CN102068549B - Detection method for Chinese medicinal preparation heat clearing and blood cooling pills - Google Patents
Detection method for Chinese medicinal preparation heat clearing and blood cooling pills Download PDFInfo
- Publication number
- CN102068549B CN102068549B CN200910228359A CN200910228359A CN102068549B CN 102068549 B CN102068549 B CN 102068549B CN 200910228359 A CN200910228359 A CN 200910228359A CN 200910228359 A CN200910228359 A CN 200910228359A CN 102068549 B CN102068549 B CN 102068549B
- Authority
- CN
- China
- Prior art keywords
- solution
- reference substance
- methyl alcohol
- thin
- clearing heat
- 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
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of Chinese patent medicaments, and relates to a quality control method for a Chinese medicinal preparation, namely heat clearing and blood cooling pills prepared from Chinese medicinal materials. The heat clearing and blood cooling pills consist of Chinese medicinal materials, namely scutellaria root and Rehmannia glutinosa. The quality control method comprises the following steps of: taking baicalin and catalpol as reference products, and identifying whether scutellaria root and Rehmannia glutinosa components are contained in the prescription of the heat clearing and blood cooling pills by thin-layer chromatography; and detecting the content of the baicalin in the prescription of the heat clearing and blood cooling pills by high performance liquid chromatography. A quantitative index and an inspection method thereof make the quality standard more perfect. The revised quality standard improves the quality control of medicaments.
Description
Technical field
The invention belongs to the technical field of Chinese patent drug, relating to the Chinese crude drug is the detection method of the Chinese medicine preparation clearing heat and cooling blood ball processed of raw material.
Background technology
The clearing heat and cooling blood ball is the 9th the 192 page of preparation that records of " the Sanitation Ministry medicine standard " Chinese traditional patent formulation preparation, the standard WS3-B-1843-94 of Ministry of Public Health record prescription and quality standard:
Prescription: root of large-flowered skullcap 500g glutinous rehmannia 500g
Method for making: above two flavors, be ground into fine powder, sieve, mixing, the water pill, drying promptly gets.
Differentiate: get these article 1g, grind, add methyl alcohol 20ml, reflux 2 hours is put coldly, filters, and filtrates as need testing solution.Other gets the baicalin reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin-layered chromatography (57 pages of appendix) test, draw each 10 μ l of need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate; (10: 7: 2: 2) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water; 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 fluorescence spot of same color.
But there is following problem in above-mentioned standard: after discrimination method point sample thin layer plate launches to dry; Put under the ultraviolet lamp (365nm) and inspect, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on; The bronzing spot is arranged, no fluorescence spot discriminating means existing problems; The assay item that does not reflect the index components of medicine inherent quality in addition for quality control standard, still is short of to some extent, for improving the product quality of Chinese medicine, need improve target level of product quality.
Summary of the invention
The objective of the invention is to the objective of the invention is to overcome the clearing heat and cooling blood ball standard weak point of prior art, a kind of detection method of clearing heat and cooling blood ball that can the qualitative and quantitative analysis drug ingedient is provided.
In order to reach the technical scheme that the object of the invention adopts be: a kind of detection method of Chinese medicine preparation clearing heat and cooling blood ball, wherein said pharmaceutical formulation are by Chinese crude drug: root of large-flowered skullcap 500g glutinous rehmannia 500g forms, more than two distinguish the flavor of; Be ground into fine powder, sieve, mixing; The water pill; Drying promptly gets, and it is characterized in that: the step of its method is:
(1) be reference substance with scutelloside, Catalpol, thin-layered chromatography differentiates in the clearing heat and cooling blood ball prescription whether contain the root of large-flowered skullcap, glutinous rehmannia composition;
(2) with the scutelloside be reference substance, high performance liquid chromatography detects content of baicalin in the clearing heat and cooling blood ball prescription.
