CN103804441A - Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol - Google Patents
Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol Download PDFInfo
- Publication number
- CN103804441A CN103804441A CN201210460448.2A CN201210460448A CN103804441A CN 103804441 A CN103804441 A CN 103804441A CN 201210460448 A CN201210460448 A CN 201210460448A CN 103804441 A CN103804441 A CN 103804441A
- Authority
- CN
- China
- Prior art keywords
- xylose
- deacetyl paclitaxel
- pressed powder
- obtains
- deacetyl
- 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
Images
Abstract
The invention discloses a method for separating and purifying 7-xylose-10-desacetyltaxol from an extract containing 7-xylose-10-desacetyltaxol, relating to the technical field of separation and purification of 7-xylose-10-desacetyltaxol. The method comprises the steps of extraction, normal-phase chromatography, crystallization, reverse-phase chromatography and crystallization. The method disclosed by the invention has the beneficial effects that high purity and high yield are guaranteed, and the organic solvent for extraction and the filler and organic solvent for chromatography can be reused.
Description
Technical field
The present invention relates to the technical field of separation and purification 7-xylose-10-deacetyl paclitaxel, be specifically related to a kind of from the medicinal extract separation containing 7-xylose-10-deacetyl paclitaxel and the method for purification 7-xylose-10-deacetyl paclitaxel.
Background technology
Taxol is the secondary metabolite of a kind of complexity in Chinese yew genus plants, is also that understand at present only a kind of can promote microtubule polymerization and the stable medicine of polymerization microtubule.Isotopic tracing shows, taxol is only attached on the microtubule of polymerization, does not react with unpolymerized tubulin dimer.Can be at the thin a large amount of microtubule of intracellular accumulation after cells contacting taxol, the various functions of cell have been disturbed in the accumulation of these microtubules, particularly make cell fission stop at m period, have blocked the proper splitting of cell.By the clinical study of II-III, taxol is mainly applicable to ovarian cancer and mammary cancer, and lung cancer, large bowel cancer, melanoma, incidence cancer, lymphoma, brain tumor are also had to certain curative effect.
7-xylose-10-deacetyl paclitaxel can be converted into taxol by biology and chemical process, so preparation 7-xylose-10-deacetyl paclitaxel is to have great meaning.
Granted publication number is the patent that CN101235022B, denomination of invention are " a kind of method of extracting purifying two kinds of taxane compound from Ramulus et folium taxi cuspidatae ", discloses and has obtained resin absorption after branches and leaves, normal-phase chromatography separates and obtains the above 7-xylose-10-deacetyl paclitaxel of content 67%.
Summary of the invention
If expect high purity 7-xylose-10-deacetyl paclitaxel in prior art, inevitable 7-xylose-10-deacetyl paclitaxel yield can reduce.Actual production moderate purity and yield are conflicts.The invention provides and a kind ofly can guarantee that high purity can guarantee again the separation of high yield and the method for purification 7-xylose-10-deacetyl paclitaxel.
The invention provides a kind ofly from separating containing the medicinal extract of 7-xylose-10-deacetyl paclitaxel and the method for purification 7-xylose-10-deacetyl paclitaxel, described method comprises the steps:
(1) dissolve the medicinal extract containing 7-xylose-10-deacetyl paclitaxel with aqueous ethanolic solution;
(2) (1) dissolve the solution of 7-xylose-10-deacetyl paclitaxel by organic solvent extraction step;
(3) dry extraction phase, obtains the pressed powder I containing 7-xylose-10-deacetyl paclitaxel;
(4) purify pressed powder I with normal-phase chromatography, collect the cut containing 7-xylose-10-deacetyl paclitaxel;
(5) the cut containing 7-xylose-10-deacetyl paclitaxel that (4) drying step obtains, obtains the pressed powder II containing 7-xylose-10-deacetyl paclitaxel;
(6) crystalline solid powder II;
(7) the crystal that (6) drying step obtains, obtains the pressed powder III containing 7-xylose-10-deacetyl paclitaxel;
(8) purify pressed powder III with reverse-phase chromatography, collect the cut containing 7-xylose-10-deacetyl paclitaxel;
(9) the cut containing 7-xylose-10-deacetyl paclitaxel that (8) drying step obtains, obtains the pressed powder IV containing 7-xylose-10-deacetyl paclitaxel;
(10) crystalline solid powder IV;
(11) the crystal that (10) drying step obtains, obtains high purity 7-xylose-10-deacetyl paclitaxel.
