CN101157676A - Method for preparing costene lactone - Google Patents
Method for preparing costene lactone Download PDFInfo
- Publication number
- CN101157676A CN101157676A CNA2007101194502A CN200710119450A CN101157676A CN 101157676 A CN101157676 A CN 101157676A CN A2007101194502 A CNA2007101194502 A CN A2007101194502A CN 200710119450 A CN200710119450 A CN 200710119450A CN 101157676 A CN101157676 A CN 101157676A
- Authority
- CN
- China
- Prior art keywords
- costunolide
- extraction
- water
- ethyl acetate
- extract
- 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.)
- Pending
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to an extraction and separation method of costunolide, which takes Southern magnolia leaves as the raw materials, the leaves are smashed into thick powder, the water-soluble impurities are removed by water extraction, the remnants are extracted by reflux of 85 percent of ethanol, after some pigment impurities are removed by petroleum ether extraction, and then the costunolide can be obtained after the silica gel column chromatography of the costunolide crude extract with the accumulation of chloroform or ethyl acetate. The extraction process of the invention has the advantages of easy operation and high yield, and is applicable to the industrialization production.
Description
Technical field
The invention belongs to effective ingredient extraction separation preparation field, be specifically related to a kind of method of from southern magnolia, carrying the extraction separation Costunolide.
Background technology
Costunolide (costunolide) is a sesquiterpenoid lactone compound, contains one alpha-methylene-gamma lactone group in the molecule, has the effect of anti-bacteria and anti-virus sample.Document (Song XK, Tu PF, et al.A newsesquiterpene lactone from Tsoongiodendron odorum Chun.J Asian Nat Prod Res.2001:3 (4): 285-291) report, Costunolide shows that leukemia cancerous cell line (HL-60) is had cytotoxic effect, can suppress the growth of leukaemia cancer cell.Document (Koo TH, Lee JH, et al.A sesquiterpenelactone, costunolide, from Magnolia grandiflora inhibits NF-kappa B by targeting Ikappa B phosphorylation.Planta Med, 2001 Mar, 67 (2): 103-107) record, Costunolide has the nuclear factor of inhibition NF-kB effect, experiment showed, that it can suppress NF-kB and combine with DNA, and be dose-dependence, effect is better than parithenolide, also can suppress the expression by the mRNA of lipopolysaccharides (LPS) inductive iNOS, when concentration is 1ug/ml; Can reach complete restraining effect.
Summary of the invention
The object of the present invention is to provide a kind of from southern magnolia the method for extraction separation Costunolide, the inventive method is fit to suitability for industrialized production, the yield height, product purity can reach more than 90%.
The present invention is raw material with the southern magnolia, extract the water soluble component of removing wherein with water logging bubble after, with higher the extracting of concentration, after column chromatography for separation is refining, can obtain Costunolide again with the miscible organic solvent system of water.
Realize that concrete technology of the present invention is as follows:
A) water extraction is abandoned assorted: the southern magnolia meal, soak extraction twice with deionized water, and each 12 hours, filter, abandon aqueous extract, residue is standby;
B) preparation of Costunolide crude extract: the southern magnolia behind water extraction, with organic solvent extraction that can be miscible with water, the concentrating under reduced pressure extracting solution, adding distilled water in the concentrated solution again disperses, thin out with sherwood oil or hexane extraction to the extraction liquid color, use chloroform or ethyl acetate extraction again instead, combined chloroform or acetic acid ethyl acetate extract, concentrating under reduced pressure get the Costunolide crude extract;
C) Costunolide is refining: with the above-mentioned Costunolide crude extract that obtains, adopt silicagel column further to separate, with petroleum ether-ethyl acetate 20: 1-4: 1 or sherwood oil-acetone 20: 1-4: 1 is the eluent gradient wash-out, collects the stream part section that contains Costunolide, concentrates.
D) will collect the Costunolide that obtains the first time and adopt silica gel column chromatography to separate once more,, collect Costunolide stream part, and concentrate, be drying to obtain with 5: 1 isocratic elutions of petroleum ether-ethyl acetate.
