CN102426205B - Detection method of thrombus sumalin capsules - Google Patents
Detection method of thrombus sumalin capsules Download PDFInfo
- Publication number
- CN102426205B CN102426205B CN 201110357472 CN201110357472A CN102426205B CN 102426205 B CN102426205 B CN 102426205B CN 201110357472 CN201110357472 CN 201110357472 CN 201110357472 A CN201110357472 A CN 201110357472A CN 102426205 B CN102426205 B CN 102426205B
- Authority
- CN
- China
- Prior art keywords
- solution
- reference substance
- methyl alcohol
- ginsenoside
- adds
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a detection method of thrombus sumalin capsules, belonging to the field of traditional Chinese medicine detection. The method comprises the following steps: carrying out developing identification on a same thin layer system of salvia miltiorrhiza, Ligusticum wallichii and borneol, carrying out gas chromatography identification on artificial musk, identifying the thin layer of artificial bezoar, and carrying out content determination of ginsenoside. The method has the advantages of strong specificity, rapidness, economy, simple operation and the like. According to the invention, by simultaneously adding the identification of artificial musk and artificial bezoar, the invention establishes a multi-index content determination; by concretely adding the content determination of Ligusticum wallichii and ginsenoside and cooperating with original content determination, the detection of components of the medicine is comprehensive, the content and stability are understood, and the method is helpful for explaining the medicinal material base of the medicine.
Description
Technical field
The invention belongs to modern Chinese traditional medicine field, be specifically related to a kind of detection method of Xueshuan Xinmaining capsule.
Background technology
Traditional Chinese medicine ingredients is complicated, and particularly Chinese medicine compound prescription often contains multiclass, Multiple components, has consisted of a very complicated system.At present, new active component Chinese medicine preparation and the research and development of modern Chinese traditional compound medicine are on the increase, and new technique and multiple different extracting mode also are used for research and the production of medicine.
Along with the fast development of Modern Testing, equipment and instrument, the assay of Chinese medicine, chromatographic identification become the important means of detection.But assay or the discriminating of at present generally only setting up a certain index chemical constitution for the simultaneously applied situation of above-mentioned multiple new technology, are only set up the assay of certain single component or are differentiated to a great extent with larger one-sidedness.
Xueshuan Xinmaining capsule has qi and activate blood circulation, and the pain-relieving functions of having one's ideas straightened out is used for apoplexy, the obstruction of qi in the chest due to the qi deficiency to blood stasis, and disease opinion is had a dizzy spell, hemiplegia, pained, palpitation uncomfortable in chest; Ishemic stroke convalescence, coronary disease and angina pectoris are seen above-mentioned disease person.
Its prescription is: Ligusticum wallichii, the red sage root, sophora flower, leech, ilex pubescens, muscone, calculus bovis factitius, ginseng stem and leave general saponin, borneol, the dried venom of toads;
Method for making: above ten flavors, get the red sage root, ilex pubescens, Ligusticum wallichii with 60% alcohol extract 3 times, add successively 8,5,5 times of amount ethanol, extracted respectively 3,2,1 hours, filter, merging filtrate, decompression recycling ethanol, concentrated, drying, pulverizing, for subsequent use; Other is ground into fine powder at the leech of fetching water, and is for subsequent use; Get sophora flower again, add 5 times of water gagings, regulate pH value to 8～9 with saturated aqua calcis, be heated to little boiling, be incubated 30 minutes, filter while hot, the dregs of a decoction as above method extract 2 times again, merging filtrate, and reduced pressure concentration, 60 ℃ of dryings of low temperature are pulverized, and are for subsequent use.The calculus bovis factitius of muscone, the dried venom of toads is ground into fine powder, presses facing-up method and borneol fine powder, ginseng stem and leave general saponin and above-mentioned two kinds of powder mixings, incapsulate, make 1000, and get final product.Every heavy 0.50g, oral, one time 4.
In the side with hot temperature, enter the liver and gall warp, activating QI to alleviate the depression dispels pathogenic wind and remove dampness, the Ligusticum wallichii of promoting blood circulation and stopping pain and bitter tepor enter the conscience warp, promoting blood circulation, calming heart and tranquilizing mind be pain, the red sage root that nourishes blood is monarch drug in a prescription altogether.Minister is with ilex pubescens, leech, sophora flower, muscone, and is promoting blood circulation and removing blood stasis, the inducing resuscitation of having one's ideas straightened out, removes obstruction in channels to relieve pain.Assistant is promoting blood circulation and removing blood stasis with borneol, calculus bovis factitius, ginseng stem and leave general saponin three medicines assistant the monarch and his subjects, and the inducing resuscitation of having one's ideas straightened out is removed obstruction in channels to relieve pain, the hard kidney water of the liver that clears away heart-fire, and it is main taking stopgap measures, and takes into account the gesture of this void and Yu Huare.Make with the dried venom of toads, pain relieving is kept fit by the priming sick institute that goes directly, current channels and collaterals.
Discriminating in the original detection method of Xueshuan Xinmaining capsule comprises the discriminating of the red sage root, the discriminating of Ligusticum wallichii and the discriminating of borneol, and above-mentioned discriminating is separately carried out, and has the shortcomings such as specificity is strong, need gradation operation.Aspect assay, the assay of tanshin polyphenolic acid B in pair red sage root is arranged at present, but the Ligusticum wallichii of monarch drug in a prescription does not have assay in the side of being all so that only to the assay of tanshin polyphenolic acid B in the monarch drug in a prescription to a great extent with larger one-sidedness; The discriminating and the assay that originally in the detection method only there are pair sophora flower discriminating and the assay aspect of ministerial drug, but the muscone of ministerial drug does not have discrimination method in the side of being all, and ministerial drug is the medicine that auxiliary monarch drug in a prescription is strengthened treatment master disease and main symptom, and because of the muscone expensive, as there is not corresponding detection means to cause easily illegal enterprise not fill out or fill out less the muscone, so that only to the discriminating of sophora flower and assay to a great extent with larger one-sidedness, discriminating to cholic acid in the calculus bovis factitius in the former capsule is to use the extracted by ether need testing solution, make and contain ether in the development system, the spot diffusion, hangover is serious, and identification result has much room for improvement; Also not to the assay of ginseng stem and leave general saponin, can not comprehensively reflect the content of the important composition of Xueshuan Xinmaining capsule simultaneously.
Summary of the invention
The invention provides a kind of detection method of Xueshuan Xinmaining capsule, with solve the specificity that present detection method exists strong, need gradation operation, incomplete problem.
The technical scheme that the present invention takes is to comprise following discrimination method and content assaying method:
(1) discrimination method
(1) get the content of 20 of this capsules, porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; Get respectively red sage root control medicinal material 1g, Ligusticum wallichii control medicinal material 1g in addition, be made in the same way of control medicinal material solution; Get again tanshinone IIA reference substance, borneol reference substance, add ethyl acetate and make the solution that every 1ml contains 1mg, in contrast product solution; According to thin-layered chromatography test, draw each 1 μ l of above-mentioned five kinds of solution, put respectively on same silica gel g thin-layer plate, take 60～90 ℃-ethyl acetate of sherwood oil 11: 2 as developping agent, launch, taking-up is dried; In the test sample chromatogram, with red sage root control medicinal material, the corresponding position of tanshinone IIA reference substance chromatogram on, the spot of aobvious same color under the daylight; Put under the ultraviolet lamp 365nm and inspect, in the test sample chromatogram, with the corresponding position of Ligusticum wallichii control medicinal material chromatogram on, the fluorescence spot of aobvious same color; Spray with 5% phosphomolybdic acid ethanol solution, it is clear to be heated to the spot colour developing at 105 ℃ again; In the test sample chromatogram, with the spot of the aobvious same color in the corresponding position of borneol reference substance chromatogram;
(2) muscone's gas chromatography is differentiated: get the content of 20 of this capsules, and porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; It is an amount of to get the muskone reference substance, adds absolute ethyl alcohol and makes the solution that every 1ml contains 0.1mg, in contrast product solution; According to vapor-phase chromatography test, the DB-FFAP capillary column: column length is 30m, and internal diameter is 0.32mm, and film thickness is 0.5 μ m, and column temperature is temperature programme: initial temperature is 200 ℃, keeps 10 minutes, is warming up to 250 ℃ with the speed of 20 ℃ of per minutes, keeps 10 minutes; Precision is drawn above-mentioned reference substance solution and each 1 μ l of need testing solution respectively, and inject gas chromatograph is measured; The chromatographic peak identical with reference substance chromatographic peak retention time in the record test sample chromatogram;
(3) get the content of 6 of this capsules, porphyrize adds methyl alcohol 50ml, adds hot reflux 1 hour, filters, filtrate evaporate to dryness, residue add water 30ml dissolving, use extracted by ether 2 times, each 20ml, merge ether solution, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrate is medicinal material solution in contrast; Get again the cholic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take normal hexane-ethyl acetate-acetic acid-methyl alcohol 20: 25: 2: 3 upper solution was as developping agent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) content assaying method:
(1) Ligusticum wallichii assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take methyl alcohol-1% acetum 25: 75 as mobile phase; The detection wavelength is 321nm, and number of theoretical plate calculates by the forulic acid peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the forulic acid reference substance, accurately weighed, puts in the brown measuring bottle, adds 70% methyl alcohol and makes the solution that every 1ml contains 20 μ g, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize, get 0.3g～0.5g, accurately weighed, put in the tool plug conical flask, accurate adding 70% methyl alcohol 20ml, weighed weight, ultrasonic processing 30 minutes, power 250W, frequency 50kHz lets cool, weighed weight is supplied the weight that subtracts mistake with 70% methyl alcohol again, shakes up, filter, get subsequent filtrate, and get final product;
Determination method: precision is drawn reference substance solution 5 μ l and need testing solution 10 μ l respectively, and the injection liquid chromatography is measured, and be get final product.