Described method steps (1) in medicinal extract be preferably and contain taxol, 7-xylose-10-deacetyl paclitaxel, 10-deacetylate taxol, 10-deacetylate Bakating III, Bakating III, the peaceful alkali of tricuspid shirt and 13-ethanoyl-9-hydroxyl Bakating III.
Described method steps (1) in the alcoholic strength of aqueous ethanolic solution be preferably 40～60, dissolve medicinal extract with this alcoholic strength aqueous ethanolic solution, advantage is easy layering in the time that step extracts in (2).
Described method steps (2) middle organic solvent is preferably methylene dichloride or ethyl acetate, 20～25 ℃ of extraction temperature, and equal-volume extraction three times, merges extraction phase, and extracting phase HPLC discards while detecting containing bearing taxanes.
(3) described method steps is preferably first uses vacuum distillation method by extraction phase, obtains pressed powder I with vacuum drying method afterwards, the recycling when organic solvent of recovery can do next time extraction.
Described method steps (4) middle normal-phase chromatography condition optimization is: stationary phase is silica gel, moving phase is methylene dichloride and dehydrated alcohol, volume ratio is methylene dichloride/dehydrated alcohol=100/10～15, and applied sample amount is the pressed powder I of silica gel single purifying 1～20g of every 100g.
Described method steps (4) middle normal-phase chromatography condition most preferably is: stationary phase is 200～300 object non-sphere silica gel; moving phase is methylene dichloride and dehydrated alcohol; volume ratio is methylene dichloride/dehydrated alcohol=100/12; applied sample amount is the pressed powder I of the silica gel single purifying 10g of every 100g; collect the cut containing 7-xylose-10-deacetyl paclitaxel; after silica gel uses, be methylene dichloride/dehydrated alcohol=100/20 regeneration by volume ratio, being eluted to ultraviolet detection does not have impurity.
(5) described method steps is preferably first uses vacuum distillation method by the cut containing 7-xylose-10-deacetyl paclitaxel; obtain pressed powder II with vacuum drying method afterwards; after the moving phase reclaiming detects with GC, the moving phase that is made into volume ratio and is methylene dichloride/dehydrated alcohol=100/2 reuses can do next chromatography time.
(6) described method steps is preferably uses methanol-water crystallization.
(7) described method steps is preferably the pressed powder III that obtains containing 7-xylose-10-deacetyl paclitaxel with vacuum drying method.
Described method steps (8) middle reverse-phase chromatography condition optimization is: stationary phase is C
18, moving phase is acetonitrile and ultrapure water, and volume ratio is acetonitrile/ultrapure water=100/80～120, and applied sample amount is the C of every 100g
18the pressed powder III of single purifying 1～10g.
Described method steps (8) middle reverse-phase chromatography condition most preferably is: stationary phase is the C of 5 μ m
18, moving phase is acetonitrile and ultrapure water, and volume ratio is acetonitrile/ultrapure water=100/100, and applied sample amount is the C of every 100g
18the pressed powder III of single purifying 2g, collects the cut containing 7-xylose-10-deacetyl paclitaxel, C
18after using, be acetonitrile/ultrapure water=100/10 regeneration by volume ratio, being eluted to ultraviolet detection does not have impurity.
(9) described method steps is preferably first uses vacuum distillation method by the cut containing 7-xylose-10-deacetyl paclitaxel; in the time being evaporated to a certain degree, spontaneous nucleation is separated out; finally obtain the pressed powder IV containing 7-xylose-10-deacetyl paclitaxel with vacuum drying method; after the moving phase reclaiming detects with GC, the moving phase that is made into volume ratio and is acetonitrile/ultrapure water=100/100 reuses can do next chromatography time.
(10) described method steps is preferably with the crystallization of acetone-sherwood oil.
(11) described method steps is preferably and obtains high purity 7-xylose-10-deacetyl paclitaxel with vacuum drying method.
Beneficial effect of the present invention is:
1, the beneficial effect of maximum of the present invention be the 7-xylose-10-deacetyl paclitaxel product purity that finally obtains high to more than 98.5%, content of impurities is less than 1%, single contaminant content is less than 0.5%.The 7-xylose-10-deacetyl paclitaxel yield finally obtaining is more than 95%.Not only guarantee high purity but also guaranteed high yield.
2, in technique, extracting the organic solvent of use, methylene dichloride/dehydrated alcohol that normal-phase chromatography is used, acetonitrile/ultrapure water that reversed phase chromatography is used can recycle, and has reduced process costs.