Wherein used can be the arbitrary combination of methyl alcohol, ethanol, propyl alcohol, Virahol, acetone and these solvents with the miscible organic solvent system of water, preferred alcohol, more preferably 75%～95% ethanol.
Extracting method among the present invention can be to soak extraction, diacolation extraction or refluxing extraction, preferred refluxing extraction.
The present invention when for the first time collecting Costunolide, when the gained Costunolide more than or equal to 90% the time, can not need the second step silica gel column chromatography refinement, less than stream part of 90%, adopt the second step silica gel column chromatography further refining to Costunolide purity.
The present invention is when adopting silica gel column chromatography to separate, used moving phase is except that being petroleum ether-ethyl acetate or the sherwood oil-acetone, also available hexane-acetone, hexane-ethyl acetate, chloroform-ethyl acetate, chloroform-acetone, chloroform-methanol system, as long as in sepn process, Costunolide can be separated getting final product with other composition, can think to be equal to technical solution of the present invention.
The resulting Costunolide of the present invention adds acceptable accessories, can be made into various required formulations, is preferably oral dosage form, as tablet, capsule, dripping pill etc.
Specific embodiment
The preparation of the thick lactone of costene
Embodiment 1
Southern magnolia 500g, be ground into meal, 8 times of water gagings soak and extract twice, each 12 hours, filter, abandon the water extract, residue is with twice of 8 times of amount alcohol reflux of 75%, each 1.5 hours, filter merging filtrate, be evaporated to the medicinal extract shape, the ethanol that adds again less than 20% disperses medicinal extract, and the petroleum ether extraction dispersion liquid is thin out to the extraction liquid color, unrecovered oil ether is removed in decompression, it is thin out to the acetic acid ethyl acetate extract color to use ethyl acetate extraction instead, and the combined ethyl acetate extraction liquid, is drying to obtain at concentrating under reduced pressure.
Embodiment 2
Southern magnolia 500g is ground into meal, and 8 times of water gagings soak and extract twice, each 12 hours, filter, abandon the water extract, residue each 1.5 hours, filters with the alcohol reflux twice of 8 times of amounts 95%, merging filtrate is evaporated to the medicinal extract shape, and the ethanol that adds again less than 20% disperses medicinal extract, the hexane extraction dispersion liquid, thin out to the extraction liquid color, residual hexane is removed in decompression, it is thin out to chloroform extraction liquid color to use chloroform extraction instead, and the combined chloroform extraction liquid, is drying to obtain at concentrating under reduced pressure.
To extract the arbitrary combination that etoh solvent makes methyl alcohol, propyl alcohol, Virahol, acetone or these solvents into, be the embodiment of other solvent extraction.
The separation of costunolide
Embodiment 3
Above-mentioned resulting costunolide crude extract is admixed in the 60-90 purpose silica gel, adopt the silica gel column chromatography separation and purification, silica gel for chromatography granularity 200-300 order, the weight ratio of medicinal extract consumption and silica gel consumption is 1: 50, with petroleum ether-ethyl acetate 20: 1-4: 1 gradient elution, Fractional Collections costunolide stream part merges costunolide purity greater than stream part of 90%; Costunolide content merges in addition less than stream part of 90%, concentrate, continue to adopt silica gel column chromatography to separate, with 4: 1 isocratic elution of petroleum ether-ethyl acetate, Fractional Collections costunolide stream part, with costunolide greater than stream part of 90% be incorporated into collect for the first time greater than in stream part of 90%, concentrate, vacuum-drying promptly gets costunolide 1.2g, and purity is 95.27%.
Embodiment 4
Resulting costunolide crude extract in embodiment 1 or 2 is admixed in the 60-90 purpose silica gel, adopt the silica gel column chromatography separation and purification, silica gel for chromatography granularity 200-300 order, the weight ratio of medicinal extract consumption and silica gel consumption is 1: 50, with sherwood oil-acetone 20: 1-4: 1 gradient elution, Fractional Collections costunolide stream part merges costunolide purity greater than stream part of 90%; Costunolide content merges in addition less than stream part of 90%, concentrate, continue to adopt silica gel column chromatography to separate, with 5: 1 isocratic elution of sherwood oil-acetone, Fractional Collections costunolide stream part, with costunolide greater than stream part of 90% be incorporated into collect for the first time greater than in stream part of 90%, concentrate, vacuum-drying promptly gets costunolide 1.0g, and purity is 97.27%.
