CN102914615A - Detection method for compound desert cistanche yizhi capsule - Google Patents
Detection method for compound desert cistanche yizhi capsule Download PDFInfo
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Abstract
The invention provides a detection method for a compound desert cistanche yizhi capsule. The detection method comprises the following steps: (A) observing the characters of contents; (B) identifying polygonum multiflorum, lotus leaves, cistanche salsa, earthworm and uniflower swisscentaury root in a formula by using thin-layer chromatography; (C) checking various related stipulations meeting the capsule item according to one appendix IL of Chinese pharmacopoeia 2010; and (D) determining the content of active components of the polygonum multiflorum in the manner by using high performance liquid chromatography. Due to the adoption of the method, the components and contents of the compound desert cistanche yizhi capsule can be qualitatively and quantitatively detected well, the quality of medicines is ensured and further the safety and the effectiveness for medicine use are improved.
Description
Technical field
The present invention relates to the drug inspection technical field.Specifically, the present invention relates to a kind of detection method of Chinese patent drug.
Background technology
Tradition Chinese patent drug detection method can only qualitative explanation Chinese patent drug composition, the composition of efficient and quantitative detection Chinese patent drug is the technical matters that modern TCM industry is badly in need of solution.
Compound desert cistanche Cognex is made through series of process such as pulverizing, extract, decoct, filter, be dry by five kinds of natural drugs, has good intelligence development nourishing the liver, activating blood circulation to eliminate turbid, the effect of brain.Be used for light, the moderate vascular dementia caused by liver and kidney deficiency phlegm stagnation in collateral of holding concurrently, to hypophrenia, thought slowness, look dull, forgetful, or angry indefinite, the soreness and weakness of waist and knees of happiness, dizziness and tinnitus, insomnia and dreamful sleep etc. have preferably curative effect.
Compound desert cistanche Cognex is hard shell capsules, and content is tan powder; Gas is little, salty, little hardship of distinguishing the flavor of.Mainly made by prepared fleece flower root, lotus leaf, saline cistanche, earthworm and Radix Rhapontici seu Radix Echinopsis, wherein, that the fleece-flower root has is anti-ageing, reducing blood lipid, promotion are erythrocyticly grown, antiatherosclerosis, the effect of increase coronary blood flow and the effect of neuroprotective unit; Saline cistanche has kidney-replenishing, benefiting essence-blood, and the effect that relaxes bowel can be adjusted endocrine, promotes metabolism, delay senility; Earthworm has stronger anticoagulation, anti-thrombosis function, and can promote established thrombus to dissolve gradually; Radix Rhapontici seu Radix Echinopsis has the hyperlipidemia of improvement, alleviates arteriosclerotic lesion, reduces lipid peroxide level in the serum, reduces the effect that the heart, brain, kidney etc. are organized lipofuscin content; Lotus leaf has the effect of reducing blood lipid.
The prescription of compound desert cistanche Cognex is as follows:
Prepared fleece flower root 833g lotus leaf 583g
Saline cistanche 417g earthworm 417g
Radix Rhapontici seu Radix Echinopsis 250g
The method for making of compound desert cistanche Cognex is as follows: the above five tastes, prepared fleece flower root is ground into meal, with 75% alcohol reflux secondary, adding for the first time 8 times of amounts extracted 1 hour, adding for the second time 6 times of amounts extracted 1 hour, merge ethanol extract, being concentrated into relative density is the clear cream of 1.28～1.30 (60 ℃).Lotus leaf, saline cistanche, Radix Rhapontici seu Radix Echinopsis boiling secondary add 12 times of water gagings for the first time, decoct 2 hours, for the second time add 10 times of water gagings, decocted collecting decoction 1 hour, filter, it is 1.04～1.05 (60 ℃) that filtrate is concentrated into relative density, lets cool, adding ethanol makes and contains alcohol amount and reach 60%, refrigerate 12 hours, filter, filtrate is concentrated into the clear cream that relative density is 1.28～1.30 (60 ℃), merge 80 ℃ of following vacuum (0.08MPa) drying with above-mentioned clear cream.Earthworm is ground into meal, adds 4 times of water gaging homogenate 5 times, each 15 minutes, merge homogenate, add ethanol and make and contain the alcohol amount and be 10%, centrifugal, get supernatant and add ethanol and make and contain alcohol amount and reach 50%, centrifugal, sediment washs secondary with acetone, centrifugal, sediment vacuum (0.08MPa) drying merges with above-mentioned dry extract, add an amount of dextrin, be ground into middle powder, incapsulate, make 1000, and get final product.
The method for making of compound desert cistanche Cognex is complicated, and its curative effect is subjected to the manufacture craft of quality, content and the medicament of medicinal material to affect larger.Yet existing detection method, qualitative and quantitative analysis are not comprehensive, can't better guarantee the quality of medicine, and then security and validity that medicine is used constitute a threat to.
