CN101554410B - Detection method of fresh fleece flower root and prepared fleece flower root - Google Patents
Detection method of fresh fleece flower root and prepared fleece flower root Download PDFInfo
- Publication number
- CN101554410B CN101554410B CN2009100590243A CN200910059024A CN101554410B CN 101554410 B CN101554410 B CN 101554410B CN 2009100590243 A CN2009100590243 A CN 2009100590243A CN 200910059024 A CN200910059024 A CN 200910059024A CN 101554410 B CN101554410 B CN 101554410B
- Authority
- CN
- China
- Prior art keywords
- polygoni multiflori
- radix polygoni
- measured
- medical material
- solution
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a quality control method of fresh fleece flower root and prepared fleece flower root which comprises the steps as follows: A. standard crude drug powder of fresh fleece flowerroot or prepared fleece flower root is weighed and dissolved in acetone; the solution undergoes ultrasonic treatment and is then filtered, brought to constant volume and evaporated to dry; obtained dry powder is dissolved in methyl alcohol and the methyl alcohol solution is filtered by Millipore, thereby obtaining standard crude drug solution; fingerprint spectrum of the standard crude drug is then worked out by high performance liquid chromatography; the same method is then employed to establish fingerprint spectrum awaiting measurement of fresh fleece flower root or prepared fleece flower root to be measured; similarity between the fingerprint spectrum awaiting measurement and the standard fingerprint spectrum are then used for deciding whether the crude drug to be measured is qualified. B. contents of two index components, including 2, 3, 4', 5-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and rheum emodin, are measured by high performance liquid chromatography. The quality control method has various judging evidences, thereby not only realizing qualitative detection of major chemical compositions of fresh and prepared fleece flower root, but also measuring contents of the two major components. The quality control method has reliable and precise measurement result, thereby reducing possibility of quality pass rate affected by manual treatment.
Description
Technical field
A kind of Chinese medicine quality control method of the present invention relates in particular to the quality discrimination and the detection method of a kind of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata;
Background technology
The Radix Polygoni Multiflori beginning is stated from " Japan hanako materia medica "; Dried root for polygonum multiflorum thunb (Polygonum multiflorum Thunb); Main product in Henan, ground such as Shandong, Hubei, Sichuan, Jiangsu, Guangxi, be mostly wild, cultivation is also arranged; Be the famous ripe different Chinese medicine of controlling of life, the branch of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata is arranged; Its property of Radix Polygoni Multiflori is flat, sweet in the mouth, hardship; GUIXIN, liver, large intestine channel; Its slightly warm in nature of Radix Polygoni Multiflori Preparata, sweet in the mouth, puckery are returned liver, kidney channel; The detoxifcation of Radix Polygoni Multiflori preventing the attack (or recurrence) of malaria, loosening bowel to relieve constipation; The Radix Polygoni Multiflori Preparata merit is apt to enrich and benefit essence and blood, and the crow of reinforcing the kidney must; Li Shizhen (1518-1593 A.D.) is put down in writing in Compendium of Material Medica: " all famous mountains, remote mountains product person, promptly big and good also "; Give birth at present, Radix Polygoni Multiflori Preparata medical material market demand grows with each passing day, shoddy phenomenon happens occasionally, this has had a strong impact on life, Radix Polygoni Multiflori Preparata at home and abroad reputation;
The main quality control method of existing Chinese medicine Radix Polygoni Multiflori is the Pharmacopoeia of the People's Republic of China prescribed shape inspection of adopting version in 2005, microscopical identification; Physicochemical identification and a kind of index components (2; 3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside) Determination on content.Appearance character in this method of quality control is differentiated (comprising microscopical identification), and subjective factors is strong, and testing result is not objective and accurate, and error is big.And the drug effect of Radix Polygoni Multiflori has more than relevant with single component, but confirms that by the various active composition there is limitation in single component content assay method, can not reflect the drug effect of Radix Polygoni Multiflori exactly.Therefore, the method for quality control of existing Chinese medicine Radix Polygoni Multiflori can not satisfy the needs of producing with utilization.
Summary of the invention
The object of the invention just provides the detection method of a kind of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata; This method judgment basis is various; Can realize qualitative detection, measure the content of two kinds of main components again, realize conduct monitoring at all levels quality of medicinal material to life, Radix Polygoni Multiflori Preparata main chemical compositions; Its testing result is reliable, accurate, and having reduced is the artificial probability of handling of requisite quality.
The present invention realizes its goal of the invention, and the technical scheme that is adopted is: the detection method of a kind of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata is made up of following steps:
A, finger printing are relatively
A1, set up Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata standard finger-print
Get the powder 0.1-0.3 gram of the standard medical material of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata, be dissolved in the acetone of 8ml, supersound process 0.5-1 hour after-filtration got filtrating; Triplicate, merging filtrate adds acetone, is settled to 25ml; The acetone soln of standard medical material, precision is measured the 5ml acetone soln, treat that acetone volatilizes after, dry powder; Get dry powder and be dissolved in methanol and get methanol solution, it is behind 0.45 micron the filtering with microporous membrane that methanol solution uses the aperture, the standard medical material solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata;
Draw the standard medical material solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata, inject high performance liquid chromatograph, set up the standard finger-print of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata standard medical material;
The finger printing of A2, making Radix Polygoni Multiflori to be measured or Radix Polygoni Multiflori Preparata
The powder of standard Radix Polygoni Multiflori in A1 step or Radix Polygoni Multiflori Preparata medical material is replaced with the powder of Radix Polygoni Multiflori to be measured or Radix Polygoni Multiflori Preparata medical material, adopt A1 to go on foot identical operation then, make the finger printing of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata medical material to be measured;
A3, collection of illustrative plates are relatively
Calculate the similarity of the finger printing and the standard finger-print in A1 step in A2 step; If the medical material to be measured of correspondence is carried out the operation of following steps in finger printing and standard finger-print similarity >=0.900; Otherwise be defective, end operation;
B, 2,3,4 ', the assay of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside index components
The preparation of B1, reference substance solution
Precision takes by weighing 2,3,4 ' of 3.2-3.4mg, and 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance places the 10ml volumetric flask, adds 50% dissolve with ethanol and is diluted to scale, shakes up, and joins to such an extent that concentration is the reference substance solution of 0.32-0.34g/L;
The preparation of B2, product to be tested solution
Take by weighing Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata powder 0.2g, place the 100ml conical flask, add 50% ethanol 25ml; Claim to decide quality, reflux 30min is put cold; Claim again to decide quality, supply the quality that subtracts mistake, shake up with 50% ethanol; Supernatant filters with 0.45 μ m microporous filter membrane, promptly gets the product to be tested solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata;
B3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume, adopts HPLC to measure in the product to be tested solution 2,3; 4 '; The content of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside, and converse 2,3 in Radix Polygoni Multiflori to be measured or the Radix Polygoni Multiflori Preparata; 4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside composition accounts for the percentage composition of whole medical material dry to be measured;
The assay of C, emodin index components
The preparation of C1, reference substance solution
Precision takes by weighing emodin 2.8-3.2mg, puts in the 100ml volumetric flask, adds dissolve with methanol and is diluted to 100ml, shakes up;
The preparation of C2, product to be tested solution
Precision takes by weighing Radix Polygoni Multiflori or each 0.15g of Radix Polygoni Multiflori Preparata powder, puts in the 100ml conical flask, and precision adds methanol 25ml, claim to decide quality, and reflux 1h, cooling is claimed to decide quality again, supplies the quality that subtracts mistake with methanol, shakes up, and filters; Precision is measured filtrating 5ml, puts in the 100ml conical flask, flings to methanol, adds 8% hydrochloric acid 10ml, and supersound process 2min adds chloroform 10ml again, reflux 1h, cooling; In the backflow dislocation separatory funnel,, incorporate in the separatory funnel, obtain the chloroform solution of lower floor with minimum of chloroform washing 100ml conical flask; The hydrochloric acid solution reuse chloroform extraction on upper strata and obtaining 3 times, each chloroform consumption 8-12ml, combined chloroform liquid moves in the 100ml conical flask, flings to chloroform to doing; Residue adds dissolve with methanol, and standardize solution in the 10ml volumetric flask filters with 0.45 μ m microporous filter membrane, promptly gets the product to be tested solution of Radix Polygoni Multiflori to be measured or Radix Polygoni Multiflori Preparata powder;
C3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume; Adopt HPLC to measure emodin content in the product to be tested solution, and converse the percentage composition that emodin composition in Radix Polygoni Multiflori to be measured or the Radix Polygoni Multiflori Preparata accounts for whole medical material dry to be measured;
D, judgement index become content
If in the Radix Polygoni Multiflori medical material to be measured 2,3,4 ', 5-tetrahydroxystilbene-percentage composition >=3.00% of 2-O-β-D-pyranglucoside and percentage composition >=0.050% of emodin judge that then Radix Polygoni Multiflori medical material to be measured is qualified, otherwise defective:
If in the Radix Polygoni Multiflori Preparata medical material to be measured 2,3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside content >=1.93% and emodin content >=0.067% judges that then Radix Polygoni Multiflori Preparata medical material to be measured is qualified, otherwise defective.
Compared with prior art, the invention has the beneficial effects as follows:
One, finger printing is relatively: adopt the room temperature ultrasonic extraction, extract the effective ingredient of Radix Polygoni Multiflori, avoided the decomposition of main effective ingredient-stilbene glucoside (SG) compounds in hot solvent of Radix Polygoni Multiflori, and to adopt acetone be solvent that its extraction effect is best:
1, chemical compound such as the main active component stilbene glycoside (SG) of Radix Polygoni Multiflori, anthraquinone class all can be extracted effectively;
2, the macromole impurity of pollution such as protein, polysaccharide chromatographic column does not get into product to be tested solution, thereby chromatographic column is not polluted, and prolongs the service life of chromatographic column.The optical purity that respectively becomes swarming of finger printing is all up to more than 98%.Like 2,3,4 ' of Radix Polygoni Multiflori, the purity of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside (THSG) main peak is up to 99%; Then be merely about 70% during with the methanol extraction of methanol and 50%.
3, make the main curative effect composition of Radix Polygoni Multiflori that the characteristic of correspondence peak all arranged in finger printing, non-characteristic components such as the protein of Radix Polygoni Multiflori, polysaccharide then do not occur in the characteristic peak of finger printing.6 characteristic peaks that peak value is high are arranged in standard finger-print, be respectively through identifying: stilbene glucoside, emodin pyranglucoside, chrysophanol pyranglucoside, physcione pyranglucoside, emodin, physcione.
Therefore the present invention has realized the qualitative detection to life, six main chemical compositions of Radix Polygoni Multiflori Preparata, and makes its six main components, becomes known compound; Its testing result is more reliable, accurate.
Two, the assay of index components:, can not embody the chemical property and concrete content of the index components of finger printing owing to the degree of finger printing similarity difference and the disproportionate relation of compounds content difference in the medical material.And the result that the variation relation experiment of the hepatotoxicity gene expression profile of Kunming mouse, drug effect change and chemical constituent is obtained shows; 2; 3; 4 ', these two kinds of materials of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside and emodin are to confirm life, Radix Polygoni Multiflori Preparata drug effect and toxic basic reason.
Therefore, the present invention carries out medical material on the basis of qualitative detection carrying out finger printing, also to two kinds of topmost index components 2; 3; 4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside and emodin content are measured, and guarantee that its content meets the requirements.Thereby method of quality control of the present invention; Both pay attention to the similarity of finger printing on overall profile, and can embody the concrete content of index components again, overcome the limitation of finger printing; Control approach and means have been increased to the Radix Polygoni Multiflori Chinese medicine quality; Reduced to the artificial probability of handling of requisite quality, made the Chinese medicine information representation ability of method of quality control of the present invention strong, effective more, stable and controlled to the quality control of Radix Polygoni Multiflori.
Above-mentioned Radix Polygoni Multiflori standard medical material produces Radix Polygoni Multiflori for the river of directly picking up from the Mount Emei, Sichuan; Radix Polygoni Multiflori Preparata standard medical material is that the river produces the steamed method of Radix Polygoni Multiflori employing or the process of preparing Chinese medicine of Semen sojae atricolor method for making gets.
Can make the peak value of the standard finger-print that makes high like this, have good representativeness; And the content of the index components that tests out is high; Thereby set up high-quality Radix Polygoni Multiflori finger printing, confirm higher index components content, thereby improve the quality of Chinese medicine Radix Polygoni Multiflori.
When above-mentioned A1 and A2 tested standard medical material solution and medical material solution to be measured with high performance liquid chromatograph in the step, all select the detection wavelength of 290nm wavelength for use as the finger printing of high performance liquid chromatograph.
The 290nm wavelength can be given full play to the function of high performance liquid chromatograph as the detection wavelength of the finger printing of high performance liquid chromatograph.Less chromatographic peak has strong absorption at the 290nm place, can demonstrate fully the chemical compound of the leading indicator in life, the Radix Polygoni Multiflori Preparata.Be that stilbene glycoside, anthraquinone analog compound chemical compound are maximum absorption wavelength with 290nm all, thereby under the 290nm wavelength, each chromatographic peak separates good, main component has reached baseline separation; Response is high, and the finger printing of formation is effective.
Through comparing chromatogram and the UV scanning maximum absorption wavelength of each quantitative composition in the 210nm-500nm scope, find that 320nm is the maximum absorption wavelength of stilbene glycosides compound, 290nm is the maximum absorption wavelength of anthraquinone analog compound.Because two ethylene glycoside content are higher, and, therefore select 290nm can embody main component living, Radix Polygoni Multiflori Preparata at 320nm and 290nm absorption no significant difference, and the main component baseline separation.On the certificate, seemingly should select 290nm.Be further refinement The above results; We have investigated 200nm, 254nm, 268nm, 296nm, 320nm, six separation spectrograms that detect under the wavelength of 290nm respectively by aforementioned chromatographic condition; Find: under the 290nm wavelength, each chromatographic peak separates good, and main component has reached baseline separation; Response is also higher under this wavelength.According to above experimental result, finally selected 290nm is as the detection wavelength of finger printing.
Below in conjunction with accompanying drawing and concrete embodiment, the present invention is done further detailed explanation;
Description of drawings
Fig. 1 is the standard finger-print of the Radix Polygoni Multiflori of the embodiment of the invention one.
Fig. 2 is the fingerprint image of the Radix Polygoni Multiflori to be measured of the embodiment of the invention one.
Fig. 3 is the standard finger-print of the Radix Polygoni Multiflori Preparata of the embodiment of the invention two.
Fig. 4 is the finger printing of the Radix Polygoni Multiflori Preparata to be measured of the embodiment of the invention two.
Among each figure, transverse axis is the time, and unit is minute (minutes); The longitudinal axis is a trap, and unit is: milli absorbance units (mAU).The numeral on characteristic peak next door is the numbering of this characteristic peak among Fig. 1,3.
The specific embodiment
Embodiment one
A kind of specific embodiment of the present invention is: the method for quality control of a kind of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata is made up of following steps:
A, finger printing are relatively
A1, set up the standard finger-print of Radix Polygoni Multiflori
Radix Polygoni Multiflori standard medical material in this example produces Radix Polygoni Multiflori for the river of directly picking up from the Mount Emei, Sichuan.
Get powder 0.2 gram of the standard medical material of Radix Polygoni Multiflori, be dissolved in the acetone of 8ml, 0.5 hour after-filtration of supersound process is got filtrating; Triplicate (containing this), merging filtrate adds acetone, is settled to 25ml; The acetone soln of standard medical material, precision is measured the 5ml acetone soln, treat that acetone volatilizes after, dry powder; Get dry powder and be dissolved in 2ml methanol and get methanol solution, it is behind 0.45 micron the filtering with microporous membrane that methanol solution uses the aperture, the standard medical material solution of Radix Polygoni Multiflori; Draw the standard medical material solution of Radix Polygoni Multiflori, inject high performance liquid chromatograph, set up the standard finger-print of 60 minutes Radix Polygoni Multiflori standard medical material.Fig. 1 is the standard finger-print of the Radix Polygoni Multiflori that this routine method makes.
The finger printing of A2, making Radix Polygoni Multiflori to be measured
The Radix Polygoni Multiflori to be measured of this example is purchased in the Deqing, Guangdong.
The powder of the standard Radix Polygoni Multiflori medical material in A1 step is replaced with the powder of Radix Polygoni Multiflori medical material to be measured, adopt A1 to go on foot identical operation then, make the finger printing of Radix Polygoni Multiflori medical material to be measured; Fig. 3 is purchasing in the finger printing of the medical material to be measured of Deqing, Guangdong of recording of this example.
A3, collection of illustrative plates are relatively
Calculate the similarity of the finger printing to be measured and the standard finger-print in A1 step in A2 step; If the medical material to be measured of correspondence is carried out the operation of following steps in finger printing to be measured and standard finger-print similarity >=0.900; Otherwise be defective, end operation.When calculating similarity in this example, directly adopt China national Bureau of Drugs Supervision specified " the Chinese medicine chromatogram is analyzed and data handling system " software to calculate.
In this example, with the similarity of standard finger-print shown in Figure 1 and finger printing to be measured shown in Figure 2 through being calculated as 0.988, greater than setting 0.900.Judge that this medical material to be measured is preliminary qualified products, carries out the test of following steps again.
In this routine finger printing detected, chromatographic column did, and hypersilC18 (5 μ m, 4.6mmx250mm); Detect wavelength, 290nm; Flow rate of mobile phase, 1mL/min, mobile phase is made up of acetonitrile and water; Sample size is 10 μ L; Gradient elution; The composition of mobile phase and gradient setting such as following table:
In the standard finger-print of the Radix Polygoni Multiflori that this routine method of the usefulness of Fig. 1 is made 6 characteristic peaks that peak value is high are arranged.Through identifying: No. 4 peaks are that stilbene glucoside, No. 5 peaks are that emodin pyranglucoside, No. 6 peaks are that chrysophanol pyranglucoside, No. 7 peaks are that physcione pyranglucoside, No. 10 peaks are that emodin, No. 11 peaks are physcione.In these peaks, No. 4 peak stilbene glucosides and No. 10 peak emodins are to produce active main component, also are to produce the property of medicine and toxic main component, and 6 main component characteristic peaks that characterize the Radix Polygoni Multiflori medical material become known compound.
B, 2,3,4 ', the assay of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside index components
The preparation of B1, reference substance solution
Precision takes by weighing 2,3,4 ' of 3.3mg, and 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance places the 10ml volumetric flask, adds 50% dissolve with ethanol and is diluted to scale, shakes up, and joins to such an extent that concentration is the reference substance solution of 0.33g/L;
The preparation of B2, product to be tested solution
Take by weighing the Radix Polygoni Multiflori powder 0.2g of Deqing, Guangdong, place the 100ml conical flask, add 50% ethanol 25ml; Claim to decide quality, reflux 30min is put cold; Claim again to decide quality, supply the quality that subtracts mistake, shake up with 50% ethanol; Supernatant filters with microporous filter membrane (0.45 μ m), promptly gets the product to be tested solution of Radix Polygoni Multiflori;
B3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume, adopts HPLC to measure in the product to be tested solution 2,3; 4 '; The content of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside, and converse 2,3 in the Radix Polygoni Multiflori to be measured; 4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside composition accounts for the percentage composition of whole medical material dry to be measured; Concrete conversion relation is: in the 0.2g Radix Polygoni Multiflori powder 2; 3; 4 ', the amount of taking by weighing * 2.5 of THSG among the content (b) of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside (THSG)=(the liquid chromatograph peak heights of the liquid chromatograph peak heights/reference substance solution of product to be tested solution) * step B; The weight percentage of THSG is in the medical material to be measured: b * 1000/0.2 * 100%.
The assay of C, emodin index components
The preparation of C1, reference substance solution
Precision takes by weighing emodin 3.0mg, puts in the 100ml volumetric flask, adds dissolve with methanol and is diluted to 100ml, shakes up;
The preparation of C2, product to be tested solution
Precision takes by weighing the Radix Polygoni Multiflori powder 0.15g to be measured that originates in the Deqing, Guangdong, puts in the 100ml conical flask, and precision adds methanol 25ml, claim to decide quality, and reflux 1h, cooling is claimed to decide quality again, supplies the quality that subtracts mistake with methanol, shakes up, and filters; Precision is measured filtrating 5ml, puts in the 100ml conical flask, flings to methanol, adds 8% hydrochloric acid 10ml, and supersound process 2min adds chloroform 10ml again, reflux 1h, cooling; In the backflow dislocation separatory funnel,, incorporate in the separatory funnel, obtain the chloroform solution of lower floor with minimum of chloroform washing 100ml conical flask; The hydrochloric acid solution reuse chloroform extraction on upper strata and obtaining 3 times, each chloroform consumption 8-12ml, combined chloroform liquid moves in the 100ml conical flask, flings to chloroform to doing; Residue adds dissolve with methanol, and standardize solution in the 10ml volumetric flask filters with 0.45 μ m microporous filter membrane, promptly gets the need testing solution of Radix Polygoni Multiflori powder;
C3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume, adopts HPLC to measure emodin content in the product to be tested solution, and converses the percentage composition that emodin composition in the Radix Polygoni Multiflori to be measured accounts for whole medical material dry to be measured; Concrete conversion relation is: the amount of taking by weighing * 10 of emodin among emodin content (c) in the 0.15g Radix Polygoni Multiflori powder=(the liquid chromatograph peak heights of the liquid chromatograph peak heights/reference substance solution of product to be tested solution)/step C; The weight percentage of emodin is in the medical material to be measured: c * 1000/0.15 * 100%.
D, judgement index components content
If in the Radix Polygoni Multiflori medical material to be measured 2,3,4 ', 5-tetrahydroxystilbene-percentage composition >=3.00% of 2-O-β-D-pyranglucoside and percentage composition >=0.050% of emodin judge that then Radix Polygoni Multiflori medical material to be measured is qualified, otherwise defective.
Chromatographic condition when carrying out the index components assay in this example is: chromatographic column: Hypersil C18 post (4.6mmx200mm, 5 μ m); Detect wavelength 254nm; Flow velocity 0.8ml/min; Column temperature: 30 ℃; Mobile phase is methanol and 0.1% phosphoric acid, and the mass ratio of the two is 85: 15; Theoretical cam curve is calculated by the emodin peak and is not less than 6000.
With above this routine method to the above-mentioned two kinds of index components 2,3,4 ' of carrying out of purchasing in the medical material to be measured of Deqing, Guangdong, 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside (THSG) and emodin index components content detection result such as following table:
| The medical material title | THSG (percentage by weight %) | Emodin (percentage by weight %) |
| Radix Polygoni Multiflori | 5.512 | 0.094 |
Become the standard of content to compare with judging index, require in the Radix Polygoni Multiflori medical material to be measured 2,3,4 ', 5-tetrahydroxystilbene-percentage composition >=3.00% of 2-O-β-D-pyranglucoside and percentage composition >=0.050% of emodin.Therefore, adopting this routine method is qualified to purchasing in the testing result of the Radix Polygoni Multiflori medical material of Deqing, Guangdong.
The present invention in the specific implementation, high performance liquid chromatograph can be selected Tianjin, island LC-10A high performance liquid chromatograph for use.
Embodiment two
This example and embodiment one are basic identical, and different only is: the standard medical material of Radix Polygoni Multiflori is replaced with the standard medical material of Radix Polygoni Multiflori Preparata, and Radix Polygoni Multiflori medical material to be measured replaces with Radix Polygoni Multiflori Preparata medical material to be measured.Radix Polygoni Multiflori Preparata standard medical material wherein adopts the river of Mount Emei, Sichuan to produce Radix Polygoni Multiflori and concocts and get with steamed method or Semen sojae atricolor method for making; The one concrete medical material to be measured of surveying in this example is the Radix Polygoni Multiflori Preparata medical material of buying in the Deqing, Guangdong.
Fig. 3 is the standard finger-print of the Radix Polygoni Multiflori Preparata that records with this routine method.Fig. 3 is visible; 6 characteristic peaks that peak value is high are arranged in the standard finger-print of Radix Polygoni Multiflori Preparata, and through identifying: No. 4 peaks are that stilbene glucoside, No. 5 peaks are that emodin pyranglucoside, No. 6 peaks are that chrysophanol pyranglucoside, No. 7 peaks are that physcione pyranglucoside, No. 10 peaks are that emodin, No. 11 peaks are physcione.In these peaks, No. 4 peak stilbene glucosides and No. 10 peak emodins are to produce active main component, also are to produce the property of medicine and toxic main component, and 6 main component characteristic peaks that characterize the Radix Polygoni Multiflori medical material become known compound.
Fig. 4 is the Radix Polygoni Multiflori Preparata medicinal materials fingerprint to be measured that this law records.Adopt China national Bureau of Drugs Supervision specified " the Chinese medicine chromatogram is analyzed and data handling system " software to carry out similarity and calculate, similarity is 0.965, finger printing to be measured and standard finger-print similarity >=0.900.
Adopt purchasing in two kinds of index components 2,3,4 ' of a concrete Radix Polygoni Multiflori Preparata to be measured of Deqing, Guangdong that this routine method measures, 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside (THSG) and emodin content are as shown in the table:
| The medical material title | THSG (percentage by weight %) | Emodin (percentage by weight %) |
| Steamed Radix Polygoni Multiflori Preparata | 3.811 | 0.119 |
| The bean Radix Polygoni Multiflori Preparata | 3.538 | 0.126 |
Become the standard of content to compare with judging index, in this Radix Polygoni Multiflori Preparata medical material to be measured 2,3,4 ', 5-tetrahydroxystilbene-percentage composition >=1.93% of 2-O-β-D-pyranglucoside and percentage composition >=0.067% of emodin.Visible by last table, the Radix Polygoni Multiflori Preparata medical material to be measured that this example institute specifically tests is qualified products.
Embodiment three
This example and embodiment one are basic identical, and different only is: the finger printing of steps A relatively in, the powder of the standard medical material of the Radix Polygoni Multiflori that takes by weighing is 0.1 gram, the time of each supersound process is 1 hour; Among the step B accurate take by weighing 2,3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance is 3.2mg, the accurate emodin that takes by weighing is 2.8mg among the step C.
Embodiment four
This example and embodiment one are basic identical, and different only is: the finger printing of steps A relatively in, the powder of the standard medical material of the Radix Polygoni Multiflori that takes by weighing is 0.3 gram, the time of each supersound process is 0.75 hour; Among the step B accurate take by weighing 2,3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance is 3.4mg, the accurate emodin that takes by weighing is 3.2mg among the step C.
Embodiment five
This example and embodiment two are basic identical, and different only is: the finger printing of steps A relatively in, the powder of the standard medical material of the Radix Polygoni Multiflori Preparata that takes by weighing is 0.1 gram, the time of each supersound process is 1 hour; Among the step B accurate take by weighing 2,3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance is 3.2mg, the accurate emodin that takes by weighing is 3.2mg among the step C.
Embodiment six
This example and embodiment two are basic identical, and different only is: the finger printing of steps A relatively in, the powder of the standard medical material of the Radix Polygoni Multiflori Preparata that takes by weighing is 0.3 gram, the time of each supersound process is 0.75 hour; Among the step B accurate take by weighing 2,3,4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance is 3.4mg, the accurate emodin that takes by weighing is 2.8mg among the step C.
Claims (3)
1. the detection method of Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata is made up of following steps:
A, finger printing are relatively
A1, set up Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata standard finger-print
Get the powder 0.1-0.3 gram of the standard medical material of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata, be dissolved in the acetone of 8ml, supersound process 0.5-1 hour after-filtration got filtrating; Triplicate, merging filtrate adds acetone, is settled to 25ml; The acetone soln of standard medical material, precision is measured the 5ml acetone soln, treat that acetone volatilizes after, dry powder; Get dry powder and be dissolved in methanol and get methanol solution, it is behind 0.45 micron the filtering with microporous membrane that methanol solution uses the aperture, the standard medical material solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata;
Draw the standard medical material solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata, inject high performance liquid chromatograph, set up the standard finger-print of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata standard medical material;
The finger printing of A2, making Radix Polygoni Multiflori to be measured or Radix Polygoni Multiflori Preparata
The powder of standard Radix Polygoni Multiflori in A1 step or Radix Polygoni Multiflori Preparata medical material is replaced with the powder of Radix Polygoni Multiflori to be measured or Radix Polygoni Multiflori Preparata medical material, adopt A1 to go on foot identical operation then, make the finger printing of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata medical material to be measured;
A3, collection of illustrative plates are relatively
Calculate the similarity of the finger printing and the standard finger-print in A1 step in A2 step; If finger printing and standard finger-print similarity >=0.900 are differentiated the medical material for supplying to carry out further quality testing with the medical material to be measured of correspondence, and are carried out the operation of following steps; Otherwise, end operation;
B, 2,3,4 ', the assay of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside index components
The preparation of B1, reference substance solution
Precision takes by weighing 2,3,4 ' of 3.2-3.4mg, and 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside reference substance places the 10ml volumetric flask, adds 50% dissolve with ethanol and is diluted to scale, shakes up, and joins to such an extent that concentration is the reference substance solution of 0.32-0.34g/L;
The preparation of B2, product to be tested solution
Take by weighing Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata powder 0.2g, place the 100ml conical flask, add 50% ethanol 25ml; Claim to decide quality, reflux 30min is put cold; Claim again to decide quality, supply the quality that subtracts mistake, shake up with 50% ethanol; Supernatant filters with 0.45 μ m microporous filter membrane, promptly gets the product to be tested solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata;
B3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume, adopts HPLC to measure in the product to be tested solution 2,3; 4 '; The content of 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside, and converse 2,3 in Radix Polygoni Multiflori to be measured or the Radix Polygoni Multiflori Preparata; 4 ', 5-tetrahydroxystilbene-2-O-β-D-pyranglucoside composition accounts for the percentage composition of whole medical material dry to be measured;
The assay of C, emodin index components
The preparation of C1, reference substance solution
Precision takes by weighing emodin 2.8-3.2mg, puts in the 100ml volumetric flask, adds dissolve with methanol and is diluted to 100ml, shakes up;
The preparation of C2, product to be tested solution
Precision takes by weighing Radix Polygoni Multiflori or each 0.15g of Radix Polygoni Multiflori Preparata powder, puts in the 100ml conical flask, and precision adds methanol 25ml, claim to decide quality, and reflux 1h, cooling is claimed to decide quality again, supplies the quality that subtracts mistake with methanol, shakes up, and filters; Precision is measured filtrating 5ml, puts in the 100ml conical flask, flings to methanol, adds 8% hydrochloric acid 10ml, and supersound process 2min adds chloroform 10ml again, reflux 1h, cooling; In the backflow dislocation separatory funnel,, incorporate in the separatory funnel, obtain the chloroform solution of lower floor with minimum of chloroform washing 100ml conical flask; The hydrochloric acid solution reuse chloroform extraction on upper strata and obtaining 3 times, each chloroform consumption 8-12ml, combined chloroform liquid moves in the 100ml conical flask, flings to chloroform to doing; Residue adds dissolve with methanol, and standardize solution in the 10ml volumetric flask filters with 0.45 μ m microporous filter membrane, promptly gets the product to be tested solution of Radix Polygoni Multiflori or Radix Polygoni Multiflori Preparata powder;
C3, product to be tested Determination on content
Precision is measured the reference substance solution and the product to be tested solution of equal volume; Adopt HPLC to measure emodin content in the product to be tested solution, and converse the percentage composition that emodin composition in Radix Polygoni Multiflori to be measured or the Radix Polygoni Multiflori Preparata accounts for whole medical material dry to be measured.
2. the detection method of a kind of Radix Polygoni Multiflori according to claim 1 and Radix Polygoni Multiflori Preparata is characterized in that: described Radix Polygoni Multiflori standard medical material produces Radix Polygoni Multiflori for the river of directly picking up from the Mount Emei, Sichuan; Radix Polygoni Multiflori Preparata standard medical material is that the river produces the steamed method of Radix Polygoni Multiflori employing or the process of preparing Chinese medicine of Semen sojae atricolor method for making gets.
