CN101352565A - Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling - Google Patents
Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling Download PDFInfo
- Publication number
- CN101352565A CN101352565A CNA2008100460387A CN200810046038A CN101352565A CN 101352565 A CN101352565 A CN 101352565A CN A2008100460387 A CNA2008100460387 A CN A2008100460387A CN 200810046038 A CN200810046038 A CN 200810046038A CN 101352565 A CN101352565 A CN 101352565A
- Authority
- CN
- China
- Prior art keywords
- solution
- granule
- reference substance
- methanol
- need testing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a quality control method for a granule with the effects of activating blood, dissolving stasis and removing toxin for detumescence. The quality control method comprises parts or the whole of the items of character identification, inspection and content determination; wherein, the identification comprises the thin-layer chromatography identification of angelica and Szechwan lovage rhizome in the granular preparation and paeoniflorin and fumitory in red paeony root; the content determination is implemented to determine the content of paeoniflorin in the red paeony root of the granular preparation. In connection with the disadvantages of simple quality control standards and hard product quality control of the original 'Gongyankang granule', the quality control method of the invention studies the quality control method of the granular preparation and draws out the thin-layer chromatography identification to the angelica and Szechwan lovage rhizome in the granular preparation and the paeoniflorin and fumitory in red paeony root and the contents determination to the paeoniflorin in the red paeony root of the granular preparation, thereby improving the quality control standards of blood-activating and stasis-dissolving and detoxification and detumescence granular medicines. The control method in the quality standards can effectively control the quality of Gongyankang granule, thus ensuring the clinical efficacy of the medicine.
Description
Technical field
The invention belongs to medical technical field, the method of quality control that relates to a kind of blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule particularly relates to the method for quality control of the granule medicament of the effect of being made by Chinese prescriptions such as Radix Angelicae Sinensis, Radix Paeoniae Rubra, Sonchus brachyotus DC., Rhizoma Cyperi, Rhizoma Zingiberis Preparatum, Herba Lycopi, Rhizoma Chuanxiong, Flos Carthami, Radix Bupleuri, Sargassum, Semen Plantaginis, Rhizoma Corydalis with blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling.
Background technology
The granule medicament of the effect with blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling that is used for the treatment of gynaecopathia is more, " Fuyankan particles " wherein is to adopt ten multi-flavor natural Chinese medicines materials such as Radix Angelicae Sinensis, Radix Paeoniae Rubra, Sonchus brachyotus DC., Rhizoma Cyperi, Rhizoma Zingiberis Preparatum, Herba Lycopi, Rhizoma Chuanxiong, Flos Carthami, Radix Bupleuri, Sargassum, Semen Plantaginis, Rhizoma Corydalis, Chinese patent medicine through being prepared from, be mainly used in the treatment chronic pelvic inflammatory disease, its determined curative effect is reliable, has no side effect." Fuyankan particles " publication in the 20 of the drug standard " Chinese traditional patent formulation preparation " of Ministry of Public Health promulgation, and in clinical practice for many years, obtaining than satisfactory therapeutic effects aspect the treatment chronic pelvic inflammatory disease.But find after deliberation, it is simple that the quality control standard of existing " Fuyankan particles " exists quality control standard, and the uppity shortcoming of product quality is in process of producing product, the quality of this medicine can not be effectively controlled with existing method of quality control, thereby its clinical efficacy will be influenced.
Summary of the invention
The objective of the invention is to, the method for quality control of a kind of blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule is provided.The present invention is directed to and have blood circulation promoting and blood stasis dispelling, the granular preparation medicine of the effect of removing toxic substances and promoting subsidence of swelling, especially " Fuyankan particles " quality control standard of recording at existing ministry standard is simple, the uppity shortcoming of product quality, method of quality control to granular preparation is studied, worked out Radix Angelicae Sinensis in the preparation and Rhizoma Chuanxiong, Radix Paeoniae Rubra, the thin layer chromatography of Rhizoma Corydalis is differentiated, and to the contained content of paeoniflorin mensuration of Radix Paeoniae Rubra in the preparation, improved blood circulation promoting and blood stasis dispelling, the quality control standard of removing toxic substances and promoting subsidence of swelling granule, control method in this quality standard can be controlled blood circulation promoting and blood stasis dispelling effectively, the quality of removing toxic substances and promoting subsidence of swelling granule " Fuyankan particles ", thus the clinical efficacy of this granular preparation guaranteed.
Blood circulation promoting and blood stasis dispelling of the present invention, removing toxic substances and promoting subsidence of swelling granule are to constitute like this:
[prescription] Radix Angelicae Sinensis 90g, Radix Paeoniae Rubra 90g, Sonchus brachyotus DC. 240g, Rhizoma Cyperi (vinegar system) 90g, Rhizoma Zingiberis Preparatum 90g, Herba Lycopi 90g, Rhizoma Chuanxiong 60g, Flos Carthami 60g, Radix Bupleuri 90g, Sargassum 90g, Semen Plantaginis (salt moxibustion) 120g, Rhizoma Corydalis 60g.
[method for making] above 12 flavors remove Rhizoma Corydalis and are ground into fine powder, sieve; Radix Angelicae Sinensis, Rhizoma Cyperi, Rhizoma Chuanxiong, Radix Bupleuri extracts volatile oil, and the aqueous solution after distillation device is in addition collected, medicinal residues and Radix Paeoniae Rubra, Rhizoma Zingiberis Preparatum, Sargassum, Semen Plantaginis decocts with water secondary, adds 8 times of water gagings for the first time and decocts 2 hours, adds 6 times of water gagings for the second time and decocts 1 hour, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, medicinal liquid is standby, all the other Sonchus brachyotus DC., Flos Carthami, after Herba Lycopi's three flavors add water boil,, add 8 times of water gaging warm macerating 2 hours for the first time in 80 ℃ of warm macerating secondaries, add for the second time 6 times of water gaging warm macerating 1 hour, filter, merge above each medicinal liquid, be concentrated into the extractum of relative density 1.32～1.38 (50～60 ℃), get whole extractum, add an amount of dextrin, with above-mentioned Rhizoma Corydalis powder mixing, adding ethanol is an amount of, make granule, drying adds above-mentioned Radix Angelicae Sinensis, Rhizoma Cyperi, Rhizoma Chuanxiong, Radix Bupleuri volatile oil, mixing, make the 450g granule, promptly.
Method of quality control of the present invention mainly comprise in character, discriminating, inspection, the assay project partly or entirely; Differentiate that wherein the thin layer chromatography comprise Radix Angelicae Sinensis in the granule and Rhizoma Chuanxiong, Radix Paeoniae Rubra, Rhizoma Corydalis differentiates; Assay is that the contained content of paeoniflorin of Radix Paeoniae Rubra in the granule is measured.
The discrimination method of Radix Angelicae Sinensis and Rhizoma Chuanxiong is to be contrast with Radix Angelicae Sinensis and Rhizoma Chuanxiong reference substance, and with normal hexane: ethyl acetate=6～12: 0.5～1.5 is the tlc identification method of developing solvent.
The discrimination method of peoniflorin is to be contrast with the peoniflorin reference substance, and with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～24: 7～13: 2～3 is the tlc identification method of developing solvent.
The discrimination method of Rhizoma Corydalis is to be contrast with the Rhizoma Corydalis reference substance, and with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is the tlc identification method of developing solvent.
Described discrimination method comprises the part or all of of following project:
(1) get this granule, porphyrize adds ethanol, and supersound process filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate makes its dissolving, as need testing solution; Other gets Radix Angelicae Sinensis respectively, the Rhizoma Chuanxiong control medicinal material is an amount of, makes control medicinal material solution respectively with method; According to Chinese Pharmacopoeia thin layer chromatography test, draw need testing solution, two kinds of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6～12: 0.5～1.5 be developing solvent, launches, and taking-up is dried, and puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(2) get this granule, porphyrize adds ethanol, reflux is handled, and puts coldly, filters, filtrate evaporate to dryness, residue add water makes its dissolving, with twice of water saturated n-butanol extraction, merge n-butyl alcohol liquid, with the saturated water washing twice of n-butyl alcohol, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes solution, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～25: 7～13: 2～3 is developing solvent, launch, take out, dry, spray is with 2～8% vanillin sulfuric acid solutions, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this granule, porphyrize adds methanol, and supersound process filters, and filtrate evaporate to dryness, residue add water makes its dissolving, transfers PH to 8 with strong ammonia solution, uses ether extraction three times, merges ether solution, and evaporate to dryness, residue add methanol makes its dissolving, as need testing solution; It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate with the sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is developing solvent, launching in the pre-saturated expansion cylinder with developing solvent, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Discrimination method comprises the part or all of of following project more specifically:
(1) get this granule 9g, porphyrize adds ethanol 20ml, and supersound process 20 minutes filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate 0.5ml makes its dissolving, as need testing solution; Get Radix Angelicae Sinensis, each 1g of Rhizoma Chuanxiong control medicinal material respectively in addition, make control medicinal material solution respectively with method; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, two kinds of each 5～10 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=9: 1 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
(2) get this granule 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, with twice of water saturated n-butanol extraction, each 25ml merges n-butyl alcohol liquid, uses twice of the saturated water washing of n-butyl alcohol, each 10ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, each 4 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=50: 20: 10: 2.5 is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this granule 18g, porphyrize adds methanol 50ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes its dissolving, transfer PH to 8 with strong ammonia solution, use ether extraction three times, each 10ml, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution; Other gets Rhizoma Corydalis control medicinal material 1g, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of described need testing solution, reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=7.5: 4: 1 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
To measure be to be contrast with the peoniflorin reference substance to the contained content of paeoniflorin of Radix Paeoniae Rubra in this granule, and with acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is the high performance liquid chromatography of mobile phase.
Described paeoniflorin content is measured according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring, comprises following:
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: take by weighing 80 ℃ of drying under reduced pressure to constant weight peoniflorin reference substance, add methanol and make the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this granule under the content uniformity item, porphyrize is got 1g, puts in the tool plug conical flask, adds 70% methanol solution 25ml, and supersound process 20 minutes filters, and measures subsequent filtrate 5ml, puts in the 10ml measuring bottle, adds water to scale, shakes up, promptly;
Algoscopy: draw reference substance solution 10 μ l and need testing solution 5～10 μ l respectively, inject chromatograph of liquid, measure, promptly;
This granule contains Radix Paeoniae Rubra with peoniflorin (C
23H
28O
11) meter, must not be less than 5mg.
Described method of quality control comprises:
Character: this granule is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of;
Differentiate:
(1) get this granule, porphyrize adds ethanol, and supersound process filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate makes its dissolving, as need testing solution; Other gets Radix Angelicae Sinensis respectively, the Rhizoma Chuanxiong control medicinal material is an amount of, makes contrast solution respectively with method; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, two kinds of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6～12: 0.5～1.5 is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
(2) get this granule, porphyrize adds ethanol, reflux is handled, and puts coldly, filters, filtrate evaporate to dryness, residue add water makes its dissolving, with twice of water saturated n-butanol extraction, merge n-butyl alcohol liquid, with the saturated water washing twice of n-butyl alcohol, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes solution, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～24: 7～13: 2～3 is developing solvent, launch, take out, dry, spray is with 2～8% vanillin sulfuric acid solutions, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this granule, porphyrize adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add water makes its dissolving, transfers PH to 8 with strong ammonia solution, uses ether extraction, merges ether solution, and evaporate to dryness, residue add methanol makes its dissolving, as need testing solution; It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate with sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is developing solvent, in with the expansion cylinder after the developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: this granule should meet pertinent regulations under the Chinese Pharmacopoeia granule item;
Assay: paeoniflorin content is measured according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: it is an amount of to the peoniflorin reference substance of constant weight to take by weighing 80 ℃ of drying under reduced pressure, adds methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this granule under the content uniformity item, porphyrize is got 1g, puts in the tool plug conical flask, adds methanol solution, and supersound process filters, and measures subsequent filtrate, puts in the measuring bottle, adds water to scale, shakes up, promptly;
Algoscopy: draw reference substance solution 10 μ l and need testing solution 5～10 μ l respectively, inject chromatograph of liquid, measure, promptly;
This granule contains Radix Paeoniae Rubra with peoniflorin (C for every bag
23H
28O
11) meter, must not be less than 5mg.
In order to ensure method of quality control science of the present invention, reasonable, feasible, the applicant studies the discriminating and the content assaying method of each medicine in the method, and concrete testing data is as follows:
One, Radix Angelicae Sinensis, Rhizoma Chuanxiong Study on Identification
This discriminating is that TLC differentiates, is the discriminating of Radix Angelicae Sinensis, Rhizoma Chuanxiong volatile oil composition, mainly contains common components such as butylene furan base lactone, ligustilide in the volatile oil of Radix Angelicae Sinensis, Rhizoma Chuanxiong, and in the ultra-violet (UV) band certain absorption is arranged.This method adopts the blank of scarce Radix Angelicae Sinensis, the blank that lacks Rhizoma Chuanxiong and two blank that lacks (promptly by the prescription consumption, do not add Radix Angelicae Sinensis, Rhizoma Chuanxiong medical material, preparation technology's preparation that analog sample is identical) contrasts respectively product in contrast as blank and Radix Angelicae Sinensis, Rhizoma Chuanxiong, with ethanol ultrasonic extraction, water-bath volatilizes, the ethyl acetate dissolving is as the offerings test solution, and is simple to operate, is easy to point sample; Respectively with normal hexane-ethyl acetate (9: 1), normal hexane-ethyl acetate (17: 3) as developing solvent, launch, dry, put under the ultra-violet lamp (365nm) and inspect.The principal spot and the sample that lack the blank of the blank of Radix Angelicae Sinensis, scarce Rhizoma Chuanxiong have common principal spot, and two scarce blank does not have this a little principal spots; Back-developing solvent, speckle slightly spreads, so adopt normal hexane-ethyl acetate (9: 1).
Two, the thin layer Study on Identification of peoniflorin
By the method that " Fuyankan particles " records my company continuous three series-produced " Fuyankan particles " is carried out identification experiment in the 20 in the drug standard Chinese traditional patent formulation preparation of Ministry of Public Health promulgation, sample is corresponding with reference substance chromatograph speckle displacement, and show identical blue spot, the result is consistent with content described in the health ministry standard, so adopt the health ministry standard.
Three, the thin layer Study on Identification of tetrahydropalmatine
The discrimination method of the Rhizoma Corydalis in " YUANHU ZHITONG PIAN " recorded with reference to Chinese Pharmacopoeia one one of version in 2000, and through experiment repeatedly, the equal show sample of result is corresponding with reference substance chromatograph speckle displacement, and show identical fluorescence speckle, so the thin layer discrimination method of Rhizoma Corydalis in this granule and the thin layer discrimination method basically identical of " YUANHU ZHITONG PIAN " middle Rhizoma Corydalis have been determined.
Four, content of paeoniflorin is measured research
1, the selection of assay composition and method
Radix Paeoniae Rubra has pharmacological actions such as antiinflammatory, vasodilation for one of this prescription principal agent, peoniflorin is the main active of Radix Paeoniae Rubra, and is consistent with the effect of this preparation for treating; Peoniflorin content in crude drug is higher, and soluble in water, be easy to change in the preparation such as the water extracted immersing paste, thus select that peoniflorin is its assay object in the Radix Paeoniae Rubra, to control the inherent quality of this preparation.Because prescription is formed complicated, do not find suitable internal standard substance, paeoniflorin content is measured and is calculated by external standard method, in actual content is measured, consider the factor affecting such as error of mobile phase preparation, take Chinese Pharmacopoeia (appendix VID of version in 2000) to calculate content of paeoniflorin in the test sample.On the basis of the quality standard after Fuyankan particles improves, Fuyankan particles has been carried out content assaying method learned research, and worked out described content assaying method.
2, experimental apparatus and reagent
High performance liquid chromatograph: HP-1100series, four pumps, automixer, VWD UV-detector, manual injector, HP chemstation for LC (Rev.A.06.01) data processor.Chromatographic column: DOS post (Tianjin); Ultrasonic washing unit: ripple is reached 004 type; Electronic analytical balance: prunus mume (sieb.) sieb.et zucc. Teller T24 (1/100000), AB-104-N (Shanghai, 1/10000); Centrifuge: table model high speed centrifuge TGL-16G type; Microsyringe: 10 μ l, U.S. HAMITON.
Experiment reagent: reference substance: peoniflorin (available from Beijing Nat'l Pharmaceutical ﹠ Biological Products Control Institute, assay usefulness, lot number is 0736-200015); Sample: Fuyankan particles, the 9g/ bag, the aluminium foil inner packing, the carton outer package, the U.S. greatly healthy pharmaceutical Co. Ltd in Sichuan produces lot number 080301,080302,080303; Lack the Radix Paeoniae Rubra negative control: U.S. greatly healthy pharmaceutical Co. Ltd provides by Sichuan, outside the deferrization Radix Paeoniae Rubra, and prescription ratio and the preparation of processing simulation finished product.
Mobile phase water: ultra-pure water; Acetonitrile: chromatographically pure; Phosphoric acid: analytical pure, content is no less than 85%.
3, content assaying method is learned research
(1) chromatographic condition and system are suitable for test: with octadecylsilane chemically bonded silica is filler, and acetonitrile-0.1% phosphoric acid solution (15: 85) is a mobile phase; The detection wavelength is 243nm.
The different experiments of comparing of flowing: other experiment conditions are constant, acetonitrile-0.1% phosphoric acid solution (15: 85) and methanol-water (34: 66), acetonitrile-water-triethylamine (13: 87: 0.2) (it is 2.3 that phosphoric acid is transferred PH) two mobile phases are compared, the result, the peoniflorin peak symmetry of acetonitrile-0.1% phosphoric acid solution (15: 85) is better, half-peak breadth is less, and retention time is moderate.
Different brands chromatographic column compatibility test: by the standard aforesaid operations, homemade post (Tianjin SNTEK Kromasil C18 (15 * 4.6mm, 5 μ) and import post (Waters Nova Pak C18 (150 * 3.9mm, 4 μ) all can make the peoniflorin peak in the sample reach baseline separation, the peak shape symmetry illustrates that this chromatographic system adaptability is good.
Specificity is investigated the selection with wavelength: lack the Radix Paeoniae Rubra negative control when measuring at the 230nm place and slightly disturb, the selected wavelength 243nm place that measures measures, the peak energy of peoniflorin reaches fully and separates in the sample chromatogram, lack in the Radix Paeoniae Rubra negative control chromatograph, the place is noiseless substantially at the peoniflorin peak, so this law possesses the specificity of experiment.
(2) drafting of standard curve: it is an amount of that precision takes by weighing the peoniflorin reference substance, prepares peoniflorin standard solution 9.82,19.64,39.28,58.92,78.56 μ gml respectively with 35% methanol
-1Precision is got 10 μ l injecting chromatographs respectively.With the peak area integrated value sample size (μ g) is returned, get standard curve and regression equation: Area=774.8936*Amount～4.2192, r=0.9997.The result shows: sample size 0.0982～0.7856 μ g scope internal linear relation is good.
(3) investigation of extraction conditions: because the granule made for the water extracted immersing paste agent of sample, consider the solubility behavior of peoniflorin, use with a collection of test sample and investigate extracting solvent and supersound extraction time respectively that measurement result sees Table 1, table 2.
Table 1 different solvents extracts peoniflorin measurement result (n=3)
Table 2 peoniflorin measurement result of different extraction time
Extract the measured value no significant difference that solvent adopts water, 50% methanol, 70% methanol and methanol, methanol extraction has more the assorted peak at next-door neighbour's peoniflorin peak, but the insoluble matter of 70% methanol extract liquid is less, and assorted effect is less; The measured value of 10～20 minutes supersound extraction time is higher.
Conclusion: the optimum extraction condition that paeoniflorin content is measured in the test sample is: with 70% methanol as extracting solvent, supersound process 10～20 minutes.Influence in conjunction with the fineness of sample powder was defined as supersound process 20 minutes in the quality standard.
(4) need testing solution and reference substance solution study on the stability: press method operation under the quality standard assay item, different time mensuration reference substance (39.28 μ g/ml) and the concentration and the content of need testing solution see Table 3 on the same day.
Table 3 reference substance solution and need testing solution study on the stability
Result of the test shows: reference substance and need testing solution were placed 27 hours at ambient temperature, and the concentration of peoniflorin does not obviously change.Once found also in experiment that reference substance solution preserved one month in refrigerator, its concentration is constant substantially.
(5) precision test: reference substance solution (39.28 μ g/ml) 10 μ l and continuous 5 sample introductions of need testing solution 10 μ l, RSD is respectively 2.42% and 2.15% (seeing Table 4).Show that according to result of the test sample introduction precision is good.
Table 4 Precision test result
(6) replica test: to pressing the method under the assay item in the quality standard, carry out parallel assay 5 times, the results are shown in Table 5, this method good reproducibility with a collection of test sample (lot number 000701).
Table 5 replica test result
(7) sample pipetting volume recovery test: get the about 0.55g of test sample, the accurate title, decide, and the adding peoniflorin is an amount of, presses the method time-and-motion study response rate under the quality standard assay item, the results are shown in Table 6.Show that according to result of the test the response rate of this method is good.
Table 6 sample pipetting volume recovery test result
The content of (8) three batch samples: measure three batch samples by method under the paeoniflorin content mensuration item in the quality standard, it the results are shown in Table 7.According to measurement result and crude drug source and production concrete condition, in the standard in the regulation Fuyankan particles content of paeoniflorin be: every bag is no less than 5mg.
Paeoniflorin content measurement result in three batches of Fuyankan particles of table 7
The specific embodiment:
Embodiment one: the method for quality control of this blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule comprises
[prescription] Radix Angelicae Sinensis 90g, Radix Paeoniae Rubra 90g, Sonchus brachyotus DC. 240g, Rhizoma Cyperi (vinegar system) 90g, Rhizoma Zingiberis Preparatum 90g, Herba Lycopi 90g, Rhizoma Chuanxiong 60g, Flos Carthami 60g, Radix Bupleuri 90g, Sargassum 90g, Semen Plantaginis (salt moxibustion) 120g, Rhizoma Corydalis 60g.
[method for making] above 12 flavors remove Rhizoma Corydalis and are ground into fine powder, sieve; Radix Angelicae Sinensis, Rhizoma Cyperi, Rhizoma Chuanxiong, Radix Bupleuri extracts volatile oil, and the aqueous solution after distillation device is in addition collected, medicinal residues and Radix Paeoniae Rubra, Rhizoma Zingiberis Preparatum, Sargassum, Semen Plantaginis decocts with water secondary, adds 8 times of water gagings for the first time and decocts 2 hours, adds 6 times of water gagings for the second time and decocts 1 hour, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, medicinal liquid is standby, all the other Sonchus brachyotus DC., Flos Carthami, after Herba Lycopi's three flavors add water boil,, add 8 times of water gaging warm macerating 2 hours for the first time in 80 ℃ of warm macerating secondaries, add for the second time 6 times of water gaging warm macerating 1 hour, filter, merge above each medicinal liquid, be concentrated into the extractum of relative density 1.32～1.38 (50～60 ℃), get whole extractum, add an amount of dextrin, with above-mentioned Rhizoma Corydalis powder mixing, adding ethanol is an amount of, make granule, drying adds above-mentioned Radix Angelicae Sinensis, Rhizoma Cyperi, Rhizoma Chuanxiong, Radix Bupleuri volatile oil, mixing, make the 450g granule, promptly.
[character] this product is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of.
[discriminating]
(1) get this product 9g, porphyrize, 20ml adds diethyl ether, supersound process 20 minutes filters, and the filtrate water-bath volatilizes, residue adds ethyl acetate 0.5ml makes its dissolving, as need testing solution: get Radix Angelicae Sinensis, each 1g of Rhizoma Chuanxiong control medicinal material respectively in addition, make control medicinal material solution respectively with method; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, Radix Angelicae Sinensis control medicinal material solution, each 5～10 μ l of Rhizoma Chuanxiong control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane: the solution of ethyl acetate=9: 1 is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
(2) get this product 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, carries with water saturated n-butyl alcohol and getting respectively twice, each 25ml merges n-butyl alcohol liquid, with the saturated water difference washed twice of n-butyl alcohol, each 10ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, each 4 μ l of peoniflorin reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=50: 20: 10: 2.5 is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this product 18g, porphyrize adds methanol 50ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes its dissolving, transfer PH to 8 with strong ammonia solution, extract respectively three times, each 10ml with ether, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution; Other gets Rhizoma Corydalis control medicinal material 1g, shines medical material solution in pairs with legal system; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, each 5 μ l of Rhizoma Corydalis control medicinal material solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=7.5: 4: 1 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[inspection] this product should meet pertinent regulations under the granule item (appendix IC of Chinese Pharmacopoeia).
[assay] measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia).
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=15: 85 is a mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500.
The preparation of reference substance solution: precision takes by weighing at 80 ℃ of drying under reduced pressure an amount of to the peoniflorin reference substance of constant weight, adds 35% methanol and makes the solution that every 1ml contains 20 μ g, promptly.
The preparation of need testing solution: get this product under the content uniformity item, porphyrize is got 0.5g, and accurate the title decides, put in the tool plug conical flask, the accurate 70% methanol solution 25ml that adds, supersound process 20 minutes filters, precision is measured subsequent filtrate 5ml, puts in the 10ml measuring bottle, adds water to scale, shakes up promptly.
Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 5～10 μ l of drawing, inject chromatograph of liquid, measure, promptly.
This product contains Radix Paeoniae Rubra with peoniflorin (C for every bag
23H
28O
11) meter, must not be less than 5mg.
[function with cure mainly] blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling; Be used for chronic pelvic inflammatory disease.
[usage and consumption] boiled water is taken after mixing it with water, a 9g, 2 times on the one.
[specification] every packed 9g.
[storage] sealing.
Embodiment two: the method for quality control of this blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule comprises
[prescription], [method for making] are with embodiment one.
[character] this product is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of.
[discriminating]
(1) get this product 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, with twice of water saturated n-butanol extraction, each 25ml merges n-butyl alcohol liquid, uses twice of the saturated water washing of n-butyl alcohol, each 10ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, each 4 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=58: 22: 12: 2.8 is developing solvent, launch, take out, dry, spray is with 7% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(2) get this product 18g, porphyrize adds methanol 50ml, and supersound process 30 minutes filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, transfers PH to 8 with strong ammonia solution, uses ether extraction three times, each 10ml, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution.It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, according to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw each 5 μ L of above-mentioned need testing solution, reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=9: 5: 1.2 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[inspection] this product should meet pertinent regulations under the granule item (appendix IC of Chinese Pharmacopoeia).
[assay] measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia).
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=19: 95 is a mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: it is an amount of to constant weight peoniflorin reference substance that precision takes by weighing 80 ℃ of drying under reduced pressure, adds 35% methanol and make the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this product under the content uniformity item, porphyrize is got 1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate 70% methanol solution 25ml that adds, supersound process 20 minutes filters, and precision is measured subsequent filtrate 5ml, put in the 1Oml measuring bottle, add water to scale, shake up, promptly;
Algoscopy: accurate respectively reference substance solution 10 μ l and the about 8 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This preparation contains Radix Paeoniae Rubra with peoniflorin (C for every bag
23H
28O
11) meter, must not be less than 5mg.
[function with cure mainly] blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling; Be used for chronic pelvic inflammatory disease.
[usage and consumption] boiled water is taken after mixing it with water, a 9g, 2 times on the one.
[specification] every packed 9g.
[storage] sealing.
Embodiment three: the method for quality control of this blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granular preparation comprises
[prescription], [method for making] are with embodiment one.
[character] this preparation is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of.
[discriminating]
(1) get this preparation or its content 9g, porphyrize, the 20ml that adds diethyl ether, supersound process 20 minutes filters, and the filtrate water-bath volatilizes, and O.5ml residue adds ethyl acetate makes its dissolving, as need testing solution; Other gets Radix Angelicae Sinensis respectively, the Rhizoma Chuanxiong control medicinal material is an amount of, makes contrast solution respectively with method; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, two kinds of each 5～10 μ L of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane: the solution of ethyl acetate=9: 1 is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
(2) get this preparation or its content 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 1Oml makes its dissolving, carries with water saturated n-butyl alcohol and getting respectively twice, each 25ml merges n-butyl alcohol liquid, with the saturated water difference washed twice of n-butyl alcohol, each 1Oml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw above-mentioned need testing solution, each 4 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=45: 22: 12: 2.5 is developing solvent, launch, take out, dry, spray is with 1～8% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this preparation or its content 18g, porphyrize adds methanol 50ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes its dissolving, transfer PH to 8 with strong ammonia solution, extract respectively three times, each 10ml with ether, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution; It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, according to appendix thin layer chromatography test of Chinese Pharmacopoeia, draw each 5 μ l of above-mentioned need testing solution, control medicinal material solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=8: 5: 1.3 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[inspection] this product should meet pertinent regulations under the granule item (appendix IC of Chinese Pharmacopoeia).
[assay] measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia).
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=16: 82 is a mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: it is an amount of to constant weight peoniflorin reference substance that precision takes by weighing 80 ℃ of drying under reduced pressure, adds 35% methanol and make the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this product under this preparation content uniformity item, porphyrize is got 1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate 70% methanol solution 25ml that adds, supersound process 20 minutes filters, and precision is measured subsequent filtrate 5ml, put in the 10ml measuring bottle, add water to scale, shake up, promptly;
Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 5～10 μ l of drawing, inject chromatograph of liquid, measure, promptly.
This preparation contains Radix Paeoniae Rubra with peoniflorin (C for every bag
23H
28O
11) meter, must not be less than 5mg.
[function with cure mainly] blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling; Be used for chronic pelvic inflammatory disease.
[usage and consumption] boiled water is taken after mixing it with water, a 9g, 2 times on the one.
[specification] every packed 9g.
[storage] sealing.
Adopt the method for quality control of above embodiment one,, more help guaranteeing the clinical efficacy of this medicine the quality of this preparation of easier control.
Claims (10)
1. the method for quality control of a blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule, described granule is prepared from by Radix Angelicae Sinensis, Radix Paeoniae Rubra, Sonchus brachyotus DC., Rhizoma Cyperi (processed with vinegar), Rhizoma Zingiberis Preparatum, Herba Lycopi, Rhizoma Chuanxiong, Flos Carthami, Radix Bupleuri, Sargassum, salt moxibustion Semen Plantaginis, Rhizoma Corydalis, it is characterized in that described method of quality control mainly comprise in character, discriminating, inspection, the assay project partly or entirely; Differentiate that wherein the thin layer chromatography comprise Radix Angelicae Sinensis in the granule and Rhizoma Chuanxiong, Radix Paeoniae Rubra, Rhizoma Corydalis differentiates; Assay is that the contained content of paeoniflorin of Radix Paeoniae Rubra in the granule is measured.
2. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 1, removing toxic substances and promoting subsidence of swelling granule, the discrimination method that it is characterized in that Radix Angelicae Sinensis and Rhizoma Chuanxiong is to be contrast with the control medicinal material of Radix Angelicae Sinensis and Rhizoma Chuanxiong respectively, and with normal hexane: ethyl acetate=6～12: 0.5～1.5 is the tlc identification method of developing solvent.
3. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 1, removing toxic substances and promoting subsidence of swelling granule, the discrimination method that it is characterized in that peoniflorin is to be contrast with the peoniflorin reference substance, and with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～24: 7～13: 2～3 is the tlc identification method of developing solvent.
4. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 1, removing toxic substances and promoting subsidence of swelling granule, the discrimination method that it is characterized in that Rhizoma Corydalis is to be contrast with the Rhizoma Corydalis control medicinal material, and with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is the tlc identification method of developing solvent.
5. according to the method for quality control of claim 1,2,3 or 4 described blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that described discrimination method comprises the part or all of of following project:
(1) get this granule, porphyrize adds ethanol, and supersound process filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate makes its dissolving, as need testing solution; Other gets Radix Angelicae Sinensis respectively, the Rhizoma Chuanxiong control medicinal material is an amount of, makes control medicinal material solution respectively with method; According to Chinese Pharmacopoeia thin layer chromatography test, draw need testing solution, two kinds of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6～12: 0.5～1.5 be developing solvent, launches, and taking-up is dried, and puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(2) get this granule, porphyrize adds ethanol, reflux is handled, and puts coldly, filters, filtrate evaporate to dryness, residue add water makes its dissolving, with twice of water saturated n-butanol extraction, merge n-butyl alcohol liquid, with the saturated water washing twice of n-butyl alcohol, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes solution, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～25: 7～13: 2～3 is developing solvent, launch, take out, dry, spray is with 2～8% vanillin sulfuric acid solutions, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this granule, porphyrize adds methanol, and supersound process filters, and filtrate evaporate to dryness, residue add water makes its dissolving, transfers PH to 8 with strong ammonia solution, uses ether extraction three times, merges ether solution, and evaporate to dryness, residue add methanol makes its dissolving, as need testing solution; It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate with the sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is developing solvent, launching in the pre-saturated expansion cylinder with developing solvent, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
6. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 5, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that discrimination method comprises the part or all of of following project more specifically:
(1) get this granule 9g, porphyrize adds ethanol 20ml, and supersound process 20 minutes filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate 0.5ml makes its dissolving, as need testing solution; Get Radix Angelicae Sinensis, each 1g of Rhizoma Chuanxiong control medicinal material respectively in addition, make control medicinal material solution respectively with method; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, two kinds of each 5～10 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=9: 1 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(2) get this granule 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, with twice of water saturated n-butanol extraction, each 25ml merges n-butyl alcohol liquid, uses twice of the saturated water washing of n-butyl alcohol, each 10ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, each 4 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=50: 20: 10: 2.5 is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this granule 18g, porphyrize adds methanol 50ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes its dissolving, transfer PH to 8 with strong ammonia solution, use ether extraction three times, each 10ml, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution; Other gets Rhizoma Corydalis control medicinal material 1g, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of described need testing solution, reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=7.5: 4: 1 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
7. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 1, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that: it is to be contrast with the peoniflorin reference substance that the contained content of paeoniflorin of Radix Paeoniae Rubra is measured, and with acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is the high performance liquid chromatography of mobile phase.
8. according to the method for quality control of claim 1 or 7 described blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that described paeoniflorin content measures according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring, comprise following:
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: take by weighing the peoniflorin reference substance of 80 ℃ of drying under reduced pressure, add methanol and make the solution that every 1ml contains 20 μ g, promptly to constant weight;
The preparation of need testing solution: get this granule under the content uniformity item, porphyrize is got 0.5g, puts in the tool plug conical flask, add 70% methanol solution 25ml, supersound process 20 minutes filters, and measures subsequent filtrate 5ml, put in the 10ml measuring bottle, add water to scale, shake up, promptly;
Algoscopy: draw reference substance solution 10 μ l and need testing solution 5～10 μ l respectively, inject chromatograph of liquid, measure, promptly;
This granule contains Radix Paeoniae Rubra in peoniflorin, must not be less than 5mg.
9. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 1, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that described method of quality control comprises:
Character: this granule is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of;
Differentiate:
(1) get this granule, porphyrize adds ethanol, and supersound process filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate makes its dissolving, as need testing solution; Other gets Radix Angelicae Sinensis respectively, the Rhizoma Chuanxiong control medicinal material is an amount of, makes contrast solution respectively with method; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, two kinds of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6～12: 0.5～1.5 is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(2) get this granule, porphyrize adds ethanol, reflux is handled, and puts coldly, filters, filtrate evaporate to dryness, residue add water makes its dissolving, with twice of water saturated n-butanol extraction, merge n-butyl alcohol liquid, with the saturated water washing twice of n-butyl alcohol, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes solution, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=40～60: 16～24: 7～13: 2～3 is developing solvent, launch, take out, dry, spray is with 2～8% vanillin sulfuric acid solutions, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this granule, porphyrize adds the methanol supersound process, filters, and filtrate evaporate to dryness, residue add water makes its dissolving, transfers PH to 8 with strong ammonia solution, uses ether extraction, merges ether solution, and evaporate to dryness, residue add methanol makes its dissolving, as need testing solution; It is an amount of that other gets the Rhizoma Corydalis control medicinal material, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw described need testing solution, reference substance solution, put respectively on same silica gel g thin-layer plate with sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=5～10: 3～5: 0.6～1.5 is developing solvent, in with the expansion cylinder after the developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Check: this granule should meet pertinent regulations under the Chinese Pharmacopoeia granule item;
Assay: paeoniflorin content is measured according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=10～20: 70～100 is mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: it is an amount of to the peoniflorin reference substance of constant weight to take by weighing 80 ℃ of drying under reduced pressure, adds methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this granule under the content uniformity item, porphyrize is got 0.5g, puts in the tool plug conical flask, adds methanol solution, and supersound process filters, and measures subsequent filtrate, puts in the measuring bottle, adds water to scale, shakes up, promptly;
Algoscopy: draw reference substance solution 10 μ l and need testing solution 5～10 μ l respectively, inject chromatograph of liquid, measure, promptly;
This granule contains Radix Paeoniae Rubra in peoniflorin for every bag, must not be less than 5mg.
10. according to the method for quality control of the described blood circulation promoting and blood stasis dispelling of claim 9, removing toxic substances and promoting subsidence of swelling granule, it is characterized in that described method of quality control comprises:
Character: this granule is brown xanchromatic granule; Sweet, little hardship of distinguishing the flavor of;
Differentiate:
(1) get this granule 9g, porphyrize adds ethanol 20ml, and supersound process 20 minutes filters, and the filtrate water-bath volatilizes, and residue adds ethyl acetate 0.5ml makes its dissolving, as need testing solution; Get Radix Angelicae Sinensis, each 1g of Rhizoma Chuanxiong control medicinal material respectively in addition, make control medicinal material solution respectively with method; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned need testing solution, two kinds of each 5～10 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=9: 1 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(2) get this granule 9g, porphyrize adds ethanol 30ml, reflux 1 hour is put coldly, filters, filtrate evaporate to dryness, residue add water 10ml makes its dissolving, with twice of water saturated n-butanol extraction, each 25ml merges n-butyl alcohol liquid, uses twice of the saturated water washing of n-butyl alcohol, each 10ml discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 1ml makes its dissolving, as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned need testing solution, each 4 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform: methanol: ethyl acetate: strong ammonia solution=50: 20: 10: 2.5 is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(3) get this granule 18g, porphyrize adds methanol 50ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes its dissolving, transfer to PH to 8 with strong ammonia solution, use ether extraction three times, each 10ml, merge ether solution, evaporate to dryness, residue add methanol 1ml makes its dissolving, as need testing solution; Other gets Rhizoma Corydalis control medicinal material 1g, shine medical material solution in pairs with legal system, test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of described need testing solution, reference substance solution, put respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation, with normal hexane: chloroform: methanol=7.5: 4: 1 is developing solvent, in with 1 hour expansion cylinder of developing solvent presaturation, launch, take out, dry, smoked clear with iodine steam to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Check: this granule should meet pertinent regulations under the Chinese Pharmacopoeia granule item;
Assay: according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile: 0.1% phosphoric acid solution=15: 85 is a mobile phase; The detection wavelength is 243nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 5500;
The preparation of reference substance solution: precision takes by weighing at 80 ℃ of drying under reduced pressure an amount of to the peoniflorin reference substance of constant weight, adds 35% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get this granule under the content uniformity item, porphyrize is got 0.5g, and accurate the title decides, put in the tool plug conical flask, the accurate 70% methanol solution 25ml that adds, supersound process 20 minutes filters, precision is measured subsequent filtrate 5ml, puts in the 10ml measuring bottle and adds water to scale, shakes up, promptly;
Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 5～10 μ l of drawing, inject chromatograph of liquid, measure, promptly;
This granule contains Radix Paeoniae Rubra in peoniflorin for every bag, must not be less than 5mg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100460387A CN101352565A (en) | 2008-09-09 | 2008-09-09 | Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100460387A CN101352565A (en) | 2008-09-09 | 2008-09-09 | Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101352565A true CN101352565A (en) | 2009-01-28 |
Family
ID=40305744
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008100460387A Pending CN101352565A (en) | 2008-09-09 | 2008-09-09 | Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101352565A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103308644A (en) * | 2013-07-03 | 2013-09-18 | 广西邦琪药业集团有限公司 | Quality detection method for miscarriage-preventing leonurus preparation |
| CN103983734A (en) * | 2014-05-26 | 2014-08-13 | 江西民济药业有限公司 | Detection method for preparing puerperal blood-stasis dispersing capsule |
| CN107179380A (en) * | 2017-06-21 | 2017-09-19 | 山西振东安特生物制药有限公司 | Naozhenning TLC Identification |
| CN113252835A (en) * | 2021-05-18 | 2021-08-13 | 段复华 | Thin-layer chromatography identification method of Yanhuweian capsules |
| CN115902084A (en) * | 2022-11-04 | 2023-04-04 | 杭州市中医院 | Method for detecting quality of spleen-tonifying miscarriage-prevention granules |
-
2008
- 2008-09-09 CN CNA2008100460387A patent/CN101352565A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103308644A (en) * | 2013-07-03 | 2013-09-18 | 广西邦琪药业集团有限公司 | Quality detection method for miscarriage-preventing leonurus preparation |
| CN103983734A (en) * | 2014-05-26 | 2014-08-13 | 江西民济药业有限公司 | Detection method for preparing puerperal blood-stasis dispersing capsule |
| CN103983734B (en) * | 2014-05-26 | 2015-12-09 | 江西民济药业有限公司 | A kind of detection method preparing Chanhou Zhuyu Capsule |
| CN107179380A (en) * | 2017-06-21 | 2017-09-19 | 山西振东安特生物制药有限公司 | Naozhenning TLC Identification |
| CN113252835A (en) * | 2021-05-18 | 2021-08-13 | 段复华 | Thin-layer chromatography identification method of Yanhuweian capsules |
| CN115902084A (en) * | 2022-11-04 | 2023-04-04 | 杭州市中医院 | Method for detecting quality of spleen-tonifying miscarriage-prevention granules |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN102539553B (en) | Method for establishing fingerprint spectrum of liver-enhancing medicine | |
| CN107478749B (en) | Detection method of medicinal preparation of Taohong Siwu decoction | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN101766771A (en) | Quality control method of medicine for gynecopathy treatment | |
| CN103285306B (en) | Preparation method and detection method of traditional Chinese medicine composition for benefiting Qi and tonifying kidney | |
| CN106370738B (en) | Fingerprint quality determination method for periplaneta americana medicinal material | |
| CN101966223A (en) | Fingerprint detection method for compound wintercreeper preparation | |
| CN105301168B (en) | The detection method of dredging collateral resolving sputum capsule | |
| CN101513467A (en) | Method for controlling quality of dermatosis toxemia preparation | |
| CN106290599B (en) | Content determination method of traditional Chinese medicine composition | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN101352565A (en) | Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling | |
| CN106370756B (en) | A kind of detection method of Chinese materia medica preparation that preventing and treating infectious bronchitis of chicken | |
| CN102145146B (en) | Method for detecting medicinal composition for treating urinary system diseases | |
| CN104407092A (en) | Quality detection method of traditional Chinese medicinal composition with efficacy of appetizing and invigorating spleen | |
| CN101926889A (en) | Method for detecting white paeony root-medlar particles | |
| CN102068598B (en) | Quality control method of Yangrong Baicao Wan for treating irregular menses caused by hemophthisis | |
| CN100401061C (en) | Quality control method of kidney beneficial bone fortifying capsule | |
| CN101816753B (en) | Method for detecting quality of compound preparation for treating cold | |
| CN101716270B (en) | Method for detecting quality of traditional Chinese herbal medicament compound preparation for invigorating blood and regulating menses | |
| CN100585401C (en) | Detection method for orally administered formulation for reducing fat and expelling toxins | |
| CN1857445B (en) | Quality control method for Desheng preparation | |
| CN108535399A (en) | A kind of detection method of Fuyankang pill | |
| CN101028474B (en) | Method for inspecting the quality of Chinese preparation with Yang-and kidney tonifying functions |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20090128 |