Summary of the invention
The present invention provides one to overlap comprehensively and effectively detection method for solving above-mentioned the deficiencies in the prior art, strictly controls the quality stability of compound desert cistanche Cognex.
The detection method of compound desert cistanche Cognex provided by the invention comprises the steps:
A) content character observation;
B) fleece-flower root, lotus leaf, saline cistanche, earthworm and Radix Rhapontici seu Radix Echinopsis in the thin-layered chromatography discriminating side;
C) according to appendix IL of Chinese Pharmacopoeia version in 2010, inspection should meet every regulation relevant under the capsule item; With
D) content of the fleece-flower root in the high effective liquid chromatography for measuring side.
In the present invention, steps A) observation is that characters powder is observed, and namely compound desert cistanche Cognex medicine is observed, and its content should be tan powder; Gas is little, salty, little hardship of distinguishing the flavor of.
Step B) discriminating comprises:
(1) get compound desert cistanche Cognex content 2g, add acetone 15ml, ultrasonic processing 40 minutes filters, and filtrate is made need testing solution.Other gets fleece-flower root control medicinal material 2g, is made in the same way of control medicinal material solution.Get again the archen reference substance, add methyl alcohol and make the solution that every 1ml contains 0.1mg, in contrast product solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2010), draw need testing solution 15 μ l, reference substance medicinal material solution 3 μ l, reference substance solution 1 μ l, put respectively on same silica gel g thin-layer plate, take the upper solution of sherwood oil (60～90 ℃)-ethyl formate-formic acid (volume ratio 15: 5: 1) as developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material, the corresponding position of reference substance chromatogram on, the fluorescence spot of aobvious same color.
(2) get compound desert cistanche Cognex content 4g, add methyl alcohol 50ml, added hot reflux 30 minutes, filter, filtrate evaporate to dryness, residue add 1% hydrochloric acid solution 10ml makes dissolving, filters, filtrate adds strong ammonia solution and regulates pH value to 9～10, extract 3 times with the methenyl choloride jolting, each 10ml merges methenyl choloride liquid, evaporate to dryness, residue makes dissolving with a small amount of methenyl choloride, by neutral alumina column (100～200 orders, 1g, internal diameter 0.8cm), use the methenyl choloride wash-out, collect eluent 5ml, evaporate to dryness, residue adds methenyl choloride 0.2ml makes dissolving, as need testing solution.Other gets lotus leaf control medicinal material 2g, adds 60% ethanol 40ml, refluxes 20 minutes, filters, and filtrate is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take methenyl choloride-acetone-strong ammonia solution (volume ratio 6: 2: 0.1) as developping agent, launch, take out, dry, spray is with the improvement bismuth potassium iodide test solution.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious same color spot.
(3) get compound desert cistanche Cognex content 1g, add methyl alcohol 10ml, ultrasonic processing 10 minutes filters, and filtrate is as need testing solution.Other gets saline cistanche control medicinal material 2g, adds methyl alcohol 5ml, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 2 μ l of above-mentioned two kinds of solution, put respectively in same polyamide film (on 8 * 8cm), take methyl alcohol-glacial acetic acid-water (volume ratio 2: 1: 7) as developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material chromatogram relevant position on, the fluorescence spot of aobvious same color.
(4) get compound desert cistanche Cognex content 1g, add 20% ethanol 10ml, ultrasonic processing 20 minutes filters, and filtrate is collected eluent, as need testing solution by granule activated carbon post (0.5g, high 4.0cm * diameter 0.8cm).Other gets earthworm control medicinal material 0.5g, puts in the mortar, adds water and grinds 2 times, each 5ml, centrifugal, merge supernatant, add absolute ethyl alcohol 15ml, shake up, left standstill 30 minutes, centrifugal, abandoning supernatant, the collecting precipitation thing, with 60% ethanol 2ml washing once, sediment adds 20% ethanol 2.5ml makes dissolving, gets in contrast medicinal material solution of supernatant.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 5 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol one glacial acetic acid one water (volume ratio 7: 2: 1) as developping agent, launch, take out, dry, spray is with ninhydrin solution, 105 ℃ of heating 5～10 minutes.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.
(5) get compound desert cistanche Cognex content 1g, add methyl alcohol 20ml, ultrasonic processing 20 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Rhapontici seu Radix Echinopsis control medicinal material 1g, be made in the same way of control medicinal material solution, test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-butanone (4: 1) as developping agent, launch, take out, dry, put under the uviol lamp (365nm) and inspect.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.
Step C) inspection: according to an appendix I of Chinese Pharmacopoeia version in 2010 L, inspection should meet every regulation relevant under the capsule item.That is, moisture, content uniformity, disintegration time limited, heavy metal inspection, arsenic salt check, limit test of microbe should be up to specification.
