CN102370721A - Method for detecting dredging particular Chinese medicinal preparation - Google Patents
Method for detecting dredging particular Chinese medicinal preparation Download PDFInfo
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- CN102370721A CN102370721A CN2010102638343A CN201010263834A CN102370721A CN 102370721 A CN102370721 A CN 102370721A CN 2010102638343 A CN2010102638343 A CN 2010102638343A CN 201010263834 A CN201010263834 A CN 201010263834A CN 102370721 A CN102370721 A CN 102370721A
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Abstract
The invention relates to a method for detecting a dredging Chinese medicinal preparation, in particular to a method for detecting dredging particles. The detecting method comprises a medicament ingredient identifying method and a ferulic acid content measuring method.
Description
Technical field
The present invention relates to normal open easypro Chinese medicine preparation detection method, the particularly detection method of normal open relaxation grain.
Background technology
The normal open relaxation grain records in drug standard (WS promulgated by the ministries or commissions of the Central Government
3-B-2031-95), form by five tastes medical materials such as Radix Angelicae Sinensis, Radix Polygoni Multiflori, Radix Paeoniae Rubra, be mainly used in nourishing YIN and benefiting blood, loosening bowel to relieve constipation.Habitual constipation, diseases such as senile constipation and puerperal constipation.These article do not have content assaying method in former ministry standard; Be the control product quality; Wherein main flavour of a drug Radix Angelicae Sinensis, Radix Polygoni Multiflori have been carried out the thin layer chromatography discriminating, and the content of ferulic acid in the normal open relaxation grain that adopted high effective liquid chromatography for measuring, with this quality control standard as product.
Summary of the invention
The detection method that the object of the present invention is to provide a kind of Chinese medicine preparation normal open to relax.
Chinese medicine preparation of the present invention is a granule.
Detection method of the present invention comprises the discrimination method and the ferulaic acid content assay method of drug ingredient.
Wherein, said discrimination method may further comprise the steps:
A. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets the paeoniflorin reference substance, adds ethanol and processes the solution that every 1ml contains 2mg, as reference substance solution.According to the thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with chloroform-methanol (4: 1), launch, take out, to dry, spray is with 5% vanillin sulfuric acid solution, about 3 minutes of 105 ℃ of bakings.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical bluish violet speckle.
B. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets Radix Polygoni Multiflori control medicinal material 0.25g and shines medical material solution in pairs with legal system.According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose; With chloroform-methanol (7: 3) is developing solvent, and exhibition is during to about 3.5cm, taking-up; Drying, is developing solvent with chloroform-methanol (20: 1) again, and exhibition is to about 7cm; Take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of identical color.
C. get these article 5g porphyrize, the 30ml that adds diethyl ether, supersound process 30 minutes filters, and filtrating volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis, each 0.5g of control medicinal material, and the 10ml that adds diethyl ether respectively shines medical material solution in pairs with legal system.According to thin layer chromatography,, draw each 5 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane-ethyl acetate (9: 1), launch; Take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
Wherein, content assaying method may further comprise the steps:
Measure according to HPLC (version appendix VID in 2005)
Chromatographic condition and system suitability condition are filler with the octadecyl silane, and methanol-1% acetum (30: 70) is a mobile phase; The detection wavelength is 316nm; Number of theoretical plate calculates by the ferulic acid peak should be not less than 5000
The preparation of reference substance solution: it is an amount of to get the ferulic acid reference substance, and accurate the title decides, and puts in the brown bottle, adds 70% methanol and processes the solution that every 1ml contains 20ug, promptly gets;
The preparation of need testing solution: get that these article powder 1g is accurate to be claimed surely, put in the tool plug conical flask, the accurate 30%-100% methanol 25-50ml that adds, close plug is claimed decide weight, and supersound process 30-70 minute, put coldly, claim again to decide weight, shake up, subsequent filtrate is got in filtration, promptly gets;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and promptly get.
These article contain Radix Angelicae Sinensis in ferulic acid (C10H10O4) for every bag, must not be less than 2.0mg (every bag in 20g).
Detection method of the present invention has not only increased the qualitative identification method but also has increased the Content Measurement of Effective Ingredient in Happiness method, has improved the specificity and the quality stability of Chinese medicine preparation quality of the present invention, has guaranteed the safety and the effectiveness of people's medications.Detection method speed of the present invention is fast, and accuracy is high, has guaranteed the quality of medicine.
