g shines medicinal material solution in pairs with legal system in addition; Test according to thin-layered chromatography; Draw each 5-15 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose; With volume ratio is that 13: 7: 2 methenyl choloride-methyl alcohol-subsurface layer liquid is developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
4. the quality determining method of dysmenorrhoea Yiganning capsule as claimed in claim 3, wherein inspection: should meet each item regulation relevant under the capsule item.
5. the quality determining method of dysmenorrhoea Yiganning capsule as claimed in claim 4; Wherein: the method for making of these article is for frying Radix Angelicae Sinensis 800g, system rhizoma cyperi 800g, stir-baked RADIX PAEONIAE ALBA 800g, fry corydalis tuber 720g, frying Ligusticum wallichii 480g, honey-fried licorice root 320g, red sage root 800g, stir-baked FRUCTUS TOOSENDAN 720g, safflower 400 g; More than nine the flavor; Except that safflower; Eight flavor boiling secondaries such as all the other Radix Angelicae Sinensis; Add for the first time 12 times of water gagings decoctions and add safflower after 2 hours; Continue to decoct 0.5 hour; Adding for the second time 10 times of water gagings decocted 1.5 hours; Collecting decoction; Filter the filtrating standing over night; Get supernatant, being concentrated into relative density is 1.10-1.12 (85 ℃), puts coldly, adds the ethanol of 2 times of amounts, stirs, and leaves standstill 24 hours; Get supernatant and reclaim ethanol, be concentrated into thick medicinal extract, drying under reduced pressure is pulverized, and adds appropriate amount of auxiliary materials, granulates, and drying incapsulates, and processes 1000, promptly gets.
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| CN106226452A (en) * | 2016-08-29 | 2016-12-14 | 贵州信邦制药股份有限公司 | The discrimination method of Radix Angelicae Sinensis in SHIQUAN DABU JIU |
| CN110398562A (en) * | 2019-07-23 | 2019-11-01 | 株洲千金药业股份有限公司 | The TLC Identification of Radix Glycyrrhizae in a kind of Fuke Tiaojing tablets |
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Application publication date: 20120215 |