The detection method of described Chinese medicine preparation clearing heat and cooling blood ball is characterized in that: described thin-layered chromatography differentiates that the method that does not contain the root of large-flowered skullcap in the clearing heat and cooling blood ball prescription is: get these article 1g, grind; Add methyl alcohol 20ml, reflux 2 hours is put cold; Filter, filtrating is as need testing solution; Other gets the scutelloside reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with ethyl acetate-butanone-formic acid-water, ethyl acetate; Butanone: formic acid: water=10: 7: 2: 2, launch, take out, to dry, spray is with 1% ferric trichloride ethanolic solution, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
Described thin-layered chromatography differentiates that the method that whether contains the glutinous rehmannia composition in the clearing heat and cooling blood ball prescription is: get these article powder 2g, add methyl alcohol 20ml, reflux 1 hour; Put coldly, filter the filtrating evaporate to dryness; Residue adds ethyl acetate 20ml sonicated 30 minutes, filters the filtrating evaporate to dryness; Residue adds methyl alcohol 5ml, as need testing solution; Other gets the Catalpol reference substance, adds methyl alcohol and processes the solution that every 1ml contains 0.5mg, as reference substance solution; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate, be developping agent with chloroform-methanol-water; Chloroform: methyl alcohol: water=14: 6: 1, launch, take out; Dry, spray is with the anisaldehyde test solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
The detection method of described Chinese medicine preparation clearing heat and cooling blood ball is characterized in that: be reference substance with the scutelloside, the method that liquid phase chromatography detects content of baicalin in the clearing heat and cooling blood ball prescription is:
(1) chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; Methyl alcohol: water: phosphoric acid=47: 53: 0.2 is moving phase; The detection wavelength is 280nm; Number of theoretical plate calculates by the scutelloside peak should be not less than 2000;
(2) preparation of reference substance solution: it is an amount of to get the scutelloside reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.1mg, promptly gets;
(3) preparation of need testing solution: sample thief 2g, porphyrize is got powder 0.2g, and accurate the title, decide; Put in the tool plug conical flask, precision is measured 70% ethanol 100ml, claims to decide weight, reflux 30 minutes; Put coldly, claim to decide weight, supply the weight that subtracts mistake, shake up with 70% ethanol; Filter, get subsequent filtrate, promptly get;
(4) determination method: inaccurate reference substance solution and each 5 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get; The every gram of these article contains scutelloside must not be less than 24mg.
Being provided with scutelloside, Catalpol in the clearing heat and cooling blood ball quality standard of the present invention is reference substance, and thin-layered chromatography differentiates in the clearing heat and cooling blood ball prescription whether contain the root of large-flowered skullcap, glutinous rehmannia composition; High performance liquid chromatography detects content of baicalin in the clearing heat and cooling blood ball prescription; The every gram of these article contains the quantitative target and the method for inspection that scutelloside must not be less than 24mg; Make quality standard comparatively perfect, the quality of strict control medicine is guaranteed to improve the quality of products; Revised quality standard, the quality control that has improved medicine.
Advantage of the present invention and good effect are:
1. thin-layered chromatography is differentiated in the scutelloside colour developing and is adopted spray with 1% ferric trichloride ethanolic solution in this detection method, and it is clear to be heated to the spot colour developing at 105 ℃, increases the thin layer spot greatly and detects effect, is convenient to observe and differentiates, has ensured the accuracy of identification result.
2. increase thin-layered chromatography in this detection method and differentiated glutinous rehmannia active component Catalpol step, can check out counterfeit drug and substandard drug more exactly, improved the security in using.
3. increase high-efficient liquid phase technique in this detection method and measured the content of baicalin step; Improved the controllability of clearing heat and cooling blood ball quality standard; Further guarantee product inherent quality and curative effect, and to promoting production marketing, increase the competitiveness of product in market, guaranteeing that patient's drug safety is significant
4. detection method of the present invention is simple, easy to operate, has eliminated the uncertainty that original scutelloside is differentiated, is a kind of revised quality standard of discrimination method that can differentiate Chinese medicine preparation clearing heat and cooling blood ball more comprehensively and accurately, the quality control that has improved medicine.
Description of drawings
Fig. 1 is that the thin layer of including in the former ministerial standard of clearing heat and cooling blood ball is differentiated the figure that inspects under the ultraviolet lamp of the root of large-flowered skullcap.
Fig. 2 is the thin-layer chromatogram of the discriminating root of large-flowered skullcap of clearing heat and cooling blood ball of the present invention.
Fig. 3 is the thin-layer chromatogram of the discriminating glutinous rehmannia of clearing heat and cooling blood ball of the present invention.
