CN108101869A - A kind of extracting method of natural Japanese yew alcohol - Google Patents
A kind of extracting method of natural Japanese yew alcohol Download PDFInfo
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- CN108101869A CN108101869A CN201711386598.2A CN201711386598A CN108101869A CN 108101869 A CN108101869 A CN 108101869A CN 201711386598 A CN201711386598 A CN 201711386598A CN 108101869 A CN108101869 A CN 108101869A
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- alcohol
- extracting method
- japanese yew
- natural japanese
- methanol
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D305/00—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms
- C07D305/14—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms condensed with carbocyclic rings or ring systems
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Abstract
The invention discloses a kind of extracting methods of natural Japanese yew alcohol.Including：(1) mixed solvent based on water is used, water-solubility impurity and lipidic compounds are removed from branches and leaves of yew；(2) analog based on natural Japanese yew alcohol is extracted from the solid slag of upper step using alcohols solvent；(3) natural Japanese yew alcohol extracting thing crude product is quickly purified by flash using macroreticular resin；(4) the natural Japanese yew alcohol of high-purity is obtained by crystal refining；This process is few, environmental-friendly using organic solvent, equipment is simple, it is easy to operate be easily achieved large-scale production, entire production process loss less, be conducive to the comprehensive utilization of resource, product purity reaches more than 98%, maximum single miscellaneous to be less than 0.1%.
Description
Technical field
The present invention relates to a kind of process for extracting of natural antitumor drug, and in particular to a kind of extraction natural Japanese yew alcohol
Preparation method.
Background technology
Taxol, is the secondary metabolite extracted from Chinese yew, and taxol mainly acts on tubulin
System, it can promote tubulin to be assembled into micro-pipe, inhibit the depolymerization of micro-pipe, and abnormal, formation is arranged so as to cause vascular bundle
Aster makes spindle lose normal function, causes cancer cell death.Clinically be mainly used for treat oophoroma, breast cancer and
Non-small cell lung cancer, antitumor activity is higher than common antitumor drugs such as thiazole furan quinoline, adriamycin and fluorouracils.Therefore because
Broad spectrum activity and high active anticancer for it, at present taxol become most widely used anticancer drug.
Americanized scholar's Giovannis (M.C.Wani) in 1963 and Wall (Monre E.Wall) are grown in U.S. from one kind for the first time
The crude extract of taxol is isolated in state's western part fully stocked wood in appellation peace red deal (Pacific Yew) bark and timber.
In screening experiment, Wani and Wall have found that taxol crude extract has very high activity to the mouse tumour cell of cultured in vitro, and start
Separate this active ingredient.Due to the active ingredient, content is extremely low in plant, until 1971, their just same Du Ke (Duke)
The chemistry professor Mu Kefaer (Andre T.McPhail) of university cooperates, and the active ingredient is determined by x-ray analysis
Chemical constitution, a kind of tricyclic diterpene compound, and it is named as taxol.
Taxol is directly extracted from ginkgo bark, tree root earliest, however content of the taxol in plant is suitable
Low, the bark of about 13.6kg could propose the taxol of 1g, and one ovarian cancer patients for the treatment of need more than 3-12 a century
Chinese yew, Japanese yew resource itself is very poor in addition, and Chinese yew genus plants are slow-growing, and this rare tree is caused to be on the point of
Face extinction.And current biological fermentation process and medicine source of Taxol plant endogenesis epiphyte cultivation method limit it is more, it is not mature enough, no
It is suitble to large-scale production.
Therefore people continue research higher paclitaxel precursor compound of searching amount in Chinese yew later, then pass through again
Chemical method is translated into taxol and is very effective solution route.Taxol is obtained by the reaction by multi-step chemical.But
This method, which obtains taxol, to be needed, by substantial amounts of organic synthesis, to cause resource and environment larger destruction.
It is of the invention then using taxusyunnanensis branches and leaves, natural Japanese yew alcohol content is high, extracts and needs not move through from branches and leaves
Chemical synthesis can obtain substantial amounts of high-purity natural taxol, you can meet the needs of people's medical treatment ＆ health, while the present invention
In order to avoid the shortcomings of traditional handicraft, reduce the use of organic solvent, improve the extraction efficiency of natural Japanese yew alcohol and red
The utilization rate of beans China fir branches and leaves reduces the content of taxol lmpurities, so in turn avoids the waste of resource and breaking for environment
It is bad, obtain comprehensive cyclic utilization well.
The content of the invention
It is an object of the present invention to provide a kind of extracting method of natural Japanese yew alcohol, to overcome present in the prior art
Disadvantages mentioned above and deficiency.
The Chinese yew of China's distribution is divided into 4 kind of 1 mutation, i.e. taxus chinensis in northeast, Xizang Taxus chinensis, taxusyunnanensis, China
Chinese yew and the mutation southerm yew of Chinese Chinese yew.
Taxusyunnanensis (Taxus yunnanensis ChengetL.K.Fu) also known as southwestern Chinese yew, are taxaceae
The aiphyllium of Taxus is one of 4 kinds of Chinese yew of China.
The Chinese yew that later Chinese Academy of Sciences plant classification scholar's Fourier Mr. state and its disciple will be considered to China's distribution is 3 kind 2
Mutation, title is also different, is respectively taxus chinensis in northeast (Taxus cuspidata) Himalaya heavy foliage Chinese yew (Taxus
Fuana), two mutation China red beans of Himalaya Chinese yew (Taxus wallichiana) and Himalaya Chinese yew
China fir (Taxus wallichianavar.chinensis) southerm yew (Taxus wallichiana var.mairei).
The principle of the present invention：
With the taxusyunnanensis branches and leaves (Classification system Taxus yunnanensis) of crushing for initial feed, by water
Mixed solution impregnates, and water-soluble and oil-soluble impurities is removed, mixed solution recovered recycling again, the slag after immersion is through alcohol
Class solvent soaking obtains taxol crude product, this crude product purity is preferable, is easy to crystal refining and obtains the finished product taxol of high-purity.
Technical problems to be solved needed for the present invention can be achieved through the following technical solutions：
A kind of extracting method of natural Japanese yew alcohol, which is characterized in that including：
(1) taxusyunnanensis branches and leaves are taken, are crushed through pulverizer；
(2) by the taxusyunnanensis branches and leaves after crushing through 0.1%~0.15% organic acid, 1.0%~5.0% acetone or alcohol
Mixed aqueous solution cook extractant, extract；
(3) slag after first time extracts, then the soak extraction through alcohol organic solvent obtain taxol crude product；
(4) taxol crude product does resin column purification by AB-8 type resins；
(5) crystallized using acetonitrile, the mixed solvent system crystallization of methanol, dichloromethane and n-hexane, so as to obtain height
The paclitaxel prodrugs of purity.
Wherein, the taxusyunnanensis branches and leaves are smashed as 100~200 mesh.
Wherein, the extractant is water-soluble for the mixing of 0.1%~0.15% organic acid, 1.0%~5.0% acetone or alcohol
Liquid, volume ratio are 0.1%~0.15%：1.0%~5.0%：98.9%~94.85%.
Wherein, it is recycled after the extracting solution is recovered.
Wherein, the alcohol organic solvent is methanol or ethyl alcohol.
Wherein, the mixed solvent be methanol, dichloromethane and n-hexane, volume ratio 1：2：6.
Wherein, the acetonitrile crystallization is crystallization when 10~15 DEG C of placements 6 are small, and it acts as removal impurity, reach and progressively carry
High natural Japanese yew alcohol purity.
Wherein, the mixed solvent system crystallizes the mixed solvent system for methanol, dichloromethane and n-hexane, volume
Ratio is 1：2：6, when stirring 4 is small, 10~15 DEG C of crystallizations of temperature control, it acts as removal impurity, reach and step up natural Japanese yew
Alcohol purity.
The characteristics of AB-8 types resin does resin column purification therefore model resin to natural Japanese yew alcohol have spy
Sign property, can better purifying natural taxol.
Wherein in step (2), carried out using Zigzag type adverse current extracting mode.
Beneficial effects of the present invention：
The present invention researches and develops a kind of process route of natural Chinese yew extraction purification, and accomplishes scale production, product purity
Reach more than 98%, maximum list is miscellaneous to be less than 0.1%.
Innovative point：
(1) own technology extracts natural Japanese yew alcohol by raw material of taxusyunnanensis branches and leaves；
(2) a large amount of impurity in branches and leaves of yew are removed using water-soluble mixed solution, is using alcohols organic molten
Agent extracts taxol crude product；
(3) resin column purification is done using AB-8 type resins, then can obtain the natural purple of high-purity through crystal refining twice
China fir alcohol；
Description of the drawings
Fig. 1 is the process flow chart of invention.
Fig. 2 is the high-efficient liquid phase chromatogram of product prepared by the present invention.
Specific embodiment
Below in conjunction with specific embodiment, the invention will be further described.It is to be understood that following embodiment is merely to illustrate this
Invention is not for restriction the scope of the present invention.
Embodiment 1
Fig. 1 is the process flow chart of the present invention, as shown in Figure 1, a kind of extracting method of natural Japanese yew alcohol, including following step
Suddenly：
The taxusyunnanensis branches and leaves that 10kg is crushed will be prepared from 10 minutes uniform continuous inputs of extraction equipment entrance point
200kg mixed aqueous solutions (acetic acid 0.1%, the aqueous solution of acetone 2.0%) extracting solution simultaneously at the uniform velocity squeeze into extraction equipment, impregnate
Liquid is transferred to lower batch extraction procedure again from 1.0 tons of storage tanks are flow to after soak is recovered, solid extracts slag and squeezed by squeezer
After the water for removing the overwhelming majority, then turn back into extraction equipment, squeeze into 60kg methanol, the speed completed according to 10 minutes into
Material, after the completion of extraction, extracting solution is concentrated under reduced pressure into dripless and is oozed, after 10L acetone dissolved clarifications with AB-8 type resin sample preparations, resin
Column is purified by flash, and eluent is concentrated to dryness, and 2L acetonitriles are heated to 60 DEG C of dissolved clarifications, and vacuum decompression removes 1L acetonitriles, 10~15 DEG C
Place 6 it is small when crystallization, obtain solid after filtering, use the methanol of 2L and dichloromethane (methanol：Dichloromethane=0.66：1.34)
Mixed solvent by solid dissolved clarification, be added with stirring the n-hexane of 4L, when stirring 4 is small, 10~15 DEG C of crystallizations of temperature control, filtering drying
Obtained solid is natural Japanese yew alcohol finished product afterwards, packs censorship.
Embodiment 2
A kind of extracting method of natural Japanese yew alcohol, comprises the following steps：
The taxusyunnanensis branches and leaves that 100kg is crushed will be prepared from 80 minutes uniform continuous inputs of extraction equipment entrance point
Good 2000kg mixed aqueous solutions (acetic acid 0.1%, the aqueous solution of acetone 2.0%) extracting solution at the uniform velocity squeezes into extraction equipment simultaneously,
Soak is transferred to lower batch extraction procedure again from 10 tons of storage tanks are flow to after soak is recovered, solid extraction slag passes through squeezer
It after squeezing removes the water of the overwhelming majority, then turns back into extraction equipment, squeezes into 600kg methanol, the speed completed according to 80 minutes
Degree charging, after the completion of extraction, extracting solution is concentrated under reduced pressure into dripless and is oozed, after 100L acetone dissolved clarifications with AB-8 type resin systems
Sample, resin column are purified by flash, and eluent is concentrated to dryness, and 20L acetonitriles are heated to 60 DEG C of dissolved clarifications, and vacuum decompression removes 10L acetonitriles,
Crystallization when 10~15 DEG C of placements 6 are small, obtains solid after filtering, uses the methanol of 20L and dichloromethane (methanol：Dichloromethane=
0.66：1.34) solid dissolved clarification is added with stirring the n-hexane of 40L by mixed solvent, when stirring 4 is small, 10~15 DEG C of analysis of temperature control
Crystalline substance, obtained solid is natural Japanese yew alcohol finished product after filtering drying, packs censorship.
Embodiment 3
A kind of extracting method of natural Japanese yew alcohol, comprises the following steps：
The taxusyunnanensis branches and leaves of 1.0 tons of crushing from extraction equipment entrance point 8 it is small when uniformly continuously put into, will prepare
20 tons of mixed aqueous solutions (acetic acid 0.1%, the aqueous solution of acetone 2.0%) extracting solution simultaneously at the uniform velocity squeeze into extraction equipment, impregnate
Liquid is transferred to lower batch extraction procedure again from 50 tons of storage tanks are flow to after soak is recovered, solid extracts slag and squeezed by squeezer
After the water for removing the overwhelming majority, then turn back into extraction equipment, squeeze into 6.0 tons of methanol, the speed completed according to 8 when small into
Material, after the completion of extraction, is concentrated under reduced pressure into dripless by extracting solution and oozes, and with AB-8 type resin sample preparations after 1000L acetone dissolved clarifications, sets
Fat column is purified by flash, and eluent is concentrated to dryness, and 200L acetonitriles are heated to 60 DEG C of dissolved clarifications, and vacuum decompression removes 100L acetonitriles, and 10
Crystallization when~15 DEG C of placements 6 are small, obtains solid after filtering, uses the methanol of 200L and dichloromethane (methanol：Dichloromethane=
0.66：1.34) solid dissolved clarification is added with stirring the n-hexane of 400L by mixed solvent, when stirring 4 is small, 10~15 DEG C of temperature control
Crystallization, obtained solid is natural Japanese yew alcohol finished product after filtering drying, packs censorship.