Step D) assay comprises:
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 D).
Chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take methyl alcohol-acetonitrile-water-phosphoric acid (volume ratio 10: 20: 110: 0.01) as mobile phase; The detection wavelength is 320nm.Number of theoretical plate by 2,3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides peak calculates and should be not less than 2500.
The preparation of reference substance solution is got the phosphorus pentoxide drying under reduced pressure to 2 of constant weight, 3,5,4 ＇-tetrahydroxystilbene-2-O-β-D-Glucose glycosides reference substance 1.0mg, accurately weighed, put the brown measuring bottle of 50ml, add the methyl alcohol dissolving and be diluted to scale, shake up, namely get and (contain 2 among every 1ml, 3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides 0.02mg) (keeping in Dark Place).
Content under the compound desert cistanche Cognex content uniformity item is got in the preparation of need testing solution, is ground into fine powder, mixing, get approximately 0.15g of fine powder, accurately weighed, put in the tool plug conical flask, accurate methyl alcohol 20ml, close plug, the weighed weight of adding, ultrasonic processing (power 100W, frequency 40KHZ) 20 minutes is taken out, let cool, weighed weight is again supplied the weight of less loss with methyl alcohol, shake up, supernatant filters with miillpore filter (0.45 μ m), namely gets (keeping in Dark Place).
Determination method is accurate reference substance solution and each 10 μ l of need testing solution of drawing respectively, and the injection liquid chromatography is measured, and be get final product.
Every of described compound desert cistanche Cognex contains prepared fleece flower root with 2,3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides (C
20H
22O
9) meter, must not be less than 0.70mg.
By above-mentioned detection method, can do good qualitative detection and quantitatively detect the quality of assurance medicine, thereby security and validity that the raising medicine uses composition and the content of compound desert cistanche Cognex.
Claims (3)
1. the detection method of a compound desert cistanche Cognex is characterized in that, the method is comprised of the following step:
A) content character observation;
B) fleece-flower root, lotus leaf, saline cistanche, earthworm and Radix Rhapontici seu Radix Echinopsis in the thin-layered chromatography discriminating side;
C) according to appendix IL of Chinese Pharmacopoeia version in 2010, inspection should meet every regulation relevant under the capsule item; With
D) content of fleece-flower root active component in the high effective liquid chromatography for measuring side.
2. the detection method of compound desert cistanche Cognex according to claim 1 is characterized in that,
Described step B) step is as follows:
(1) get compound desert cistanche Cognex content 2g, add acetone 15ml, ultrasonic processing 40 minutes filters, and filtrate is made need testing solution; Other gets fleece-flower root control medicinal material 2g, is made in the same way of control medicinal material solution; Get again the archen reference substance, add methyl alcohol and make the solution that every 1ml contains 0.1mg, in contrast product solution; Test according to thin-layered chromatography, draw need testing solution 15 μ l, reference substance medicinal material solution 3 μ l, reference substance solution 1 μ l, put respectively on same silica gel g thin-layer plate, take the sherwood oil-ethyl formate of 15: 5: 1 ratios of volume ratio-formic acid upper solution as developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material, the corresponding position of reference substance chromatogram on, the fluorescence spot of aobvious same color;
(2) get compound desert cistanche Cognex content 4g, add methyl alcohol 50ml, add hot reflux 30 minutes, and filtered the filtrate evaporate to dryness, residue adds 1% hydrochloric acid solution 10ml makes dissolving, filter, filtrate adds strong ammonia solution and regulates pH value to 9～10, extracts 3 times with the methenyl choloride jolting, each 10ml, merge methenyl choloride liquid, evaporate to dryness, residue makes dissolving with a small amount of methenyl choloride, pass through neutral alumina column, use the methenyl choloride wash-out, collect eluent 5ml, evaporate to dryness, residue adds methenyl choloride 0.2ml makes dissolving, as need testing solution; Other gets lotus leaf control medicinal material 2g, adds 60% ethanol 40ml, refluxes 20 minutes, filters, and filtrate is made in the same way of control medicinal material solution; According to thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the methenyl choloride-acetone of 6: 2: 0.1 ratios of volume ratio-strong ammonia solution as developping agent, launch, take out, to dry, spray is with the improvement bismuth potassium iodide test solution; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious same color spot;
(3) get compound desert cistanche Cognex content 1g, add methyl alcohol 10ml, ultrasonic processing 10 minutes filters, and filtrate is as need testing solution; Other gets saline cistanche control medicinal material 2g, adds methyl alcohol 5ml, is made in the same way of control medicinal material solution; According to thin-layered chromatography test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, take the methyl alcohol-glacial acetic acid of 2: 1: 7 ratios of volume ratio-water as developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material chromatogram relevant position on, the fluorescence spot of aobvious same color;
(4) get compound desert cistanche Cognex content 1g, add 20% ethanol 10ml, ultrasonic processing 20 minutes filters, and filtrate is collected eluent, as need testing solution by the granule activated carbon post; Other gets earthworm control medicinal material 0.5g, puts in the mortar, adds water and grinds 2 times, each 5ml, centrifugal, merge supernatant, add absolute ethyl alcohol 15ml, shake up, left standstill 30 minutes, centrifugal, abandoning supernatant, the collecting precipitation thing, with 60% ethanol 2ml washing once, sediment adds 20% ethanol 2.5ml makes dissolving, gets in contrast medicinal material solution of supernatant; Test according to thin-layered chromatography, draw need testing solution 5 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol one glacial acetic acid one water of 7: 2: 1 ratios of volume ratio as developping agent, launch, take out, dry, spray is with ninhydrin solution, 105 ℃ of heating 5～10 minutes; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;
(5) get compound desert cistanche Cognex content 1g, add methyl alcohol 20ml, ultrasonic processing 20 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Rhapontici seu Radix Echinopsis control medicinal material 1g, be made in the same way of control medicinal material solution, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take cyclohexane-butanone of 4: 1 of volume ratio as developping agent, launch, take out, dry, put under the uviol lamp (365nm) and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.
3. the detection method of compound desert cistanche Cognex according to claim 1 and 2 is characterized in that,
Described step D) step is as follows:
Chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take 10: 20: 110: the methyl alcohol-acetonitrile-water of 0.01 volume ratio-phosphoric acid was mobile phase; The detection wavelength is 320nm; Number of theoretical plate by 2,3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides peak calculates and should be not less than 2500;
The preparation of reference substance solution is got the phosphorus pentoxide drying under reduced pressure to 2,3,5 of constant weight, 4 ＇-tetrahydroxystilbene-2-O-β-D-Glucose glycosides reference substance 1.0mg, accurately weighed, put the brown measuring bottle of 50ml, add the methyl alcohol dissolving and be diluted to scale, shake up, and get final product, contain 2 among every 1ml, 3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides 0.02mg, keep in Dark Place;
Content under the compound desert cistanche Cognex content uniformity item is got in the preparation of need testing solution, is ground into fine powder, mixing, get approximately 0.15g of fine powder, accurately weighed, put in the tool plug conical flask, accurate methyl alcohol 20ml, close plug, the weighed weight of adding, ultrasonic processing 20 minutes is taken out, and lets cool, weighed weight is again supplied the weight of less loss with methyl alcohol, shake up, supernatant filters with miillpore filter, and get final product, and keeps in Dark Place;
Determination method is accurate reference substance solution and each 10 μ l of need testing solution of drawing respectively, and the injection liquid chromatography is measured, and be get final product;
Every of described compound desert cistanche Cognex contains prepared fleece flower root with 2,3,5,4 '-tetrahydroxystilbene-2-O-β-D-Glucose glycosides meter, must not be less than 0.70mg.
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Cited By (5)
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| CN107929374A (en) * | 2017-12-04 | 2018-04-20 | 辽源誉隆亚东药业有限责任公司 | preparation method of traditional Chinese medicine preparation for treating senile dementia |
| CN108802260A (en) * | 2018-08-17 | 2018-11-13 | 四川新绿色药业科技发展有限公司 | A kind of thin-layer identification method for lotus leaf medicinal material, medicine materical crude slice and granule quality control |
| 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 |
| CN114306491A (en) * | 2021-12-22 | 2022-04-12 | 江苏七○七天然制药有限公司 | Fleece-flower root hair growth capsule and preparation, detection and component identification methods thereof |
| CN114720600A (en) * | 2022-04-07 | 2022-07-08 | 广东安诺药业股份有限公司 | Method for detecting stilbene glucoside component in polygonum multiflorum prepared by bone-protecting capsule |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107929374A (en) * | 2017-12-04 | 2018-04-20 | 辽源誉隆亚东药业有限责任公司 | preparation method of traditional Chinese medicine preparation for treating senile dementia |
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| CN108802260A (en) * | 2018-08-17 | 2018-11-13 | 四川新绿色药业科技发展有限公司 | A kind of thin-layer identification method for lotus leaf medicinal material, medicine materical crude slice and granule quality control |
| 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 |
| CN114306491A (en) * | 2021-12-22 | 2022-04-12 | 江苏七○七天然制药有限公司 | Fleece-flower root hair growth capsule and preparation, detection and component identification methods thereof |
| CN114720600A (en) * | 2022-04-07 | 2022-07-08 | 广东安诺药业股份有限公司 | Method for detecting stilbene glucoside component in polygonum multiflorum prepared by bone-protecting capsule |
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Application publication date: 20130206 |