Fig. 4 is the liquid chromatogram of the confession test agent solution of clearing heat and cooling blood ball of the present invention.
Fig. 5 is the liquid chromatogram of the scutelloside standard solution of clearing heat and cooling blood ball of the present invention.
Fig. 6 is the liquid chromatogram of the blank article solution of clearing heat and cooling blood ball of the present invention.
Fig. 7 is the linear curve map of investigating of the assay of clearing heat and cooling blood ball of the present invention.
Embodiment
The present invention has increased the thin layer discriminating of glutinous rehmannia and the assay of the root of large-flowered skullcap to Chinese medicine preparation clearing heat and cooling blood ball detection method; Simultaneously,, revise again,, the present invention further specified, the clearing heat and cooling blood ball quality standard after embodiment 1 improves below in conjunction with embodiment to root of large-flowered skullcap discrimination method in the primary standard:
Prescription: root of large-flowered skullcap 500g glutinous rehmannia 500g
Method for making: above two flavors, be ground into fine powder, sieve, mixing, the water pill, drying promptly gets.
Proterties: these article are the yellowish-brown water-bindered pill to pitchy; It is sweet, bitter to distinguish the flavor of.
Differentiate: these article 1g is got in (1), grinds, and adds methyl alcohol 20ml, and reflux 2 hours is put coldly, filters, and filtrates as need testing solution.Other gets the scutelloside reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With ethyl acetate-butanone-formic acid-water (10: 7: 2: 2) be developping agent; Launch, take out, dry; Spray is with 1% ferric trichloride ethanolic solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
(2) get these article powder 2g, add methyl alcohol 20ml, reflux 1 hour is put coldly, filter, and the filtrating evaporate to dryness, residue adds ethyl acetate 20ml sonicated 30 minutes, filters, the filtrating evaporate to dryness, residue adds methyl alcohol 5ml, as need testing solution.Other gets the Catalpol reference substance, adds methyl alcohol and processes the solution that every 1ml contains 0.5mg, as reference substance solution.According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate; With chloroform-methanol-water (14: 6: 1) is developping agent; Launch, take out, dry; Spray is with the anisaldehyde test solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Inspection: should meet each item relevant under pill item regulation (" appendix IA of Chinese pharmacopoeia version in 2005).
Assay: the photograph high performance liquid chromatography (" appendix VID of Chinese pharmacopoeia version in 2005) measure.
Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filling agent; Methanol-water-phosphoric acid (47: 53: 0.2) is a moving phase; The detection wavelength is 280nm.Number of theoretical plate calculates by the scutelloside peak should be not less than 2000.
It is an amount of that the scutelloside reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.1mg, promptly gets.
The preparation sample thief 2g of need testing solution, porphyrize is got powder 0.2g, and accurate the title, decide, and puts in the tool plug conical flask; Precision is measured 70% ethanol 100ml, claims decide weight, and reflux 30 minutes is put coldly, and weight decided in title; Supply the weight that subtracts mistake with 70% ethanol, shake up, filter, get subsequent filtrate, promptly get.
Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and promptly get.
The every gram of these article contains scutelloside must not be less than 24mg.
Function with cure mainly: enriching yin, heat-clearing, cool blood.Be used for pregnant woman's part of the body cavity above the diaphragm housing the heart and lungs fire and contain, have a dizzy spell, aphthae, tinnitus toothache, pregnant woman's blood-head restlessness during pregnancy.
Usage and consumption: oral, a 6g, 1～2 time on the one.
Attention: the strongly fragrant restlessness during pregnancy person of phlegm moisture avoids clothes.Specification: every bottled 6g storage: airtight, protection against the tide.
There is the root of large-flowered skullcap to differentiate in the clearing heat and cooling blood ball proper mass standard, no assay item, this time the raising standard has increased the thin layer discriminating of glutinous rehmannia and the assay of the root of large-flowered skullcap; Simultaneously, to root of large-flowered skullcap discrimination method in the primary standard, revise again.