3, purification on normal-phase silica gel, anti-phase C in technique
18can regeneration, reduce process costs.
Accompanying drawing explanation
Fig. 1 is from the medicinal extract separation containing 7-xylose-10-deacetyl paclitaxel and the process flow sheet of purification 7-xylose-10-deacetyl paclitaxel.
Embodiment
Following non-limiting example can make the present invention of those of ordinary skill in the art's comprehend, but does not limit the present invention in any way.
Embodiment 1
(1) dissolve the medicinal extract containing taxol, 7-xylose-10-deacetyl paclitaxel, 10-deacetylate taxol, 10-deacetylate Bakating III, Bakating III, the peaceful alkali of tricuspid shirt and 13-ethanoyl-9-hydroxyl Bakating III, 7-xylose-10-deacetyl paclitaxel content 2% in medicinal extract with the aqueous ethanolic solution of alcoholic strength 50;
(2) (1) dissolve the solution of 7-xylose-10-deacetyl paclitaxel by dichloromethane extraction step, 25 ℃ of extraction temperature, equal-volume extraction three times, merges three times methylene dichloride phase, and HPLC detects extracting phase, waste treatment in the time that extraction phase does not contain taxane compounds;
(3) evaporate to dryness methylene dichloride phase, the condition of underpressure distillation is 35 ℃ of vacuum tightness 0.4MPa, bath temperatures, evaporate to dryness is loss of weight extremely no longer, vacuum drying condition is 60 ℃ of vacuum tightness 0.8MPa, drying temperatures, drying products is loss of weight extremely no longer, obtain the pressed powder I containing 7-xylose-10-deacetyl paclitaxel, 7-xylose-10-deacetyl paclitaxel content 20% in pressed powder I;
(4) normal-phase chromatography purifying, chromatographic condition is that preparative column specification is Φ 200 × 300, stationary phase is 200～300 order non-sphere silica gel of 4kg, moving phase is methylene dichloride and the dehydrated alcohol of 60L, volume ratio is methylene dichloride/dehydrated alcohol=100/12, and applied sample amount is the pressed powder I of 400g, and elution flow rate is 1L/min, ultraviolet detection wavelength is 227nm, collects the cut containing 7-xylose-10-deacetyl paclitaxel;
(5) evaporate to dryness is containing the cut of 7-xylose-10-deacetyl paclitaxel, the condition of underpressure distillation is 35 ℃ of vacuum tightness 0.4MPa, bath temperatures, evaporate to dryness is loss of weight extremely no longer, vacuum drying condition is 60 ℃ of vacuum tightness 0.8MPa, drying temperatures, drying products is loss of weight extremely no longer, obtain the pressed powder II containing 7-xylose-10-deacetyl paclitaxel, 7-xylose-10-deacetyl paclitaxel content 50% in pressed powder II;
(6) use methanol-water crystalline solid powder II;
(7) the crystal that (6) drying step obtains, vacuum drying condition is 60 ℃ of vacuum tightness 0.8MPa, drying temperatures, drying products is loss of weight extremely no longer, obtains the pressed powder III containing 7-xylose-10-deacetyl paclitaxel, 7-xylose-10-deacetyl paclitaxel content 70% in pressed powder III;
(8) reversed phase chromatography separation, chromatographic condition is that preparative column specification is Φ 200 × 300, stationary phase is the C of the 5 μ m of 4kg
18, moving phase is 80L acetonitrile and ultrapure water, and volume ratio is acetonitrile/ultrapure water=100/100, and applied sample amount is the pressed powder III of 80g, and elution flow rate is 1L/min, ultraviolet detection wavelength is 227nm, collects the cut containing 7-xylose-10-deacetyl paclitaxel;
(9) evaporation is containing the cut of 7-xylose-10-deacetyl paclitaxel, the condition of underpressure distillation is 55 ℃ of vacuum tightness 0.8MPa, bath temperatures, being evaporated to crystal separates out, vacuum-drying crystallize out, vacuum drying condition is 60 ℃ of vacuum tightness 0.8MPa, drying temperatures, drying products is loss of weight extremely no longer, obtains the pressed powder IV containing 7-xylose-10-deacetyl paclitaxel, 7-xylose-10-deacetyl paclitaxel content 95% in pressed powder IV;
(10) use acetone-sherwood oil crystalline solid powder IV;
(11) the crystal that (10) drying step obtains, vacuum drying condition is 60 ℃ of vacuum tightness 0.8MPa, drying temperatures, drying products is loss of weight extremely no longer, obtains high purity 7-xylose-10-deacetyl paclitaxel content 99%, yield 90%.
