CN103626829A - Method for separating and preparing lupeol from diospyros rhombifolia hemsl. - Google Patents
Method for separating and preparing lupeol from diospyros rhombifolia hemsl. Download PDFInfo
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- CN103626829A CN103626829A CN201310619326.8A CN201310619326A CN103626829A CN 103626829 A CN103626829 A CN 103626829A CN 201310619326 A CN201310619326 A CN 201310619326A CN 103626829 A CN103626829 A CN 103626829A
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Abstract
The invention provides a method for separating and preparing lupeol from diospyros rhombifolia hemsl., belonging to the technical field of plant purification methods. The method comprises the following processing steps: 1) extraction: grinding branches or roots of diospyros rhombifolia hemsl., extracting the ground branches or roots of diospyros rhombifolia hemsl. with 10-15 times amount of solvent, and recovering the solvent from the extracting solution to obtain a crude extract; 2) separation: carrying out silica column or alumina column chromatography on the crude extract, carrying out gradient elution with petroleum ether-ethyl acetate or n-hexane-chloroform, collecting the eluent, and recovering the solvent to obtain crude lupeol; 3) purification: repeatedly re-crystallizing the crude lupeol by adopting absolute ethyl alcohol or ethyl acetate to obtain pure lupeol. The wild plant diospyros rhombifolia hemsl. adopted by the method is rich in resources. The adopted extraction and purification method has a simple process, is simple and convenient to operate and is suitable for large-scale preparation of lupeol. The obtained lupeol is measured by adopting high performance liquid chromatography (HPLC) and has purity more than 95%.
Description
Technical field
The invention belongs to plant method of purification technical field, be specifically related to a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol.
Background technology
Lupeol is a kind of triterpene compound, has the multiple biological activitys such as antitumor, step-down, anti-inflammatory.Patent CN 101692822A " the new purposes of Lupeol " introduced Lupeol as botanical pesticide for miticide.Root or twig of Diamondleaf persimmon (Diospyros rhombifolia Hemsl.) is Ebenaceae Diospyros defoliation small arbor or shrub, and another name mountain persimmon, wild mountain persimmon, wild persimmon, be distributed in the ground such as Zhejiang Province, China, Jiangsu, Anhui, Jiangxi, Fujian.Wild resource is abundant.
The inventor studies and finds that the content of Lupeol in root or twig of Diamondleaf persimmon is larger, is about 0.2～0.4%.Patent CN 1793161A " extracts the method for Lupeol " and discloses a kind of method of extracting purifying Lupeol from Betula plant in Betula plant.Betula plant wild resource is less, the invention provides a kind of plant origin new and that wild resource is abundant for extracting purifying Lupeol.
Summary of the invention
The problem existing for prior art, the object of the invention is to design provide a kind of from root or twig of Diamondleaf persimmon the separated technical scheme of preparing the method for Lupeol.
Described a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that comprising following processing step:
1) extract: get root or twig of Diamondleaf persimmon branch or root, pulverize, with the solvent extraction of 10～15 times of amounts, extracting solution reclaims solvent, obtains crude extract;
2) separation: crude extract is crossed to silica gel or alumina column chromatography, with petroleum ether-ethyl acetate or normal hexane-chloroform gradient elution, collect elutriant, reclaim solvent, obtain thick Lupeol;
3) purifying: adopt dehydrated alcohol or ethyl acetate to carry out recrystallization repeatedly to thick Lupeol, obtain pure Lupeol.
Described a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that in described step 1), solvent is 95% ethanol, methyl alcohol, benzene or chloroform.
Described a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that being extracted as refluxing extraction in described step 1), extract twice, extract for the first time 2 hours, extract for the second time 1 hour.
Described a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, the wash-out ratio of the petroleum ether-ethyl acetate the step 2 described in it is characterized in that) is 10:0,9:1,8:2, elution volume is 3～5 times of column volume, collects petroleum ether-ethyl acetate 8:2 wash-out part; The wash-out ratio of normal hexane-chloroform is 9:1,7:3,5:5, elution volume is column volume 3～5 times, collect normal hexane-chloroform 5:5 wash-out part.
Described a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that in described step 3) that the number of times of recrystallization is 2～3 times repeatedly.
Above-mentioned a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, reasonable in design, root or twig of Diamondleaf persimmon wild plant resource of the present invention is abundant, extracting and purifying method process of the present invention is simple, easy and simple to handle, be applicable to preparing on a large scale Lupeol, the resulting Lupeol of the present invention adopts high performance liquid chromatography (HPLC method) to measure, and purity is all greater than 95%.
Embodiment
Below in conjunction with embodiment, further illustrate the present invention, but the present invention is not limited to this.
Embodiment 1:
Get root or twig of Diamondleaf persimmon root 1Kg, pulverize, with 15 times, measure 95% alcohol reflux 2 hours for the first time, with 10 times, measure 95% extraction using alcohols 1 hour for the second time; Merge extracted twice liquid, decompression and solvent recovery, obtains crude extract; Crude extract is crossed to silica gel column chromatography, with petroleum ether-ethyl acetate (10:0,9:1,8:2), carry out gradient elution, 3 times of amount column volumes of the first two gradient elution, 5 times of amount column volumes of last gradient elution, reclaim respectively elutriant, obtain eluate; By appropriate dehydrated alcohol recrystallization 3 times for the eluate of last gradient, obtain Lupeol sterling 2.3g(content 97.1%).
Embodiment 2:
Get root or twig of Diamondleaf persimmon root 1Kg, pulverize, with 15 times of amount methanol eddies, extract 2 hours for the first time, with 10 times, measure methanol extraction 1 hour for the second time; Merge extracted twice liquid, decompression and solvent recovery, obtains crude extract; Crude extract is crossed to silica gel column chromatography, with petroleum ether-ethyl acetate (10:0,9:1,8:2), carry out gradient elution, 3 times of amount column volumes of the first two gradient elution, 5 times of amount column volumes of last gradient elution, reclaim respectively elutriant, obtain eluate; By appropriate dehydrated alcohol recrystallization 3 times for the eluate of last gradient, obtain Lupeol sterling 2.7g(content 98.3%).
Embodiment 3:
Get root or twig of Diamondleaf persimmon root 1Kg, pulverize, with 15 times, measure benzene refluxing extraction 2 hours for the first time, with 10 times, measure benzene extraction 1 hour for the second time; Merge extracted twice liquid, decompression and solvent recovery, obtains crude extract; Crude extract is crossed to silica gel column chromatography, with normal hexane-chloroform (9:1,7:3,5:5), carry out gradient elution, 4 times of amount column volumes of the first two gradient elution, 5 times of amount column volumes of last gradient elution, reclaim respectively elutriant, obtain eluate.By appropriate re-crystallizing in ethyl acetate 2 times for the eluate of last gradient, obtain Lupeol sterling 2.6g(content 96.7%).
Embodiment 4:
Get root or twig of Diamondleaf persimmon root 1Kg, pulverize, with 15 times, measure chloroform refluxing extraction 2 hours for the first time, with 10 times, measure chloroform extraction 1 hour for the second time; Merge extracted twice liquid, decompression and solvent recovery, obtains crude extract; Crude extract is crossed to silica gel column chromatography, with normal hexane-chloroform (9:1,7:3,5:5), carry out gradient elution, 4 times of amount column volumes of the first two gradient elution, 5 times of amount column volumes of last gradient elution, reclaim respectively elutriant, obtain eluate.By appropriate re-crystallizing in ethyl acetate 2 times for the eluate of last gradient, obtain Lupeol sterling 2.1g(content 97.4%).
In above-described embodiment, the purity of Lupeol adopts high performance liquid chromatography (HPLC method) to measure chromatographic condition: chromatographic column is Kromasil C18(250 * 4.6mm, 5 μ m), moving phase is methanol-water (99:1), flow velocity is 1mlmin
-1, detection wavelength is 210nm, column temperature is 30 ℃.
Claims (5)
1. a separated method of preparing Lupeol from root or twig of Diamondleaf persimmon, is characterized in that comprising following processing step:
1) extract: get root or twig of Diamondleaf persimmon branch or root, pulverize, with the solvent extraction of 10～15 times of amounts, extracting solution reclaims solvent, obtains crude extract;
2) separation: crude extract is crossed to silica gel or alumina column chromatography, with petroleum ether-ethyl acetate or normal hexane-chloroform gradient elution, collect elutriant, reclaim solvent, obtain thick Lupeol;
3) purifying: adopt dehydrated alcohol or ethyl acetate to carry out recrystallization repeatedly to thick Lupeol, obtain pure Lupeol.
As claimed in claim 1 a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that in described step 1), solvent is 95% ethanol, methyl alcohol, benzene or chloroform.
As claimed in claim 1 a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that being extracted as refluxing extraction in described step 1), extract twice, extract for the first time 2 hours, extract for the second time 1 hour.
As claimed in claim 1 a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, the wash-out ratio of the petroleum ether-ethyl acetate the step 2 described in it is characterized in that) is 10:0,9:1,8:2, elution volume is 3～5 times of column volume, collects petroleum ether-ethyl acetate 8:2 wash-out part; The wash-out ratio of normal hexane-chloroform is 9:1,7:3,5:5, elution volume is column volume 3～5 times, collect normal hexane-chloroform 5:5 wash-out part.
As claimed in claim 1 a kind of from root or twig of Diamondleaf persimmon the separated method of preparing Lupeol, it is characterized in that in described step 3) that the number of times of recrystallization is 2～3 times repeatedly.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310619326.8A CN103626829B (en) | 2013-11-29 | 2013-11-29 | A kind of separation from root or twig of Diamondleaf persimmon prepares the method for Lupeol |
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| CN201310619326.8A CN103626829B (en) | 2013-11-29 | 2013-11-29 | A kind of separation from root or twig of Diamondleaf persimmon prepares the method for Lupeol |
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| CN103626829A true CN103626829A (en) | 2014-03-12 |
| CN103626829B CN103626829B (en) | 2015-08-19 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108836968A (en) * | 2018-09-20 | 2018-11-20 | 广西壮族自治区药用植物园 | The purposes of subprostrate sophora lupin alkanol |
| CN109180769A (en) * | 2018-08-16 | 2019-01-11 | 陕西嘉禾生物科技股份有限公司 | A method of lupeol is isolated and purified from sand pear wood |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1793161A (en) * | 2005-12-31 | 2006-06-28 | 中国海洋大学 | Process for extracting lupinol from betulin category plant |
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2013
- 2013-11-29 CN CN201310619326.8A patent/CN103626829B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1793161A (en) * | 2005-12-31 | 2006-06-28 | 中国海洋大学 | Process for extracting lupinol from betulin category plant |
Non-Patent Citations (3)
| Title |
|---|
| WERNER HERZ ET AL: ""A new tetralone from Diospyros cauiflora"", 《BIOCHEMICAL SYSTEMATICS AND ECOLOGY》 * |
| WITCHUDA THANAKIJCHAROENPATH ET AL: ""Triterpenoids from the stem of Diospyros glandulosa"", 《THAI JOURNAL OF PHARMACEUTICAL SCIENCES》 * |
| 张荣庆 等: ""柿叶化学成分的研究(Ⅱ)"", 《中草药》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109180769A (en) * | 2018-08-16 | 2019-01-11 | 陕西嘉禾生物科技股份有限公司 | A method of lupeol is isolated and purified from sand pear wood |
| CN109180769B (en) * | 2018-08-16 | 2021-03-12 | 陕西嘉禾生物科技股份有限公司 | Method for separating and purifying lupeol from pyrus ussuriensis maxim |
| CN108836968A (en) * | 2018-09-20 | 2018-11-20 | 广西壮族自治区药用植物园 | The purposes of subprostrate sophora lupin alkanol |
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| CN103626829B (en) | 2015-08-19 |
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