The specific embodiment
Through following specific embodiment the present invention is described further, but does not play restriction.
Experimental example 1The research of ferulaic acid content assay method
(1) instrument and reagent
High performance liquid chromatograph comprises Tianjin, island LC-10AT pump, SPD-10AVP dual pathways UV-detector; Tianjin, island column oven; Zhejiang University's liquid chromatograph work station, ferulic acid reference substance (lot number: 110773-20061108 supplies assay usefulness, Nat'l Pharmaceutical & Biological Products Control Institute) methanol is U.S. Tedia chromatographically pure; Water is ultra-pure water, and other reagent is analytical pure.Drug sample to be measured is a normal open relaxation grain (lot number: 090101 Jiangxi Jimin Kexin Drug Industry Co., Ltd)
2) chromatographic condition is with reference to Radix Angelicae Sinensis medical material assay
Chromatographic column: Penomenex Luna C18 (2) 4.6mm * 150mm column temperature: 30 ℃ of mobile phases: methanol-1% acetum; The detection wavelength is 316nm; Flow velocity: 1.0ml/min
3) selection of detection wavelength
Get the ferulic acid reference substance solution, in 200nm～400nm scope interscan, the result has absorption maximum at the 316nm place, so select 316nm as detecting wavelength.
4) supersound process choice of Solvent: respectively with 50% methanol, methanol as solvent, according to the operation down of assay item, (lot number: content of ferulic acid is measured 090101), and the result sees table 1 to sample.
Table 1 solvent is to measuring result's influence
Therefore, select for use methanol as solvent
5) extraction conditions selection: take by weighing testing sample respectively and carry out supersound process, water-bath reflow treatment, measure all article (lot number: 090101) in content of ferulic acid, the result sees table 2
Table 2 extraction conditions is to measuring result's influence
Therefore, select for use supersound process as extraction conditions.
6) selection of supersound process time: according to the operation down of assay item, the supersound process time was respectively 30,40,50,60,70 minutes, and (lot number does working sample; 090101) content of ferulic acid in, the result sees table 3
The table 3 supersound process time is to measuring result's influence
Therefore, the supersound process time selected for use 60 minutes.
7) blank assay
In the ratio of prescription taste of Chinese medicine, be furnished with the blank preparation that contains the normal open relaxation grain certainly, process blank solution as stated above, measure in accordance with the law, blank solution is not seen apparent chromatographic peak at the place of identical retention time with the ferulic acid reference substance as a result, so think no interference.
8) linear relationship is investigated
Accurate above-mentioned reference substance solution (0.02126mg/ml) 2,4,6,8, the 10ul of drawing injects chromatograph of liquid, measures peak area, is horizontal vertical mark with the sample introduction (ug) of reference substance, is vertical coordinate with the peak area, the drawing standard curve.The result sees table 4
Table 4 linear relationship experimental data (n=5)
Y＝300000X+17249
R＝0.9992
The result shows: in 0.0424～0.212ug scope, be good linear relationship.
9) stability test
The accurate reference substance solution 10ul that draws, interval certain hour sample introduction, the peak area of mensuration ferulic acid, BSD is: 0.23%, the result shows that the mensuration result is stable in 24 hours, the result sees table 5.
Table 5 stability test data (n=5)
10) precision test
The accurate need testing solution 10ul that draws injects chromatograph of liquid, measures the peak area of ferulic acid, repeats sample introduction 5 times, and RSD is 0.34% as a result, sees table 6
Table 6 precision test data (n=5)
11) repeatability test
According to the operation down of assay item, (lot number: 090101) sample is measured for 5 parts, and trying to achieve relative standard deviation RSD is 0.29%, and the result sees table 7 to same lot number
Table 7 repeatability test data
12) recovery test
Reclaim with application of sample; Precision take by weighing known ferulaic acid content same lot number (lot number: the about 0.5g of the test sample 0901012.61mg/ bag), it is a certain amount of that precision adds ferulic acid reference substance solution (0.206mg/ml) respectively, according to the operation down of assay item; Be calculated as follows the response rate, the result sees table 8
Table 8 average recovery experimental data (n=6)
Average recovery rate is: 98.38%RSD is 0.66%
Experimental example 2,Content of ferulic acid is measured in the normal open relaxation grain
According to the method operation of above-mentioned assay, surveyed content of ferulic acid in 10 batches of normal open relaxation grain grains, the result sees
Table 9
Table 9 normal open relaxation grain content of ferulic acid is measured
Record normal open and relax in the preparation content of ferulic acid all more than the 2.6mg/ bag; Consider the loss error in the industry amplification; Therefore on the basis of minimum content, float downward 20%, therefore obtain examination criteria and be normal open and relax and contain ferulaic acid content in every bag in the preparation and should be not less than 2.0mg.