1. the discriminating of the root of large-flowered skullcap
1.1 material and reagent
Methyl alcohol, ethyl acetate, butanone, formic acid: analyze pure
Scutelloside reference substance: Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110715-200514
Clearing heat and cooling blood ball sample: Lerentang Pharmaceutical Factory, Zhongxin Pharmaceutical Group Co., Ltd., Tianj produces;
Lot number: D081001
Silica G plate: Qingdao Haiyang chemical industry subsidiary factory
1.2 the preparation of need testing solution
These article of getting 1.0g adds methyl alcohol 20ml, and reflux 2 hours is put coldly, filters, and filtrating is as need testing solution.
1.3 the preparation of reference substance solution
Get the scutelloside reference substance, add methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution.
1.4 the preparation of blank article solution
By prescription, technology, except that the root of large-flowered skullcap, prepare the blank article, again the blank article are ground into fine powder, prepare blank article solution according to the need testing solution preparation method.
1.5 experimental technique
Method 1: with reference to the root of large-flowered skullcap thin layer discrimination method of including in the former ministerial standard of clearing heat and cooling blood ball.
Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005); Draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-butanone-formic acid-water (10: 7: 2: 2) be developping agent; Launch; Take out, dry, put under the ultraviolet lamp (254nm) and inspect.
The result: in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, no fluorescence spot has the bronzing spot.The result sees Fig. 1.Among Fig. 11: test sample 2: reference substance 3: blank article
Root of large-flowered skullcap thin layer discrimination method to including in the former ministerial standard of clearing heat and cooling blood ball is made amendment, and changes
Coloration method:
Method 2: according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With ethyl acetate-butanone-formic acid-water (10: 7: 2: 2) be developping agent; Launch, take out, dry; Spray is with 1% ferric trichloride ethanolic solution, and it is clear to be heated to the spot colour developing at 105 ℃.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color, blank article chromatogram is noiseless.The result sees Fig. 2.Among Fig. 21: test sample 2: reference substance 3: blank article
1.6 confirm root of large-flowered skullcap thin layer discrimination method be:
These article of getting 1g grinds, and adds methyl alcohol 20ml, and reflux 2 hours is put coldly, filters, and filtrating is as need testing solution.Other gets the scutelloside reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With ethyl acetate-butanone-formic acid-water (10: 7: 2: 2) be developping agent; Launch, take out, dry; Spray is with 1% ferric trichloride ethanolic solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
2. the discriminating of glutinous rehmannia
2.1 material and reagent
Methyl alcohol, ethyl acetate, chloroform: analyze pure
Catalpol reference substance: Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110808-200407
Clearing heat and cooling blood ball sample: Lerentang Pharmaceutical Factory, Zhongxin Pharmaceutical Group Co., Ltd., Tianj produces;
Lot number: D081001
High-efficient silica gel G plate:
2.2 the preparation of need testing solution
These article of getting powder 2g adds methyl alcohol 20ml, and reflux 1 hour is put coldly, filter, and the filtrating evaporate to dryness, residue adds ethyl acetate 20ml sonicated 30 minutes, filters, the filtrating evaporate to dryness, residue adds methyl alcohol 5ml, as need testing solution.
2.3 the preparation of reference substance solution
Get the Catalpol reference substance, add methyl alcohol and process the solution that every 1ml contains 0.5mg, as reference substance solution.
2.4 the preparation of blank article solution
By prescription, technology, except that glutinous rehmannia, prepare the blank article, again the blank article are ground into fine powder, prepare blank article solution according to the need testing solution preparation method.
2.5 experimental technique
According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate; With chloroform-methanol-water (14: 6: 1) is developping agent; Launch, take out, dry; Spray is with the anisaldehyde test solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.The blank article are noiseless.The result sees Fig. 3.Among Fig. 31: test sample 2: reference substance 3: blank article.
2.6 confirm the glutinous rehmannia discrimination method be: get these article powder 2g, add methyl alcohol 20ml, reflux 1 hour is put coldly, filter, and the filtrating evaporate to dryness, residue added the ethyl acetate sonicated 30 minutes, filtered, the evaporate to dryness of filtrating, residue adds methyl alcohol 5ml, as need testing solution.Other gets the Catalpol reference substance, adds methyl alcohol and processes the solution that every 1ml contains 0.5mg, as reference substance solution.According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate; With chloroform-methanol-water (14: 6: 1) is developping agent; Launch, take out, dry; Spray is with the anisaldehyde test solution, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Two. content of baicalin is measured
1. instrument and material
Instrument: Waters2695 high performance liquid chromatograph; Detecting device: Waters2487; Sartorius CP224S. electronic balance.