(2) ginseng stem and leave general saponin assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-water 20: 80 as mobile phase; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately weighed, add methyl alcohol and make the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.4～0.6g, and is accurately weighed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic extraction 30 minutes filters, and discards filtrate; The dregs of a decoction volatilize solvent together with filter paper, accurate methyl alcohol 50ml, weighed weight, the refluxing extraction 1 hour of adding, let cool, weighed weight is supplied the weight that subtracts mistake with methyl alcohol again, shakes up, filter, get subsequent filtrate 25ml, evaporate to dryness, residue add water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discard the ammonia solution layer, normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol makes dissolving in right amount, is transferred in the 10ml measuring bottle, adds methyl alcohol to scale and makes dissolving, shake up, filter, and get final product.
Advantage of the present invention is: by the extraction of easy steps, same thin layer system launches, and reaches the purpose of differentiating three kinds of compositions, has the characteristics such as specificity is strong, quick, economic, easy and simple to handle; By setting up the discriminating to the muscone, and cooperate discriminating and the assay of original sophora flower, can more fully detect composition and its compatibility of ministerial drug, the present invention sets up many indexs assay, the concrete assay that passes through to increase to Ligusticum wallichii cooperates existing assay to the red sage root, sophora flower, can more fully detect the composition of monarch drug in a prescription, understand its content and stability, also help to illustrate the medical substance basis of medicine.
Embodiment
Xueshuan Xinmaining capsule is provided by prescription and explained hereafter in the background technology by Jilin Huakang Pharmaceutical Co., Ltd.
Embodiment 1
Comprise following discrimination method and content assaying method:
(1) discrimination method
Get the content of 20 of this capsules, porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; Get respectively red sage root control medicinal material 1g, Ligusticum wallichii control medicinal material 1g in addition, be made in the same way of control medicinal material solution; Get again tanshinone IIA reference substance, borneol reference substance, add ethyl acetate and make the solution that every 1ml contains 1mg, in contrast product solution; According to thin-layered chromatography test, draw each 1 μ l of above-mentioned five kinds of solution, put respectively on same silica gel g thin-layer plate, take 60～90 ℃-ethyl acetate of sherwood oil 11: 2 as developping agent, launch, taking-up is dried; In the test sample chromatogram, with red sage root control medicinal material, the corresponding position of Tanshinone I I A reference substance chromatogram on, the spot of aobvious same color under the daylight; Put under the ultraviolet lamp 365nm and inspect, in the test sample chromatogram, with the corresponding position of Ligusticum wallichii control medicinal material chromatogram on, the fluorescence spot of aobvious same color; Spray with 5% phosphomolybdic acid ethanol solution, it is clear to be heated to the spot colour developing at 105 ℃ again; In the test sample chromatogram, with the spot of the aobvious same color in the corresponding position of borneol reference substance chromatogram;
(2) muscone's gas chromatography is differentiated: get the content of 20 of this capsules, and porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; It is an amount of to get the muskone reference substance, adds absolute ethyl alcohol and makes the solution that every 1ml contains 0.1mg, in contrast product solution; According to vapor-phase chromatography test, the DB-FFAP capillary column: column length is 30m, and internal diameter is 0.32mm, and film thickness is 0.5 μ m, and column temperature is temperature programme: initial temperature is 200 ℃, keeps 10 minutes, is warming up to 250 ℃ with the speed of 20 ℃ of per minutes, keeps 10 minutes; Precision is drawn above-mentioned reference substance solution and each 1 μ l of need testing solution respectively, and inject gas chromatograph is measured; The chromatographic peak identical with reference substance chromatographic peak retention time in the record test sample chromatogram;
(3) get the content of 6 of this capsules, porphyrize adds methyl alcohol 50ml, adds hot reflux 1 hour, filters, filtrate evaporate to dryness, residue add water 30ml dissolving, use extracted by ether 2 times, each 20ml, merge ether solution, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrate is medicinal material solution in contrast; Get again the cholic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take normal hexane-ethyl acetate-acetic acid-methyl alcohol 20: 25: 2: 3 upper solution was as developping agent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) content assaying method:
The Ligusticum wallichii assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take methyl alcohol-1% acetum 25: 75 as mobile phase; The detection wavelength is 321nm, and number of theoretical plate calculates by the forulic acid peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the forulic acid reference substance, accurately weighed, puts in the brown measuring bottle, adds 70% methyl alcohol and makes the solution that every 1ml contains 20 μ g, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize, get 0.5g, accurately weighed, put in the tool plug conical flask, accurate adding 70% methyl alcohol 20ml, weighed weight, ultrasonic processing 30 minutes, power 250W, frequency 50kHz lets cool, weighed weight is supplied the weight that subtracts mistake with 70% methyl alcohol again, shakes up, filter, get subsequent filtrate, and get final product;
Determination method is accurate reference substance solution 5 μ l and the need testing solution 10 μ l of drawing respectively, and the injection liquid chromatography is measured, and be get final product.
The ginseng stem and leave general saponin assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-water 20: 80 as mobile phase; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately weighed, add methyl alcohol and make the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.5g, and is accurately weighed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic extraction 30 minutes filters, and discards filtrate; The dregs of a decoction volatilize solvent together with filter paper, accurate methyl alcohol 50ml, weighed weight, the refluxing extraction 1 hour of adding, let cool, weighed weight is supplied the weight that subtracts mistake with methyl alcohol again, shakes up, filter, get subsequent filtrate 25ml, evaporate to dryness, residue add water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discard the ammonia solution layer, normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol makes dissolving in right amount, is transferred in the 10ml measuring bottle, adds methyl alcohol to scale and makes dissolving, shake up, filter, and get final product.
Embodiment 2
Discrimination method is with embodiment 1.
In the content assaying method, the preparation of need testing solution in the Ligusticum wallichii assay: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.3g, and is accurately weighed, and all the other are with embodiment 1.
The preparation of need testing solution in the ginseng stem and leave general saponin assay: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.4g, and is accurately weighed, and all the other are with embodiment 1.
Embodiment 3
Discrimination method is with embodiment 1.
In the content assaying method, the preparation of need testing solution in the Ligusticum wallichii assay: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.4g, and is accurately weighed, and all the other are with embodiment 1.
The preparation of need testing solution in the ginseng stem and leave general saponin assay: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.6g, and is accurately weighed, and all the other are with embodiment 1.
Every of this product contains Ligusticum wallichii with forulic acid (C
10H
10O
4) meter, be no less than 0.10mg.
Every of this product contains ginseng stem and leave general saponin with ginsenoside Rg1 (C
42H
72O
14) and ginsenoside Re (C
48H
82O
18) the total amount meter, be no less than 4.0mg.
Test example 1 further specifies Ligusticum wallichii content assaying method of the present invention below by test example.
Instrument, reagent and sample
Agilent 1200 high performance liquid chromatographs; Forulic acid reference substance (lot number: 110773-200611), provided by Chinese pharmaceutical biological product check, for assay; Reagent: methyl alcohol is chromatographically pure, and water is purified water, and it is pure that other reagent is analysis.Sample is provided by Jilin Huakang Pharmaceutical Co., Ltd.
1 precision test
The instrument precision test is pressed the chromatographic condition of the embodiment of the invention 1 to a need testing solution, and METHOD FOR CONTINUOUS DETERMINATION 6 times, results peaks area average are that 359.1244, RSD is 0.12% (n=6), the results are shown in Table 1.
Table 1 forulic acid instrument precision test findings
| Sequence number | 1 | 2 | 3 | 4 | 5 | 6 | Average | RSD ％ |
| Peak area | 359.7123 | 358.7959 | 359.0851 | 359.1065 | 358.5632 | 359.4838 | 359.1244 | 0.12 |
The result shows that the precision of instrument is good.
2 replica tests are to same lot number sample, measure 6 times by method parallel extraction of the present invention, and ferulaic acid content average out to 0.140mg/ sheet as a result, RSD=0.31% (n=6) the results are shown in Table 2.
Table 2 forulic acid reappearance experimental result
| Sequence number | 1 | 2 | 3 | 4 | 5 | 6 | Mean value | RSD(％) |
| Content (mg/ sheet) | 0.140 | 0.140 | 0.140 | 0.140 | 0.141 | 0.141 | 0.140 | 0.37 |
The result shows that the repeatability of this law is good.
The test of 3 specificities
For further investigating the rationality of test, make the negative control solution that does not contain Ligusticum chuanxiong Hort according to recipe quantity, measure in accordance with the law, as a result negative controls with the corresponding retention time in reference substance peak place without chromatographic peak, show noiselessly, specificity is good.
4 linear relationships and scope
Precision is drawn forulic acid reference substance solution (lot number: 110773-200611 respectively, concentration 0.02046mg/ml) 1 μ l, 3 μ l, 5 μ l, 7 μ l, 9 μ l injection liquid chromatographies, chromatographic condition of the present invention is measured, take the sample size (μ g) of reference substance as horizontal ordinate, chromatographic peak area is ordinate drawing standard curve.The results are shown in Table 3.
Table 3 forulic acid linear relationship is investigated the result
| Sample size (μ g) | 0.02046 | 0.06138 | 0.1023 | 0.14322 | 0.18414 |
| The forulic acid peak area | 82.37376 | 245.863 | 412.1724 | 576.1788 | 738.6967 |
Regression equation is: y=4015.1x+0.3165, and R=0.99999, the result shows that forulic acid is good log-linear relation in 0.02046 μ g～0.18414 μ g scope.
5 accuracy tests
Precision is measured test sample (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201) 6 parts of measuring known content with method, every part of 0.25g, accurate reference substance solution (0.0043mg/ml) 20ml that adds, extract, measure by method of the present invention, calculate recovery rate, average recovery rate=99.07%, RSD=0.20% (n=6).Meet the requirement of quantitative test.The results are shown in Table 4.
Table 4 forulic acid accuracy test result
Test example 2 adopts the muskone among the vapor-phase chromatography discriminating muscone.
Agilent 7890A gas chromatograph, (column length is 30m to the DB-FFAP capillary column, and internal diameter is 0.32mm, film thickness is 0.5 μ m), column temperature is temperature programme: initial temperature is 200 ℃, keeps 10 minutes, speed with 20 ℃ of per minutes is warming up to 250 ℃, keeps 10 minutes.Fid detector.Prepare need testing solution and muskone reference substance solution by the inventive method, with the standby negative control solution that does not contain the muscone of legal system, in 5 batches of sample test sample chromatograms, all present the chromatographic peak identical with reference substance chromatographic peak retention time, negative noiseless.
Test example 3:
Embodiment 1 is seen in the preparation of test sample, control medicinal material and reference substance, and other gets the negative control that does not contain calculus bovis factitius, is made in the same way of negative control solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 3 μ l of above-mentioned four kinds of solution, put respectively in same with silica gel g thin-layer plate on, take normal hexane-ethyl acetate-acetic acid-methyl alcohol 20: 25: 2: 3 as developping agent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃.In the test sample chromatogram, with reference substance, the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.Negative control is noiseless.Tested 5 batch samples, all can reappear.
Test example 4: further specify ginseng stem-leaf total saponin content assaying method of the present invention below by test example.
1, stability test
To with a need testing solution, by chromatographic condition of the present invention, measure once at regular intervals, investigate altogether 25 hours, measured 6 times, measure ginsenoside Rg1 and ginsenoside Re's content and average out to 4.3mg/ sheet 6 times as a result, RSD=0.63% (n=6) sees table 5 for details.
Table 5 ginsenoside stability test result
The result shows that need testing solution is stable in 25 hours.