Embodiment 5 technology pilot scale study on the stability
According to the extraction separation program of embodiment 1 and embodiment 3, do three batches of products, every batch of medicinal material consumption is 10kg, it the results are shown in Table 1
Three batches of medicinal material pilot scales of table 1 Costunolide extraction process study on the stability
From last table result as can be known, this process stabilizing, favorable reproducibility is fit to suitability for industrialized production.
Embodiment 6
Costunolide 10g, starch 20g, mixing, in the hard capsule of packing into, shape is 100 altogether, not contain costunolide 100mg in the unit capsule.
Embodiment 7
Costunolide 10g, Microcrystalline Cellulose 42g, sodium starch glycolate 3.2g, croscarmellose sodium 3.2g, Magnesium Stearate 1.6g, 75% ethanol adopts the medicament common process, is pressed into tablet, and every 0.3g contains costunolide 50mg.
Claims (5)
1. the method for extraction separation Costunolide from southern magnolia is characterized in that comprising the steps:
A) water extraction is abandoned assorted: the southern magnolia meal, soak extraction twice with deionized water, and each 12 hours, filter, abandon aqueous extract, residue is standby;
B) preparation of Costunolide crude extract: the southern magnolia behind water extraction, with organic solvent extraction that can be miscible with water, the concentrating under reduced pressure extracting solution, adding distilled water in the concentrated solution again disperses, thin out with sherwood oil or hexane extraction to the extraction liquid color, use chloroform or ethyl acetate extraction again instead, combined chloroform or acetic acid ethyl acetate extract, concentrating under reduced pressure get the Costunolide crude extract;
C) Costunolide is refining: with the above-mentioned Costunolide crude extract that obtains, adopt silicagel column further to separate, with petroleum ether-ethyl acetate 20: 1-4: 1 or sherwood oil-acetone 20: 1-4: 1 is the eluent gradient wash-out, collects the stream part section that contains Costunolide, concentrates.
D) will collect the Costunolide that obtains the first time and adopt silica gel column chromatography to separate once more,, collect Costunolide stream part, and concentrate, be drying to obtain with 4: 1 isocratic elutions of petroleum ether-ethyl acetate.
2. the method for claim 1, it is characterized in that described can meaning with the miscible organic solvent of water: any of methyl alcohol, ethanol, propyl alcohol, Virahol, acetone and these solvents mixes.
3. method as claimed in claim 2 is characterized in that described organic solvent preferred alcohol that can be miscible with water.
4. method as claimed in claim 3 is characterized in that described ethanol is preferably the ethanol of 75%-95%.