Fig. 2 is the high-efficient liquid phase chromatogram of product prepared by the present invention, as shown in Figure 2.It is detected using conventional high-performance liquid phase
Product prepared by the present invention.
1 high performance liquid chromatography detection of table
As seen from Table 1, the present invention researches and develops a kind of extracting method of natural Japanese yew alcohol, and accomplishes scale production, and product is pure
Degree reaches more than 98%, and maximum list is miscellaneous to be less than 0.1%.Embody the novelty and creativeness of the present invention.
The specific embodiment of the present invention is illustrated above, but the present invention is not limited thereto, without departing from
Spirit of the invention, the present invention can also have various change.
Claims (9)
1. a kind of extracting method of natural Japanese yew alcohol, which is characterized in that including：
(1) taxusyunnanensis branches and leaves are taken, are crushed through pulverizer；
(2) it is the taxusyunnanensis branches and leaves after crushing are mixed through 0.1%~0.15% organic acid, 1.0%~5.0% acetone or alcohol
Heshui solution cooks extractant, extraction；
(3) slag after first time extracts, then the soak extraction through alcohol organic solvent obtain taxol crude product；
(4) taxol crude product does resin column purification by AB-8 type resins；
(5) crystallized using acetonitrile, the mixed solvent system crystallization of methanol, dichloromethane and n-hexane, so as to obtain high-purity
Paclitaxel prodrugs.
2. extracting method according to claim 1, which is characterized in that the taxusyunnanensis branches and leaves are smashed as 100~200
Mesh.
3. extracting method according to claim 1, which is characterized in that the extractant for 0.1%~0.15% organic acid,
The mixed aqueous solution of 1.0%~5.0% acetone or alcohol, volume ratio are 0.1%~0.15%：1.0%~5.0%：
98.9%~94.85%.
4. extracting method according to claim 3, which is characterized in that recycled after the extracting solution is recovered.
5. extracting method according to claim 1, which is characterized in that the alcohol organic solvent is methanol or ethyl alcohol.
6. extracting method according to claim 1, which is characterized in that the mixed solvent is methanol, dichloromethane and just
Hexane, volume ratio 1：2：6.
7. extracting method according to claim 1, which is characterized in that when the acetonitrile crystallization is that 10~15 DEG C of placements 6 are small
Crystallization, it acts as removal impurity, reach and step up natural Japanese yew alcohol purity.
8. extracting method according to claim 1, which is characterized in that the mixed solvent system crystallization is methanol, dichloro
The mixed solvent system of methane and n-hexane, volume ratio 1：2：6, when stirring 4 is small, 10~15 DEG C of crystallizations of temperature control are made
With to remove impurity, reach and step up natural Japanese yew alcohol purity.
9. extracting method according to claim 1, which is characterized in that wherein in step (2), extracted using Zigzag type adverse current
Mode carries out.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711386598.2A CN108101869A (en) | 2017-12-20 | 2017-12-20 | A kind of extracting method of natural Japanese yew alcohol |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711386598.2A CN108101869A (en) | 2017-12-20 | 2017-12-20 | A kind of extracting method of natural Japanese yew alcohol |
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| CN108101869A true CN108101869A (en) | 2018-06-01 |
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ID=62211455
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| CN201711386598.2A Pending CN108101869A (en) | 2017-12-20 | 2017-12-20 | A kind of extracting method of natural Japanese yew alcohol |
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| CN110845451A (en) * | 2019-11-28 | 2020-02-28 | 云南皓瑞逸生物科技有限公司 | Method for extracting paclitaxel from taxus chinensis |
| CN110857289A (en) * | 2018-08-22 | 2020-03-03 | 台江县吉阳生物科技有限公司 | Paclitaxel extraction method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110857289A (en) * | 2018-08-22 | 2020-03-03 | 台江县吉阳生物科技有限公司 | Paclitaxel extraction method |
| CN110845451A (en) * | 2019-11-28 | 2020-02-28 | 云南皓瑞逸生物科技有限公司 | Method for extracting paclitaxel from taxus chinensis |
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