CN105153076A - Method for extracting paclitaxel from yew roots - Google Patents
Method for extracting paclitaxel from yew roots Download PDFInfo
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- CN105153076A CN105153076A CN201510492051.5A CN201510492051A CN105153076A CN 105153076 A CN105153076 A CN 105153076A CN 201510492051 A CN201510492051 A CN 201510492051A CN 105153076 A CN105153076 A CN 105153076A
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- 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 relates to a method for extracting paclitaxel from Yunnan yew roots and belongs to the field of refining processing. According to the method, the Yunnan yew roots are used as a raw material, broken, dissolved in an organic solvent and subjected to extraction and degreasing, and then gradient elution is performed, a leachate containing the paclitaxel is concentrated and dried, purification is performed again, and the high-purity paclitaxel is obtained. The extraction and purification rate of the paclitaxel is increased, the extraction and purification cost is reduced, the yew roots are regenerative, the obtained raw material is richer, operation is easy, separation is thorough, and the medicinal standard can be reached directly.
Description
Technical field
The present invention relates to a kind of method extracting taxol from Ramulus et folium taxi cuspidatae tree root, belong to refining manufacture field.
Background technology
Taxol is a kind of compound extracted from Chinese yew, mainly be applicable to treatment ovarian cancer and mammary cancer, and lung cancer, large bowel cancer, melanoma, incidence cancer, lymphoma, brain tumor etc., be one of the most effective PTS after Zorubicin, total effective rate reaches more than 75%.For a long time, taxol extracts from the bark of Ramulus et folium taxi cuspidatae, because the bark of Ramulus et folium taxi cuspidatae is Nonrenewable resources, causes the acquisition raw material of taxol very limited; In addition; Ramulus et folium taxi cuspidatae is the endangered natural rare plants for anticancer of generally acknowledging in the world; the speed of growth is slow under field conditions (factors); regenerative power is poor; the Ramulus et folium taxi cuspidatae raw material forestbase of advising greatly and touching also is not formed in world wide; China has been classified as one-level rare endangered plants, and United Nations also prohibites felling, thus makes the acquisition raw material of taxol more deficient.
Extremely limited amount taxol is all contained in the Ramulus et folium taxi cuspidatae of all kinds.Such as, in the bark of yewtree and taxusyunnanensis tree containing have an appointment 0.02% taxol, containing the taxol of 0.005% to about 0.1% of having an appointment in media hicksii ground needle and root.Therefore, the effective method extracting high-purity taxol from vegetable material of exploitation has caused the great interest of people.
Summary of the invention
The technical problem that the present invention mainly solves: though for current technology can from the branches and leaves of Ramulus et folium taxi cuspidatae effective separation and Extraction taxol, but its purification effect is very undesirable, directly cannot reach medicinal standard, need to carry out follow-up purification process, thus increase the extraction cost of taxol, extraction efficiency is low, can not the present situation of proper scale mass production, provides a kind of method extracting taxol from taxusyunnanensis tree root.The method for raw material, smashes rear organic solvent dissolution with taxusyunnanensis tree root, extraction also degreasing, then carries out gradient elution, by the leacheate concentrate drying containing taxol, again purifies, obtains highly purified taxol.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
(1) get 1 ~ 5kg Ramulus et folium taxi cuspidatae tree root, put into the fragment that pulverizer is ground into 1 ~ 2cm, add the aqueous isopropanol that concentration is 70 ~ 80%, put into fermentor tank, set temperature is 28 ~ 38 DEG C, sealing and fermenting 24 ~ 36h;
(2) by above-mentioned filtering fermentation liquor, allow the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing;
(3) by ethanol and sherwood oil volume ratio be 1:1 ~ 1:3 prepare mixed solution, again extract is carried out mixing and stirring than for 1:0.1 ~ 1:0.5 with mixeding liquid volume, put into high pressure degreasing tank, solvent degreasing process 2 ~ 5h, skimming temp is 50 ~ 80 DEG C, pressure is 0.1 ~ 0.5MPa, room temperature is cooled to after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 120 ~ 130 DEG C, process 1 ~ 2h, then be cooled to room temperature release to take out, filter, put into baking oven, dry, oven temperature is 105 ~ 110 DEG C, obtain taxol crude product,
(4) taxol crude product and its silica gel of 10 ~ 20 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 200 ~ 250L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:5:2, use chromatography isolation of taxol, its retention time is 14min, flows out in 13 ~ 15min and collects taxol, the taxol collected is decompressed to 900 ~ 1000Pa and carries out concentrating under reduced pressure in moving phase, enriched material is put into baking oven, dry, oven temperature is 100 ~ 105 DEG C, obtains taxol work in-process;
(5) Virahol of work in-process by its 5 ~ 10 times of weight is dissolved, adds sherwood oil while stirring, until have crystal separate out till, leave standstill 15 ~ 20h, filter, put into oven for drying, oven temperature is 105 ~ 110 DEG C, namely obtain purity more than 80% taxol.
