CN1994995A - Method for extracting and purifying sequoyitol - Google Patents
Method for extracting and purifying sequoyitol Download PDFInfo
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- CN1994995A CN1994995A CN 200610023151 CN200610023151A CN1994995A CN 1994995 A CN1994995 A CN 1994995A CN 200610023151 CN200610023151 CN 200610023151 CN 200610023151 A CN200610023151 A CN 200610023151A CN 1994995 A CN1994995 A CN 1994995A
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Abstract
The invention discloses an extracting and purifying method of sequoia alcohol, which comprises the following steps: 1. extracting effective component in the sequoia based on alcohol, acetone or water as extract; obtaining rough raw material with 0.3-5%(wt) sequoia alcohol; 2. adopting rough raw material of sequoia alcohol as raw material to separate through chromatographic system; obtaining the raw material with over 10% sequoia alcohol; 3. crystallizing the sequoia alcohol for 2-4 times; obtaining pure sequoia alcohol with purity over 99%.
Description
Technical field
The present invention relates to the traditional Chinese medicine extraction field, relate in particular to a kind of method of extraction and purification sequoyitol.
Background technology
Sequoyitol (Sequoyitol) is an isolated natural organic-compound from Ramulus et folium taxi cuspidatae, can be used to treat indications such as diabetes.Its molecular formula is as follows:
From Ramulus et folium taxi cuspidatae, can extract taxol, Cephalomannine, 10-DAB III and other important organism, and utilize the 10-DAB III can semi-synthetic Docetaxel.Wherein, taxol and Docetaxel are one of the most effective cancer therapy drug of generally acknowledging in the world.Research starts from 1984 to one of Ramulus et folium taxi cuspidatae effective constituent taxol antitumor action in China, listed " the Seventh Five-Year Plan " tackling key problem scientific effort of national medical general bureau in 1986 in, carry out clinical study through Ministry of Health's approval to domestic paclitaxel February nineteen ninety-five, obtains license licensed licenser licence September in the same year.Taxol is described as one of great achievement of China's antitumour drug nineties.
But up to this point, no matter be in the world or domestic patent or document, all also do not have to find about extracting the relevant report of sequoyitol from plant Ramulus et folium taxi cuspidatae (comprising other homology species) or in other corresponding plants.Wherein, close with it patent documentation, as on April 14th, 2004, " a kind of plant milk extract of preventing and treating diabetes and preparation method thereof and the pharmaceutical use " of China Medicine University's application, publication number: CN1488344A, its document also relates to and extract a kind of plant milk extract of preventing and treating diabetes from Ramulus et folium taxi cuspidatae, and its main component is a myo-inositol derivative compounds, and its content is more than 60%.Though, sequoyitol also belongs to the inositol derivative compounds, but the invention of China Medicine University is meant myo-inositol derivative compounds, and since in the different and space structure of the position of methyl in 6 hydroxyls the chiral structure of hydroxyl differ, caused myo-inositol derivative compounds to comprise one-component more than at least 10 kinds.So, say on the stricti jurise that the invention of China Medicine University mainly is meant a kind of mixture but not a kind of one-component; In addition, 60% content is meant that the total content of myo-inositol derivative compounds is more than 60% in the patent of China Medicine University, and its yield is very low.
Summary of the invention
Technical problem to be solved by this invention is to provide the method for a kind of extraction and purifying sequoyitol, to solve defective of the prior art.
The invention provides a kind of from the medicinal plant Ramulus et folium taxi cuspidatae, especially southern china Ramulus et folium taxi cuspidatae (Taxus chinensis var.mairei), taxusyunnanensis (Taxus Yunnanensis), Europe berry Ramulus et folium taxi cuspidatae (Taxus Baccata), T. canadensis (Taxus Canadensis), with extraction, isolating sequoyitol in the Pacific Ocean Ramulus et folium taxi cuspidatae Ramulus et folium taxi cuspidatae such as (Taxus Pacific, Taxus Brevifolia), content can reach more than 99%.This compound can be directly used in antidiabetic medicines such as preparation injection, capsule, tablet.
