CN101757049A - Ginkgo leaf extractive and method for extracting total ginkgoic acid in preparation thereof - Google Patents
Ginkgo leaf extractive and method for extracting total ginkgoic acid in preparation thereof Download PDFInfo
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- CN101757049A CN101757049A CN200910079854A CN200910079854A CN101757049A CN 101757049 A CN101757049 A CN 101757049A CN 200910079854 A CN200910079854 A CN 200910079854A CN 200910079854 A CN200910079854 A CN 200910079854A CN 101757049 A CN101757049 A CN 101757049A
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Abstract
The invention provides a ginkgo leaf extractive and a method for extracting total ginkgoic acid in a preparation thereof. The method comprises the following steps of: adding a certain amount of ethyl acetate or toluene or chloroform in a proper amount of ginkgo leaf extractives or the preparation thereof for heating and refluxing, cooling and then weighing the weight of the mixture, complementing the reduced weight, filtering and measuring subsequent filtrate, recovering a solvent in reduced pressure to the dry state, adding water to dissolve the solvent and transferring the solvent to a separating funnel, extracting by using petroleum ether (60-90 DEG C) or normal hexane or the chloroform, merging the extractive liquid, drying by distillation and then determining the volume by using methyl alcohol. The method is simple, can be used for effectively extracting the trace amount of total ginkgoic acid (less than 1 ppm) in the ginkgo leaf extractive or the preparation thereof, achieve the effective separation and enrichment, simultaneously detect five monomers of the ginkgoic acid and quantitatively detect the five monomers.
Description
Technical field
The present invention relates to the process for extracting of total ginkgolic acids in Folium Ginkgo extract and its preparation.
Background technology
Ginkgoic acid is the component that another except flavone and lactone has important physiologically active in the Folium Ginkgo, it is 6-alkyl or the salicylic derivant of 6-thiazolinyl, side chain carbon number on 6 can from 13 to 19, the pendant double bonds number can be 0 to 3, therefore ginkgoic acid is a homology mixture, and common ginkgoic acid structure is:
That multinomial studies show that, ginkgoic acid have is antibiotic, antiinflammatory, antiallergic, anti-tumor activity, has important medicinal exploitation and is worth, and becomes one of focus of current people's research.But according to another report, ginkgoic acid also has immunotoxicity and fetal toxicity effect, can cause dermatitis rhus sample dermatitis, is the toxic component in the Semen Ginkgo, and human beings'health is had potential harm.
The content of ginkgoic acid must be less than 5 μ g/g in Germany's Ministry of Public Health regulation Folium Ginkgo extract (EGb) preparation.The limit of total ginkgolic acids must not be 10ppm in middle Semen Ginkgo extrac of version Chinese Pharmacopoeia in 2005, and this limit can satisfy Quality Control and purpose of safety in the Folium Ginkgo oral formulations.But for the high YINXINGYE ZHUSHEYE of security requirement, this limit can not satisfy far away.Therefore in the Folium Ginkgo injection type, limit requires be 1ppm.Therefore how the ginkgoic acid that content in the Folium Ginkgo is atomic extracts enrichment has just become primary task so that carry out effective analyzing and testing.
Folium Ginkgo extract is to be raw material with the Folium Ginkgo, is solvent with the acetone or alcohol, products obtained therefrom behind multistep extraction, separation and purification.Relative Folium Ginkgo, impurity component wherein significantly reduces, and the content of ginkgoic acid also reduces.Usually the analysis of ginkgoic acid needs to measure after with ginkgoic acid separation and concentration wherein by pretreatment again in the Folium Ginkgo extract, and performance liquid chromatographic column itself can't be realized effective separation of ginkgoic acid from Folium Ginkgo extract.Owing to the used preprocess method difference of domestic different departments, the measurement result difference of same sample is very big at present.Therefore, accurately measure ginkgoic acid content, one of key is to set up the appropriate pretreatment method, and ginkgoic acid is separated and enrichment from Folium Ginkgo extract and preparation thereof fully.
The analytical method report to ginkgoic acid in Folium Ginkgo extract and the preparation thereof was less in the document in the past, and the separation method of mentioning has supercritical fluid extraction, the mode that extraction and post eluting combine; The extraction of total ginkgolic acids is with petroleum ether (60～90 ℃) reflux under middle Folium Ginkgo extract item of Chinese Pharmacopoeia version in 2005.Preceding two kinds of method complicated operations, the response rate is low, and can only detect 2 to 4 kinds of monomeric substances of ginkgoic acid.Though it is easy and simple to handle that official method is planted in the back, Semen Ginkgo extrac in the time of also can only detecting total ginkgolic acids content greater than 1ppm and preparation thereof.
Show through relevant patent and prior art literature search result, there is no the identical content report of the present invention.
Summary of the invention
The present invention overcomes the deficiencies in the prior art exactly, has proposed a kind of extracting method of easy and simple to handle, total ginkgolic acids that the response rate is high.Use conventional extraction means, to the trace total ginkgolic acids in Folium Ginkgo extract and the preparation thereof (＜1ppm) reached effective separation and enrichment, and can detect 5 kinds of monomers of ginkgoic acid simultaneously.
The technical scheme that adopts for solving the problems of the technologies described above may further comprise the steps:
(a) get Folium Ginkgo extract or its preparation is an amount of, add a certain amount of organic reagent reflux certain hour; Put coldly, claim decide weight, supply the weight that subtracts mistake, filtration; Get subsequent filtrate, decompression and solvent recovery is to doing; (b) add water and dissolve in right amount and be transferred in the separatory funnel, with an amount of organic reagent extraction; (c) combining extraction liquid, evaporate to dryness, with methanol constant volume to 2ml.
Among the step a of the present invention, the organic reagent that adds in the reflux vessel can be ethyl acetate or cyclohexane extraction or toluene or chloroform, considers from economy and environmental angle, is preferably ethyl acetate;
The amount that adds the organic solvent in the reflux vessel is adjusted according to the amount of Folium Ginkgo extract or its preparation, is generally 15～30 times that are carried thing, is preferably 18-25 doubly; Return time 1.5～4 hours is preferably 2～3 hours.
Add water among the step b of the present invention and dissolve in right amount, the water yield can be 15～30ml, is preferably 20ml.
Among the present invention among the step b selection of Extractant can be petroleum ether (60～90 ℃), or be normal hexane, or chloroform.Consider from economy and environmental angle, be preferably petroleum ether (60～90 ℃).
The consumption of above-mentioned extractant can be 15～30ml, is preferably 20ml.
Above-mentioned extraction times can be 2～4 times, be preferably 3 times.
Description of drawings
Fig. 1 is the HPLC figure of total ginkgolic acids in the embodiment 1 gained sample Folium Ginkgo extract.
Fig. 2 is the HPLC figure of total ginkgolic acids in the embodiment 5 gained sample ginkgo leaf extract preparations.
Specific embodiment
Embodiment 1
Get Folium Ginkgo extract 3g, the accurate ethyl acetate 50ml that adds claims decide weight, and reflux, extract, 2 hours is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with ethyl acetate, shakes up filtration.Get subsequent filtrate, decompression and solvent recovery is to doing, and adds the about 20ml dissolving of water and is transferred in the separatory funnel, extract 3 times with petroleum ether (60～90 ℃), and each 20ml, merging petroleum ether liquid, evaporate to dryness to 2ml, promptly gets specimen solution with methanol constant volume.
Get Folium Ginkgo extract 3g, the accurate chloroform 50ml that adds claims decide weight, and reflux, extract, 2.5 hours is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shakes up filtration.Get subsequent filtrate, decompression and solvent recovery is to doing, and adds the about 25ml dissolving of water and is transferred in the separatory funnel, extract 3 times with petroleum ether (60～90 ℃), and each 20ml, merging petroleum ether liquid, evaporate to dryness to 2ml, promptly gets specimen solution with methanol constant volume.
Embodiment 3
Get Folium Ginkgo extract 3g, the accurate toluene 75ml that adds claims decide weight, and reflux, extract, 3 hours is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with toluene, shakes up filtration.Get subsequent filtrate, decompression and solvent recovery is to doing, and add the about 30ml dissolving of water and be transferred in the separatory funnel, with normal hexane extraction 3 times, each 30ml, merging normal hexane liquid, evaporate to dryness to 2ml, promptly gets specimen solution with methanol constant volume.
Get Folium Ginkgo extract 3g, the accurate cyclohexane extraction 60ml that adds claims decide weight, and reflux, extract, 3 hours is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with cyclohexane extraction, shakes up filtration.Get subsequent filtrate, decompression and solvent recovery is to doing, and add the about 20ml dissolving of water and be transferred in the separatory funnel, with chloroform extraction 4 times, 20ml at every turn, combined chloroform liquid, evaporate to dryness to 2ml, promptly gets specimen solution with methanol constant volume.
Get ginkgo leaf extract preparation freeze-dried powder 21g, the accurate ethyl acetate 350ml that adds claims decide weight, and reflux, extract, 3 hours is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with ethyl acetate, shakes up filtration.Get subsequent filtrate, decompression and solvent recovery is to doing, and add the about 25ml dissolving of water and be transferred in the separatory funnel, with chloroform extraction 4 times, 25ml at every turn, combined chloroform liquid, evaporate to dryness to 2ml, promptly gets specimen solution with methanol constant volume.
The Folium Ginkgo extract that will obtain respectively by the extracting method of embodiment 1 and embodiment 5 and the specimen solution of preparation are measured according to the testing conditions of total ginkgolic acids in Folium Ginkgo extract of Chinese Pharmacopoeia version in 2005.
Detection method:
Instrument: island Tianjin-high performance liquid chromatograph (LC-10AT
VPThe solvent delivery pump, SPD-10AT
VPThe UV, visible light detector)
Reagent: glacial acetic acid (analytical pure), methanol (chromatographically pure), water are the secondary redistilled water
Chromatographic column: KromasilC18,250mm * 4.6mm, 5 μ m
Mobile phase: methanol-1% glacial acetic acid solution (90: 10)
Detect wavelength: 310nm, column temperature: 40 ℃
Flow velocity: 1.0ml/min
Sample introduction: 20 μ l
Measurement result sees Table 1,2.
Table 1 embodiment 1 gained sample solution testing result table
Table 2 embodiment 5 gained sample solution testing result tables
Claims (7)
1. the extracting method of total ginkgolic acids in a Folium Ginkgo extract and its preparation is characterized in that it mainly may further comprise the steps:
(a) get Folium Ginkgo extract or its preparation is an amount of, add a certain amount of ethyl acetate or cyclohexane extraction or toluene or chloroform reflux certain hour; Put coldly, claim decide weight, supply the weight that subtracts mistake, filtration; Get subsequent filtrate, decompression and solvent recovery is to doing; (b) add water and dissolve in right amount and be transferred in the separatory funnel, with petroleum ether (60～90 ℃) or normal hexane or chloroform extraction; (c) combining extraction liquid, evaporate to dryness, with methanol constant volume to 2ml.
2. according to claims 1 described extracting method, it is characterized in that: the amount of ethyl acetate that adds among the step a or cyclohexane extraction or toluene or chloroform is that the ratio of Folium Ginkgo extract or preparation is 15-30 times, is preferably 18-25 doubly.
3. extracting method according to claim 1 is characterized in that: the reflux time is 1.5～4 hours among the step a, is preferably 2～3 hours.
4. extracting method according to claim 1 is characterized in that: amount of water is 15～30ml among the step b, is preferably 20ml.
5. extracting method according to claim 1 is characterized in that: petroleum ether that adds among the step b (60～90 ℃) or normal hexane or chloroform extraction agent consumption are 15～30ml, are preferably 20ml.
6. extracting method according to claim 1 is characterized in that: extraction times can be 2～4 times among the step b, is preferably 3 times.
7. extracting method according to claim 1 is characterized in that it is undertaken by following step: (a) get Folium Ginkgo extract or its preparation is an amount of, add the ethyl acetate reflux 2 hours of 25 times of amounts; Put coldly, claim decide weight, supply the weight that subtracts mistake, filtration; Get subsequent filtrate, decompression and solvent recovery is to doing; (b) add water 20ml dissolving and being transferred in the separatory funnel, with petroleum ether (60～90 ℃) extraction 3 times, each consumption 20ml; (c) combining extraction liquid, evaporate to dryness, with methanol constant volume to 2ml.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103163252A (en) * | 2012-04-23 | 2013-06-19 | 成都百裕科技制药有限公司 | Determination method for total ginkgolic acid in ginkgolide composition |
| CN103211845A (en) * | 2013-04-26 | 2013-07-24 | 南京林业大学 | Method for removing ginkgolic acid from ginkgo biloba extract |
| CN106074630A (en) * | 2016-06-23 | 2016-11-09 | 吴迪 | A kind of Flavones from Ginkgo Leaves extracting method |
-
2009
- 2009-03-13 CN CN200910079854A patent/CN101757049A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103163252A (en) * | 2012-04-23 | 2013-06-19 | 成都百裕科技制药有限公司 | Determination method for total ginkgolic acid in ginkgolide composition |
| CN103163252B (en) * | 2012-04-23 | 2014-09-24 | 成都百裕科技制药有限公司 | Determination method for total ginkgolic acid in ginkgolide composition |
| CN103211845A (en) * | 2013-04-26 | 2013-07-24 | 南京林业大学 | Method for removing ginkgolic acid from ginkgo biloba extract |
| CN103211845B (en) * | 2013-04-26 | 2015-05-27 | 南京林业大学 | Method for removing ginkgolic acid from ginkgo biloba extract |
| CN106074630A (en) * | 2016-06-23 | 2016-11-09 | 吴迪 | A kind of Flavones from Ginkgo Leaves extracting method |
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Open date: 20100630 |