CN102426206A - Quality control method of traditional Chinese medicine preparation for treating rheumatism - Google Patents
Quality control method of traditional Chinese medicine preparation for treating rheumatism Download PDFInfo
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- CN102426206A CN102426206A CN2011103027394A CN201110302739A CN102426206A CN 102426206 A CN102426206 A CN 102426206A CN 2011103027394 A CN2011103027394 A CN 2011103027394A CN 201110302739 A CN201110302739 A CN 201110302739A CN 102426206 A CN102426206 A CN 102426206A
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Abstract
The invention discloses a quality control method of a traditional Chinese medicine preparation for treating rheumatism, and specifically provides a quality control method of an activating and pain-relieving capsule. According to the invention, discrimination or content determination is carried out on one or a plurality of bulk drugs in a prescription, such as acanthopanax senticosus, radix clematidis, angelica sinensis, prepared aconite root, prepared kusnezoff monkshood root, radde anemone rhizome, root of red-rooted salvia, processed frankincense, processed myrrh and herbal ephedrae to realize purpose of effective control on product quality. An abstract figure is shown as below. The invention has advantage that the method can carry out discrimination and content determination on a plurality of bulk drugs in the prescription to effectively guarantee drug effect and stability of the produced activating and pain-relieving capsule, thereby better protecting drug usage safety of patients.
Description
The application is for dividing an application, and the application number of its female case application is 200910226650.7, and the applying date is on Dec 16th, 2009, and denomination of invention is " a kind of method of quality control of treating the Chinese medicine preparation of rheumatism ".
Technical field
The present invention relates to a kind of method of quality control of Chinese medicine preparation, relate in particular to a kind of method of quality control of Chinese medicine preparation of rheumatism.
Background technology
Rheumatism is a kind of systemic disease, and full name is " rheumatism ", is a kind of common disease frequently-occurring disease.It is of a great variety; Comprise kind surplus rheumatoid arthritis, lupus erythematosus, dermatomyositis, sclerderm, ankylosing spondylitis, rheumatic fever, infectional arthritis, the soft tissue rheumatism etc. 100; Wherein occurred frequently with several kinds of diseases such as ankylosing spondylitis, rheumatoid arthritis, Osteoarthritis especially; And prognosis is relatively poor, and disability rate is than higher.The cause of disease of these diseases is different, and common performance is but arranged: limbs, joint, myalgia, or with swelling, inconvenient activity.Patient's pain is unbearably, and is handicapped, and be difficult to cure, and many patients lack rheumatismal understanding, do not know how to prevent and go to a doctor, to such an extent as to the state of an illness finally is difficult to reverse by light to heavy, and serious deformity even the life-threatening of causing.
So, very multiple type rheumatism class medicine has appearred on the market, and the validity of rheumatism class medicine and the stable content that has become day by day for everybody paid close attention to.
The active pain preparation that disappears is the Chinese medicine patent medicine that is used to treat illnesss such as wind-cold-dampness arthralgia, channels and collaterals obturation, arthralgia and myalgia and numb limb.List marketing at present active pain relieving tablet and two kinds of formulations of active pain-eliminating capsule are arranged.Wherein the standard of tablet institute foundation is numbered: WS3-B-0355-90, in this standard, what apply to is existing current techique, can reach the purpose of the most basic control drug quality.
But along with the continuous progress of pharmaceutical technology, reach the purpose of safer medication, must constantly improve the drug standards.Especially contain Chinese ephedra in the preparation raw material medicine of the present invention; Ephedrine hydrochloride in the Chinese ephedra is in a single day excessive; Will cause anxious, have a sleepless night, tremble, the serious side effects of cardiovascular and cerebrovascular such as palpitaition, headache, dizzy, nauseating, vomiting, poor appetite, elevation of blood pressure, renal shutdown, perspiration and fash and nervous system aspect; Entail dangers to is to medication person's life security, and the strictness that therefore its content in medicine also received State Food and Drug Administration limits.
Just be based on this, the present invention puts forth effort on the better drug quality of the active pain preparation that disappears of control of research.
Summary of the invention
The purpose of this invention is to provide a kind of method of quality control of treating the Chinese medicine preparation of rheumatism; It is the method for quality control by the prepared active pain-eliminating capsule of bulk drug Extractum Acanthopanacis Senticosi, the root of Chinese clematis, Radix Angelicae Sinensis, aconiti preparata,radix, wild aconite root, Radde Anemone Rhizome, the red sage root, frankincense, myrrh, Chinese ephedra; Come better to guarantee the stability of medicine with this; Quality controllability makes medication safer, effective.