The method of from the medicinal plant Ramulus et folium taxi cuspidatae, extracting sequoyitol provided by the present invention, summary comprises as follows: use solvent extraction, concentrate and obtain medicinal extract, two kinds of methods of medicinal extract branch dry method upper prop and wet method upper prop are carried out column chromatography for separation, collect the flow point that contains the sequoyitol component, through concentrating, again with alcohols or repeatedly crystallization of acetone, after filtration, drying, obtain the sequoyitol bulk drug intermediate more than 99%.
The invention provides the method for a kind of extraction and purifying sequoyitol, this method comprises the steps: that A. extracts Ramulus et folium taxi cuspidatae; Specifically comprise the steps:
(1) the Ramulus et folium taxi cuspidatae raw material is pulverized, made its particle diameter below 1mm;
(2) with extraction agent above-mentioned raw materials is extracted, remove by filter filter residue after, obtain extracting solution, the processing condition during extraction are:
Solid-liquid ratio: material (w): extraction agent (V)=1: 2～5;
Extract temperature: 40～100 ℃;
Extraction time: 3～5 hours;
Mixing speed: 60～240 rev/mins;
Extraction time: 1～3 time.
Wherein used Ramulus et folium taxi cuspidatae comprises: southerm yew (TAXUS CHINENSIS VARMAIREI (LEMEE ET L ' EVL) CHENG ET L.K.FU), taxusyunnanensis (TAXUS YUENNAN ENSIS CHENG ET L.K.FU), Xizang Taxus chinensis (TAXUS WALL ICHIANA IUCC), taxus chinensis in northeast (TAXUS CUSPIDATASTEB.ET IUCC), China Ramulus et folium taxi cuspidatae (TAXUS CHINENSIS (PILGER) ROHD), Taxus x media (TAXUS MEDIA), Europe berry Ramulus et folium taxi cuspidatae (TAXUSBACCATA), T. canadensis (TAXUS CANADENSIS), Pacific Ocean Ramulus et folium taxi cuspidatae (TAXUS PACIFIC TAXUS BREVIFOLIA).
Said Ramulus et folium taxi cuspidatae raw material comprises: leaf, tender shoots, branch, bark, trunk, tree root etc., can be wet also can be dried.
Extraction agent described in the step (2) is one or more of alcohols, acetonitrile, acetone or water, and wherein alcohols comprises methyl alcohol, ethanol, Virahol or propyl carbinol; Preferred extraction agent is methyl alcohol, ethanol or water.
The method of extraction provided by the present invention and purifying sequoyitol also comprises the steps B: the Ramulus et folium taxi cuspidatae crude extract is carried out purifying, comprise the steps: specifically
(1) with gained extracting solution in the steps A through concentrating under reduced pressure, obtain sequoyitol content and be 0.3%～5% concentrated extracting solution coarse raw materials, be called for short crude product A;
(2) with methyl alcohol or dissolve with ethanol crude product A, making the crude product A (W) and the ratio of methyl alcohol or ethanol (V) is 1: 2-10 makes it become uniform solution;
(3) be that the silica gel of 10～50 μ m is made sorbent material with adding particle diameter in the solution of gained in the step (2), stir, revaporization is removed methyl alcohol or ethanol is made " silica-gel mixture ", then this " silica-gel mixture " is placed the top (dry method upper prop) of the glass or the stainless steel chromatography post of filling gel; Or the solution in the step (2) is incorporated in fills macroporous resin or the glass of gac or the top (wet method upper prop) of stainless steel chromatography post;
(4) with 0.1～0.8: the mixed solvent that 1 alcohols and acetone are formed carries out elution chromatography, the Fractional Collections cut, and sampling analysis is removed qualified cut evaporation the sequoyitol " crude product B " that obtains content 〉=10% (wt) behind the mixed solvent.
Wherein the macroporous resin described in the step (3) is strongly-acid macroporous resin or slightly acidic macroporous resin; Alcohols described in the step (4) is methyl alcohol, ethanol, Virahol or propyl carbinol.
The method of extraction provided by the present invention and purifying sequoyitol can also comprise the steps C: sequoyitol is carried out crystallization treatment, specifically comprise the steps:
With crude product B is raw material, and in the alcohols or acetone solvent that 50 ℃～100 ℃ are dissolved in, raw material (W) is 1 with the ratio of solvent (V): 2-10, and freezing and crystallizing filters then; So repeat crystallization 2-3 time, dry after filtration, obtain the pure product sequoyitol more than 99%.
The product that finally obtains is the white crystals sprills, and main component is sequoyitol (Sequoyitol), and content can reach more than 99%, even more than 99.9%.
Content to the main component sequoyitol in the method for the present invention can detect with following high performance liquid phase (HPLC) method:
Assay: get the about 25mg of this product, it is fixed closely to claim, in the 25ml volumetric flask, is diluted to scale marks with moving phase, shakes up, and gets need testing solution.The HPLC chromatographic condition: the C18 post, 5 μ m, 4.6 * 250mm, refractive power or evaporation scatter detector, sample size 20 μ l, moving phase: methyl alcohol: water=50: 50 filters and outgases flow velocity 1.0ml/min through the organic worry film of 0.45 μ m before using.
Description of drawings
Fig. 1: sequoyitol preparation flow figure
Embodiment
Be exactly the embodiment that concrete extraction of the present invention and purge process provide some below, further to illustrate content of the present invention.But should be understood that these embodiment only for the purpose of illustration, get scope but not be used to limit the present invention.
Embodiment 1:
Get 200 kilograms in the bright branches and leaves of southerm yew, extracted 3 times in 3 hours with 80% methanol eddy of 2.5 times of volumes, united extraction liquid is concentrated into thick solution, adds the silica gel of 1: 1 (volume/volume), stir, and vacuum drying, the raw material dry sample is made in pulverizing.In material: the ratio dress post of silica gel=1: 20, last sample.With methyl alcohol: acetone=15: 85 and methyl alcohol: the ratio gradient elution of acetone=40: 60, detect segmentation with the HPLC method, merge product section, be concentrated into dried.With dehydrated alcohol crystallization 3 times, filter, drying obtains 124.1 gram white crystals dress powder, detects with the HPLC method, and wherein the content of sequoyitol is 99.68%.
Embodiment 2:
Get 200 kilograms in the dried branches and leaves of European berry Ramulus et folium taxi cuspidatae, reflux in 50 ℃ with 95% ethanol of 2.5 times of volumes and extracted 3 times in 3 hours, united extraction liquid, be concentrated into can not concentrate till.Use mobile phase methanol: water=dissolving fully in 70: 30, go up acid macroporous resin chromatography column, with methyl alcohol: water=70: 30～50 are the eluent gradient wash-out, with the monitoring of HPLC method, merge the sequoyitol cut, be evaporated to dried, with acetone crystallization 3 times, obtain 212.5 gram white crystals sprills, detect through the HPLC method, wherein the content of sequoyitol is 99.98%.
Embodiment 3:
Get Pacific Ocean Ramulus et folium taxi cuspidatae and do 2 kilograms of bark fines, reflux in 45 ℃ with 2.5 times of amount 80% acetonitriles and extracted 3 times in 3 hours, united extraction liquid, be concentrated into can not concentrate till.Use mobile phase methanol: water=dissolving fully in 70: 30, last activated carbon chromatography post, with methyl alcohol: water=70: 30～60 are the eluent gradient wash-out, with the monitoring of HPLC method, merge the sequoyitol cut, be evaporated to dried, with methanol crystallization 3 times, obtain 1.64 gram white crystals sprills, detect through the HPLC method, wherein the content of sequoyitol is 99.12%.