Claims (6)
1. from the medicinal extract separation containing 7-xylose-10-deacetyl paclitaxel and a method for purification 7-xylose-10-deacetyl paclitaxel, described method comprises the steps:
(1) dissolve the medicinal extract containing 7-xylose-10-deacetyl paclitaxel with aqueous ethanolic solution;
(2) (1) dissolve the solution of 7-xylose-10-deacetyl paclitaxel by organic solvent extraction step;
(3) dry extraction phase, obtains the pressed powder I containing 7-xylose-10-deacetyl paclitaxel;
(4) purify pressed powder I with normal-phase chromatography, collect the cut containing 7-xylose-10-deacetyl paclitaxel;
(5) the cut containing 7-xylose-10-deacetyl paclitaxel that (4) drying step obtains, obtains the pressed powder II containing 7-xylose-10-deacetyl paclitaxel;
(6) crystalline solid powder II;
(7) the crystal that (6) drying step obtains, obtains the pressed powder III containing 7-xylose-10-deacetyl paclitaxel;
(8) purify pressed powder III with reverse-phase chromatography, collect the cut containing 7-xylose-10-deacetyl paclitaxel;
(9) the cut containing 7-xylose-10-deacetyl paclitaxel that (8) drying step obtains, obtains the pressed powder IV containing 7-xylose-10-deacetyl paclitaxel;
(10) crystalline solid powder IV;
(11) the crystal that (10) drying step obtains, obtains high purity 7-xylose-10-deacetyl paclitaxel.
2. method according to claim 1, is characterized in that: described method steps (1) middle medicinal extract contains taxol, 7-xylose-10-deacetyl paclitaxel, 10-deacetylate taxol, 10-deacetylate Bakating III, Bakating III, the peaceful alkali of tricuspid shirt and 13-ethanoyl-9-hydroxyl Bakating III.
3. method according to claim 1, is characterized in that: the described method steps (1) alcoholic strength of middle aqueous ethanolic solution is 40～60.
4. method according to claim 1, is characterized in that: described method steps (2) middle organic solvent is methylene dichloride or ethyl acetate.
5. method according to claim 1, it is characterized in that: described method steps is middle normal-phase chromatography condition (4): stationary phase is silica gel, moving phase is methylene dichloride and dehydrated alcohol, volume ratio is methylene dichloride/dehydrated alcohol=100/10～15, and applied sample amount is the pressed powder I of silica gel single purifying 1～20g of every 100g.
6. method according to claim 1, is characterized in that: described method steps is middle reverse-phase chromatography condition (8): stationary phase is C
18, moving phase is acetonitrile and ultrapure water, and volume ratio is acetonitrile/ultrapure water=100/80～120, and applied sample amount is the C of every 100g
18the pressed powder III of single purifying 1～10g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210460448.2A CN103804441A (en) | 2012-11-15 | 2012-11-15 | Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210460448.2A CN103804441A (en) | 2012-11-15 | 2012-11-15 | Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103804441A true CN103804441A (en) | 2014-05-21 |
Family
ID=50701870
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210460448.2A Pending CN103804441A (en) | 2012-11-15 | 2012-11-15 | Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103804441A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992007842A1 (en) * | 1990-11-02 | 1992-05-14 | University Of Florida | Method for the isolation and purification of taxane derivatives |
| WO2005105767A1 (en) * | 2004-04-23 | 2005-11-10 | Chatham Biotec Ltd. | Semi-synthesis and isolation of taxane intermediates from a mixture of taxanes |
| CN101143883A (en) * | 2006-09-15 | 2008-03-19 | 中国科学院大连化学物理研究所 | Mixture containing taxanes xyloside compounds and preparation method thereof |
| CN101235022A (en) * | 2007-11-01 | 2008-08-06 | 东北林业大学 | Method for extracting and purifying two kinds of taxane compound from yew branches and leaves |
| CN101397284A (en) * | 2007-09-30 | 2009-04-01 | 桂林市振达生物科技有限责任公司 | Method for extracting and separating paclitaxel and taxones compounds from yew |
| CN101575356A (en) * | 2009-06-09 | 2009-11-11 | 东北林业大学 | Method for extracting and purifying 7-xylose-10-deacetyl paclitaxel from taxus cuspidata |
| CN102260227A (en) * | 2011-06-21 | 2011-11-30 | 沈阳化工大学 | Method for separating paclitaxel and related taxane substances |
-
2012
- 2012-11-15 CN CN201210460448.2A patent/CN103804441A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992007842A1 (en) * | 1990-11-02 | 1992-05-14 | University Of Florida | Method for the isolation and purification of taxane derivatives |
| WO2005105767A1 (en) * | 2004-04-23 | 2005-11-10 | Chatham Biotec Ltd. | Semi-synthesis and isolation of taxane intermediates from a mixture of taxanes |
| CN101143883A (en) * | 2006-09-15 | 2008-03-19 | 中国科学院大连化学物理研究所 | Mixture containing taxanes xyloside compounds and preparation method thereof |
| CN101397284A (en) * | 2007-09-30 | 2009-04-01 | 桂林市振达生物科技有限责任公司 | Method for extracting and separating paclitaxel and taxones compounds from yew |
| CN101235022A (en) * | 2007-11-01 | 2008-08-06 | 东北林业大学 | Method for extracting and purifying two kinds of taxane compound from yew branches and leaves |
| CN101575356A (en) * | 2009-06-09 | 2009-11-11 | 东北林业大学 | Method for extracting and purifying 7-xylose-10-deacetyl paclitaxel from taxus cuspidata |
| CN102260227A (en) * | 2011-06-21 | 2011-11-30 | 沈阳化工大学 | Method for separating paclitaxel and related taxane substances |
Non-Patent Citations (1)
| Title |
|---|
| 张沿军，等: "东北紫杉茎皮化学成分的研究", 《中草药》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103172598B (en) | Method for extracting paclitaxel from taxus chinensis branch and leaf | |
| CN102260227B (en) | Method for separating paclitaxel and related taxane substances | |
| CN101397284A (en) | Method for extracting and separating paclitaxel and taxones compounds from yew | |
| CN103588785B (en) | The process for purification of a kind of Ginkgolide A and Ginkgolide B | |
| CN106967137B (en) | Method for separating high-purity oleuropein by liquid chromatography through macroporous resin combined preparation | |
| CN105481711B (en) | Hydroxy-beta-sanshool monomer preparation method | |
| CN102653528B (en) | Method for separating purified paclitaxel and 10 diaminobenzidine (DAB) III by utilizing macroporous resin | |
| CN103804327A (en) | Method for separating and purifying paclitaxel from paclitaxel-containing extract | |
| CN102731592A (en) | Method for extracting cleupin and amentoflavone from olive leaf | |
| CN103804325A (en) | Method for separating and purifying 10-deacetyltaxol from 10-deacetyltaxol-containing extract | |
| CN104817445A (en) | Method for separating purified physcion and emodin from giant knot weed | |
| CN104693249A (en) | Method for simultaneously separating and preparing syringin and oleuropein from syringa oblate lindl. | |
| CN102417492A (en) | Method for separating and purifying paclitaxel | |
| CN103788032A (en) | Preparation method of paclitaxel | |
| CN103467262A (en) | Method for preparing 9-oxonerolidol from camphor tree plants | |
| CN103804441A (en) | Method for separating and purifying 7-xylose-10-desacetyltaxol from extract containing 7-xylose-10-desacetyltaxol | |
| CN103351373B (en) | Method for extracting Schisandrin B from Schisandra chinensis | |
| CN103804326A (en) | Method for separating 10-Deacetylbaccatin III from 10-Deacetylbaccatin III-containing extract and purifying 10-Deacetylbaccatin III | |
| CN105646424A (en) | A method of extracting luteolin | |
| CN103804324A (en) | Method for separating and purifying baccatin III from baccatin III containing extract | |
| CN104402949A (en) | Method for simultaneously separating and preparing syringoside and oleuropein from syringa oblata lindl | |
| CN103626829B (en) | A kind of separation from root or twig of Diamondleaf persimmon prepares the method for Lupeol | |
| CN102241658A (en) | Method for purifying gamma-mangostin by using high-speed countercurrent chromatography | |
| CN104072504A (en) | Method for extracting and purifying hernandia nymphaeifolia alkali from hernandia nymphaeifolia | |
| CN102190665B (en) | Method for separating and purifying artemisinin by activated charcoal column chromatography employing nonaqueous system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140521 |
|
| WD01 | Invention patent application deemed withdrawn after publication |