The invention has the beneficial effects as follows:
(1) taxol extraction purification rate increases, and reduces cost for purification;
(2) taxol root can regenerate, and obtains raw material abundanter;
(3) processing ease, be separated thoroughly, directly can reach medicinal standard.
Specific embodiments
Get 1 ~ 5kg Ramulus et folium taxi cuspidatae tree root, put into the fragment that pulverizer is ground into 1 ~ 2cm, add the aqueous isopropanol that concentration is 70 ~ 80%, put into fermentor tank, set temperature is 28 ~ 38 DEG C, sealing and fermenting 24 ~ 36h;
By above-mentioned filtering fermentation liquor, allow the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing; By ethanol and sherwood oil volume ratio be 1:1 ~ 1:3 prepare mixed solution, again extract is carried out mixing and stirring than for 1:0.1 ~ 1:0.5 with mixeding liquid volume, put into high pressure degreasing tank, solvent degreasing process 2 ~ 5h, skimming temp is 50 ~ 80 DEG C, and pressure is 0.1 ~ 0.5MPa, is cooled to room temperature after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 120 ~ 130 DEG C, process 1 ~ 2h, then be cooled to room temperature release to take out, filter, put into baking oven, dry, oven temperature is 105 ~ 110 DEG C, obtains taxol crude product; Taxol crude product and its silica gel of 10 ~ 20 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 200 ~ 250L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:5:2, use chromatography isolation of taxol, its retention time is 14min, flows out in 13 ~ 15min and collects taxol, the taxol collected is decompressed to 900 ~ 1000Pa and carries out concentrating under reduced pressure in moving phase, enriched material is put into baking oven, dry, oven temperature is 100 ~ 105 DEG C, obtains taxol work in-process; The Virahol of work in-process by its 5 ~ 10 times of weight is dissolved, adds sherwood oil while stirring, until have crystal separate out till, leave standstill 15 ~ 20h, filter, put into oven for drying, oven temperature is 105 ~ 110 DEG C, namely obtain purity more than 80% taxol.
After testing, the content of taxol extracted can reach more than 80%.
Get 1kg taxusyunnanensis tree root, put into the fragment that pulverizer is ground into 1cm, add the aqueous isopropanol that concentration is 70%, soak 15h; Above-mentioned soak solution is filtered, allows the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing; Be that 1:1 prepares mixed solution by ethanol and sherwood oil volume ratio, then extract and mixeding liquid volume are carried out mixing and stirring than for 1:0.1, put into high pressure degreasing tank, solvent degreasing process 2h, skimming temp is 50 DEG C, and pressure is 0.1MPa, be cooled to room temperature after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 120 DEG C, process 1h, be then cooled to room temperature release and take out, filter, put into baking oven, dry, oven temperature is 105 DEG C, obtains taxol crude product; Thick taxol and its silica gel of 10 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 200L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:5:1, the taxol in eluted fraction is detected by high performance liquid chromatography, when taxol flow point being detected, collecting the drip washing flow point containing taxol, the drip washing flow point containing taxol being decompressed to 900Pa and carrying out concentrating under reduced pressure, enriched material is put into baking oven, dry, oven temperature is 100 DEG C, obtains taxol work in-process; Dissolved by the Virahol of work in-process by its 5 times of weight, add sherwood oil while stirring, till having crystal to separate out, leave standstill 15h, filter, put into oven for drying, oven temperature is 105 DEG C, namely obtains highly purified taxol.
After testing, the content of taxol extracted reaches 82%.
Get 3kg taxusyunnanensis tree root, put into the fragment that pulverizer is ground into 1.5cm, add the aqueous isopropanol that concentration is 75%, soak 18h; Above-mentioned soak solution is filtered, allows the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing; Be that 1:2 prepares mixed solution by ethanol and sherwood oil volume ratio, then extract and mixeding liquid volume are carried out mixing and stirring than for 1:0.3, put into high pressure degreasing tank, solvent degreasing process 3h, skimming temp is 60 DEG C, and pressure is 0.3MPa, be cooled to room temperature after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 125 DEG C, process 1.5h, be then cooled to room temperature release and take out, filter, put into baking oven, dry, oven temperature is 108 DEG C, obtains taxol crude product; Thick taxol and its silica gel of 15 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 230L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:6:1, the taxol in eluted fraction is detected by high performance liquid chromatography, when taxol flow point being detected, collecting the drip washing flow point containing taxol, the drip washing flow point containing taxol being decompressed to 950Pa and carrying out concentrating under reduced pressure, enriched material is put into baking oven, dry, oven temperature is 103 DEG C, obtains taxol work in-process; Dissolved by the Virahol of work in-process by its 8 times of weight, add sherwood oil while stirring, till having crystal to separate out, leave standstill 18h, filter, put into oven for drying, oven temperature is 108 DEG C, namely obtains highly purified taxol.