Embodiment 3,The method that the easypro quality of the pharmaceutical preparations of normal open detects
1, differentiates detection method
A. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets the paeoniflorin reference substance, adds ethanol and processes the solution that every 1ml contains 2mg, as reference substance solution.According to the thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with chloroform-methanol (4: 1), launch, take out, to dry, spray is with 5% vanillin sulfuric acid solution, about 3 minutes of 105 ℃ of bakings.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical bluish violet speckle.
B. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets Radix Polygoni Multiflori control medicinal material 0.25g and shines medical material solution in pairs with legal system.According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose; With chloroform-methanol (7: 3) is developing solvent, and exhibition is during to about 3.5cm, taking-up; Drying, is developing solvent with chloroform-methanol (20: 1) again, and exhibition is to about 7cm; Take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of identical color.
C. get these article 5g porphyrize, the 30ml that adds diethyl ether, supersound process 30 minutes filters, and filtrating volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis, each 0.5g of control medicinal material, and the 10ml that adds diethyl ether respectively shines medical material solution in pairs with legal system.According to thin layer chromatography,, draw each 5 μ l of above-mentioned two kinds of solution; Put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane-ethyl acetate (9: 1), launch; Take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
2, assay: measure according to HPLC (version appendix VID in 2005)
Chromatographic condition and system suitability condition are filler with the octadecyl silane, and methanol-1% acetum (30: 70) is a mobile phase; The detection wavelength is 316nm; Number of theoretical plate calculates the preparation that should be not less than 5000 reference substance solution by the ferulic acid peak: it is an amount of to get the ferulic acid reference substance, and accurate the title decides, and puts in the brown bottle, adds 70% methanol and processes the solution that every 1ml contains 20ug, promptly gets; The preparation of need testing solution: get that these article powder 1g is accurate to be claimed surely, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug is claimed decide weight, and supersound process 60 minutes is put coldly, claims to decide weight again, shakes up, and subsequent filtrate is got in filtration, promptly gets; Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and promptly get.These article contain Radix Angelicae Sinensis with ferulic acid (C for every bag
10H
10O
4) meter, must not be less than 2.0mg (every bag in 20g).
Claims (5)
1. the detection method that the Chinese medicine preparation normal open relaxes is characterized in that said detection method comprises the discrimination method and the ferulaic acid content assay method of drug ingredient.
2. detection method according to claim 1 is characterized in that, said Chinese medicine preparation granule.
3. detection method according to claim 1 is characterized in that, the discrimination method of said drug ingredient may further comprise the steps:
A. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the paeoniflorin reference substance, adds ethanol and processes the solution that every 1ml contains 2mg, as reference substance solution; According to the thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1 chloroform-methanols of volume ratio, launch, take out, to dry, spray is with 5% vanillin sulfuric acid solution, about 3 minutes of 105 ℃ of bakings.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical bluish violet speckle;
B. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.25g and shines medical material solution in pairs with legal system; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose; With 7: 3 chloroform-methanol of volume ratio is developing solvent, and exhibition is during to about 3.5cm, taking-up; Drying, is developing solvent with 20: 1 chloroform-methanol of volume ratio again, opens up to 7cm; Take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of identical color;
C. get these article 5g porphyrize, the 30ml that adds diethyl ether, supersound process 30 minutes filters, and filtrating volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis, each 0.5g of control medicinal material, and the 10ml that adds diethyl ether respectively shines medical material solution in pairs with legal system; According to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 9: 1 normal hexane-ethyl acetates of volume ratio is developing solvent, launches, and takes out; Dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
4. detection method according to claim 1 is characterized in that, said ferulaic acid content assay method may further comprise the steps:
According to version appendix VID high effective liquid chromatography for measuring in 2005
Chromatographic condition and system suitability condition are filler with the octadecyl silane, are mobile phase with 30: 70 methanol-1% of volume ratio acetum; The detection wavelength is 316nm; Number of theoretical plate calculates by the ferulic acid peak should be not less than 5000;
The preparation of reference substance solution: it is an amount of to get the ferulic acid reference substance, and accurate the title decides, and puts in the brown bottle, adds 70% methanol and processes the solution that every 1ml contains 20ug, promptly gets;
The preparation of need testing solution: get that these article powder 1g is accurate to be claimed surely, put in the tool plug conical flask, the accurate 30%-100% methanol 25-50ml that adds, close plug is claimed decide weight, and supersound process 30-70 minute, put coldly, claim again to decide weight, shake up, subsequent filtrate is got in filtration, promptly gets;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and promptly get;
These article contain Radix Angelicae Sinensis in ferulic acid for every bag, must not be less than 2.0mg.