2, precision test
The instrument precision test is pressed chromatographic condition of the present invention to a need testing solution, continuous sample introduction 6 times, and ginsenoside Rg1 and ginsenoside Re's peak area in the working sample, the RSD value is respectively 1.00% and 0.47%, the results are shown in Table 6.
Table 6 instrument precision test findings
The result shows that the precision of instrument is good.
3, replica test is to same sample, by 6 parts of method replicate determinations of the present invention, calculate this product contain ginsenoside Rg1 and ginsenoside Re and, mean value is the 4.3mg/ sheet, the RSD value is 1.39%, the results are shown in Table 6.
Table 6 ginsenoside reappearance experimental result
The result shows that the repeatability of this law is good.
4, specificity test
Be further to investigate the specificity of test, do not contain the negative control solution of ginsenoside according to the recipe quantity preparation, measure in accordance with the law, as a result negative control solution with the corresponding retention time of reference substance chromatographic peak place without chromatographic peak, show noiselessly, specificity is good.
5, linear relationship and scope
Precision is drawn ginsenoside Rg1's reference substance solution (lot number: 110703-200425 respectively, concentration 0.159mg/ml) and ginsenoside Re's reference substance solution (lot number: 110754-200421, concentration 0.2406mg/ml) injection liquid chromatography, measure by chromatographic condition of the present invention, the results are shown in following table.Take the sample size (μ g) of reference substance as horizontal ordinate, chromatographic peak area is ordinate drawing standard curve.The results are shown in Table 7, table 8.
Table 7 ginsenoside Rg1 linear relationship is investigated the result
| Sample size (μ g) | 0.159 | 0.477 | 0.795 | 1.113 | 1.431 |
| The Rg1 peak area | 66605 | 213226 | 349093 | 496380 | 633770 |
Regression equation is: Y=-2556.2+445749.7X, and R=0.9999, the result shows that the ginsenoside Rg1 is good log-linear relation (ginsenoside Rg1's linear relationship is seen assay Fig. 4 .12) in 0.159 μ g～1.431 μ g scopes
Table 8 ginsenoside Re linear relationship is investigated the result
| Sample size (μ g) | 0.7218 | 1.203 | 2.406 | 3.609 | 4.812 |
| The Re peak area | 250029 | 421690 | 816128 | 1220516 | 1638418 |
Regression equation is: Y=8238.5+337645.5X, and R=0.9999, the result shows that the ginsenoside Re is good linear relation (ginsenoside Re's linear relationship is seen assay Fig. 4 .13) in 0.7218 μ g～4.812 μ g scopes.
6, accuracy test
6.1 ginsenoside Rg1's accuracy test
Precision takes by weighing 6 parts of the test samples of known content, every part of 0.25g, accurate ginsenoside Rg1's reference substance solution (0.0324mg/ml that adds, purity 96.3%) 20ml, extract, measure by method of the present invention, calculate recovery rate, average recovery rate=96.5%, RSD=2.32% (n=6).Meet the requirement of quantitative test.The results are shown in Table 9.
Table 9 ginsenoside Rg1 accuracy test result
6.2 ginsenoside Re's accuracy test
Precision takes by weighing 6 parts of test samples measuring known content with method, every part of 0.25g, accurate ginsenoside Re's reference substance solution (0.2256mg/ml that adds, purity 88.8%) 20ml, extract, measure by the method for working out standard, calculate recovery rate, average recovery rate=96.4%, RSD=1.34% (n=6).Meet the requirement of quantitative test.The results are shown in Table 10.
Table 10 ginsenoside Re accuracy test result
Claims (1)
1. the detection method of an Xueshuan Xinmaining capsule is characterized in that: comprise following discrimination method and content assaying method:
(1) discrimination method
(1) get the content of 20 of this capsules, porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; Get respectively red sage root control medicinal material 1g, Ligusticum wallichii control medicinal material 1g in addition, be made in the same way of control medicinal material solution; Get again tanshinone IIA reference substance, borneol reference substance, add ethyl acetate and make the solution that every 1ml contains 1mg, in contrast product solution; According to the thin-layered chromatography test, draw each 1 μ l of above-mentioned five kinds of solution, put respectively on same silica gel g thin-layer plate, take 60～90 ℃-ethyl acetate of sherwood oil 11:2 as developping agent, launch, take out, dry; In the test sample chromatogram, with red sage root control medicinal material, the corresponding position of tanshinone IIA reference substance chromatogram on, the spot of aobvious same color under the daylight; Put under the ultraviolet lamp 365nm and inspect, in the test sample chromatogram, with the corresponding position of Ligusticum wallichii control medicinal material chromatogram on, the fluorescence spot of aobvious same color; Spray with 5% phosphomolybdic acid ethanol solution, it is clear to be heated to the spot colour developing at 105 ℃ again; In the test sample chromatogram, with the spot of the aobvious same color in the corresponding position of borneol reference substance chromatogram;
(2) muscone's gas chromatography is differentiated: get the content of 20 of this capsules, and porphyrize, the 40ml cold soaking 1 hour of adding diethyl ether filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol 2ml makes dissolving, as need testing solution; It is an amount of to get the muskone reference substance, adds absolute ethyl alcohol and makes the solution that every 1ml contains 0.1mg, in contrast product solution; According to vapor-phase chromatography test, the DB-FFAP capillary column: column length is 30m, and internal diameter is 0.32mm, and film thickness is 0.5 μ m, and column temperature is temperature programme: initial temperature is 200 ℃, keeps 10 minutes, is warming up to 250 ℃ with the speed of 20 ℃ of per minutes, keeps 10 minutes; Precision is drawn above-mentioned reference substance solution and each 1 μ l of need testing solution respectively, and inject gas chromatograph is measured; The chromatographic peak identical with reference substance chromatographic peak retention time in the record test sample chromatogram;
(3) get the content of 6 of this capsules, porphyrize adds methyl alcohol 50ml, adds hot reflux 1 hour, filters, filtrate evaporate to dryness, residue add water 30ml dissolving, use extracted by ether 2 times, each 20ml, merge ether solution, evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrate is medicinal material solution in contrast; Get again the cholic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, take the upper solution of normal hexane-ethyl acetate-acetic acid-methyl alcohol 20:25:2:3 as developping agent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) content assaying method:
(1) Ligusticum wallichii assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take methyl alcohol-1% acetum 25:75 as mobile phase; The detection wavelength is 321nm, and number of theoretical plate calculates by the forulic acid peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the forulic acid reference substance, accurately weighed, puts in the brown measuring bottle, adds 70% methyl alcohol and makes the solution that every 1ml contains 20 μ g, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize, get 0.3g ~ 0.5g, accurately weighed, put in the tool plug conical flask, accurate adding 70% methyl alcohol 20ml, weighed weight, ultrasonic processing 30 minutes, power 250W, frequency 50kHz lets cool, weighed weight is supplied the weight that subtracts mistake with 70% methyl alcohol again, shakes up, filter, get subsequent filtrate, and get final product;
Determination method: precision is drawn reference substance solution 5 μ l and need testing solution 10 μ l respectively, and the injection liquid chromatography is measured, and be get final product;
(2) ginseng stem and leave general saponin assay
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-water 20:80 as mobile phase; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately weighed, add methyl alcohol and make the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, and get final product;
The preparation of need testing solution: get the content of 20 of this capsules, accurately weighed, porphyrize is got 0.4 ~ 0.6g, and is accurately weighed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic extraction 30 minutes filters, and discards filtrate; The dregs of a decoction volatilize solvent together with filter paper, accurate methyl alcohol 50ml, weighed weight, the refluxing extraction 1 hour of adding, let cool, weighed weight is supplied the weight that subtracts mistake with methyl alcohol again, shakes up, filter, get subsequent filtrate 25ml, evaporate to dryness, residue add water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discard the ammonia solution layer, normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol makes dissolving in right amount, is transferred in the 10ml measuring bottle, adds methyl alcohol to scale and makes dissolving, shake up, filter, and get final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110357472 CN102426205B (en) | 2011-11-13 | 2011-11-13 | Detection method of thrombus sumalin capsules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110357472 CN102426205B (en) | 2011-11-13 | 2011-11-13 | Detection method of thrombus sumalin capsules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102426205A CN102426205A (en) | 2012-04-25 |
| CN102426205B true CN102426205B (en) | 2013-01-09 |
Family
ID=45960210
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110357472 Active CN102426205B (en) | 2011-11-13 | 2011-11-13 | Detection method of thrombus sumalin capsules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102426205B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507844B (en) * | 2011-11-13 | 2013-10-16 | 吉林华康药业股份有限公司 | Testing method for Xueshuan xinmaining tablet |
| CN103884676B (en) * | 2014-02-27 | 2016-06-22 | 浙江大学 | A kind of rapid assay methods of Chinese crude drug multi-target ingredient content |
| CN106483260A (en) * | 2015-10-09 | 2017-03-08 | 厦门中药厂有限公司 | A kind of pill of Eight Treasures capsule quality standard control method |
| CN109085284A (en) * | 2018-08-28 | 2018-12-25 | 王智森 | A kind of quick thin-layer identification method of multiple medicine taste of Ershiwuwei zhenzhu wan |
| CN110632240B (en) * | 2019-11-08 | 2021-11-02 | 亳州职业技术学院 | Identification kit and identification method for traditional Chinese medicine ground beetles |
| CN113720958A (en) * | 2021-08-25 | 2021-11-30 | 国药集团精方（安徽）药业股份有限公司 | Quality detection method of Shenyanshu capsule |
-
2011
- 2011-11-13 CN CN 201110357472 patent/CN102426205B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN102426205A (en) | 2012-04-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN102426205B (en) | Detection method of thrombus sumalin capsules | |
| CN103197027A (en) | Quality control method of astragalus-leech capsules capable of regulating collaterals | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN102100818A (en) | Quality control method for lophanthus antifebrile tablets | |
| CN101708208B (en) | Detection method of capsule preparation for treating painful swollen joint | |
| CN105301168B (en) | The detection method of dredging collateral resolving sputum capsule | |
| CN102353735B (en) | Quality detection method for Tongmai Tangyanming capsule | |
| CN103344737A (en) | Quality control method of traditional Chinese medicine tablet for treating nasosinusitis | |
| CN102218122A (en) | Quality control and detection method for sea dragon and gecko oral liquid | |
| CN104391072A (en) | Quality control method of traditional Chinese medicine compound preparation for treating osteoporosis | |
| CN102507844B (en) | Testing method for Xueshuan xinmaining tablet | |
| CN102068627A (en) | Quality control method for Chinese medicine preparation Xinnaojing tabelets | |
| CN104922587A (en) | Preparation method of swelling-removing and pain-relieving paste and quality-detecting method of swelling-removing and pain-relieving paste | |
| CN102879516A (en) | Method for identifying Buyang Huanwu soup and measuring content of Buyang Huanwu soup | |
| CN106124668B (en) | A kind of fingerprint atlas detection method of Yixinshu tablet | |
| CN106290643B (en) | A kind of method of quality control of Chinese medicine schizonepeta anti-lung cancer activity ingredient | |
| CN105929080B (en) | A kind of detection method of Chinese materia medica preparation that treating nerve root cervical vertebra sickness | |
| CN102507842A (en) | Testing method for Xueshuan xinmaining tablet | |
| CN108226325A (en) | Roripa montana gives birth to the method for building up of arteries and veins oral liquid composition finger-print | |
| CN101269203A (en) | Quality control testing method for brain-invigorating and heart-soothing capsule | |
| CN105169234B (en) | A kind of quality determining method of Chinese materia medica preparation that treating diabetic retinopathy | |
| CN102507845B (en) | Detection method of Xueshuan Xinmaining capsule | |
| CN105866289A (en) | Method for quickly extracting and measuring various phenolic acid components in compound radix salviae miltiorrhizae tablets | |
| CN105259264A (en) | Method for measuring content of multiple active ingredients in xiaoshuan tongluo tablets |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent of invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wang Xinhua Inventor after: Han Hongxiang Inventor after: Jin Liqun Inventor after: Liu Chuangui Inventor after: Yan Zhili Inventor after: Wang Li Inventor before: Wang Xinhua Inventor before: Jin Liqun Inventor before: Liu Chuangui Inventor before: Yan Zhili Inventor before: Wang Li |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: WANG XINHUA JIN LIQUN LIU CHUANGUI YAN ZHILI WANG LI TO: WANG XINHUA HAN HONGXIANG JIN LIQUN LIU CHUANGUI YAN ZHILI WANG LI |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230523 Address after: 133700 No. 667, Kangping street, Dunhua Economic Development Zone, Yanbian Korean Autonomous Prefecture, Jilin Province Patentee after: Jilin Ruilong Pharmaceutical Co.,Ltd. Address before: 133700 No. 10, Huakang street, Dunhua City, Yanbian Korean Autonomous Prefecture, Jilin Province Patentee before: Jilin Huakang Pharmaceutical Co.,Ltd. |
|
| TR01 | Transfer of patent right |