Material: chromatographic column: Diamonsil C18,5 μ, 250 * 4.6mm; Scutelloside reference substance: lot number: 110715-200514 is produced by Chinese pharmaceutical biological product check; Methyl alcohol: chromatographically pure, Tianjin Concord Technology Co., Ltd. produces; Glacial acetic acid, phosphoric acid: analyze pure;
Clearing heat and cooling blood ball sample: Lerentang Pharmaceutical Factory, Zhongxin Pharmaceutical Group Co., Ltd., Tianj produces;
Lot number: D081001
2. experimental technique
2.1 the preparation of reference substance solution: it is an amount of to get the scutelloside reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.1mg, promptly gets.See Fig. 4
2.2 the selection of moving phase: with reference to " moving phase in radix scutellariae medicinal materials content assaying method of Chinese pharmacopoeia version in 2005: methanol-water-phosphoric acid (50: 50: 0.2).
Chromatographic condition: 30 ℃ of column temperatures, detect wavelength: 280nm, flow velocity: 0.8ml/min
2.3 the selection of method for distilling
The result: employing method 2 scutellosides extract fully, confirm that employing method 2 is need testing solution extraction solvent.
2.4 the selection of extraction time
With these article porphyrize, it is fixed to get the accurate title of fine powder 0.2g, puts in the tool plug Erlenmeyer flask, and precision is measured 70% ethanol 100ml, weigh, and the heating and refluxing extraction different time, it is heavy to put cold-patch, filters, and injects high performance liquid chromatograph and measures.Result such as table 2
The result: employing method 1 scutelloside extracts fully, confirms that employing method 1 is need testing solution extraction time.
The selection of table 1 method for distilling
The selection of table 2 extraction time
2.5 confirm the need testing solution preparation method
Sample thief 2g, porphyrize is got powder 0.2g, accurate claims surely, puts in the tool plug conical flask, and precision is measured 70% ethanol 100ml, claims decide weight, and reflux 30 minutes is put coldly, claims to decide weight, supplies the weight that subtracts mistake with 70% ethanol, shakes up, and subsequent filtrate is got in filtration, promptly gets.See Fig. 5
3. blank article formulations prepared from solutions
Except that the root of large-flowered skullcap, prepare the blank article, prepare blank article solution by the need testing solution preparation method by formulation and technology.See Fig. 6
4. methodology checking
4.1 linear the investigation
Get scutelloside reference substance solution (concentration 340.8 μ g/ml) and be diluted to the reference substance solution that concentration is 34.08 μ g/ml, 68.16 μ g/ml, 136.32 μ g/ml, 204.48 μ g/ml, 272.64 μ g/ml, 340.8 μ g/ml respectively; Draw 5 μ l respectively and inject high performance liquid chromatograph; The record chromatogram is done linear regression with peak area to content.Result such as table 3.
Table 3 is linear to be investigated
Getting regression equation by data is: y=A+Bx=19255.25x-55946.54, r=0.9999 result show that scutelloside is good in 0.1704～1.704 μ g scope internal linear relation.
4.2 precision is got same need testing solution, continuous sample introduction 6 times is measured relative standard deviation RSD=0.3%, shows that precision is good.Result such as table 4
4.3 reappearance
The fixed 6 parts of same lot sample article powder of accurate title, according to preparation method's preparation of need testing solution, the RSD that measures content of baicalin is 0.7%, shows that reappearance is good.Result such as table 5.
The experiment of table 4 precision
The experiment of table 5 reappearance
4.4 stability experiment
Get same need testing solution, whenever make an experiment for 1 time at a distance from 2 hours sample introductions, continuous sample introduction 6 times, RSD=0.2% shows that these article are basicly stable in 10h.Result such as table 6.
Table 6 stability experiment
4.5 recovery experiment
Get with a collection of known content (content: 6 parts in sample 58.7mg/g), every part of 0.1g accurate claims surely, adds the scutelloside reference substance respectively, according to preparation method's preparation of need testing solution, calculating average recovery rate is 95.78%, RSD=0.7% again.The show sample recovery is good as a result.Result such as table 7.