5. detection method according to claim 1 is characterized in that, said discrimination method may further comprise the steps:
A. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the paeoniflorin reference substance, adds ethanol and processes the solution that every 1ml contains 2mg, as reference substance solution; According to the thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; With 4: 1 chloroform-methanols of volume ratio is developing solvent, launches, and takes out; Dry, spray is with 5% vanillin sulfuric acid solution, about 3 minutes of 105 ℃ of bakings; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical bluish violet speckle;
B. get these article 10g, porphyrize adds ethanol 20ml, and reflux 1 hour filters, the filtrating evaporate to dryness, and residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets Radix Polygoni Multiflori control medicinal material 0.25g and shines medical material solution in pairs with legal system; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose; With 7: 3 chloroform-methanol of volume ratio is developing solvent, and exhibition is during to about 3.5cm, taking-up; Drying, is developing solvent with 20: 1 chloroform-methanol of volume ratio again, opens up to 7cm; Take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of identical color;
C. get these article 5g porphyrize, the 30ml that adds diethyl ether, supersound process 30 minutes filters, and filtrating volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis, each 0.5g of control medicinal material, and the 10ml that adds diethyl ether respectively shines medical material solution in pairs with legal system; According to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose; With 9: 1 normal hexane-ethyl acetates of volume ratio is developing solvent, launches, and takes out; Dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
Said content assaying method may further comprise the steps:
According to version appendix VID high effective liquid chromatography for measuring in 2005
Chromatographic condition and system suitability condition: with 30: 70 octadecyl silanes of volume ratio is filler, and methanol-1% acetum is a mobile phase; The detection wavelength is 316nm; Number of theoretical plate calculates by the ferulic acid peak should be not less than 5000;
The preparation of reference substance solution: it is an amount of to get the ferulic acid reference substance, and accurate the title decides, and puts in the brown bottle, adds 70% methanol and processes the solution that every 1ml contains 20ug, promptly gets;
The preparation of need testing solution: get that these article powder 1g is accurate to be claimed surely, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug is claimed decide weight, and supersound process 60 minutes is put coldly, claims to decide weight again, shakes up, and subsequent filtrate is got in filtration, promptly gets;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and promptly get, and these article contain Radix Angelicae Sinensis in ferulic acid for every bag, must not be less than 2.0mg.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102680593A (en) * | 2011-04-19 | 2012-09-19 | 四川川大华西药业股份有限公司 | Method for detecting quality of Lemai granules |
| CN106727295A (en) * | 2016-12-28 | 2017-05-31 | 江西济民可信药业有限公司 | A kind of normal open oral liquid for relaxation and preparation method thereof |
-
2010
- 2010-08-26 CN CN2010102638343A patent/CN102370721A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102680593A (en) * | 2011-04-19 | 2012-09-19 | 四川川大华西药业股份有限公司 | Method for detecting quality of Lemai granules |
| CN102680593B (en) * | 2011-04-19 | 2013-11-27 | 四川川大华西药业股份有限公司 | Method for detecting medicine composition promoting qi to activate blood and dispersing blood stasis |
| CN106727295A (en) * | 2016-12-28 | 2017-05-31 | 江西济民可信药业有限公司 | A kind of normal open oral liquid for relaxation and preparation method thereof |
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Application publication date: 20120314 |