The objective of the invention is to realize, comprise that Determination of Ephedrine Hydrochloride is measured in following discriminating and/or microscopical identification and/or the Chinese ephedra through following technical scheme:
The discriminating of wilsonii
These article of getting content 0.35-1g, porphyrize adds methyl alcohol 10ml-30ml, sonicated 10-30 minute, filters; Filtrating is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 0.5-1.5ml, and as need testing solution, other gets wilsonii control medicinal material 2.5-7.5g, adds 75% ethanol 25-75ml; Reflux 0.5-1.5 hour, put coldly, filter, filtrating is put evaporate to dryness in the water-bath, and residue adds water 5-15ml makes dissolving; Add chloroform and extract 1-3 time, each 5-15ml, combined chloroform liquid is put evaporate to dryness in the water-bath; Residue adds methyl alcohol 0.5-1.5ml makes dissolving, as control medicinal material solution, gets the isofraxidin reference substance again, adds methyl alcohol and processes the solution that every 0.5-1.5ml contains 0.5-1.5mg; As reference substance solution, according to the thin-layered chromatography test, draw above-mentioned three kinds of each 5-15ul of solution, put respectively on same silica gel g thin-layer plate; The chloroform-methanol that with the ratio is 9-29: 0.5-1.5 is a developping agent, launches, and takes out, and dries; Put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color; With the position of reference substance chromatogram on, show the spot of same color;
Microscopical identification
These article of getting content, put microscopically and observe: lithocyte is colourless, with apparent brown, type square, rectangle like, similar round, class prismatic or the strip that metaderm links, diameter 20-133um; Long 50-190um, wall are thick slightly, and wall thickness person laminated striation is obvious, and the many bunchys of fiber are faint yellow; Be elongated strip, terminal short point, the visible short branch that has, edge out-of-flatness, diameter 22-56um; Wall thickness 4-8um, tool edge line cell rufous, polygon, wall is thin, the phloem parenchyma cell spindle; Wall is slightly thick, and atomic thin oblique cross lamination is arranged, and the epidermal cell fragment is faint yellow, and the cell rectangle includes small calcium oxalate crystal.
Assay
Assay is according to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; Acetonitrile-0.1% phosphoric acid solution that with the ratio is 3.5-10: 46-140 is a moving phase: the detection wavelength is 207nm, and number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 3000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and process and contain 20ug solution among every 1ml, promptly get,
These article content 1.7-5.2g is got in the preparation of need testing solution, stable precision, and porphyrize is got about 0.5-1.5g, and accurate the title, decide; Put conical flask, add 30-90% ethanol 25-75ml, ultrasonic 15-45 minute, put coldly, filter; With a small amount of 30-90% ethanol gradation washing filter residue and filter, merging filtrate and washing lotion are put and are steamed in the water-bath to there not being the alcohol flavor, add water 15-45ml, are transferred in the separating funnel; Shake up, add strong ammonia solution and regulate pH value, add diethyl ether again and extract 2-7 time, merge ether solution, put low temperature evaporate to dryness in the water-bath to 10-11; Residue adds dissolve with methanol and is transferred in the 12-37ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets
Accurate respectively reference substance solution and each 2.5-7.5ul of need testing solution of drawing of determination method injects liquid chromatograph, measures, promptly get,
Determination of Ephedrine Hydrochloride experiment in the starting material Chinese ephedra medicinal material of active pain-eliminating capsule, concrete test method and result are following:
1, instrument and reagent
Instrument: Autech high performance liquid chromatograph; C
18(Hypersil ODS
2250 * 4.6mm, 5u) chromatographic column, Cschrom data processing software.Reagent: acetonitrile is a chromatorgaphy reagent, and water is redistilled water, and it is pure that other reagent is analysis.
Reference substance:, supply assay to use for Nat'l Pharmaceutical & Biological Products Control Institute provides.
2, chromatographic condition
Moving phase: acetonitrile-0.1% phosphoric acid solution (7: 93)
Detect wavelength: 207nm; Flow velocity: 1.0nm/min.
Under above-mentioned selected condition, to measure through the HPLC method, other component chromatographic peak can reach baseline separation in ephedrine hydrochloride peak and the sample, and adjacent chromatographic peak degree of separation with other is greater than 1.5; Press the ephedrine hydrochloride peak and calculate, number of theoretical plate is more than 3000; In the negative solution chromatogram with ephedrine hydrochloride chromatographic peak relevant position on do not have chromatographic peak, explain that feminine gender is noiseless, reference substance, test sample, negative sample chromatogram are seen Fig. 1,2,3 respectively.