After testing, the content of taxol extracted reaches 90%.
Get 5kg taxusyunnanensis tree root, put into the fragment that pulverizer is ground into 2cm, add the aqueous isopropanol that concentration is 80%, soak 20h; Above-mentioned soak solution is filtered, allows the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing; Be that 1:3 prepares mixed solution by ethanol and sherwood oil volume ratio, then extract and mixeding liquid volume are carried out mixing and stirring than for 1:0.5, put into high pressure degreasing tank, solvent degreasing process 5h, skimming temp is 80 DEG C, and pressure is 0.5MPa, be cooled to room temperature after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 130 DEG C, process 2h, be then cooled to room temperature release and take out, filter, put into baking oven, dry, oven temperature is 110 DEG C, obtains taxol crude product; Thick taxol and its silica gel of 20 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 250L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:7:2, the taxol in eluted fraction is detected by high performance liquid chromatography, when taxol flow point being detected, collecting the drip washing flow point containing taxol, the drip washing flow point containing taxol being decompressed to 1000Pa and carrying out concentrating under reduced pressure, enriched material is put into baking oven, dry, oven temperature is 105 DEG C, obtains taxol work in-process; Dissolved by the Virahol of work in-process by its 10 times of weight, add sherwood oil while stirring, till having crystal to separate out, leave standstill 20h, filter, put into oven for drying, oven temperature is 110 DEG C, namely obtains highly purified taxol.
After testing, the content of taxol extracted reaches 88%.
Claims (1)
1. from Ramulus et folium taxi cuspidatae tree root, extract a method for taxol, it is characterized in that concrete preparation process is:
(1) get 1 ~ 5kg Ramulus et folium taxi cuspidatae tree root, put into the fragment that pulverizer is ground into 1 ~ 2cm, add the aqueous isopropanol that concentration is 70 ~ 80%, put into fermentor tank, set temperature is 28 ~ 38 DEG C, sealing and fermenting 24 ~ 36h;
(2) by above-mentioned filtering fermentation liquor, allow the filtrate obtained by activated carbon column, decolour, obtain destainer, then with methylene dichloride, destainer is extracted, after normal pressure rising temperature reclamation solvent, be extracted thing;
(3) by ethanol and sherwood oil volume ratio be 1:1 ~ 1:3 prepare mixed solution, again extract is carried out mixing and stirring than for 1:0.1 ~ 1:0.5 with mixeding liquid volume, put into high pressure degreasing tank, solvent degreasing process 2 ~ 5h, skimming temp is 50 ~ 80 DEG C, pressure is 0.1 ~ 0.5MPa, room temperature is cooled to after degreasing completes, discharge degreasing wasting liquid, after drained waste liquid, pass into the superheated vapour of 120 ~ 130 DEG C, process 1 ~ 2h, then be cooled to room temperature release to take out, filter, put into baking oven, dry, oven temperature is 105 ~ 110 DEG C, obtain taxol crude product,
(4) taxol crude product and its silica gel of 10 ~ 20 times are stirred, dress post, carry out gradient elution, rate of flow in rinse is 200 ~ 250L/h, leacheate is trichloromethane, ethanol, espeleton, three's mixed volume is than being 100:5:2, use chromatography isolation of taxol, its retention time is 14min, flows out in 13 ~ 15min and collects taxol, the taxol collected is decompressed to 900 ~ 1000Pa and carries out concentrating under reduced pressure in moving phase, enriched material is put into baking oven, dry, oven temperature is 100 ~ 105 DEG C, obtains taxol work in-process;
(5) Virahol of work in-process by its 5 ~ 10 times of weight is dissolved, adds sherwood oil while stirring, until have crystal separate out till, leave standstill 15 ~ 20h, filter, put into oven for drying, oven temperature is 105 ~ 110 DEG C, namely obtain purity more than 80% taxol.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107400104A (en) * | 2017-04-11 | 2017-11-28 | 浙江大德龙生物技术有限公司 | A kind of taxol ultrasonic purification method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107400104A (en) * | 2017-04-11 | 2017-11-28 | 浙江大德龙生物技术有限公司 | A kind of taxol ultrasonic purification method |
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Application publication date: 20151216 |