The experiment of table 7 recovery
5.6 sample size is measured
Get five lot sample article, prepare according to the need testing solution preparation method respectively, measure content of baicalin, the result sees table 8.
Table 8 five lot sample article assay results
30.2×80％＝24mg/g
According to above five batches testing result, confirm that content of baicalin must not be lower than 24mg/g in the clearing heat and cooling blood ball.
Claims (1)
1. the detection method of a Chinese medicine preparation clearing heat and cooling blood ball, wherein said pharmaceutical formulation are by Chinese crude drug: root of large-flowered skullcap 500g glutinous rehmannia 500g forms, more than two flavors, be ground into fine powder; Sieve mixing, water pill; Drying promptly gets, and it is characterized in that: the step of its method is:
(1) be reference substance with scutelloside, Catalpol, thin-layered chromatography differentiates in the clearing heat and cooling blood ball prescription whether contain the root of large-flowered skullcap, glutinous rehmannia composition;
Described thin-layered chromatography differentiates that the method that does not contain the root of large-flowered skullcap in the clearing heat and cooling blood ball prescription is: get these article 1g, grind, add methyl alcohol 20ml, reflux 2 hours is put coldly, filters, and filtrates as need testing solution; Other gets the scutelloside reference substance, adds methyl alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with ethyl acetate-butanone-formic acid-water, ethyl acetate; Butanone: formic acid: water=10: 7: 2: 2, launch, take out, to dry, spray is with 1% ferric trichloride ethanolic solution, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
Described thin-layered chromatography differentiates that the method that whether contains the glutinous rehmannia composition in the clearing heat and cooling blood ball prescription is: get these article powder 2g, add methyl alcohol 20ml, reflux 1 hour; Put coldly, filter the filtrating evaporate to dryness; Residue adds ethyl acetate 20ml sonicated 30 minutes, filters the filtrating evaporate to dryness; Residue adds methyl alcohol 5ml, as need testing solution; Other gets the Catalpol reference substance, adds methyl alcohol and processes the solution that every 1ml contains 0.5mg, as reference substance solution; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate, be developping agent with chloroform-methanol-water; Chloroform: methyl alcohol: water=14: 6: 1, launch, take out; Dry, spray is with the anisaldehyde test solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
(2) with the scutelloside be reference substance, the method that high performance liquid chromatography detects content of baicalin in the clearing heat and cooling blood ball prescription is:
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filling agent; Methyl alcohol: water: phosphoric acid=47: 53: 0.2 is moving phase; The detection wavelength is 280nm; Number of theoretical plate calculates by the scutelloside peak should be not less than 2000;
The preparation of reference substance solution: it is an amount of to get the scutelloside reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.1mg, promptly gets;
The preparation of need testing solution: sample thief 2g, porphyrize is got powder 0.2g, and accurate the title, decide, and puts in the tool plug conical flask; Precision is measured 70% ethanol 100ml, claims decide weight, and reflux 30 minutes is put coldly, and weight decided in title; Supply the weight that subtracts mistake with 70% ethanol, shake up, filter, get subsequent filtrate, promptly get;
Determination method: inaccurate reference substance solution and each 5 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly get.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910228359A CN102068549B (en) | 2009-11-19 | 2009-11-19 | Detection method for Chinese medicinal preparation heat clearing and blood cooling pills |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910228359A CN102068549B (en) | 2009-11-19 | 2009-11-19 | Detection method for Chinese medicinal preparation heat clearing and blood cooling pills |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102068549A CN102068549A (en) | 2011-05-25 |
| CN102068549B true CN102068549B (en) | 2012-09-05 |
Family
ID=44027428
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910228359A Active CN102068549B (en) | 2009-11-19 | 2009-11-19 | Detection method for Chinese medicinal preparation heat clearing and blood cooling pills |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102068549B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526356B (en) * | 2012-03-14 | 2013-11-27 | 青岛汉河动植物药业有限公司 | Traditional Chinese medicine compound for clearing Ying heat and cooling blood of poultry |
| CN103070971A (en) * | 2013-02-28 | 2013-05-01 | 吕忠绪 | Traditional Chinese medicament composition for treating hemodia, odontoseisis and toothache |
| CN110646521A (en) * | 2018-06-26 | 2020-01-03 | 北京紫云腾中药饮片有限公司 | Detection method of prepared rehmannia root decoction pieces |
| CN115166128A (en) * | 2022-06-22 | 2022-10-11 | 四川逢春制药有限公司 | Method for identifying Scutellaria Yunnanensis and Scutellaria baicalensis |
| CN115420816A (en) * | 2022-07-21 | 2022-12-02 | 广东万年青制药股份有限公司 | Content determination method of traditional Chinese medicine composition for nourishing yin and cooling blood |
| CN115389366B (en) * | 2022-08-24 | 2024-07-19 | 临沂市检验检测中心 | Detection method of traditional Chinese medicine preparation heat-clearing and blood-cooling pill |
-
2009
- 2009-11-19 CN CN200910228359A patent/CN102068549B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| 药典委员会.清热凉血丸.《药品标准中药成方制剂第9册》.1994,z9－192. * |
| 金佩芬,张关顺,崔波.高效液相色谱法测定清热凉血丸中黄芩苷的含量.《基层中药杂志》.2001,第15卷(第1期),20-21. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102068549A (en) | 2011-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102068549B (en) | Detection method for Chinese medicinal preparation heat clearing and blood cooling pills | |
| CN102100818B (en) | Quality control method for lophanthus antifebrile tablets | |
| CN102269751B (en) | Detection method of Liuweinengxiao preparation | |
| CN102749401B (en) | Inspection method of traditional Chinese medicine composition twenty-five-ingredient lung disease preparation | |
| CN102854281A (en) | Detection method of sugar-free strong loquat syrup | |
| CN102068600B (en) | Quality control method of venation relaxing and heart-nourishing pill as Chinese herbal preparation | |
| CN101028460B (en) | Method for inspecting throat-clearing Chinese medicinal pills | |
| CN102552496A (en) | Quality detection method of compound stomachache treating capsules | |
| CN102068573B (en) | Quality control method for stomach reinforcing and digestion promoting tangerine peel, pinellia, immature bitter orange and atractylodes macrocephala pill | |
| CN101982189A (en) | Method for detecting salvia heart-soothing capsules | |
| CN102038908B (en) | Detection method of tambac depression-alleviating pill as traditional Chinese medical preparation | |
| CN104407092A (en) | Quality detection method of traditional Chinese medicinal composition with efficacy of appetizing and invigorating spleen | |
| CN101744946A (en) | Chinese medicinal composition and detection method for Chinese medicinal composition preparation | |
| CN102068598B (en) | Quality control method of Yangrong Baicao Wan for treating irregular menses caused by hemophthisis | |
| CN102068657B (en) | Quality control method of venation relaxing pill as Chinese herbal preparation | |
| CN102068673B (en) | Quality control method of stomach-strengthening and chest-relieving pill as Chinese herbal preparation | |
| CN102068649B (en) | Quality control method for gastric condition-regulating pill as traditional Chinese preparation | |
| CN102507840A (en) | Sanwei sandalwood granule and quality control method of preparation thereof | |
| CN102707006B (en) | Quality detection method of cudrania tricuspidata formula granules | |
| CN105004833A (en) | Detection method for traditional Chinese medicine preparation for treating acute gouty arthritis and gout | |
| CN102091297A (en) | Quality control method for liver health care medicine | |
| CN102068656A (en) | Quality control method for traditional Chinese medicinal preparation epilepsy pills | |
| CN101239117A (en) | Quality control method of traditional Chinese medicine formulations for treating gynaecologic disease | |
| CN101385808B (en) | Detection method of Zhubai tranquilizing pill | |
| CN100570358C (en) | Treat the detection method of asthenopic pharmaceutical preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 300380 Yumen Road, Xiqing District, Tianjin Patentee after: Lerentang Pharmaceutical Factory of Jinyao Darentang Group Co.,Ltd. Address before: 300380 Yumen Road, Xiqing District, Tianjin Patentee before: Letrentang pharmaceutical factory of Tianjin Zhongxin Pharmaceutical Group Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder |