CN101274036A - Method for testing quality criteria of conghuang preparation for tonifying kidney - Google Patents
Method for testing quality criteria of conghuang preparation for tonifying kidney Download PDFInfo
- Publication number
- CN101274036A CN101274036A CNA2008100280007A CN200810028000A CN101274036A CN 101274036 A CN101274036 A CN 101274036A CN A2008100280007 A CNA2008100280007 A CN A2008100280007A CN 200810028000 A CN200810028000 A CN 200810028000A CN 101274036 A CN101274036 A CN 101274036A
- Authority
- CN
- China
- Prior art keywords
- reference substance
- solution
- preparation
- kidney
- echinacoside
- 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.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a method for inspecting the quality standards of a Conghuang kidney-reinforcing preparation, which used for controlling the quality of the Conghuang kidney reinforcing preparation, wherein, the preparation refers to granules, tablets and hard gelatin capsules. When disclosing the method for inspecting the quality standards of a new Conghuang kidney reinforcing preparation, relatively great improvement is made to the existing methods for inspecting the quality standards of medical pills by the invention, which is particularly characterized in that: thin layers qualitative differential diagnosis of prepared rhizome of rehmannia, seed of Chinese dodder and fruit of Chinese magnolia vine (steaming with wine) in a prescription is added, the preparation method of desertliving cistache differential diagnosis sample solution is changed and determination of echinacoside with little interference but high content by high performance liquid chromatography is modified. The quality controllability of the new Conghuang kidney-reinforcing preparation can be improved by using the inspecting method of the invention, which is favorable to the quality stability of products.
Description
Technical field
The invention belongs to the Chinese patent medicine field of quality control.
Background technology
The yellow the kidney invigorating prescription of desert is made up of Herba Cistanches, Radix Rehmanniae Preparata, Semen Cuscutae, Fructus Schisandrae Chinensis (wine steaming) four Chinese medicine material, has the effect of nourishing kidney yin, bone and muscle strengthening.Can make dosage forms such as ball, granule, sheet, hard capsule herein.At present, only recorded pill standard (" country of National Drug Administration standard for traditional Chinese medicines compilation " internal medicine kidney is a fascicle) in the disclosed drug standard, yellow other dosage form standards of the kidney invigorating of desert are not appeared in the newspapers.Have only microscopical identification in the pill standard, Herba Cistanches TLC differentiates and verbascoside HPLC assay.This medicine prescription is made up of four Chinese medicine, and obvious pill standard and imperfection can not effectively guarantee the quality of product.
Simultaneously, we also find in test: the yellow the kidney invigorating novel form of these three kinds of deserts of the granule that relates to, sheet and hard capsule Herba Cistanches in the pill standard is differentiated the test liquid that the test liquid preparation method obtains under the item, and there are a certain distance in speckle on its thin layer chromatography and the stratographic speckle of betanin reference substance on the position.
In addition, the yellow the kidney invigorating novel forms of these three kinds of deserts of granule, sheet and hard capsule are when doing the HPLC assay of verbascoside, and the negative test liquid peak that disturbs exists.Testing different chromatographic columns and mobile phase composition all can not well separate the peak of Interference Peaks with verbascoside.
Summary of the invention
The object of the invention provides the detection method of the yellow kidney-tonifying preparation quality standard of a kind of desert, thereby improves the yellow the kidney invigorating novel form of desert quality controllability, guarantees the stable of product quality.Wherein preparation is meant granule, sheet and hard capsule.
The present invention has carried out bigger improvement to the pill standard detecting method, be specially: increased the thin-layer qualitative discriminating of Radix Rehmanniae Preparata, Semen Cuscutae, Fructus Schisandrae Chinensis (wine steaming) in the prescription, changed the preparation method of Herba Cistanches discriminating test liquid, changed high performance liquid chromatography assay composition for disturbing the echinacoside little, that content is high.
Why change that to measure echinacoside be because discovery has bigger interference during with containing the survey method and detect the verbascoside content of the yellow the kidney invigorating novel form of desert in the former pill standard into.Tested methanol-acetonitrile-1% acetic acid (13: 10: 77,11: 9: 80,15: 10: 75), methanol-0.1% formic acid (28: 72,35: 65) mobile phase, and kromasil-C18 (250mm * 4.6mm, 5 μ m) and Ultimate-C18 (250mm * 4.6mm, 5 μ m) chromatographic column, the result all can not make the peak of Interference Peaks and verbascoside separate.And do not find obvious Interference Peaks when detecting echinacoside.Because the complexity of Chinese medicine, and can not obtain former pill standard and declare the research of comparing of the pill sample of producer, so exist the reason of bigger interference problem can't determine now when producing this detection verbascoside, may be that used medical material place of production difference or the equal reason of the concrete operations parameter in the manufacturing process causes.
Bibliographical information is arranged, and the main constituent that plays the sexual function improving effect in the Herba Cistanches is the phenethyl alcohol glycoside compounds.Echinacoside, verbascoside all belong to the higher phenethyl alcohol glycoside compounds of content.Through our detection to commercially available 5 different places of production Herba Cistanches medical materials (wherein 2 Cistanche Tubulosas for the Xinjiang product), the content of finding 4 place of production echinacoside is all greater than verbascoside content, and the content of two kinds of compositions is suitable in the medical material in another place of production.So changing into detecting disturbs the content of the echinacoside little, that content is high also can reflect product quality preferably.
The method for making of the yellow the kidney invigorating novel form of desert involved in the present invention, except that adjuvant kind and consumption, make total amount, drying process, final dosage form different, other is all according to prescription in the pill standard and method for making manufacturing.Whole manufacture method and pill relatively do not have change big, essence.So according to the classification of medicine registration management way, the yellow the kidney invigorating granule of desert involved in the present invention, sheet or hard capsule preparation should belong to 8 kind new medicines (being the imitation medicine dosage changing form).
The invention provides yellow kidney-tonifying preparation---granule, sheet, hard capsule---detection method of quality standard of desert, comprise with thin layer chromatography (TLC) Herba Cistanches, Radix Rehmanniae Preparata, Semen Cuscutae, Fructus Schisandrae Chinensis (wine steaming) are carried out qualitative identification and measure active component---the content of echinacoside (echinacoside) in the Herba Cistanches with high performance liquid chromatography (HPLC).
1, the thin layer of Herba Cistanches is differentiated
It is an amount of to get the yellow kidney-tonifying preparation of desert (granule, sheet or hard capsule), and porphyrize takes by weighing powder 0.5～1.0g, add water 20ml, ultrasonic 10min makes molten diffusing, centrifugal (4500r/min) 5min, get supernatant vacuum filtration again, filtrate is crossed strong acid cation exchange resin column, washes post with 20ml, reuse 0.5～4mol/l ammonia spirit 20～60ml eluting, collect eluent, evaporate to dryness is with ethanol 2ml dissolving, leave standstill, get supernatant as need testing solution.Other gets the negative sample of misrun Herba Cistanches, makes negative test liquid with method.The betanin reference substance adds ethanol and makes the reference substance solution that every 1ml contains 5mg.Draw each 10 μ l of need testing solution and negative test liquid, reference substance solution 5 μ l, point sample is on same silica gel g thin-layer plate respectively, with methanol-glacial acetic acid-water (10～20: 0.5～2: 2～8) be developing solvent, launch, take out, dry, spray is with the improvement bismuth potassium iodide test solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.The negative sample chromatograph does not have corresponding speckle.
2, the thin layer of Radix Rehmanniae Preparata is differentiated
It is an amount of to get the yellow kidney-tonifying preparation of desert (granule, sheet or hard capsule), porphyrize takes by weighing powder 1.0～3.0g, adds water 30ml, supersound process 10min, centrifugal (4500r/min) 5min gets supernatant and extracts 2 times with n-butyl alcohol (chloroform, ethyl acetate), each 15ml, merge extractive liquid,, extracting solution is concentrated into dried, and residue adds ethanol 1ml dissolving, as need testing solution.Other gets the negative sample that lacks Radix Rehmanniae Preparata, makes negative test liquid with method.The 5 hydroxymethyl furfural reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.5mg.Draw each 10 μ l of need testing solution and negative test liquid, reference substance solution 5 μ l, point sample is in same silica gel G F respectively
254On the lamellae, (1～8: 2～10) upper strata liquid is developing solvent, launches, and takes out, and dries, and places under the 254nm uviol lamp and inspects with petroleum ether II-ethyl acetate.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color.The negative sample chromatograph does not have corresponding skin dark stain point.
3, the thin layer of Semen Cuscutae is differentiated
It is an amount of to get the yellow kidney-tonifying preparation of desert (granule, sheet or hard capsule), and porphyrize takes by weighing powder 1.0～3.0g, add chloroform (dehydrated alcohol, methanol, ethyl acetate, n-butyl alcohol) 10ml, supersound process 15min filters, filtrate is concentrated into dried, adds ethanol 1ml dissolved residue, as need testing solution.Other gets the negative sample that lacks Semen Cuscutae, makes negative test liquid with method.The kaempferol reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.2mg.Draw each 10 μ l of need testing solution and negative test liquid, reference substance solution 5 μ l, point sample is on same silica gel g thin-layer plate respectively, with toluene-ethyl acetate-methanol (4～10: 4～8: 1～6) be developing solvent, launch, take out, dry, spray places under the 365nm uviol lamp and inspects with 3% aluminum chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.The negative sample chromatograph does not have corresponding speckle.
4, the thin layer of Fructus Schisandrae Chinensis (wine steaming) is differentiated
It is an amount of to get the yellow kidney-tonifying preparation of desert (granule, sheet or hard capsule), and porphyrize takes by weighing powder 8.0～15.0g, add methanol (dehydrated alcohol, chloroform, ethyl acetate, n-butyl alcohol) 20ml, supersound process or reflux 30min filter, filtrate is concentrated into about 1ml, as need testing solution.Other gets the negative sample that lacks Fructus Schisandrae Chinensis, makes negative test liquid with method.The schisandrin B reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.4mg.Draw each 10 μ l of need testing solution, negative test liquid and reference substance solution, point sample is in same silica gel G F respectively
254On the lamellae, (8～20: 2～8: 0.5～2) upper strata liquid is developing solvent, launches, and takes out, and dries, and places under the 254nm uviol lamp and inspects with petroleum ether I-Ethyl formate-formic acid.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color.The negative sample chromatograph does not have corresponding skin dark stain point.
5, echinacoside assay in the Herba Cistanches
Chromatographic condition and system suitability test chromatographic column adopting octadecylsilane chemically bonded silica are filler; Mobile phase is methanol-acetonitrile-1% acetum (15: 10: 75); The detection wavelength is 334nm.Theoretical cam curve is calculated by the echinacoside peak should be not less than 4000.
Reference substance solution prepares precision, and to take by weighing the echinacoside reference substance an amount of, puts in the brown bottle, adds mobile phase and be made into the reference substance solution that every 1ml contains 0.16mg
It is an amount of that the yellow kidney-tonifying preparation of desert (granule, sheet or hard capsule) is got in the need testing solution preparation, and porphyrize is got 0.5g, and accurate title is fixed, put in the brown measuring bottle of 50ml, precision adds mobile phase 20ml, and close plug shakes up, claim to decide weight, soaked 0.5 hour, supersound process 40min takes out, put coldly, claim again to decide weight, add mobile phase and supply the weight that subtracts mistake, shake up, centrifugal, get supernatant, micro-filtration membrane with 0.45 μ m filters, and filtrate is put in the brown bottle, promptly.
Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure.
Calculating is calculated as follows:
In the formula:
W: echinacoside content, %;
Cs: the concentration of echinacoside in the reference substance, mg/ml;
M: the sample weighting amount of test sample, mg;
As: the chromatographic peak area of echinacoside in the reference substance;
Ai: the chromatographic peak area of echinacoside in the test sample.
The content assaying method of echinacoside has passed through the methodological study test (because tablet and hard capsule involved in the present invention all is to form by machine pressing or filling from granule in the yellow kidney-tonifying preparation of desert of the present invention, the just variation of profile, there is not composition to change, so if the subjects that following summary of the invention adopted does not have explanation to be granule.)：
(1) instrument and reagent
SPD-10Avp Ultraviolet Detector day island proper Tianjin company; P4000 quaternary pump U.S. power ﹠ light company; AT-130 chromatographic column calorstat Tianjin Hengao Technology Development Co., Ltd.; HN1006 ultrasonic cleaning machine China south China ultrasonic device factory; HM202 electronic analytical balance Japan AND company
Echinacoside (111670-200502) Nat'l Pharmaceutical ﹠ Biological Products Control Institute; The yellow kidney-tonifying preparation granule of desert (lot number 070801,070802,070803, specification are the 2.0g/ bag) provides for Guangzhou Meichen Pharmaceutical Co., Ltd.; The second eyeball is a chromatographically pure Di Ma company.All the other reagent are analytical pure.
(2) chromatographic condition adopts kromasil-C18 post (250mm * 4.6mm, 5 μ m) chromatographic column, 30 ℃ of column temperatures; Mobile phase is methanol-acetonitrile-1% acetum (15: 10: 75); Flow velocity is 1ml/min; The detection wavelength is 334nm.
(3) reference substance solution prepares precision to take by weighing the echinacoside reference substance an amount of, puts in the brown bottle, adds mobile phase and is made into the reference substance solution that every 1ml contains 0.16mg
(4) it is an amount of that this product is got in the need testing solution preparation, and porphyrize is got 0.5g, and accurate the title decides, put in the brown measuring bottle of 50ml, precision adds mobile phase 20ml, and close plug shakes up, claim to decide weight, soaked 0.5 hour, supersound process 40min takes out, put coldly, claim again to decide weight, add mobile phase and supply the weight that subtracts mistake, shake up, centrifugal, get supernatant, micro-filtration membrane with 0.45 μ m filters, and filtrate is put in the brown bottle, promptly.Get the negative sample 0.3g of misrun Herba Cistanches, make negative test liquid with method.
(5) the above-mentioned reference substance solution 3 μ l of the accurate absorption of linear relationship investigation, 5 μ l, 7 μ l, 10 μ l, 12 μ l inject chromatograph of liquid respectively, with peak area A is abscissa, with sample size W (mg) is vertical coordinate, calculates regression equation: W=-3.3143 * 10
-5+ 7.97149 * 10
-10A, R=0.99996.Echinacoside is at 0.48 μ g---between the 1.92 μ g good linear relationship is arranged.
(6) accurate each the 5 μ l of above-mentioned reference substance solution, test liquid and negative test liquid that draw of negative control test inject chromatograph of liquid, and result's negative test liquid chromatograph on echinacoside retention time position is not obviously disturbed.
(7) average recovery test precision suitably takes by weighing 9 parts of this product, and per 3 parts is 1 group, and every group all adds above-mentioned reference substance solution 3ml, 2ml, 1ml respectively by high, medium and low three concentration.According to above-mentioned need testing solution preparation method preparation and mensuration.Average recovery rate is 99.1% as a result, RSD%=1.52%.
(8) the accurate contrast liquid 5 μ l that draw of precision test inject chromatograph of liquid, repeat the record peak area 6 times.Its RSD=1.03% shows that this method has good precision.
(9) stability test injects chromatograph of liquid, the record peak area respectively at 0,2,4,6,8 hour accurate each 5 μ l of test liquid that draw.Its RSD=1.41% shows that test liquid is stable in 0-8 hour.
(10) replica test is to same batch sample, according to the said determination method, and replication 6 times.Its RSD=1.01% shows that this method has good repeatability.
(11) sample determination is measured three batch samples according to the method described above, the results are shown in following table (by formula calculate gained mass percentage content multiply by the preparation specification be listed unit content)
The content (n=3) of echinacoside in the yellow the kidney invigorating granule of table 1 desert
The present invention has the following advantages:
1, the present invention carries out the thin layer chromatography qualitative identification respectively to the middle four Chinese medicine material of the yellow kidney-tonifying preparation---granule, sheet, hard capsule---of desert, and contained a kind of active component is carried out assay.Compare with the pill standard detecting method, the present invention is more perfect, more scientific and reasonable.。
2, the present invention propose first in the yellow kidney-tonifying preparation of desert except that Herba Cistanches other three thin layer discrimination methods of distinguishing the flavor of medical materials, and the part that is not suitable for detecting the yellow the kidney invigorating novel form of desert (granule, sheet, hard capsule) in the pill standard detecting method carried out bigger modification (changed the preparation method that Herba Cistanches is differentiated test liquid, changed high performance liquid chromatography assay composition and be the echinacoside that disturbs less, content is high).Thereby improved the quality control level of the yellow the kidney invigorating novel form of desert, ensured the stable of product quality
3, strong, the good reproducibility of specificity of the present invention can be used for the daily quality control of this medicine.
Description of drawings
Provide accompanying drawing below and be used to further specify the present invention, but the present invention is not construed as limiting.
Fig. 1 is the TLC figure of Herba Cistanches of the present invention;
Fig. 2 is the TLC figure of Radix Rehmanniae Preparata of the present invention;
Fig. 3 is the TLC figure of Semen Cuscutae of the present invention;
Fig. 4 is the TLC figure of Fructus Schisandrae Chinensis of the present invention;
Fig. 5 is the yellow capsule for tonifying kidney HPLC of a desert of the present invention chromatogram.
The specific embodiment
The embodiment that enumerates detection below further describes the present invention, but to not restriction of the present invention.
Embodiment one
1. instrument
Booth Electronic Instruments Plant in Hai'an on the ultraviolet point sample instrument; SPD-10Avp Ultraviolet Detector day island proper Tianjin company; P4000 quaternary pump U.S. power ﹠ light company; AT-130 chromatographic column calorstat Tianjin Hengao Technology Development Co., Ltd.; HN1006 ultrasonic cleaning machine China south China ultrasonic device factory; HM202 electronic analytical balance Japan AND company; The 4K15C High speed refrigerated centrifuge; Germany SIGMA company.
2. reagent
Silica gel G Qingdao Marine Chemical Co., Ltd.; Betanin (894-200202), 5 hydroxymethyl furfural (111626-200402), schisandrin B (765-200104), kaempferol (0861-200002), echinacoside (111670-200502) Nat'l Pharmaceutical ﹠ Biological Products Control Institute; The yellow the kidney invigorating granule (lot number 070801, specification 2.0g/ bag) of desert, the yellow kidney invigorating tablet (lot number 070811, specification 0.5g/ sheet) of desert, the yellow the kidney invigorating hard capsule of desert (lot number 070821, specification 0.4g/ grain) provides for Guangzhou Meichen Pharmaceutical Co., Ltd.; The second eyeball is a chromatographically pure Di Ma company.All the other reagent are analytical pure.
3. thin layer is differentiated (with the granule is example, and tablet is identical with it with hard capsule)
3.1. Herba Cistanches
It is an amount of to get the yellow the kidney invigorating granule of desert, and porphyrize takes by weighing powder 0.5～1.0g, add water 20ml, ultrasonic 10min makes molten diffusing, centrifugal (4500r/min) 5min, get supernatant vacuum filtration again, filtrate is crossed strong acid cation exchange resin column, washes post with 20ml, reuse 0.5～4mol/l ammonia spirit 20～60ml eluting, collect eluent, evaporate to dryness is with ethanol 2ml dissolving, leave standstill, get supernatant as need testing solution.The betanin reference substance adds ethanol and makes the reference substance solution that every 1ml contains 5mg.Draw need testing solution 10 μ l, reference substance solution 5 μ l, point sample is on same silica gel g thin-layer plate, with methanol-glacial acetic acid-water (10～20: 0.5～2: 2～8) be developing solvent, launch, take out, dry that spray is with the improvement bismuth potassium iodide test solution respectively.As a result in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
3.2. Radix Rehmanniae Preparata
It is an amount of to get the yellow the kidney invigorating granule of desert, porphyrize takes by weighing powder 1.0～3.0g, adds water 30ml, supersound process 10min, centrifugal (4500r/min) 5min gets supernatant and extracts 2 times with n-butyl alcohol (chloroform, ethyl acetate), each 15ml, merge extractive liquid,, extracting solution is concentrated into dried, and residue adds ethanol 1ml dissolving, as need testing solution.The 5 hydroxymethyl furfural reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.5mg.Draw need testing solution 10 μ l, reference substance solution 5 μ l, point sample is in same silica gel G F respectively
254On the lamellae, (1～8: 2～10) upper strata liquid is developing solvent, launches, and takes out, and dries, and places under the 254nm uviol lamp and inspects with petroleum ether II-ethyl acetate.As a result in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color.
3.3. Semen Cuscutae
It is an amount of to get the yellow the kidney invigorating granule of desert, and porphyrize takes by weighing powder 1.0～3.0g, adds chloroform (dehydrated alcohol, methanol, ethyl acetate, n-butyl alcohol) 10ml, and supersound process 15min filters, and filtrate is concentrated into dried, adds ethanol 1ml dissolved residue, as need testing solution.The kaempferol reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.2mg.Draw need testing solution 10 μ l, reference substance solution 5 μ l, point sample is on same silica gel g thin-layer plate respectively, with toluene-ethyl acetate-methanol (4～10: 4～8: 1～6) be developing solvent, launch, take out, dry, spray places under the 365nm uviol lamp and inspects with 3% aluminum chloride alcoholic solution.As a result in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
3.4. Fructus Schisandrae Chinensis (wine steaming)
It is an amount of to get the yellow the kidney invigorating granule of desert, and porphyrize takes by weighing powder 8.0～15.0g, adds methanol (dehydrated alcohol, chloroform, ethyl acetate, n-butyl alcohol) 20ml, and supersound process or reflux 30min filter, and filtrate is concentrated into about 1ml, as need testing solution.The schisandrin B reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.4mg.Draw each 10 μ l of need testing solution and reference substance solution, point sample is in same silica gel G F respectively
254On the lamellae, (8～20: 2～8: 0.5～2) upper strata liquid is developing solvent, launches, and takes out, and dries, and places under the 254nm uviol lamp and inspects with petroleum ether I-Ethyl formate-formic acid.As a result in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color.
" 1 " of indication is a test liquid among Fig. 1, and " 2 " are contrast liquid, and " 3 " are negative test liquids.
" 1 " of indication is a test liquid among Fig. 2, and " 2 " are contrast liquid, and " 3 " are negative test liquids.
To reach " 3 " be reference substance in " 1 " of indication among Fig. 3, and " 2 " reach " 5 " is test liquid, and " 4 " are negative test liquids.
Among Fig. 4 " 1 " of indication reach " 4 " be the contrast liquid, " 2 " reach " 5 " is test liquid, " 3 " are negative test liquids.
4. assay (with the granule is example, and tablet and hard capsule division result are outer, and other are identical with it).
4.1. chromatographic condition chromatographic column adopting kromasil-C18 post (250mm * 4.6mm, 5 μ m); 30 ℃ of column temperatures; Mobile phase is methanol-acetonitrile-1% acetum (15: 10: 75); Flow velocity is 1ml/min; The detection wavelength is 334nm.
To take by weighing the echinacoside reference substance an amount of 4.2. reference substance solution prepares precision, puts in the brown bottle, adds mobile phase and be made into the reference substance solution that every 1ml contains 0.16mg
4.3. the need testing solution preparation is got, and the yellow the kidney invigorating granule of desert is an amount of, porphyrize is got 0.5g, and accurate the title decides, put in the brown measuring bottle of 50ml, precision adds mobile phase 20ml, and close plug shakes up, claim to decide weight, soaked 0.5 hour, supersound process 40min takes out, put coldly, claim again to decide weight, add mobile phase and supply the weight that subtracts mistake, shake up, centrifugal, get supernatant, micro-filtration membrane with 0.45 μ m filters, and filtrate is put in the brown bottle, promptly.
4.4. accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure.(by formula calculating the gained mass percentage content multiply by the preparation specification and is listed unit content.)
4.5. granule contains echinacoside 18.1mg for every bag as a result
Every in tablet contains echinacoside 4.5mg
Every of hard capsule contains echinacoside 3.9mg
" I " of indication is test liquid among Fig. 5, and " II " is contrast liquid, and " III " is negative test liquid, and " a " is echinacoside.
Negative test result shown in Fig. 1,2,3,4,5 is in order to verify and illustrate the present invention.Detection method is being carried out to do negative test when specificity is investigated.After detection method is determined, in daily detection, generally do not need to do negative test.
Claims (3)
1, the detection method of the yellow kidney-tonifying preparation quality standard of desert is characterized in that with thin layer chromatography Herba Cistanches, Radix Rehmanniae Preparata, Semen Cuscutae, wine being steamed and concocts that Fructus Schisandrae Chinensis carries out qualitative identification and with the content of active component echinacoside in the high-performance liquid chromatogram determination Herba Cistanches.
2, according to the detection method of the yellow kidney-tonifying preparation quality standard of the described desert of claim 1, it is characterized in that described preparation refers to granule, sheet and hard capsule.
3,, it is characterized in that described Herba Cistanches, Radix Rehmanniae Preparata, Semen Cuscutae, wine steam the qualitative identification of concocting Fructus Schisandrae Chinensis and may further comprise the steps according to the detection method of the yellow kidney-tonifying preparation quality standard of the described desert of claim 1:
(1) it is an amount of that Herba Cistanches is got the yellow the kidney invigorating granule of desert, sheet or hard capsule dosage form preparation, porphyrize, take by weighing powder 0.5～1.0g, add water 20ml, ultrasonic 10min makes molten loosing, centrifugal commentaries on classics 5min gets supernatant vacuum filtration again, and filtrate is crossed strong acid cation exchange resin column, wash post with 20ml, reuse 0.5～4mol/l ammonia spirit 20～60ml eluting is collected eluent, evaporate to dryness, dissolve with ethanol 2ml, leave standstill, get supernatant as need testing solution, the betanin reference substance adds ethanol and makes the reference substance solution that every 1ml contains 5mg; Draw need testing solution 10 μ l, reference substance solution 5 μ l, point sample is developing solvent with methanol-glacial acetic acid-water on same silica gel g thin-layer plate respectively, ratio is 10～20: 0.5～2: 2～8, launch, take out, to dry, spray is with the improvement bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(2) it is an amount of that Radix Rehmanniae Preparata is got the yellow the kidney invigorating granule of desert, sheet or hard capsule dosage form preparation, porphyrize takes by weighing powder 1.0～3.0g, adds water 30ml, supersound process 10min, centrifugal commentaries on classics 5min gets supernatant with n-butyl alcohol, chloroform or ethyl acetate extraction 2 times, each 15ml, merge extractive liquid,, extracting solution is concentrated into dried, and residue adds ethanol 1ml dissolving, as need testing solution; The 5 hydroxymethyl furfural reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.5mg, draws need testing solution 10 μ l, reference substance solution 5 μ l, and point sample is in same silica gel G F respectively
254On the lamellae, be that 1～8: 2～10 petroleum ether II-ethyl acetate upper strata liquid are developing solvent, launch, take out, dry, place under the 254nm uviol lamp and inspect with ratio; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color;
(3) Semen Cuscutae is got the yellow the kidney invigorating granule of desert, sheet or hard capsule dosage form preparation are an amount of, porphyrize, take by weighing powder 1.0～3.0g, add chloroform, dehydrated alcohol, methanol, ethyl acetate or n-butyl alcohol 10ml, supersound process 15min filters, filtrate is concentrated into dried, add ethanol 1ml dissolved residue, as need testing solution, the kaempferol reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.2mg, draw need testing solution 10 μ l, reference substance solution 5 μ l, point sample is on same silica gel g thin-layer plate respectively, and be 4～10: 4～8 with ratio: 1～6 toluene-ethyl acetate-methanol is developing solvent, launch, take out, dry, spray is with 3% aluminum chloride alcoholic solution, place under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(4) it is an amount of that wine steaming process of preparing Chinese medicine Fructus Schisandrae Chinensis is got the yellow the kidney invigorating granule of desert, sheet or hard capsule dosage form preparation, porphyrize, take by weighing powder 8.0～15.0g, add methanol, dehydrated alcohol, chloroform, ethyl acetate or n-butyl alcohol 20ml, supersound process or reflux 30min filter, and filtrate is concentrated into about 1ml, as need testing solution, the schisandrin B reference substance adds ethanol and makes the reference substance solution that every 1ml contains 0.4mg; Draw each 10 μ l of need testing solution reference substance solution, point sample is in same silica gel G F respectively
254On the lamellae, be 8～20: 2～8 with ratio: 0.5～2 petroleum ether I-Ethyl formate-formic acid upper strata liquid is developing solvent, launches, and takes out, and dries, and places under the 254nm uviol lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the skin dark stain point of same color.
4, according to the detection method of the yellow kidney-tonifying preparation quality standard of the described desert of claim 1, it is characterized in that the assay of active component echinacoside in the described Herba Cistanches may further comprise the steps:
Chromatographic condition and system suitability test chromatographic column adopting octadecylsilane chemically bonded silica are filler; Mobile phase is that ratio is 15: 10: 75 methanol-acetonitrile-1% acetums; The detection wavelength is 334nm, and theoretical cam curve is calculated by the echinacoside peak should be not less than 4000;
Reference substance solution prepares precision, and to take by weighing the echinacoside reference substance an amount of, puts in the brown bottle, adds mobile phase and be made into the reference substance solution that every 1ml contains 0.16mg;
Need testing solution preparation gets that this product is an amount of, and porphyrize is got 0.5g, accurate claim fixed, put in the brown measuring bottle of 50ml, precision adds mobile phase 20ml, and close plug shakes up, claim to decide weight, soaked 0.5 hour, supersound process 40min takes out, put coldly, claim again to decide weight, add mobile phase and supply the weight that subtracts mistake, shake up, centrifugal, get supernatant, micro-filtration membrane with 0.45 μ m filters, and filtrate is put in the brown bottle, promptly;
Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure;
Calculating is calculated as follows:
In the formula:
W: echinacoside content, %;
Cs: the concentration of echinacoside in the reference substance, mg/ml;
M: the sample weighting amount of test sample, mg;
As: the chromatographic peak area of echinacoside in the reference substance;
Ai: the chromatographic peak area of echinacoside in the test sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100280007A CN101274036B (en) | 2008-05-12 | 2008-05-12 | Method for testing conghuang preparation for tonifying kidney |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100280007A CN101274036B (en) | 2008-05-12 | 2008-05-12 | Method for testing conghuang preparation for tonifying kidney |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101274036A true CN101274036A (en) | 2008-10-01 |
| CN101274036B CN101274036B (en) | 2012-06-06 |
Family
ID=39994172
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100280007A Expired - Fee Related CN101274036B (en) | 2008-05-12 | 2008-05-12 | Method for testing conghuang preparation for tonifying kidney |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101274036B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175629A (en) * | 2010-11-23 | 2011-09-07 | 河南中医学院 | Biological activity detection-based evaluation method of quality of prepared radix rehmanniae |
| CN107543893A (en) * | 2017-08-24 | 2018-01-05 | 重庆市畜牧科学院 | The discrimination method of Semen Cuscutae in a kind of female's Huang Zheng oral liquid |
| CN109541044A (en) * | 2017-09-22 | 2019-03-29 | 广州白云山中药业有限公司 | The quality determining method of glutinous rehmannia medicinal material |
| CN109541116A (en) * | 2018-11-23 | 2019-03-29 | 北京三和药业有限公司 | A kind of discrimination method of agalloch eaglewood |
-
2008
- 2008-05-12 CN CN2008100280007A patent/CN101274036B/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175629A (en) * | 2010-11-23 | 2011-09-07 | 河南中医学院 | Biological activity detection-based evaluation method of quality of prepared radix rehmanniae |
| CN102175629B (en) * | 2010-11-23 | 2014-01-15 | 河南中医学院 | Biological activity detection-based evaluation method of quality of prepared radix rehmanniae |
| CN107543893A (en) * | 2017-08-24 | 2018-01-05 | 重庆市畜牧科学院 | The discrimination method of Semen Cuscutae in a kind of female's Huang Zheng oral liquid |
| CN109541044A (en) * | 2017-09-22 | 2019-03-29 | 广州白云山中药业有限公司 | The quality determining method of glutinous rehmannia medicinal material |
| CN109541116A (en) * | 2018-11-23 | 2019-03-29 | 北京三和药业有限公司 | A kind of discrimination method of agalloch eaglewood |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101274036B (en) | 2012-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| 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 | |
| CN105259295B (en) | Quality detection method for ginseng, cassia twig and poria cocos oral solution | |
| CN104398642A (en) | Preparation and quality detection method of compound prescription cortex phellodendri chinensis fluid | |
| CN105606734A (en) | Method for detecting honeysuckle flower and lonicerae flos medicinal materials through rapid resolution liquid chromatography | |
| CN101766771A (en) | Quality control method of medicine for gynecopathy treatment | |
| CN102119961A (en) | Detection method of compound danshen dripping pills | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN107064322A (en) | Gallic acid and the HPLC wavelength of sanguisorbin I contents switching method in garden burnet or garden burnet class preparation are determined simultaneously | |
| CN106370756B (en) | A kind of detection method of Chinese materia medica preparation that preventing and treating infectious bronchitis of chicken | |
| CN101982189A (en) | Method for detecting salvia heart-soothing capsules | |
| CN103822975B (en) | A kind of detection method of formulation of ' Sheng Mai ' of Radix Codonopsis side | |
| CN101274036B (en) | Method for testing conghuang preparation for tonifying kidney | |
| CN109085285B (en) | Quality control method of changyanning granules | |
| CN102670951B (en) | The content assaying method of Hesperidin in a kind of Chinese medicine composition | |
| CN102068573B (en) | Quality control method for stomach reinforcing and digestion promoting tangerine peel, pinellia, immature bitter orange and atractylodes macrocephala pill | |
| CN103822888B (en) | Catch up with the quality determining method of yellow grass | |
| CN110376291A (en) | A kind of coptis reference extract and its preparation method and application | |
| CN102552515A (en) | Quality detection method for blood-nourishing Chinese angelica syrup | |
| CN100543470C (en) | A kind of quality determining method of compound pyrrosia leaf particle | |
| CN108037234B (en) | Quality detection method of abrus herb hepatitis granules | |
| CN1857445B (en) | Quality control method for Desheng preparation | |
| CN1877322B (en) | High-efficiency liquid chromatography method for detecting stachydrine content in motherwort | |
| CN101816753A (en) | Method for detecting quality of compound preparation for treating cold | |
| CN101352565A (en) | Quality control method of granular formulation for activating blood and resolving stasis, detoxifying and dispersing swelling | |
| CN113759057B (en) | Characteristic spectrum of allium macrostemon white water extract and preparation thereof and construction 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 | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120606 Termination date: 20130512 |