3, need testing solution preparation
It is tolerant to get these article 10 intragranulars, stable precision, and porphyrize is got about 1g, stable precision; Put conical flask, add 60% ethanol 50ml, ultrasonic 30 minutes, put coldly, filter; With a small amount of 60% ethanol gradation washing filter residue and filter, merge filter night and washing lotion, put and steam in the water-bath to there not being the alcohol flavor, add water 30ml, be transferred in the separating funnel; Shake up, add strong ammonia solution and transfer to pH value, add diethyl ether again and extract (25,20,20,10,10ml) 5 times, merge ether solution, put low temperature evaporate to dryness in the water-bath to 10-11; Residue adds dissolve with methanol and is transferred in the 25ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets.
4, the preparation of negative solution
Get other recipe quantity medicinal materials except that Chinese ephedra, be prepared into the negative sample 1g that lacks Chinese ephedra, process the negative solution that lacks Chinese ephedra by the need testing solution preparation method by preparation technology.
5, the preparation of reference substance solution
Precision takes by weighing ephedrine hydrochloride reference substance 5.21mg, puts in the 50ml measuring bottle, adds dissolve with methanol and is diluted to scale; Shake up (0.1042mg/ml), the accurate 3ml that draws puts in the 20ml measuring bottle; Add methyl alcohol and be diluted to scale, shake up, promptly get (hydrochloric ephedrine 0.01536mg among every 1ml).
6, linear relationship is investigated
Accurate absorption concentration is ephedrine hydrochloride reference substance solution 1,3,5,7, the 9ul sample introduction of 0.01536mg/ml; Measure its peak area integrated value; With concentration is horizontal ordinate, and the peak area integrated value is an ordinate, the drawing standard curve; Its regression equation is: A=54564576.1C+172066.7, r=0.9996. result show that ephedrine hydrochloride has good linear relationship in the 0.02048-0.18432ug scope.Data are seen table 1, and typical curve is seen Fig. 4.
Table 1 linear relationship is investigated table as a result
| Concentration (ug) | 0.01536 | 0.04608 | 0.0768 | 0.10752 | 0.13824 |
| Peak area | 934622 | 2723959 | 4480878 | 5991898 | 7681770 |
7, precision test
The same need testing solution 5ul of accurate absorption repeats sample introduction 5 times, measures its peak area integrated value, and its result sees table 2.
Table 2 Precision test result table
The result shows: this law precision is good.
8, stability test
Get same need testing solution respectively at 0,2,4,6,8 hour sample introduction, measure its peak area integrated value, the result sees table 3.
Table 3 stability test is table as a result
| Time (hour) | ?0 | 2 | 4 | 6 | 8 |
| Peak area value | ?4409154 | 4393798 | 4392978 | 4373959 | 4385798 |
The result shows that need testing solution peak area value in 12 hours is basicly stable.
9, reappearance test
Get same lot number sample by above-mentioned condition, measure 5 times (n=2), the result sees table 4.
Table 4 reproducible test results table
The result shows that average content is 0.1372mg/ grain (0.3916mg/g), this law favorable reproducibility.
10, recovery test
Precision takes by weighing known content and is the about 0.5g of 0.3916mg/g sample; Respectively accurate ephedrine hydrochloride reference substance solution (0.0942mg/ml) 2ml that adds, i.e. 0.1884mg is from " adding 60% ethanolic solution 50ml "; Measure content by the method under the above-mentioned assay item, the result sees table 5.
Table 5 determination of recovery rates is table as a result
The result shows that this method average recovery is good.
11, sample determination method and result
By the above-mentioned content assaying method working sample of drafting, the result sees table 6.
Table 6 sample size is measured table as a result
| Lot number | 20030518 | 20030520 | 20030522 | 20041020 | 20041022 |
| Content (mg/ grain) | 0.09 | 0.09 | 0.09 | 0.12 | 0.12 |
| Lot number | 20041024 | 20050812 | 20050814 | 20050816 | |
| Content (mg/ grain) | 0.12 | 0.14 | 0.14 | 0.14 |
Above-mentioned sample size is pressed 70% conversion of average content 0.117mg/ grain in 0.09-0.14mg/ grain scope, Determination of Ephedrine Hydrochloride must not be less than 0.08mg in every of these article of regulation.
Specific embodiment
The discriminating of specific embodiment one wilsonii
These article of getting content 0.7g, porphyrize adds methyl alcohol 20ml, and sonicated 20 minutes filters; Filtrating is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml, and as need testing solution, other gets wilsonii control medicinal material 5g, adds 75% ethanol 50ml; Reflux 1 hour is put coldly, filters, and filtrating is put evaporate to dryness in the water-bath, and residue adds water 10ml makes dissolving; Add chloroform and extract 2 times, each 10ml, combined chloroform liquid is put evaporate to dryness in the water-bath, and residue adds methyl alcohol 1ml makes dissolving; As control medicinal material solution, get the isofraxidin reference substance again, add methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution, according to the thin-layered chromatography test; Drawing above-mentioned three kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, is that 19: 1 chloroform-methanol is a developping agent with ratio, launches, and takes out; Dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color; With the position of reference substance chromatogram on, show the spot of same color.
Specific embodiment two microscopical identification
Get content of the present invention, put microscopically and observe: lithocyte is colourless, with apparent brown, type square, rectangle like, similar round, class prismatic or the strip that metaderm links, diameter 60um; Long 100um, wall are thick slightly, and wall thickness person laminated striation is obvious, and the many bunchys of fiber are faint yellow; Be elongated strip, terminal short point, the visible short branch that has, edge out-of-flatness, diameter 40um; Wall thickness 6um, tool edge line cell rufous, polygon, wall is thin, the phloem parenchyma cell spindle; Wall is slightly thick, and atomic thin oblique cross lamination is arranged, and the epidermal cell fragment is faint yellow, and the cell rectangle includes small calcium oxalate crystal.
Specific embodiment three assays
Assay is according to high effective liquid chromatography for measuring;
The test of chromatographic condition and system suitability: uses octadecylsilane chemically bonded silica to be filling agent, be that 7: 93 acetonitrile-0.1% phosphoric acid solution is a moving phase with ratio: the detection wavelength is 207nm, and number of theoretical plate should be not less than 3000 by the calculating of ephedrine hydrochloride peak,
It is an amount of that the preparation precision of reference substance solution takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and process and contain 20ug solution among every 1ml, promptly get,
Content 3.5g of the present invention is got in the preparation of need testing solution, stable precision, and porphyrize is got about 1g, and accurate the title, decide; Put conical flask, add 60% ethanol 50ml, ultrasonic 30 minutes, put coldly, filter; With a small amount of 60% ethanol gradation washing filter residue and filter, merging filtrate and washing lotion are put and are steamed in the water-bath to there not being the alcohol flavor, add water 30ml, are transferred in the separating funnel; Shake up, add strong ammonia solution and regulate pH value, add diethyl ether again and extract 5 times, merge ether solution, put low temperature evaporate to dryness in the water-bath to 10-11; Residue adds dissolve with methanol and is transferred in the 25ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets
Accurate respectively reference substance solution and each 5ul of need testing solution of drawing of determination method injects liquid chromatograph, measures, and promptly gets.
Description of drawings:
1, Fig. 1 is an ephedrine hydrochloride reference substance chromatogram;
2, Fig. 2 is active pain-eliminating capsule test sample chromatogram;
3, Fig. 3 is active pain-eliminating capsule negative sample chromatogram;
4, Fig. 4 is the ephedrine hydrochloride linear relationship chart.
Claims (3)
1. detection method of treating the Chinese medicine preparation of rheumatism; This Chinese medicine preparation is by bulk drug Extractum Acanthopanacis Senticosi, the root of Chinese clematis, Radix Angelicae Sinensis, aconiti preparata,radix, wild aconite root, Radde Anemone Rhizome, the red sage root, frankincense, myrrh, the prepared active pain-eliminating capsule of Chinese ephedra, it is characterized in that Determination of Ephedrine Hydrochloride determination method in the interior Chinese ephedra of active pain-eliminating capsule.