5. the method for claim 1, the purity that it is characterized in that resulting Costunolide for more than or equal to 90% less than 100%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101194502A CN101157676A (en) | 2007-07-25 | 2007-07-25 | Method for preparing costene lactone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101194502A CN101157676A (en) | 2007-07-25 | 2007-07-25 | Method for preparing costene lactone |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101157676A true CN101157676A (en) | 2008-04-09 |
Family
ID=39305925
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101194502A Pending CN101157676A (en) | 2007-07-25 | 2007-07-25 | Method for preparing costene lactone |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101157676A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102133247A (en) * | 2011-01-28 | 2011-07-27 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN104673732A (en) * | 2013-11-28 | 2015-06-03 | 中国科学院天津工业生物技术研究所 | Escherichia coli expression strain and method for biologically synthesizing costundide in Escherichia coli |
| CN105153086A (en) * | 2015-10-26 | 2015-12-16 | 沈健龙 | Novel sesquiterpenoids compound and preparation method and medical application thereof |
| CN114960230A (en) * | 2022-05-13 | 2022-08-30 | 常州纺织服装职业技术学院 | Preparation method of southern magnolia leaf pigment cotton cloth and southern magnolia leaf pigment cotton cloth with antibacterial property |
-
2007
- 2007-07-25 CN CNA2007101194502A patent/CN101157676A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102133247A (en) * | 2011-01-28 | 2011-07-27 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN102133247B (en) * | 2011-01-28 | 2013-03-13 | 西南大学 | Costustoot extractive with blood sugar reduction activity and application thereof |
| CN104673732A (en) * | 2013-11-28 | 2015-06-03 | 中国科学院天津工业生物技术研究所 | Escherichia coli expression strain and method for biologically synthesizing costundide in Escherichia coli |
| CN104673732B (en) * | 2013-11-28 | 2018-10-30 | 中国科学院天津工业生物技术研究所 | A kind of E. coli expression strains and in Escherichia coli biosynthesis costunolide method |
| CN105153086A (en) * | 2015-10-26 | 2015-12-16 | 沈健龙 | Novel sesquiterpenoids compound and preparation method and medical application thereof |
| CN114960230A (en) * | 2022-05-13 | 2022-08-30 | 常州纺织服装职业技术学院 | Preparation method of southern magnolia leaf pigment cotton cloth and southern magnolia leaf pigment cotton cloth with antibacterial property |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100595183C (en) | Method for preparing 6-gingerol and 8-gingerol from ginger | |
| CN102976909B (en) | Method for extracting and purifying 6-gingerol from ginger | |
| CN105106556A (en) | Chemical component separation and identification method for Parisforrestii (Takht.) H. Li antitumor activity part PFE-PT3 | |
| CN102924416A (en) | Method for separating and purifying monomeric compounds from ash bark | |
| CN105503786B (en) | Split Lignanoids compounds-smartweed diffractive ring lignan glucoside E and preparation method thereof | |
| CN101157676A (en) | Method for preparing costene lactone | |
| CN108186474B (en) | Total flavone extract of Loranthus parasiticus of Loranthus, preparation method and application | |
| CN1269831C (en) | Method for preparing polygonin and resveratrol | |
| CN102731592A (en) | Method for extracting cleupin and amentoflavone from olive leaf | |
| CN104910216B (en) | It is a kind of with preparing liquid phase method while obtaining the separation method of a variety of epimedium flavones | |
| CN101830892B (en) | Method for separating glycoside chemical components from tibetan capillaris | |
| CN102503996B (en) | Method for extracting active constituent from Swertia mussotii plant | |
| CN101955504A (en) | Method for preparing specnuezhenide, nuezhenoside and oleanolic acid by high-speed countercurrent chromatography | |
| CN102464693B (en) | Ginsenoside Re extraction and separation method | |
| CN111171104A (en) | Method for preparing ursolic acid from rosemary oil paste by-product | |
| CN101190259A (en) | Southern magnolia total lactone extract and preparation method and application thereof | |
| CN103880895B (en) | A kind of method of utilizing high speed adverse current chromatogram separation and purification to prepare harpagoside and Wyrmslayer glycosides A | |
| CN101024604B (en) | Novel dihydrochalcone compound separated and purified from drgon blood and preparation method thereof | |
| CN101575356A (en) | Method for extracting and purifying 7-xylose-10-deacetyl paclitaxel from taxus cuspidata | |
| Hamil et al. | Optimisation of asarone removal from Piper sarmentosum Roxburgh leaves using supercritical carbon dioxide extraction: the Box-Behnken design | |
| CN105384784A (en) | Screening and separating preparation method for three stilbene polyphenol substances with antioxidant activity in polygonum multiflorum polygonum multiflorumcultivated in Qinghai | |
| CN1194985C (en) | Diterpene compound of veronicastrum sibiricum and its extraction separation method | |
| CN102477023A (en) | Preparation method of high purity 5-hydroxy-3, 7, 4'-trimethoxyflavone | |
| KR101227156B1 (en) | Manufacturing method of xanthorrhizol having oral antibacterial | |
| CN103288898A (en) | Method for extracting, purifying and separating new mangiferin reference substance |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20080409 |