3. the detection method of a kind of Radix Polygoni Multiflori according to claim 1 and Radix Polygoni Multiflori Preparata; It is characterized in that: when described A1 and A2 tested standard medical material solution and medical material solution to be measured with high performance liquid chromatograph in the step, all select the detection wavelength of 290nm wavelength for use as the finger printing of high performance liquid chromatograph.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100590243A CN101554410B (en) | 2009-04-22 | 2009-04-22 | Detection method of fresh fleece flower root and prepared fleece flower root |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100590243A CN101554410B (en) | 2009-04-22 | 2009-04-22 | Detection method of fresh fleece flower root and prepared fleece flower root |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101554410A CN101554410A (en) | 2009-10-14 |
| CN101554410B true CN101554410B (en) | 2012-06-20 |
Family
ID=41172792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100590243A Expired - Fee Related CN101554410B (en) | 2009-04-22 | 2009-04-22 | Detection method of fresh fleece flower root and prepared fleece flower root |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101554410B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156868A (en) * | 2011-12-09 | 2013-06-19 | 天津市国际生物医药联合研究院 | Application of stilbene glucosides in treating and preventing AIDS |
| CN103018367B (en) * | 2012-12-13 | 2014-05-14 | 贵州大学 | Method for determining content of lecithin component in polygonum multiflorum medicinal material |
| CN105675741A (en) * | 2015-12-30 | 2016-06-15 | 成都九芝堂金鼎药业有限公司 | Method for determining content of effective components in hair growing pill |
| CN105726550B (en) * | 2016-05-17 | 2018-08-21 | 吉林大学 | The purposes of TSG |
| CN107490645A (en) * | 2017-07-03 | 2017-12-19 | 中国人民解放军第三〇二医院 | A kind of safe mass control method of prepared fleece flower root and application |
| CN109900827B (en) * | 2019-03-26 | 2022-02-11 | 陕西省食品药品监督检验研究院 | Method for identifying radix polygoni multiflori preparata based on one-test-multiple evaluation method |
| CN109991341B (en) * | 2019-05-17 | 2022-04-01 | 无限极（中国）有限公司 | Quality detection method of radix polygoni multiflori preparata |
| CN111307971A (en) * | 2020-03-06 | 2020-06-19 | 西南交通大学 | Quality detection method of polygonum multiflorum preparata decoction pieces without hepatotoxicity |
-
2009
- 2009-04-22 CN CN2009100590243A patent/CN101554410B/en not_active Expired - Fee Related
Non-Patent Citations (5)
| Title |
|---|
| 刘振丽，等.制何首乌高效液相指纹图谱分析.《中成药》.2005,第27卷(第4期),全文. * |
| 周洪波.何首乌质量控制与安全性初步研究.《中国优秀硕士学位论文全文数据库医药卫生科技辑》.中国学术期刊(光盘版)电子杂志社,2009,(第1期),E057-129. * |
| 房志坚，等.何首乌的HPLC指纹图谱.《华西药学杂志》.2008,第23卷(第5期),第513-515页. * |
| 李林福.制何首乌化学成分及其质量标准研究.《中国优秀硕士学位论文全文数据库医药卫生科技辑》.中国学术期刊(光盘版)电子杂志社,2009,(第2期),E057-23. * |
| 杨仙凌.HPLC法测定生何首乌和制何首乌中大黄素的含量.《浙江中医药大学学报》.2008,第32卷(第3期),第399-400页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101554410A (en) | 2009-10-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101554410B (en) | Detection method of fresh fleece flower root and prepared fleece flower root | |
| CN101167788B (en) | Quality control method of traditional Chinese medicine 'zhenqi fuzheng' containing glossy privet fruit and radix astragali for strengthening the body resistance for aeipathia deficiency damage and qi | |
| CN101062294B (en) | Method for detecting the quality of the medicine combination for treating cerebral apoplexy and vasculitis | |
| CN100457139C (en) | Method for preparing a Shuanhuanglian injection and the component detecting method | |
| CN106370749A (en) | Quality detection method of ginseng basis-consolidating oral solution | |
| CN113759020B (en) | Preparation process and quality control method of channel warming soup | |
| CN107356691A (en) | Build the detection method of bent finger-print | |
| CN102749401B (en) | Inspection method of traditional Chinese medicine composition twenty-five-ingredient lung disease preparation | |
| CN102274401B (en) | Traditional Chinese medicine preparation for treating gastropathy and preparation method and detection method thereof | |
| CN101637567A (en) | Rhizoma ligustici wallichii quality control method in Chinese medicine preparation | |
| CN102579861A (en) | Method for detecting quality of An'erning granules | |
| CN113759017A (en) | Preparation process and evaluation method of angelica sinensis Liuhuang decoction | |
| CN102579734B (en) | Traditional Chinese medicine composition of bone healing medicine, preparing method thereof and detecting method thereof | |
| CN102441057B (en) | High performance liquid chromatography (HPLC) fingerprint detection method for blood-nourishing brain-refreshing grain | |
| CN101708208A (en) | Quality control method of capsule preparation for treating a painful swollen joint | |
| CN105510488B (en) | A kind of finger-print and its quality determining method for relieving pain patch | |
| CN101204434A (en) | Quality standard for thrombus dispelling pill and test method thereof | |
| CN103472148A (en) | Fingerprint spectrum detection method of Chinese medicine composition preparation | |
| CN102068657B (en) | Quality control method of venation relaxing pill as Chinese herbal preparation | |
| CN108982738B (en) | Construction method and detection method of intermediate characteristic spectrum of Zhenqi Fuzheng granule or Zhenqi Fuzheng granule fingerprint | |
| CN102138963B (en) | Chinese angelica and astragalus extract, and preparation method and application thereof | |
| CN102068549A (en) | Quality control method for Chinese medicinal preparation heat clearing and blood cooling pills | |
| CN103293261A (en) | Content determination method of rhizoma bletillae | |
| CN1939472B (en) | Detection method of compound preparation for treating diabetes | |
| CN105616946A (en) | Preparation for treating cough, preparation method and quality control method thereof |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120620 Termination date: 20160422 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |