CN102759598A - Quality detection method for 29-componnet stagnation dissipation powder as Tibetan medicinal composition and preparations thereof - Google Patents
Quality detection method for 29-componnet stagnation dissipation powder as Tibetan medicinal composition and preparations thereof Download PDFInfo
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- CN102759598A CN102759598A CN2012102722431A CN201210272243A CN102759598A CN 102759598 A CN102759598 A CN 102759598A CN 2012102722431 A CN2012102722431 A CN 2012102722431A CN 201210272243 A CN201210272243 A CN 201210272243A CN 102759598 A CN102759598 A CN 102759598A
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Abstract
The invention discloses a quality detection method for 29-componnet stagnation dissipation powder as a Tibetan medicinal composition and preparations thereof. IN the quality detection method, thin layer chromatography is used to carry out qualitative identification on frankincense, piperine and myristica fragrans in the 29-componnet stagnation dissipation powder, high performance liquid chromatography (HPLC) is used to carry out limit test on aconitine and costustoot aristolochic acid in the 29-componnet stagnation dissipation powder, and simultaneously, the high performance liquid chromatography is used to carry out quantitative detection on aloe-emodin, chrysophanol, rhein, emodin, emodin monomethyl ether and hydroxysafflor yellow A in the 29-componnet stagnation dissipation powder. According to the invention, the thin-layer identification method for the frankincense, the piperine and the myristica fragrans, the limit test method for the aconitine and the costustoot aristolochic acid, and the content determination method for the aloe-emodin, the chrysophanol, the rhein, the emodin, the emodin monomethyl ether and the hydroxysafflor yellow A are developed, so that the quality standard of a product is increased, and drug safety and effectiveness are guaranteed while the 29-componnet stagnation dissipation powder and the preparations thereof are used for treating diseases.
Description
Technical field
The present invention relates to the quality determining method of a kind of Tibetan medicine composition and preparation thereof, particularly a kind of Tibetan medicine composition 29 flavors can dissipate and the quality determining method of preparation.
Background technology
29 flavors can dissipate and be recorded in " the clinical reading notes of Tibetan medicine ", and good the arranging of author's sea urchin roe paste is the famous Tibetan medicine in 13rd century, and he sums up through clinical practice for many years, and on the basis of forefathers' prescription, concocting out 29 flavors can dissipate.Because of its good efficacy, praised highly by successive dynasties Tibetan medicine scholar to hysteromyoma and ovarian cyst.29 flavors can dissipate and distinguished the flavor of by Tibet inula root, gypsum rubrum (forging), myrobalan, hot millet, alkali flower, nutmeg, the Bi roots of grass, rheum officinale, iron staff hammer, safflower, banksia rose birthwort, Semen seu folium abelmoschi moschati etc. 29 that medicines form, and have the dispelling cold ruffian, help digestion, the effect of liver-regulating and kidney-nourishing.Be mainly used in accumulation of food in the stomach and intes tine due to indigestion, the stomach and intestine hepatalgia, ephrosis, enteropathy is poisoned, hepatopathy, hysteropathy, yellow water spills into sering, and the stomach and intestine ruffian is sick, and the courage ruffian is sick, the ruffian knurl that chill causes etc.
29 flavors can dissipate and record in the national drug standards, standard numbering: WS3-BC-0140-95.The iron staff hammer contains aconitine in the side, has higher pharmacologically active and medical value, also has stronger toxicity simultaneously, though obviously reduce through concocting the back toxic component, can not guarantee medicinal security; Banksia rose birthwort contains aristolochic acid in the side, and aristolochic acid can cause bad reactions such as kidney damage.Differentiate not and the assay item in the proper mass standard that the aconitine that the iron staff hammer is not contained, the aristolochic acid that banksia rose birthwort contains carry out limit test, make said preparation have potential safety hazard.Simultaneously, said preparation do not differentiate and assay its effective constituent yet, thereby can not guarantee effective use of said preparation.Document and patent are not seen can the dissipate report of quality testing aspect of relevant 29 flavors yet.Therefore, safe and effective for medication in order to guarantee the patient, the proper mass standard is demanded urgently and need be improved.
Summary of the invention
To the deficiency of prior art, the purpose of this invention is to provide that a kind of Tibetan medicine composition 29 flavors can dissipate and the quality determining method of preparation.
Summary of the invention
The invention provides that a kind of Tibetan medicine composition 29 flavors can dissipate and the limit test method of preparation mesaconitine; The limit test method of aristolochic acid A; Frankincense, pipering, myristic thin-layer chromatography discrimination method are provided simultaneously; The high performance liquid chromatography content assaying method of aloe-emodin, Chrysophanol, Rhein, archen, Physcion, hydroxyl radical carthamin yellow carthamus A; Make the said preparation quality testing more accurately, reliably, make said preparation in the treatment disease, guaranteed the safe and effective of medication.
The term explanation:
29 flavors can dissipate, and are the nomenclature of drugs of national standard for traditional Chinese medicines compilation (Chinese patent drug provincial standard rising national standard part) record.
29 flavors can dissipate and preparation, comprise the 29 Tibetan medicine compositions and with 29 preparations of bulk drug formulation of dissipating of distinguishing the flavor of of dissipating of distinguishing the flavor of.
Gypsum rubrum (forging), radish (charcoal), shellfish tooth (charcoal), lime (system), vulture excrement (stir-fry): the routine techniques term that is Chinese medicine (Tibetan medicine); " forging " in the bracket, " charcoal ", " system ", " stir-fry " etc. are all concocted according to the method for " Qinghai Province's Tibetan medicine concocted specification " (food and medicine Surveillance Authority in Qinghai Province's compiles, the versions in 2010 that the Qinghai People's Press publishes) lining.
Technical scheme of the present invention is following:
A kind of bulk drug consists of Tibet inula root 25 weight portions, gypsum rubrum (forging) 125 weight portions, myrobalan's 75 weight portions, hot millet 25 weight portions, alkali and spends 125 weight portions, nutmeg 25 weight portions, the Bi roots of grass 25 weight portions, cassia seed 25 weight portions, Amomum cardamomum 25 weight portions, the rhizome of davallia 25 weight portions, pepper 25 weight portions, radish (charcoal) 2 weight portions; Tsaoko 25 weight portions, natural salt 25 weight portions, asafoetide 2 weight portions, sal ammoniac 25 weight portions, kaempferia galamga 25 weight portions, shellfish tooth (charcoal) 25 weight portions, rheum officinale 100 weight portions, wide muscle rattan 13 weight portions, safflower 25 weight portions, iron staff are hammered 15 weight portions, lime (system) 25 weight portions, vulture excrement (stir-fry) 40 weight portions, banksia rose birthwort 25 weight portions, Semen seu folium abelmoschi moschati 25 weight portions, HALITUM PURPUREUM 25 weight portions, frankincense 25 weight portions, slag into shape and are tamed and dociled that Tibetan medicine composition 29 flavors of cream 25 weight portions can dissipate and the quality determining method of preparation, and this method comprises one or more in following discriminating and/or inspection and/or the content assaying method:
Differentiate:
A. the discriminating of frankincense
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 2 ~ 4g, the 20 ~ 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the xylene-ethyl acetate of 8:1 ~ 2 is a developping agent; Launch, take out, dry; Spray is with mass and size portion rate 1% vanillic aldehyde 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;
B. the discriminating of pipering
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 2 ~ 4g, add ethyl acetate 20 ~ 30ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the cyclohexane-ethyl acetate of 3:1 ~ 3 is a developping agent; Launch, take out, dry; 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. myristic discriminating
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 4 ~ 6g, put in the 250ml round-bottomed flask, add water 100ml; Decoct 1h; Collect volatile oil with volatile oil determination apparatus, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished; Collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml, decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished, collect the ethyl acetate part, as control medicinal material solution with volatile oil determination apparatus; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the xylene-ethyl acetate of 20:0.1 ~ 0.5 is a developping agent; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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;
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 65 ~ 75:25 ~ 35:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 10 ~ 15g, the accurate title, decide, and puts in the conical flask of tool plug, adds ammonia solution 10ml; The accurate absolute ethyl alcohol 80～120ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ℃ ~ 40 ℃ decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ℃ ~ 40 ℃ decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 10 ~ 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g.
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts of 60～70:30～40 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 6 ~ 10g, the accurate title, decide, and puts in the tool plug conical flask, accurate methyl alcohol 40 ~ 60ml, the close plug of adding; Claim decide weight, ultrasonic 20 ~ 40 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times, 20ml at every turn; Merge ether extraction liquid, volatilize, with dissolve with methanol and be transferred in the 5ml volumetric flask, add methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 ~ 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
The present invention's 29 flavors can dissipate or its preparation contains banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g.
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the volume parts ratio is that the methyl alcohol-volume parts of 80 ~ 90:10 ~ 20 is a moving phase than the phosphate aqueous solution that is 0.1%; The detection wavelength is 430nm.Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Be to contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 1 ~ 3g, the accurate title, decide, and puts in the conical flask of tool plug, the accurate methyl alcohol 20 ~ 30ml that adds, and close plug claims to decide weight; Ultrasonic 20 ~ 40min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 5 ~ 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
The present invention's 29 flavors can dissipate or its preparation contains rheum officinale with aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) the total content meter, must not be less than 1.20mg/g.
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than the methyl alcohol-volume parts ratio of 20～30:70～80; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 3 ~ 5g, the accurate title, decide, and puts in the tool plug conical flask, and it is 25% methanol aqueous solution, 20 ~ 30ml that precision adds the volume parts ratio; Close plug is claimed to decide weight, ultrasonic 30 ~ 50 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 5 ~ 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
The present invention 29 flavor can dissipate or its preparation in hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
Described preparation is meant that getting can dissipate bulk drug Tibet inula root 25 weight portions, gypsum rubrum (forging) 125 weight portions, myrobalan's 75 weight portions, hot millet 25 weight portions, alkali of Tibetan medicine composition 29 flavor spends 125 weight portions, nutmeg 25 weight portions, the Bi roots of grass 25 weight portions, cassia seed 25 weight portions, Amomum cardamomum 25 weight portions, the rhizome of davallia 25 weight portions, pepper 25 weight portions, radish (charcoal) 2 weight portions; Tsaoko 25 weight portions, natural salt 25 weight portions, asafoetide 2 weight portions, sal ammoniac 25 weight portions, kaempferia galamga 25 weight portions, shellfish tooth (charcoal) 25 weight portions, rheum officinale 100 weight portions, wide muscle rattan 13 weight portions, safflower 25 weight portions, iron staff are hammered 15 weight portions, lime (system) 25 weight portions, vulture excrement (stir-fry) 40 weight portions, banksia rose birthwort 25 weight portions, Semen seu folium abelmoschi moschati 25 weight portions, HALITUM PURPUREUM 25 weight portions, frankincense 25 weight portions, tame and docile cream 25 weight portions of slag into shape; Press common process; Add conventional auxiliary material and be prepared into clinical acceptable any formulation, comprise micropill, dripping pill, tablet, capsule, particle or dispersing tablet.
Preferred a kind of bulk drug consists of Tibet inula root 25g, gypsum rubrum (forging) 125g, myrobalan 75g, hot millet 25g, alkali flower 125g, nutmeg 25g, Bi roots of grass 25g, cassia seed 25g, Amomum cardamomum 25g, rhizome of davallia 25g, pepper 25g, radish (charcoal) 2g; The Tibetan medicine composition 29 of tsaoko 25g, natural salt 25g, asafoetide 2g, sal ammoniac 25g, kaempferia galamga 25g, shellfish tooth (charcoal) 25g, rheum officinale 100g, wide muscle rattan 13g, safflower 25g, iron staff hammer 15g, lime (system) 25g, vulture excrement (stir-frys) 40g, banksia rose birthwort 25g, Semen seu folium abelmoschi moschati 25g, HALITUM PURPUREUM 25g, frankincense 25g, the tame and docile cream 25g of slag is distinguished the flavor of and can be dissipated and the quality determining method of preparation, and this method comprises one or more in following discriminating and/or inspection and/or the content assaying method:
Differentiate:
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 8:1.5; Launch, take out, dry; Spray is with 1% vanillic aldehyde 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.
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the cyclohexane-ethyl acetate that is 3:2; Launch, take out, dry; 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. myristic discriminating
Get the Tibetan medicine composition 29 flavor powder 5g that can dissipate, put in the 250ml round-bottomed flask, add water 100ml; Decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml, decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished, collect the ethyl acetate part, as control medicinal material solution with volatile oil determination apparatus; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 20:0.3; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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.
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 68:32:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 12g that can dissipate, accurately claim surely, put in the conical flask of tool plug, add ammonia solution 10ml; The accurate absolute ethyl alcohol 100ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g.
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than methyl alcohol-volume parts of 66:34 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 8g that can dissipate, accurately claim surely, put in the tool plug conical flask accurate methyl alcohol 50ml, the close plug of adding; Claim decide weight, ultrasonic 30 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g.
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts that is 86:14 than the phosphate aqueous solution that is 0.1% with volume parts; The detection wavelength is 430nm.Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 2g that can dissipate, accurately claim surely, put in the conical flask of tool plug, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Ultrasonic 30min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
The present invention 29 flavor can dissipate or its preparation in aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g.
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than methyl alcohol-volume parts ratio of 24:76; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 4g that can dissipate, accurately claim surely, put in the tool plug conical flask, accurate to add the volume parts ratio be 25% methanol aqueous solution 25ml; Close plug is claimed to decide weight, ultrasonic 40 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
The present invention 29 flavor can dissipate or its preparation in hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
The unit corresponding relation of weight portion described in this instructions and parts by volume is g/ml or kg/l.
The invention discloses that a kind of Tibetan medicine composition 29 flavors can dissipate and the quality determining method of preparation; Frankincense, pipering, nutmeg had carried out qualitative identification during the use thin-layered chromatography can dissipate to 29 flavors; Use high performance liquid chromatography (HPLC method) that can dissipate mesaconitine, banksia rose aristolochic acid of 29 flavors carried out limit test, adopt simultaneously high performance liquid chromatography (HPLC method) can dissipate to 29 flavors in aloe-emodin, Chrysophanol, Rhein, archen, Physcion, hydroxyl radical carthamin yellow carthamus A carried out detection by quantitative.The inventive method has been set up frankincense, pipering, myristic thin layer discrimination method; The limit test method of aconitine, banksia rose aristolochic acid; The content assaying method of aloe-emodin, Chrysophanol, Rhein, archen, Physcion, hydroxyl radical carthamin yellow carthamus A; Improve target level of product quality, made said preparation in the treatment disease, guaranteed the safe and effective of medication.Simultaneously this law also can be used for other preparations that Tibetan medicine composition 29 flavors can dissipate, like 29 flavors particle, 29 ball, 29 sheet etc. that can disappear of distinguishing the flavor of that can disappear of distinguishing the flavor of that can disappear.
Following experimental example and embodiment are used to further specify but are not limited to the present invention.
Experimental example 1: thin layer identification experiment
Tibetan medicine composition 29 flavor can dissipate and scold Tibetan medicine medicine company incorporated company by the Qinghai gold and provide.
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; Get in the fragrant negative control sample of prescription ratio preparation hypogalactia, be equipped with the negative control sample solution with legal system according to the need testing solution preparation method; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test; Draw each 5 μ l of above-mentioned three kinds of solution; Putting respectively on same silica gel g thin-layer plate, is developping agent with volume parts than the xylene-ethyl acetate for (8:1), (8:1.5), (8:2) respectively, launches; Take out, dry.Spray is with 1% vanillic aldehyde ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear.
The result: in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color, negative control is noiseless.Under volume parts compares for the xylene of 8:1 ~ 2-ethyl acetate developping agent condition, all have and launch effect preferably.Wherein, volume parts is more best than launching effect for the xylene of 8:1.5-ethyl acetate developping agent.The result shows that this discrimination method specificity is strong, can be used as 29 flavors can dissipate in the thin layer discrimination method of frankincense.
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test; Draw each 5 μ l of above-mentioned two kinds of solution; Putting respectively on same silica gel g thin-layer plate, is developping agent with volume parts than the cyclohexane-ethyl acetate for (3:1), (3:2), (1:1) respectively, launches; Take out, dry.Put under the ultraviolet lamp (365nm) and inspect.
The result: in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.Under volume parts compares for the cyclohexane of 3:1 ~ 3-ethyl acetate developping agent condition, all have and launch effect preferably.Wherein, volume parts is more best than launching effect for the cyclohexane of 3:2-ethyl acetate developping agent.The result shows, this discrimination method chromatogram spot colour developing is clear, and degree of separation is good, can be used as 29 flavors can dissipate in the thin layer discrimination method of pipering.
C. myristic discriminating
Get the Tibetan medicine composition 29 flavor powder 5g that can dissipate; Put in the 250ml round-bottomed flask, add water 100ml, decoct 1h; Collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus) with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml; Decoct 1h, collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus), after decoction is accomplished with volatile oil determination apparatus; Collect the ethyl acetate part, as control medicinal material solution; Get negative control sample, be equipped with the negative control sample solution with legal system according to the need testing solution preparation method in prescription ratio preparation misrun cardamom; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; Be developping agent with volume parts than xylene-ethyl acetate respectively for (20:0.1), (20:0.3), (20:0.5); Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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 result: in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color, negative control is noiseless.Under volume parts compares for the xylene of 20:0.1 ~ 0.5-ethyl acetate developping agent condition, all have and launch effect preferably.Wherein, volume parts is more best than launching effect for the xylene of 20:0.3-ethyl acetate developping agent.The result shows that this discrimination method specificity is strong, can be used as 29 the flavor can dissipate in myristic thin layer discrimination method.
Experimental example 2: the limit test experiment of aconitine
1. instrument, reagent and confession test agent
Instrument: L-2100 type Hitachi high performance liquid chromatograph; Tianjin, AuW220D island electronic analytical balance.
Reference substance: aconitine reference substance (Nat'l Pharmaceutical & Biological Products Control Institute) lot number: MUST-11031101.
Sample: 29 flavors can dissipate (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), and lot number is respectively: 20110815,20110816,20110817.
2. detect the selection of wavelength
Get the aconitine reference substance solution, in 190 ~ 400nm wavelength coverage, scan, aconitine has absorption maximum in the 235nm wavelength, so be the detection wavelength according to the selected 235nm of ultraviolet absorpting spectrum.
3. moving phase is selected
Discovering, is moving phase with volume parts than methanol-water-triethylamine-methylene chloride system of 65 ~ 75:25 ~ 35:0.2:5, and aconitine all can reach good chromatographic resolution effect.Wherein, the volume parts ratio with methanol-water-triethylamine-methylene chloride is that 68:32:0.2:5 is that moving phase is optimum.
4. system suitability test
Under above-mentioned chromatographic condition, accurate respectively reference substance solution, each 20 μ l of need testing solution of drawing inject liquid chromatograph, the record chromatogram.The result shows that test sample chromatogram mesaconitine is adjacent the degree of separation of chromatographic peak all greater than 1.5, good separating effect.
5. reference substance solution preparation
It is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets.
6. need testing solution preparation
6.1 the investigation of extraction time
By method under the inspection item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, and reflux is 0.5 hour, 1 hour, 1.5 hours respectively.Content with every gram medicine mesaconitine is that index is confirmed extraction time.The result sees table 1.
Table 1 extraction time investigation test findings
The result shows, refluxed 1 hour and the content of the 1.5 hours every gram medicine of gained mesaconitine basic identical, be 1 hour so select reflux extracting time for use.
6.2 extract the investigation of solvent
By method under the inspection item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, extracts than 80% ethanol water, absolute ethyl alcohol with methyl alcohol, volume parts respectively.Content with every gram medicine mesaconitine is that index confirms to extract solvent.The result sees table 2.
Table 2 extracts solvent and investigates test findings
The result shows, methyl alcohol, volume parts are more suitable basically than 80% ethanol water, three kinds of content that solvent is measured of absolute ethyl alcohol, but the test sample solvent that extracts with absolute ethyl alcohol, impurity peaks is minimum, so adopt absolute ethyl alcohol to be the extraction solvent.
6.3 the investigation of method for distilling
By method under the inspection item need testing solution is detected.According to the preparation of the test solution was prepared under the three copies, respectively Oscillating vibration, ultrasound, reflux extraction for 1 hour.Content with every gram medicine mesaconitine is that index is confirmed method for distilling.The result sees table 3.
Table 3 method for distilling is investigated test findings
The result shows, the measured aconitine content of refluxing extraction is the highest, is method for distilling so adopt to reflux.
7. the investigation of the preparation of typical curve and linear relationship
Precision is measured aconitine reference substance stock solution solution (20.1 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml; Put respectively in the 10ml volumetric flask, methyl alcohol is diluted to scale, shakes up; Each accurate sample introduction 20 μ l; With peak area (A) reference substance concentration (C) is carried out linear regression, get the regression equation of aconitine: A=12953C+249.14, related coefficient: R=0.9996.The result shows that aconitine is in 2.01 μ g/ml ~ 20.10 μ g/ml scopes, and the peak area of aconitine (A) is good with reference substance concentration (C) linear relationship.The result sees table 4.
Table 4 aconitine linear relationship is investigated the result
8. precision test
The accurate aconitine reference substance solution 20 μ l that draw inject liquid chromatograph, each METHOD FOR CONTINUOUS DETERMINATION 6 times, and the record peak area also calculates relative standard deviation.The result shows that instrument precision is good.The result sees table 5.
Table 5 aconitine Precision test result
9. quantitative limit
The accurate absorption in aconitine contrast solution (10.06 μ g/ml) 1ml to the 25ml volumetric flask with the methyl alcohol dilution and be settled to scale, shakes up, and the accurate 20 μ l that draw inject liquid chromatograph.The signal to noise ratio (S/N ratio) of aconitine chromatographic peak is about 10 as a result.The result shows: when aconitine concentration was 0.4024 μ g/ml, signal to noise ratio (S/N ratio) was about 10, and promptly aconitine quantitatively is limited to 8.05ng.
10. replica test
Get with a collection of 29 flavors can dissipate sample (product batch number: 20110815) 12g, accurate claim fixed, totally 6 parts; The method for preparing under the item by need testing solution prepares need testing solution; The accurate respectively 20 μ l that draw inject liquid chromatograph, the content of calculation sample mesaconitine.The result shows that this analytical approach repeatability is good.The result sees table 6.
Table 6 aconitine replica test result
11. recovery test
Precision takes by weighing and can dissipate with a collection of 29 flavors that (product batch number: 20110815) 6 parts, each accurate aconitine alkali reference substance that adds is measured its content, calculate recovery rate to sample.The result shows that it is accurate that this assay method is measured the result.The result sees table 7.
Table 7 aconitine recovery test result
12. sample determination
Get Tibetan medicine composition 29 flavors and can dissipate three batches, measure the also content of calculation sample mesaconitine.The result sees table 8.
Table 8 sample size is measured the result
Experimental example 3: the limit test experiment of aristolochic acid
1. instrument, reagent and confession test agent
Instrument: L-2100 type Hitachi high performance liquid chromatograph; Tianjin, AuW220D island electronic analytical balance.
Reference substance: aristolochic acid A reference substance (Nat'l Pharmaceutical & Biological Products Control Institute) lot number: 110746-200806.
Sample: 29 flavors can dissipate (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), and lot number is respectively: 20110815,20110816,20110817.
2. detect the selection of wavelength
Get the aristolochic acid A reference substance solution, in 190 ~ 400nm wavelength coverage, scan, aristolochic acid A has absorption maximum in the 315nm wavelength, so be the detection wavelength according to the selected 315nm of ultraviolet absorpting spectrum.
3. moving phase is selected
Discovering, is moving phase than the methyl alcohol-volume parts of 60～70:30～40 than 1% glacial acetic acid aqueous solution with volume parts, and aristolochic acid A all can reach good chromatographic resolution effect.Wherein, be that 66:34 is that moving phase is optimum with methyl alcohol-volume parts than the volume parts ratio of 1% glacial acetic acid aqueous solution.
4. system suitability test
Under above-mentioned chromatographic condition, accurate respectively reference substance solution, each 20 μ l of need testing solution of drawing inject liquid chromatograph, the record chromatogram.The result shows, the degree of separation that aristolochic acid A is adjacent chromatographic peak in the test sample chromatogram is all greater than 1.5, good separating effect.
5. reference substance solution preparation
It is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets.
6. need testing solution preparation
6.1 the investigation of extraction time
By method under the inspection item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, and ultrasonic Extraction is 20 minutes, 30 minutes, 40 minutes respectively.Content with aristolochic acid A in every gram medicine is that index is confirmed extraction time.The result sees table 9.
Table 9 extraction time investigation test findings
The result shows, when ultrasonic 30 minutes and 40 minutes in the every gram medicine of gained the content of aristolochic acid A basic identical, be 30 minutes so select the ultrasonic Extraction time for use.
6.2 extract the investigation of solvent
By method under the inspection item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, extracts with methyl alcohol, absolute ethyl alcohol respectively.Content with aristolochic acid A in every gram medicine is that index confirms to extract solvent.The result sees table 10.
Table 10 extracts solvent and investigates test findings
The result shows that the measured aristolochic acid A content of methyl alcohol is to extract solvent apparently higher than absolute ethyl alcohol so select methyl alcohol for use.
6.3 the investigation of method for distilling
By method under the inspection item need testing solution is detected.According to the preparation of the test solution was prepared under the three copies, respectively Oscillating vibration, ultrasound, reflux extraction for 30 minutes.Content with aristolochic acid A in every gram medicine is that index is confirmed method for distilling.The result sees table 11.
Table 11 method for distilling is investigated test findings
The result shows, the ultrasonic and measured aconitine content basically identical of refluxing extraction considers that ultrasonic Extraction is easy and simple to handle, so select the ultrasonic method for distilling that is for use.
7. the investigation of the preparation of typical curve and linear relationship
Precision is measured aristolochic acid A reference substance stock solution solution (40.6 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml; Put respectively in the 10ml volumetric flask, methyl alcohol is diluted to scale, shakes up; Each accurate sample introduction 20 μ l; With peak area (A) reference substance concentration (C) is carried out linear regression, get the regression equation of aristolochic acid A: A=17077C+5543.3, related coefficient: R=0.9998.The result shows that aristolochic acid A is in 4.06 μ g/ml ~ 40.60 μ g/ml scopes, and the peak area of aristolochic acid A (A) is good with reference substance concentration (C) linear relationship.The result sees table 12.
Table 12 aristolochic acid A linear relationship is investigated the result
8. precision test
The accurate aristolochic acid A reference substance solution 20 μ l that draw inject liquid chromatograph, each METHOD FOR CONTINUOUS DETERMINATION 6 times, and the record peak area also calculates relative standard deviation.The result shows that instrument precision is good.The result sees table 13.
Table 13 aristolochic acid A Precision test result
9. quantitative limit
The accurate absorption in aristolochic acid A contrast solution (17.68 μ g/ml) 1ml to the 50ml volumetric flask with the methyl alcohol dilution and be settled to scale, shakes up, and the accurate 20 μ l that draw inject liquid chromatograph.The signal to noise ratio (S/N ratio) of aristolochic acid A chromatographic peak is about 10 as a result.The result shows: when aristolochic acid A concentration was 0.3536 μ g/ml, signal to noise ratio (S/N ratio) was about 10, and promptly aristolochic acid A quantitatively is limited to 7.07ng.
10. replica test
Get with a collection of 29 flavors can dissipate sample (product batch number: 20110815) 8g, accurate claim fixed, totally 6 parts; The method for preparing under the item by need testing solution prepares need testing solution; The accurate respectively 20 μ l that draw inject liquid chromatograph, the content of aristolochic acid A in the calculation sample.The result shows that this analytical approach repeatability is good.The result sees table 14.
Table 14 aristolochic acid A replica test result
11. recovery test
Precision takes by weighing and can dissipate with a collection of 29 flavors that (product batch number: 20110815) 6 parts, each accurate aristolochic acid A reference substance that adds is measured its content, calculate recovery rate to sample.The result shows that it is accurate that this assay method is measured the result.The result sees table 15.
Table 15 aristolochic acid A recovery test result
12. sample determination
Get Tibetan medicine composition 29 flavors and can dissipate three batches, the content of aristolochic acid A in mensuration and the calculation sample.The result sees table 16.
Table 16 sample size is measured the result
Experimental example 4: the assay experiment of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
1. instrument, reagent and confession test agent
Instrument: L-2100 type Hitachi high performance liquid chromatograph; Tianjin, island AuW220D electronic balance.
Reference substance: aloe-emodin reference substance (lot number: 110795-201007), Rhein reference substance (lot number: 110757-200206), archen reference substance (110756-200100), Chrysophanol reference substance (110796-201017), Physcion reference substance (110758-201013).
Sample: 29 flavors can dissipate (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), and lot number is respectively: 20110815,20110816,20110817.
2. detect the selection of wavelength
Get the mixing reference substance solution of aloe-emodin, Rhein, archen, Chrysophanol, Physcion; In 190 ~ 500nm wavelength coverage, scan; Mixing has absorption maximum to impinging upon the 430nm wavelength, so according to absorbing the selected 430nm of collection of illustrative plates for detecting wavelength.
3. moving phase is selected
Discover that when being moving phase than the methyl alcohol-volume parts of 80 ~ 90:10 ~ 20 than the phosphate aqueous solution that is 0.1% with volume parts, aloe-emodin, Rhein, archen, Chrysophanol, Physcion all can reach good chromatographic resolution effect.Be that 86:14 is that moving phase is optimum than the volume parts ratio that is 0.1% phosphate aqueous solution wherein with methyl alcohol-volume parts.
4. system suitability test
Under above-mentioned chromatographic condition, accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing inject liquid chromatograph, the record chromatogram.The result shows that the degree of separation that aloe-emodin, Rhein, archen, Chrysophanol, Physcion are adjacent chromatographic peak in the test sample chromatogram is all greater than 1.5.
5. mix the reference substance solution preparation
It is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Be to contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g.
6. need testing solution preparation
6.1 the investigation of method for distilling
By method under the assay item need testing solution is detected.According to the preparation of the test solution was prepared under the three copies, respectively, ultrasound, reflux, vibration rockers for 30 minutes.With aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone component total contents of Physcion in every gram medicine is that index is confirmed method for distilling.The result sees table 17.
Table 17 method for distilling is investigated test findings
The result shows, the total content basically identical of anthraquinone component considers that ultrasonic Extraction is easy, easy to operate in ultrasonic and the every gram medicine of refluxing extraction gained, is ultrasonic Extraction so select method for distilling for use.
6.2 extract the investigation of solvent
By method under the assay item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 2 parts, accurate respectively methyl alcohol, each 25ml of ethanol of adding.With aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone component total contents of Physcion in every gram medicine is that index confirms to extract solvent.The result sees table 18.
Table 18 extracts solvent and investigates test findings
The result shows that methyl alcohol is measured the anthraquinone component total content apparently higher than ethanol, is methyl alcohol so select the extraction solvent for use.
6.3 the investigation of extraction time
By method under the assay item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, and sonicated is 20 minutes, 30 minutes, 40 minutes respectively.With aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone component total contents of Physcion in every gram medicine is that index is confirmed extraction time.The result sees table 19.
Table 19 extraction time investigation test findings chemical drug
The result shows, the total content basically identical of anthraquinone component is 30 minutes so select extraction time for use in ultrasonic Extraction 30 minutes and the 40 minutes every gram medicines of gained.
7. the investigation of the preparation of typical curve and linear relationship
Precision is measured and is mixed reference substance stock solution solution (aloe-emodin 32.6 μ g/ml, Rhein 33.8mg/ml, archen 33.2mg/ml, each 32.5 μ g/ml of Chrysophanol; Contain Physcion 17.2/ml) 1ml, 3ml, 5ml, 8ml, 10ml; Put respectively in the 10ml volumetric flask, methyl alcohol is diluted to scale, shakes up; Each accurate sample introduction 10 μ l; With peak area (A) reference substance concentration (C) is carried out linear regression, the regression equation that gets aloe-emodin is: A=8966.4C+364.89, related coefficient: R=0.9995; The regression equation of Rhein is: A=10015C+120.03, related coefficient: R=0.9996; The regression equation of archen is: A=9087.1C+326.52, related coefficient: R=0.9995; The regression equation of Chrysophanol is: A=13338C+534.91, related coefficient: R=0.9998; The regression equation of Physcion is: A=5033.8C+68.985, related coefficient: R=0.9997.The result shows that aloe-emodin is in 3.26 μ g/ml ~ 32.60 μ g/ml scopes, and the peak area of aloe-emodin (A) is good with reference substance concentration (C) linear relationship; Rhein is in 3.38 μ g/ml ~ 33.80 μ g/ml scopes, and the peak area of Rhein (A) is good with reference substance concentration (C) linear relationship; Archen is in 3.32 μ g/ml ~ 33.20 μ g/ml scopes, and the peak area of archen (A) is good with reference substance concentration (C) linear relationship; Chrysophanol is in 3.25 μ g/ml ~ 32.50 μ g/ml scopes, and the peak area of Chrysophanol (A) is good with reference substance concentration (C) linear relationship; Physcion is in 1.72 μ g/ml ~ 17.20 μ g/ml scopes, and the peak area of Physcion (A) is good with reference substance concentration (C) linear relationship.The results are shown in Table 20, table 21, table 22, table 23, table 24.
Table 20 aloe-emodin linear relationship is investigated the result
Table 21 Rhein linear relationship is investigated the result
Table 22 archen linear relationship is investigated the result
Table 23 Chrysophanol linear relationship is investigated the result
Table 24 Physcion linear relationship is investigated the result
8. precision test
The accurate absorption mixed reference substance solution 10 μ l, injects liquid chromatograph, each METHOD FOR CONTINUOUS DETERMINATION 6 times, and the record peak area also calculates relative standard deviation.The result shows that instrument precision is good.The result sees table 25.
Table 25 mixes the contrast Precision test result
9. stability test
After the need testing solution preparation was accomplished, the accurate 10 μ l that draw injected liquid chromatograph, and the record peak area was whenever measured once at a distance from 2 hours later on, investigated 8 hours, and the record peak area also calculates relative standard deviation.The result shows, in 8 hours, to measure the result stable for aloe-emodin, Rhein, archen, Chrysophanol, Physcion in the need testing solution.The result sees table 26.
Table 26 mixes contrast stability test result
10. replica test
Get with a collection of 29 flavor sample (product batch numbers: 20110815) 2g that can dissipate; The accurate title, decide; Totally 6 parts, prepare need testing solution by the method for preparing under the item of need testing solution, the accurate respectively 10 μ l that draw; Inject liquid chromatograph, aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone component total contents of Physcion in the calculation sample.The result shows that this analytical approach repeatability is good.The result sees table 27.
Table 27 anthraquinone component total content replica test result
11. recovery test
Precision takes by weighing and can dissipate with a collection of 29 flavors that (product batch number: 20110815) 6 parts, each accurate aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone component reference substance of Physcion of adding is measured its content, calculate recovery rate to sample.The result shows that it is accurate that this assay method is measured the result.The result sees table 28.
Table 28 anthraquinone component recovery test result
12. sample determination
Get Tibetan medicine composition 29 flavors and can dissipate three batches, measure and calculate the total content of aloe-emodin, Rhein, archen, Chrysophanol, five kinds of anthraquinone components of Physcion.The result sees table 29.
Table 29 sample size is measured the result
Experimental example 5: the assay experiment of hydroxyl radical carthamin yellow carthamus A
1. instrument, reagent and confession test agent
Instrument: L-2100 type Hitachi high performance liquid chromatograph; Tianjin, island AuW220D electronic balance.
Reference substance: hydroxyl radical carthamin yellow carthamus A reference substance (Chinese pharmaceutical biological product is identified institute), lot number: 111637-200905.
Sample: 29 flavors can dissipate (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company and provided), and lot number is respectively: 20110815,20110816,20110817.
2. detect the selection of wavelength
Get the hydroxyl radical carthamin yellow carthamus A reference substance solution, in 190 ~ 500nm wavelength coverage, scan, hydroxyl radical carthamin yellow carthamus A has absorption maximum in the 403nm wavelength, so be the detection wavelength according to the selected 403nm of ultraviolet absorpting spectrum.
3. moving phase is selected
Discovering, is 1% glacial acetic acid aqueous solution when being moving phase with volume parts than the methyl alcohol-volume parts ratio of 20～30:70～80, and hydroxyl radical carthamin yellow carthamus A all can reach good chromatographic resolution effect.Be that 24:76 is that moving phase is optimum than the volume parts ratio that is 1% glacial acetic acid aqueous solution wherein with methyl alcohol-volume parts.
4. system suitability test
Under above-mentioned chromatographic condition, accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing inject liquid chromatograph, the record chromatogram.The result shows that the degree of separation that hydroxyl radical carthamin yellow carthamus A is adjacent chromatographic peak in reference substance, the test sample chromatogram is all greater than 1.5.
5. reference substance preparation
It is an amount of to get hydroxyl safflower anthocyanidin A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts and processes the solution that every 1ml contains 50 μ g than 25% methanol aqueous solution, promptly gets.
6. test sample preparation
6.1 the investigation of method for distilling
By method under the assay item need testing solution is detected.According to the preparation of the test solution was prepared under the three copies, respectively, ultrasound, reflux, vibration rockers for 40 minutes.Content with hydroxyl radical carthamin yellow carthamus A in every gram medicine is that index is confirmed method for distilling.The result sees table 30.
Table 30 method for distilling is investigated test findings
The result shows that the content of hydroxyl radical carthamin yellow carthamus A is the highest in the every gram medicine of ultrasonic Extraction gained, is ultrasonic Extraction so select method for distilling for use.
6.2 extract the investigation of solvent
By method under the assay item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, and precision adds entry, volume parts than 25% methanol aqueous solution, each 25ml of methyl alcohol respectively.Content with hydroxyl radical carthamin yellow carthamus A in every gram medicine is that index confirms to extract solvent.The result sees table 31.
Table 31 extracts solvent and investigates test findings
The result shows, the hydroxyl radical carthamin yellow carthamus A content basically identical that water, volume parts are surveyed than 25% methanol aqueous solution and methyl alcohol extracts the solvent filter difficulty owing to use water as, and pure methyl alcohol toxicity is big, is that volume parts is than 25% methanol aqueous solution so select the extraction solvent for use.
6.3 the investigation of extraction time
By method under the assay item need testing solution is detected.The method for preparing under the item of pressing need testing solution prepares 3 parts, and sonicated is 30 minutes, 40 minutes, 50 minutes respectively.Content with hydroxyl radical carthamin yellow carthamus A in every gram medicine is that index is confirmed extraction time.The result sees table 32.
Table 32 extraction time investigation test findings
The result shows, the content basically identical of hydroxyl radical carthamin yellow carthamus A is 40 minutes so select extraction time for use in ultrasonic Extraction 40 minutes and the 50 minutes every gram medicines of gained.
7. the investigation of the preparation of typical curve and linear relationship
Precision is measured hydroxyl radical carthamin yellow carthamus A reference substance stock solution solution (hydroxyl radical carthamin yellow carthamus A content is 107.8 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml; Put respectively in the 10ml volumetric flask, volume parts is diluted to scale than 25% methanol aqueous solution, shakes up; Each accurate sample introduction 10 μ l; With peak area (A) reference substance concentration (C) is carried out linear regression, get hydroxyl radical carthamin yellow carthamus A regression equation: A=6605.5C+454.39, related coefficient: R=0.9998.The result shows that hydroxyl radical carthamin yellow carthamus A is in 10.78 μ g/ml ~ 107.80 μ g/ml scopes, and the peak area of hydroxyl radical carthamin yellow carthamus A (A) is good with reference substance concentration (C) linear relationship.The result sees table 33.
Table 33 hydroxyl radical carthamin yellow carthamus A linear relationship is investigated the result
8. precision test
The accurate hydroxyl radical carthamin yellow carthamus A reference substance solution 10 μ l that draw inject liquid chromatograph, each METHOD FOR CONTINUOUS DETERMINATION 6 times, and the record peak area also calculates relative standard deviation.The result shows that instrument precision is good.The result sees table 34.
Table 34 hydroxyl radical carthamin yellow carthamus A Precision test result
9. stability test
After the need testing solution preparation was accomplished, the accurate 10 μ l that draw injected liquid chromatograph, and the record peak area was whenever measured once at a distance from 2 hours later on, investigated 8 hours, and the record peak area also calculates relative standard deviation.The result shows that hydroxyl radical carthamin yellow carthamus A measurement result in 8 hours is stable in the need testing solution.The result sees table 35.
Table 35 hydroxyl radical carthamin yellow carthamus A stability test result
10. replica test
Get with a collection of 29 flavors can dissipate sample (product batch number: 20110815) 4g, accurate claim fixed, totally 6 parts; The method for preparing under the item by need testing solution prepares need testing solution; The accurate respectively 10 μ l that draw inject liquid chromatograph, the content of hydroxyl radical carthamin yellow carthamus A in the calculation sample.The result shows that this analytical approach repeatability is good.The result sees table 36.
Table 36 hydroxyl radical carthamin yellow carthamus A replica test result
11. recovery test
Precision takes by weighing and can dissipate with a collection of 29 flavors that (product batch number: 20110815) 6 parts, each accurate hydroxyl radical carthamin yellow carthamus A reference substance that adds is measured its content, calculate recovery rate to sample.The result shows that it is accurate that this assay method is measured the result.The result sees table 37.
Table 37 hydroxyl radical carthamin yellow carthamus A recovery test result
12. sample determination
Get Tibetan medicine composition 29 flavors and can dissipate three batches, measure and calculate hydroxyl radical carthamin yellow carthamus A content.The result sees table 38.
Table 38 sample size is measured the result
Following embodiment all can realize the described effect of above-mentioned experimental example.
Embodiment
Below in conjunction with embodiment the present invention is done detailed elaboration, but be not limited to the embodiment of these concrete records.Tibetan medicine composition that embodiment 1 is detected 29 flavors can be dissipated as the Qinghai gold and scold Tibetan medicine medicine company incorporated company and produce and sell.
The quality determining method that embodiment distinguished the flavor of and can dissipate in 1: two 19
Differentiate:
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 8:1.5; Launch, take out, dry; Spray is with 1% vanillic aldehyde 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.
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the cyclohexane-ethyl acetate that is 3:2; Launch, take out, dry; 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. myristic discriminating
Get the Tibetan medicine composition 29 flavor powder 5g that can dissipate, put in the 250ml round-bottomed flask, add water 100ml; Decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml, decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished, collect the ethyl acetate part, as control medicinal material solution with volatile oil determination apparatus; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 20:0.3; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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.
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 68:32:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 12g that can dissipate, accurately claim surely, put in the conical flask of tool plug, add ammonia solution 10ml; The accurate absolute ethyl alcohol 100ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
These article contain the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g.
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than methyl alcohol-volume parts of 66:34 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 8g that can dissipate, accurately claim surely, put in the tool plug conical flask accurate methyl alcohol 50ml, the close plug of adding; Claim decide weight, ultrasonic 30 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
These article contain banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g.
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts that is 86:14 than the phosphate aqueous solution that is 0.1% with volume parts; The detection wavelength is 430nm.Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 2g that can dissipate, accurately claim surely, put in the conical flask of tool plug, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Ultrasonic 30min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
These article aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g.
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than methyl alcohol-volume parts ratio of 24:76; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 4g that can dissipate, accurately claim surely, put in the tool plug conical flask, accurate to add the volume parts ratio be 25% methanol aqueous solution 25ml; Close plug is claimed to decide weight, ultrasonic 40 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
These article hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
Embodiment 2: the two 19 flavor ball quality determining method that can disappear
Differentiate:
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor ball powder 3g that can disappear, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 8:1.5; Launch, take out, dry.Spray is with 1% vanillic aldehyde 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.
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor ball powder 3g that can disappear, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the cyclohexane-ethyl acetate that is 3:2; Launch, take out, dry.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. myristic discriminating
Get the Tibetan medicine composition 29 flavor ball powder 5g that can disappear; Put in the 250ml round-bottomed flask, add water 100ml, decoct 1h; Collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus) with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml; Decoct 1h, collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus), after decoction is accomplished with volatile oil determination apparatus; Collect the ethyl acetate part, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 20:0.3; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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.
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 68:32:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor ball powder 12g that can disappear, accurately claim surely, put in the conical flask of tool plug, add ammonia solution 10ml; The accurate absolute ethyl alcohol 100ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
These article contain the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g.
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than methyl alcohol-volume parts of 66:34 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor ball powder 8g that can disappear, accurately claim surely, put in the tool plug conical flask accurate methyl alcohol 50ml, the close plug of adding; Claim decide weight, ultrasonic 30 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
These article contain banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g.
Assay:
A. the assay of aloe-emodin, Rhein, archen, Chrysophanol, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts that is 86:14 than the phosphate aqueous solution that is 0.1% with volume parts; The detection wavelength is 430nm.Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively; Mixing promptly gets (be to contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g);
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor ball powder 2g that can disappear, accurately claim surely, put in the conical flask of tool plug, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Ultrasonic 30min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
These article aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g.
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than methyl alcohol-volume parts ratio of 24:76; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor ball powder 4g that can disappear, accurately claim surely, put in the tool plug conical flask, accurate to add the volume parts ratio be 25% methanol aqueous solution 25ml; Close plug is claimed to decide weight, ultrasonic 40 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
These article hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
The described Tibetan medicine composition 29 flavors ball that can disappear is meant with 29 flavors can dissipate bulk drug prescription Tibet inula root 25g, gypsum rubrum (forging) 125g, myrobalan 75g, hot millet 25g, the colored 125g of alkali, nutmeg 25g, Bi roots of grass 25g, cassia seed 25g, Amomum cardamomum 25g, rhizome of davallia 25g, pepper 25g, radish (charcoal) 2g; Tsaoko 25g, natural salt 25g, asafoetide 2g, sal ammoniac 25g, kaempferia galamga 25g, shellfish tooth (charcoal) 25g, rheum officinale 100g, wide muscle rattan 13g, safflower 25g, iron staff hammer 15g, lime (system) 25g, vulture excrement (stir-fry) 40g, banksia rose birthwort 25g, Semen seu folium abelmoschi moschati 25g, HALITUM PURPUREUM 25g, frankincense 25g, slag are tamed and dociled cream 25g; Add conventional auxiliary material, according to the pill of common process preparation.
Embodiment 3: the two 19 flavor tablet quality detection method that can disappear
Differentiate:
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor sheet powder 3g that can disappear, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 8:1.5; Launch, take out, dry.Spray is with mass volume ratio 1% vanillic aldehyde 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.
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor sheet powder 3g that can disappear, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the cyclohexane-ethyl acetate that is 3:2; Launch, take out, dry.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. myristic discriminating
Get the Tibetan medicine composition 29 flavor sheet powder 5g that can disappear; Put in the 250ml round-bottomed flask, add water 100ml, decoct 1h; Collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus) with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml; Decoct 1h, collect volatile oil (beginning to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus), after decoction is accomplished with volatile oil determination apparatus; Collect the ethyl acetate part, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 20:0.3; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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.
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 68:32:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor sheet powder 12g that can disappear, accurately claim surely, put in the conical flask of tool plug, add ammonia solution 10ml; The accurate absolute ethyl alcohol 100ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
These article contain the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g.
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than methyl alcohol-volume parts of 66:34 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor sheet powder 8g that can disappear, accurately claim surely, put in the tool plug conical flask accurate methyl alcohol 50ml, the close plug of adding; Claim decide weight, ultrasonic 30 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
These article contain banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g.
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts that is 86:14 than the phosphate aqueous solution that is 0.1% with volume parts; The detection wavelength is 430nm.Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively; Mixing promptly gets (be to contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g);
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor sheet powder 2g that can disappear, accurately claim surely, put in the conical flask of tool plug, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Ultrasonic 30min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
These article aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g.
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than methyl alcohol-volume parts ratio of 24:76; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor sheet powder 4g that can disappear, accurately claim surely, put in the tool plug conical flask, accurate to add the volume parts ratio be 25% methanol aqueous solution 25ml; Close plug is claimed to decide weight, ultrasonic 40 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get.
These article hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
The described Tibetan medicine composition 29 flavors sheet that can disappear is meant with 29 flavors can dissipate bulk drug prescription Tibet inula root 25g, gypsum rubrum (forging) 125g, myrobalan 75g, hot millet 25g, the colored 125g of alkali, nutmeg 25g, Bi roots of grass 25g, cassia seed 25g, Amomum cardamomum 25g, rhizome of davallia 25g, pepper 25g, radish (charcoal) 2g; Tsaoko 25g, natural salt 25g, asafoetide 2g, sal ammoniac 25g, kaempferia galamga 25g, shellfish tooth (charcoal) 25g, rheum officinale 100g, wide muscle rattan 13g, safflower 25g, iron staff hammer 15g, lime (system) 25g, vulture excrement (stir-fry) 40g, banksia rose birthwort 25g, Semen seu folium abelmoschi moschati 25g, HALITUM PURPUREUM 25g, frankincense 25g, slag are tamed and dociled cream 25g; Add conventional auxiliary material, according to the tablet of common process preparation.
Claims (3)
1. a bulk drug consists of Tibet inula root 25 weight portions, gypsum rubrum (forging) 125 weight portions, myrobalan's 75 weight portions, hot millet 25 weight portions, alkali and spends 125 weight portions, nutmeg 25 weight portions, the Bi roots of grass 25 weight portions, cassia seed 25 weight portions, Amomum cardamomum 25 weight portions, the rhizome of davallia 25 weight portions, pepper 25 weight portions, radish (charcoal) 2 weight portions; Tsaoko 25 weight portions, natural salt 25 weight portions, asafoetide 2 weight portions, sal ammoniac 25 weight portions, kaempferia galamga 25 weight portions, shellfish tooth (charcoal) 25 weight portions, rheum officinale 100 weight portions, wide muscle rattan 13 weight portions, safflower 25 weight portions, iron staff are hammered 15 weight portions, lime (system) 25 weight portions, vulture excrement (stir-fry) 40 weight portions, banksia rose birthwort 25 weight portions, Semen seu folium abelmoschi moschati 25 weight portions, HALITUM PURPUREUM 25 weight portions, frankincense 25 weight portions, slag into shape and are tamed and dociled that Tibetan medicine composition 29 flavors of cream 25 weight portions can dissipate and the quality determining method of preparation, and this method comprises one or more in following discriminating and/or inspection and/or the content assaying method:
Differentiate:
A. the discriminating of frankincense
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 2 ~ 4g, the 20 ~ 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the xylene-ethyl acetate of 8:1 ~ 2 is a developping agent; Launch, take out, dry; Spray is with mass and size portion rate 1% vanillic aldehyde 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;
B. the discriminating of pipering
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 2 ~ 4g, add ethyl acetate 20 ~ 30ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the cyclohexane-ethyl acetate of 3:1 ~ 3 is a developping agent; Launch, take out, dry; 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. myristic discriminating
Get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 4 ~ 6g, put in the 250ml round-bottomed flask, add water 100ml; Decoct 1h; Collect volatile oil with volatile oil determination apparatus, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished; Collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml, decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished, collect the ethyl acetate part, as control medicinal material solution with volatile oil determination apparatus; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With the volume parts ratio is that the xylene-ethyl acetate of 20:0.1 ~ 0.5 is a developping agent; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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;
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 65 ~ 75:25 ~ 35:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 10 ~ 15g, the accurate title, decide, and puts in the conical flask of tool plug, adds ammonia solution 10ml; The accurate absolute ethyl alcohol 80～120ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ℃ ~ 40 ℃ decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ℃ ~ 40 ℃ decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 10 ~ 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g;
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts of 60～70:30～40 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 6 ~ 10g, the accurate title, decide, and puts in the tool plug conical flask, accurate methyl alcohol 40 ~ 60ml, the close plug of adding; Claim decide weight, ultrasonic 20 ~ 40 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 ~ 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g;
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the volume parts ratio is that the methyl alcohol-volume parts of 80 ~ 90:10 ~ 20 is a moving phase than the phosphate aqueous solution that is 0.1%; The detection wavelength is 430nm; Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Be to contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 1 ~ 3g, the accurate title, decide, and puts in the conical flask of tool plug, the accurate methyl alcohol 20 ~ 30ml that adds, and close plug claims to decide weight; Ultrasonic 20 ~ 40min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 5 ~ 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention 29 flavor can dissipate or its preparation in aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g;
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than the methyl alcohol-volume parts ratio of 20～30:70～80; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get Tibetan medicine composition 29 flavors and can dissipate or its preparation powder 3 ~ 5g, the accurate title, decide, and puts in the tool plug conical flask, and it is 25% methanol aqueous solution, 20 ~ 30ml that precision adds the volume parts ratio; Close plug is claimed to decide weight, ultrasonic 30 ~ 50 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 5 ~ 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
The present invention 29 flavor can dissipate or its preparation in hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
2. a kind of Tibetan medicine composition as claimed in claim 1 29 flavors can dissipate and the quality determining method of preparation; It is characterized in that described preparation is meant gets the Tibetan medicine composition 29 flavor bulk drug that can dissipate; Press common process; Add conventional auxiliary material and be prepared into clinical acceptable any formulation, comprise micropill, dripping pill, tablet, capsule, particle or dispersing tablet.
Like claim 1 or/and 2 described a kind of bulk drugs consist of Tibet inula root 25g, gypsum rubrum (forging) 125g, myrobalan 75g, hot millet 25g, alkali flower 125g, nutmeg 25g, Bi roots of grass 25g, cassia seed 25g, Amomum cardamomum 25g, rhizome of davallia 25g, pepper 25g, radish (charcoal) 2g; The Tibetan medicine composition 29 of tsaoko 25g, natural salt 25g, asafoetide 2g, sal ammoniac 25g, kaempferia galamga 25g, shellfish tooth (charcoal) 25g, rheum officinale 100g, wide muscle rattan 13g, safflower 25g, iron staff hammer 15g, lime (system) 25g, vulture excrement (stir-frys) 40g, banksia rose birthwort 25g, Semen seu folium abelmoschi moschati 25g, HALITUM PURPUREUM 25g, frankincense 25g, the tame and docile cream 25g of slag is distinguished the flavor of and can be dissipated and the quality determining method of preparation, and this method comprises one or more in following discriminating and/or inspection and/or the content assaying method:
Differentiate:
A. the discriminating of frankincense
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, the 25ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Get frankincense control medicinal material 0.5g, the 30ml that adds diethyl ether, sonicated 15min filters, and filtrating volatilizes, and residue adds methyl alcohol 2ml makes dissolving, as control medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 8:1.5; Launch, take out, dry; Spray is with 1% vanillic aldehyde 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;
B. the discriminating of pipering
Get the Tibetan medicine composition 29 flavor powder 3g that can dissipate, add ethyl acetate 25ml, sonicated 30min filters, and filtrating is concentrated into 2ml, as need testing solution; It is an amount of to get pipering, adds methyl alcohol and processes the reference substance solution that every 1ml contains 0.5mg; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the cyclohexane-ethyl acetate that is 3:2; Launch, take out, dry; 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. myristic discriminating
Get the Tibetan medicine composition 29 flavor powder 5g that can dissipate, put in the 250ml round-bottomed flask, add water 100ml; Decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus with volatile oil determination apparatus; After decocting completion, collect the ethyl acetate part, as need testing solution; Get nutmeg 1g, put in the 100ml round-bottomed flask, add water 50ml, decoct 1h, collect volatile oil, begin to add low amounts of water and 1ml ethyl acetate in the volatile oil determination apparatus, after decoction is accomplished, collect the ethyl acetate part, as control medicinal material solution with volatile oil determination apparatus; According to thin-layered chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Is developping agent with volume parts than the xylene-ethyl acetate that is 20:0.3; Launch, take out, dry; The long-pending portion rate of sprinkler body is 1% vanillic aldehyde 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;
Inspection:
A. the limit test of aconitine
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase with volume parts than methanol-water-triethylamine-methylene chloride of 68:32:0.2:5; The detection wavelength is 235nm, and number of theoretical plate calculates by the aconitine peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the aconitine reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 10 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 12g that can dissipate, accurately claim surely, put in the conical flask of tool plug, add ammonia solution 10ml; The accurate absolute ethyl alcohol 100ml that adds, close plug claims to decide weight; Reflux 1h behind the cold soaking 1h is put coldly, claims to decide weight again; Supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, filter; Precision is measured subsequent filtrate 50ml, and 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the accurate dilute hydrochloric acid solution 20ml that adds of residue dissolves, and places 30min in the ice bath, filters; Transfer pH to 10 with ammoniacal liquor, filtrating is transferred in the separating funnel, add ethyl acetate extraction 5 times, each 20ml, combined ethyl acetate liquid, 25 ~ 40 ℃ of decompression and solvent recoveries are to doing, and the residue precision adds methyl alcohol 2ml dissolving, filters, and gets subsequent filtrate, promptly gets;
Determination method: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains the iron staff hammer with aconitine (C
34H
47NO
11) meter, must not be higher than 17 μ g/g;
B. the limit test of aristolochic acid A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than methyl alcohol-volume parts of 66:34 than 1% glacial acetic acid aqueous solution with volume parts; The detection wavelength is 315nm; Number of theoretical plate calculates by banksia rose aristolochic acid peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the aristolochic acid A reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 8g that can dissipate, accurately claim surely, put in the tool plug conical flask accurate methyl alcohol 50ml, the close plug of adding; Claim decide weight, ultrasonic 30 minutes, put coldly, weight decided in title again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate 25ml, cryoconcentration is to doing, and it is that 0.5% sodium hydroxide solution 20ml dissolves it fully and is transferred in the separating funnel that residue adds the volume parts ratio; With extracted with diethyl ether 2 times, each 20ml, extraction back alkali lye service property (quality) mark 3% hydrochloric acid is regulated pH to 2-3, with extracted with diethyl ether 5 times; Each 20ml merges ether extraction liquid, volatilizes, and with dissolve with methanol and be transferred in the 5ml volumetric flask, adds methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get;
Determination method: draw each 20 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
The present invention's 29 flavors can dissipate or its preparation contains banksia rose birthwort with aristolochic acid A (C
17H
11NO
7) meter, must not be higher than 30 μ g/g;
Assay:
A. the assay of aloe-emodin, Chrysophanol, Rhein, archen, Physcion
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is moving phase than the methyl alcohol-volume parts that is 86:14 than the phosphate aqueous solution that is 0.1% with volume parts; The detection wavelength is 430nm; Number of theoretical plate calculates by the archen peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance, Physcion reference substance; Adding methyl alcohol processes every 1ml respectively and contains each 80 μ g of aloe-emodin reference substance, Rhein reference substance, archen reference substance, Chrysophanol reference substance; The solution of Physcion 40 μ g; Precision is measured each 2ml of above-mentioned reference substance solution respectively, and mixing promptly gets; Contain aloe-emodin, Rhein, archen, each 16 μ g of Chrysophanol among every 1ml, contain Physcion 8 μ g;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 2g that can dissipate, accurately claim surely, put in the conical flask of tool plug, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Ultrasonic 30min is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and precision is measured and got subsequent filtrate 5ml; Put in the flask, fling to solvent, adding percent by volume is 8% aqueous hydrochloric acid solution 10ml, ultrasonic 2 minutes, adds methenyl choloride 10ml again; Reflux 1 hour is put coldly, puts in the separating funnel, obtains the methenyl choloride layer, and acid solution is extracted 3 times with methenyl choloride again; Each 10ml merges methenyl choloride liquid, and recovered under reduced pressure solution is to doing, and residue adds methyl alcohol makes dissolving, is transferred in the 10ml measuring bottle; Add methyl alcohol to scale, shake up, filter, get subsequent filtrate, promptly get;
Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get;
The present invention 29 flavor can dissipate or its preparation in aloe-emodin (C
15H
10O
5), Rhein (C
15H
8O
6), archen (C
15H
10O
5), Chrysophanol (C
15H
10O
4), Physcion (C
16H
12O
5) total content must not be less than 1.20mg/g;
B. the assay of hydroxyl radical carthamin yellow carthamus A
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; Is that 1% glacial acetic acid aqueous solution is a moving phase with volume parts than methyl alcohol-volume parts ratio of 24:76; The detection wavelength is 403nm; Number of theoretical plate calculates by the hydroxyl radical carthamin yellow carthamus A peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of to get the hydroxyl radical carthamin yellow carthamus A reference substance, and accurate the title decides, and puts in the brown bottle, adds volume parts than being that 25% methanol aqueous solution is processed the solution that every 1ml contains 50 μ g, promptly gets;
The preparation of need testing solution: get the Tibetan medicine composition 29 flavor powder 4g that can dissipate, accurately claim surely, put in the tool plug conical flask, accurate to add the volume parts ratio be 25% methanol aqueous solution 25ml; Close plug is claimed to decide weight, ultrasonic 40 minutes; Put coldly, use volume parts, shake up than being that 25% methanol aqueous solution is supplied the weight that subtracts mistake; Filter, get subsequent filtrate, promptly get;
Determination method: draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure, promptly get;
The present invention 29 flavor can dissipate or its preparation in hydroxyl radical carthamin yellow carthamus A (C
27H
30O
15) content must not be less than 0.20mg/g.
Priority Applications (1)
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102879518A (en) * | 2012-11-02 | 2013-01-16 | 广西万寿堂药业有限公司 | Quality detection method of blood pressure-lowering and lipid-lowering drugs |
| CN105582256A (en) * | 2015-09-18 | 2016-05-18 | 张祖宇 | Traditional Chinese medicine for treating ovarian cyst |
| CN106596806A (en) * | 2016-12-19 | 2017-04-26 | 山东省中医药研究院 | Method of simultaneously preparing emodin and physcion from radix polygoni multiflori praeparata intelligence reinforcing capsule |
| CN112415157A (en) * | 2020-11-24 | 2021-02-26 | 内蒙古祈蒙药业股份有限公司 | Quality control method of Anxiao six-ingredient granules |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101313936A (en) * | 2007-06-01 | 2008-12-03 | 北京亚东生物制药有限公司 | Medicament composition for dispelling wind and relieving pain, preparation method and quality control method thereof |
| CN101417020A (en) * | 2007-10-23 | 2009-04-29 | 北京亚东生物制药有限公司 | Medicine composition capable of eliminating the mass and relieving swelling, absorbing clots and alleviating pain, preparation method and quality control method thereof |
| CN101444571A (en) * | 2008-12-31 | 2009-06-03 | 太极集团有限公司 | Jiuweiqianghuo soft capsule and preparation method thereof |
| CN102353733A (en) * | 2011-07-11 | 2012-02-15 | 山东阿如拉药物研究开发有限公司 | Quality detection method for Ruyi Zhenbao preparation |
| CN102397517A (en) * | 2011-07-05 | 2012-04-04 | 山东阿如拉药物研究开发有限公司 | Quality control method of traditional Tibetan medicinal lipid lowering preparation |
| CN102507834A (en) * | 2011-09-27 | 2012-06-20 | 山东阿如拉药物研究开发有限公司 | Quality control method for eight-flavor agilawood preparations |
-
2012
- 2012-07-31 CN CN201210272243.1A patent/CN102759598B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101313936A (en) * | 2007-06-01 | 2008-12-03 | 北京亚东生物制药有限公司 | Medicament composition for dispelling wind and relieving pain, preparation method and quality control method thereof |
| CN101417020A (en) * | 2007-10-23 | 2009-04-29 | 北京亚东生物制药有限公司 | Medicine composition capable of eliminating the mass and relieving swelling, absorbing clots and alleviating pain, preparation method and quality control method thereof |
| CN101444571A (en) * | 2008-12-31 | 2009-06-03 | 太极集团有限公司 | Jiuweiqianghuo soft capsule and preparation method thereof |
| CN102397517A (en) * | 2011-07-05 | 2012-04-04 | 山东阿如拉药物研究开发有限公司 | Quality control method of traditional Tibetan medicinal lipid lowering preparation |
| CN102353733A (en) * | 2011-07-11 | 2012-02-15 | 山东阿如拉药物研究开发有限公司 | Quality detection method for Ruyi Zhenbao preparation |
| CN102507834A (en) * | 2011-09-27 | 2012-06-20 | 山东阿如拉药物研究开发有限公司 | Quality control method for eight-flavor agilawood preparations |
Non-Patent Citations (7)
| Title |
|---|
| 中华人民共和国国家药品监督管理局: "《中华人民共和国药品标准2002ZD-1311》", 26 November 2002, article "八味三香散", pages: 344-348 * |
| 刘志勤 等: "胃痛七味散胶囊的色谱鉴别和含量测定", 《西北药学杂志》, vol. 19, no. 1, 29 February 2004 (2004-02-29), pages 15 - 16 * |
| 吴树鸣: "鼻窦炎口服液的研制", 《山西医科大学学报》, vol. 35, no. 4, 3 August 2004 (2004-08-03), pages 378 - 379 * |
| 国家药典委员会: "《中华人民共和国药典2010年版1部》", 31 January 2010, article "伤痛宁片", pages: 691 * |
| 孙绪丁 等，: "二十九味能消散中乌头碱的限量检测", 《中国保健营养》, no. 01, 31 January 2012 (2012-01-31), pages 333 - 334 * |
| 崔晓红 等，: "RP-HPLC法测定朱砂莲胶囊中马兜铃酸A的含量", 《中草药》, vol. 35, no. 10, 31 October 2004 (2004-10-31), pages 1118 - 1119 * |
| 韩娜 等: "RP- HPLC法测定14 种中药材中马兜铃酸A的含量", 《沈阳药科大学学报》, vol. 25, no. 2, 29 February 2008 (2008-02-29), pages 115 - 118 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102879518A (en) * | 2012-11-02 | 2013-01-16 | 广西万寿堂药业有限公司 | Quality detection method of blood pressure-lowering and lipid-lowering drugs |
| CN102879518B (en) * | 2012-11-02 | 2014-07-09 | 广西万寿堂药业有限公司 | Quality detection method of blood pressure-lowering and lipid-lowering drugs |
| CN105582256A (en) * | 2015-09-18 | 2016-05-18 | 张祖宇 | Traditional Chinese medicine for treating ovarian cyst |
| CN106596806A (en) * | 2016-12-19 | 2017-04-26 | 山东省中医药研究院 | Method of simultaneously preparing emodin and physcion from radix polygoni multiflori praeparata intelligence reinforcing capsule |
| CN112415157A (en) * | 2020-11-24 | 2021-02-26 | 内蒙古祈蒙药业股份有限公司 | Quality control method of Anxiao six-ingredient granules |
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