2. detection method as claimed in claim 1, the Determination of Ephedrine Hydrochloride determination method may further comprise the steps in the interior Chinese ephedra of wherein active pain-eliminating capsule:
Determination of high performance liquid chromatography;
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; Acetonitrile-0.1% phosphoric acid solution that with the ratio is 3.5-10: 46-140 is a moving phase: the detection wavelength is 207nm, and number of theoretical plate calculates by the ephedrine hydrochloride peak should be not less than 3000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and process and contain 20ug solution among every 1ml, promptly gets;
The preparation of need testing solution: get active pain-eliminating capsule content 1.7-5.2g, stable precision, porphyrize is got about 0.5-1.5g, and accurate the title, decide; Put conical flask, add 30-90% ethanol 25-75ml, ultrasonic 15-45 minute, put coldly, filter; With a small amount of 30-90% ethanol gradation washing filter residue and filter, merging filtrate and washing lotion are put and are steamed in the water-bath to there not being the alcohol flavor, add water 15-45ml, are transferred in the separatory funnel; Shake up, add the strong ammonia solution regulated value, add diethyl ether again and extract 2-7 time, merge ether solution, put low temperature evaporate to dryness in the water-bath to 10-11; Residue adds the methyl alcohol dissolving and is transferred in the 12-37ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;
Determination method: accurate respectively reference substance solution and each 2.5-7.5ul of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
3. detection method as claimed in claim 2, the Determination of Ephedrine Hydrochloride determination method may further comprise the steps in the interior Chinese ephedra of wherein active pain-eliminating capsule:
Determination of high performance liquid chromatography;
The test of chromatographic condition and system suitability: uses octadecylsilane chemically bonded silica to be filling agent, be that 7: 93 acetonitrile-0.1% phosphoric acid solution is a moving phase with ratio: the detection wavelength is 207nm, and number of theoretical plate should be not less than 3000 by the calculating of ephedrine hydrochloride peak;
The preparation of reference substance solution: it is an amount of that precision takes by weighing the ephedrine hydrochloride reference substance, adds methyl alcohol and process and contain 20ug solution among every 1ml, promptly gets;
The preparation of need testing solution: get active pain-eliminating capsule content 3.5g, stable precision, porphyrize is got about 1g, and accurate the title, decide; Put conical flask, add 60% ethanol 50ml, ultrasonic 30 minutes, put coldly, filter; With a small amount of 60% ethanol gradation washing filter residue and filter, merging filtrate and washing lotion are put and are steamed in the water-bath to there not being the alcohol flavor, add water 30ml, are transferred in the separatory funnel; Shake up, add the strong ammonia solution regulated value, add diethyl ether again and extract 5 times, merge ether solution, put low temperature evaporate to dryness in the water-bath to 10-11; Residue adds the methyl alcohol dissolving and is transferred in the 25ml measuring bottle, adds methyl alcohol to scale, shakes up, and promptly gets;
Determination: precision drawing reference solution and the test solution 5ul, into the liquid chromatograph to measure, that is.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011103027394A CN102426206A (en) | 2009-12-16 | 2009-12-16 | Quality control method of traditional Chinese medicine preparation for treating rheumatism |
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| CN2011103027394A CN102426206A (en) | 2009-12-16 | 2009-12-16 | Quality control method of traditional Chinese medicine preparation for treating rheumatism |
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| CN2009102266507A Division CN101766708B (en) | 2009-12-16 | 2009-12-16 | Quality control method of Chinese herbal preparation for rheumatism treatment |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106214770A (en) * | 2016-09-18 | 2016-12-14 | 湖南天地恒制药有限公司 | Huoluo Xiaotong Capsules is as the application of diabetic peripheral neuropathy medicine |
| CN110221001A (en) * | 2019-07-10 | 2019-09-10 | 青岛大学附属医院 | A kind of method of monoester alkaloid in extraction aconiti preparata,radix |
-
2009
- 2009-12-16 CN CN2011103027394A patent/CN102426206A/en active Pending
Non-Patent Citations (2)
| Title |
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| 孙雪妹等: "良园枇杷叶膏中盐酸麻黄碱的HPLC测定", 《中国医药工业杂志》 * |
| 赵大成等: "高效液相色谱测定止嗽定喘口服液中盐酸麻黄碱的方法研究", 《肿瘤研究与临床》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106214770A (en) * | 2016-09-18 | 2016-12-14 | 湖南天地恒制药有限公司 | Huoluo Xiaotong Capsules is as the application of diabetic peripheral neuropathy medicine |
| CN106214770B (en) * | 2016-09-18 | 2020-04-10 | 天地恒一制药股份有限公司 | Application of collateral activating and pain relieving capsule in preparation of medicine for treating diabetic peripheral neuropathy |
| CN110221001A (en) * | 2019-07-10 | 2019-09-10 | 青岛大学附属医院 | A kind of method of monoester alkaloid in extraction aconiti preparata,radix |
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Application publication date: 20120425 |