Embodiment 4:
Get 200 kilograms in the dried tree root powder of Taxus x media, reflux in 20 ℃ with 2.5 times of amount 98% acetone and extracted 3 times in 3 hours, united extraction liquid is concentrated into the thickness state, adds 1: 1 (volume/volume) diatomite, stirs, and vacuum drying is pulverized, and makes dry sample.In material: the ratio upper prop dress post of diatomite=1: 20, use moving phase ethanol: acetone=20～50: 80 are the eluent gradient wash-out, monitor with the HPLC method, merge the sequoyitol cut, be evaporated to driedly,, obtain 84.8 gram white crystals sprills with Virahol crystallization 4 times, detect through the HPLC method, wherein the content of sequoyitol is 99.35%.
Claims (6)
1, a kind of method of extraction and purification sequoyitol, the A that it is characterized in that the method comprising the steps of:
Ramulus et folium taxi cuspidatae is extracted, and concrete steps are:
(1) the Ramulus et folium taxi cuspidatae raw material is pulverized, made its particle diameter below 1mm;
(2) with extraction agent above-mentioned raw materials is extracted, remove by filter filter residue after, obtain extracting solution, the processing condition during extraction are:
Solid-liquid ratio: material (w): extraction agent (V)=1: 2～5;
Extract temperature: 40～100 ℃;
Extraction time: 3～5 hours;
Mixing speed: 60～240 rev/mins;
Extraction time: 1～3 time.
2, method according to claim 1 is characterized in that described extraction agent is one or more of alcohols, acetonitrile, acetone or water, and wherein alcohols comprises methyl alcohol, ethanol, Virahol or propyl carbinol.
3, method according to claim 2 is characterized in that described extraction agent is methyl alcohol, ethanol or water.
4, according to each described method of claim 1-3, it is characterized in that this method also comprises step B: the Ramulus et folium taxi cuspidatae crude extract is carried out purifying, and concrete steps are:
(1) with gained extracting solution in the steps A through concentrating under reduced pressure, obtain sequoyitol content and be 0.3%～5% concentrated extracting solution coarse raw materials, be called for short crude product A;
(2) with methyl alcohol or dissolve with ethanol crude product A, making the crude product A (W) and the ratio of methyl alcohol or ethanol (V) is 1: 2-10 makes it become uniform solution;
(3) be that the silica gel of 10～50 μ m is made sorbent material with adding particle diameter in the solution of gained in the step (2), stir, revaporization is removed methyl alcohol or ethanol is made " silica-gel mixture ", then this " silica-gel mixture " is placed the top of the glass or the stainless steel chromatography post of filling gel; Or the solution in the step (2) is incorporated in fills macroporous resin or the glass of gac or the top of stainless steel chromatography post;
(4) with 0.1～0.8: the mixed solvent that 1 alcohols and acetone are formed carries out elution chromatography, the Fractional Collections cut, and sampling analysis obtains sequoyitol " crude product B " after qualified cut evaporation removed mixed solvent;
Wherein the macroporous resin described in the step (3) is strongly-acid macroporous resin or slightly acidic macroporous resin; Alcohols described in the step (4) is methyl alcohol, ethanol, Virahol or propyl carbinol.
5, method according to claim 4 is characterized in that this method also comprises step C: described sequoyitol is carried out crystallization treatment, concrete steps are:
With crude product B is raw material, and in the alcohols or acetone solvent that 50 ℃～100 ℃ are dissolved in, raw material (W) is 1 with the ratio of solvent (V): 2-10, and freezing and crystallizing filters then; So repeat crystallization 2-3 time, and filtration drying;
Wherein said alcohols is methyl alcohol, ethanol, Virahol or propyl carbinol.
6, method according to claim 5 is characterized in that this method comprises the steps:
A, Ramulus et folium taxi cuspidatae is extracted:
(1) the Ramulus et folium taxi cuspidatae raw material is pulverized, made its particle diameter below 1mm;
(2) with methyl alcohol, ethanol or water above-mentioned raw materials is extracted, remove by filter filter residue after, obtain extracting solution, the processing condition during extraction are:
Solid-liquid ratio: material (w): extraction agent (V)=1: 2～5;
Extract temperature: 40～100 ℃;
Extraction time: 3～5 hours;
Mixing speed: 60～240 rev/mins;
Extraction time: 1～3 time;
B, the Ramulus et folium taxi cuspidatae crude extract is carried out purifying:
(1) with gained extracting solution in the steps A through concentrating under reduced pressure, obtain sequoyitol content and be 0.3%～5% concentrated extracting solution coarse raw materials, be called for short crude product A;
(2) with methyl alcohol or dissolve with ethanol crude product A, making the crude product A (W) and the ratio of methyl alcohol or ethanol (V) is 1: 2-10 makes it become uniform solution;
(3) be that the silica gel of 10～50 μ m is made sorbent material with adding particle diameter in the solution of gained in the step (2), stir, revaporization is removed methyl alcohol or ethanol is made " silica-gel mixture ", then this " silica-gel mixture " is placed the top of the glass or the stainless steel chromatography post of filling gel; Or the solution in the step (2) is incorporated in fills strongly-acid macroporous resin or the glass of slightly acidic macroporous resin or gac or the top of stainless steel chromatography post;
(4) with 0.1～0.8: the mixed solvent that 1 alcohols and acetone are formed carries out elution chromatography, the Fractional Collections cut, and sampling analysis obtains sequoyitol " crude product B " after qualified cut evaporation removed mixed solvent;
C: sequoyitol is carried out crystallization treatment:
With " crude product B " is raw material, and in the alcohols or acetone solvent that 50 ℃～100 ℃ are dissolved in, raw material (W) is 1 with the ratio of solvent (V): 2-10, and freezing and crystallizing filters then; So repeat crystallization 2-3 time, and filtration drying;
Wherein said alcohols is methyl alcohol, ethanol, Virahol or propyl carbinol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610023151 CN1994995A (en) | 2006-01-06 | 2006-01-06 | Method for extracting and purifying sequoyitol |
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| CN 200610023151 CN1994995A (en) | 2006-01-06 | 2006-01-06 | Method for extracting and purifying sequoyitol |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560197B (en) * | 2009-06-01 | 2011-07-06 | 西北农林科技大学 | Extraction method of taxol from branches and leaves of artificially cultivated yew |
| CN107879904A (en) * | 2017-11-08 | 2018-04-06 | 长沙爱扬医药科技有限公司 | The method that sequoyitol and oleanolic acid are extracted from orpiment skin |
| CN108069837A (en) * | 2017-12-28 | 2018-05-25 | 云南汉德生物技术有限公司 | A kind of method that sequoyitol is extracted from Chinese yew |
| CN113845496A (en) * | 2021-10-12 | 2021-12-28 | 华中科技大学 | Method for synchronously and efficiently separating various taxanes |
-
2006
- 2006-01-06 CN CN 200610023151 patent/CN1994995A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560197B (en) * | 2009-06-01 | 2011-07-06 | 西北农林科技大学 | Extraction method of taxol from branches and leaves of artificially cultivated yew |
| CN107879904A (en) * | 2017-11-08 | 2018-04-06 | 长沙爱扬医药科技有限公司 | The method that sequoyitol and oleanolic acid are extracted from orpiment skin |
| CN108069837A (en) * | 2017-12-28 | 2018-05-25 | 云南汉德生物技术有限公司 | A kind of method that sequoyitol is extracted from Chinese yew |
| CN108069837B (en) * | 2017-12-28 | 2020-08-07 | 云南汉德生物技术有限公司 | Method for extracting sequoyitol from taxus chinensis |
| CN113845496A (en) * | 2021-10-12 | 2021-12-28 | 华中科技大学 | Method for synchronously and efficiently separating various taxanes |
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