CN103808853A - Thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis - Google Patents
Thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis Download PDFInfo
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Abstract
The invention relates to a thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis, and relates to a detection method. The method comprises the following steps of preparing 7-xylosyl-10-deacetyltaxol reference solution; (2) preparing a sample solution; and (3) carrying out the thin-layer chromatography detection. The method has the beneficial effects of simplicity, practicability and economy.
Description
Technical field
The present invention relates to a kind of detection method, be specifically related to the thin-layered chromatography detection method of 7-xylose-10-deacetyl paclitaxel in a kind of Chinese yew.
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.Tagging 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 division stop at m period, have blocked the proper splitting of cell.By the clinical research of II-III, taxol is mainly applicable to oophoroma and breast cancer, and lung cancer, colorectal cancer, melanoma, incidence cancer, lymthoma, brain tumor are also had to certain curative effect.
7-xylose-10-deacetyl paclitaxel can be taxol by bio-transformation and chemosynthesis, is important so detect 7-xylose-10-deacetyl paclitaxel.
Conventional 7-xylose-10-deacetyl paclitaxel detection method is that HPLC detects, and filler is reverse phase filler.Mobile phase is acetonitrile-water, and detecting device is UV-detector.
Summary of the invention
Conventional HPLC detects 7-xylose-10-deacetyl paclitaxel method and requires high:
(1) need a HPLC, a HPLC price is common more than 100,000, and HPLC price is high, and popularity rate is low;
(2) HPLC need to have professional personnel's operation, maintenance;
(3) HPLC detects needs a large amount of solvents, and the use of a large amount of solvents has also caused great pollution to environment;
(4) HPLC detects and can only detect a sample at every turn, and detection time is also long.
The invention provides a kind of easy operating, the qualitative sxemiquantitative of energy and economic thin-layered chromatography detection method, the method solves the part drawback that HPLC detects.
A thin-layered chromatography detection method for 7-xylose-10-deacetyl paclitaxel in Chinese yew, described method comprises the steps:
(1) take 7-xylose-10-deacetyl paclitaxel 0.1～1mg, use 1ml organic solvent dissolution, obtain 7-xylose-10-deacetyl paclitaxel reference substance solution;
(2) Chinese yew raw material is pulverized, with alcoholic solution lixiviate three times, merge three times extracting solution, concentrated extracting solution, to 1/3～1/2 of original volume, with equal-volume organic solvent I extraction three times, merges three times extraction phase, evaporate to dryness extraction phase, take evaporate to dryness powder 0.1～1mg, dissolve by 1ml organic solvent II, obtain sample solution;
(3) draw step (1) and (2) each 5～10 μ l of solution of preparation, point is on same silica gel g thin-layer plate, developping agent is halogenated hydrocarbons and alcohol, the volume ratio of halogenated hydrocarbons and alcohol is 100:10～1, launch, take out, dry, with 5～10% concentrated sulphuric acid ethanolic solution colour developings, dry thin layer plate to spot with 105 ℃ and develop the color.
The organic solvent of described method step in is (1) preferably methyl alcohol, absolute ethyl alcohol or N, N-METHYLFORMAMIDE, and N, it is high that N-METHYLFORMAMIDE dissolves 7-xylose-10-deacetyl paclitaxel solubleness, and methyl alcohol and absolute ethyl alcohol boiling point are low.
Described method step (1) Chinese yew raw material is preferably bark, branches and leaves and tree root.
Described method step (2) alcoholic solution is preferably methyl alcohol, absolute ethyl alcohol, methanol aqueous solution or ethanol water.
Described method step (2) organic solvent I is preferably methylene chloride, methenyl choloride or ethyl acetate.
Described method step (2) organic solvent II is preferably methyl alcohol, absolute ethyl alcohol or DMF, and DMF dissolving 7-xylose-10-deacetyl paclitaxel solubleness is high, and methyl alcohol and absolute ethyl alcohol boiling point are low.
Described method step (3) halogenated hydrocarbons is preferably methylene chloride or methenyl choloride.
Described method step (3) alcohol is preferably methyl alcohol or absolute ethyl alcohol.
Beneficial effect of the present invention is:
(1) 7-xylose-10-deacetyl paclitaxel detection method is simple, practical and economical.
(2) one-time detection can detect multiple samples simultaneously.
(3) testing result can quantitative and semi-quantitative.
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
A thin-layered chromatography detection method for 7-xylose-10-deacetyl paclitaxel in Chinese yew, described method comprises the steps:
(1) take 7-xylose-10-deacetyl paclitaxel standard items 0.5mg, with 1ml methyl alcohol ultrasonic dissolution, obtain 7-xylose-10-deacetyl paclitaxel reference substance solution, concentration 0.5mg/ml, as the standard of quantitative and semi-quantitative;
(2) bark of Ramulus et folium taxi cuspidatae, branches and leaves and tree root are pulverized, after pulverizing, with absolute ethyl alcohol lixiviate three times, merged three times extracting solution, distillation under vacuum concentrates extracting solution, vacuum tightness 0.8MPa, 55 ℃ of bath temperatures, be concentrated into 1/3 of original volume, use equal-volume dichloromethane extraction concentrate three times, merge three times extraction phase, distillation under vacuum evaporate to dryness extraction phase, vacuum tightness 0.4MPa, 35 ℃ of bath temperatures, take evaporate to dryness powder 0.5mg, with 1ml methyl alcohol ultrasonic dissolution, obtain sample solution;
(3) the each 10 μ l of solution that draw step (1) and (2) prepare with glass point sample kapillary, point is at same silica gel g thin-layer plate, on 1cm place same level line at the bottom of plate, developping agent is methenyl choloride and absolute ethyl alcohol, and the volume ratio of methenyl choloride and absolute ethyl alcohol is 100:5, balance 5～10min, launch, be expanded to apart from 1cm place, plate top and take out, take out, with hot blast drying, with 10% concentrated sulphuric acid ethanolic solution colour developing, dry thin layer plate to spot with 105 ℃ and develop the color.
Claims (8)
1. a thin-layered chromatography detection method for 7-xylose-10-deacetyl paclitaxel in Chinese yew, described method comprises the steps:
(1) take 7-xylose-10-deacetyl paclitaxel 0.1～1mg, use 1ml organic solvent dissolution, obtain 7-xylose-10-deacetyl paclitaxel reference substance solution;
(2) Chinese yew raw material is pulverized, with alcoholic solution lixiviate three times, merge three times extracting solution, concentrated extracting solution, to 1/3～1/2 of original volume, with equal-volume organic solvent I extraction three times, merges three times extraction phase, evaporate to dryness extraction phase, take evaporate to dryness powder 0.1～1mg, dissolve by 1ml organic solvent II, obtain sample solution;
(3) draw step (1) and (2) each 5～10 μ l of solution of preparation, point is on same silica gel g thin-layer plate, developping agent is halogenated hydrocarbons and alcohol, the volume ratio of halogenated hydrocarbons and alcohol is 100:10～1, launch, take out, dry, with 5～10% concentrated sulphuric acid ethanolic solution colour developings, dry thin layer plate to spot with 105 ℃ and develop the color.
2. method according to claim 1, is characterized in that: described method step (1) organic solvent is methyl alcohol, absolute ethyl alcohol or DMF.
3. method according to claim 1, is characterized in that: described method step (1) Chinese yew raw material is bark, branches and leaves and tree root.
4. method according to claim 1, is characterized in that: described method step (2) alcoholic solution is methyl alcohol, absolute ethyl alcohol, methanol aqueous solution or ethanol water.
5. method according to claim 1, is characterized in that: described method step (2) organic solvent I is methylene chloride, methenyl choloride or ethyl acetate.
6. method according to claim 1, is characterized in that: described method step (2) organic solvent II is methyl alcohol, absolute ethyl alcohol or DMF.
7. method according to claim 1, is characterized in that: described method step (3) halogenated hydrocarbons is methylene chloride or methenyl choloride.
8. method according to claim 1, is characterized in that: described method step (3) alcohol is methyl alcohol or absolute ethyl alcohol.
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| CN201210460881.6A CN103808853A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis |
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| CN201210460881.6A CN103808853A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109942515A (en) * | 2019-04-28 | 2019-06-28 | 云南汉德生物技术有限公司 | A method of extracting 10- deacetylate taxol |
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| US5750709A (en) * | 1993-03-12 | 1998-05-12 | Aphios Corporation | Method and apparatus for isolating therapeutic compositions from source materials |
| JP2002502613A (en) * | 1998-02-06 | 2002-01-29 | インデナ・ソチエタ・ペル・アツィオーニ | Production method of baccatin |
| US20060281914A1 (en) * | 2005-06-10 | 2006-12-14 | Florida State University Research Foundation, Inc. | Beta-lactam synthesis |
| CN101684451A (en) * | 2008-09-28 | 2010-03-31 | 中国医学科学院药物研究所 | Microorganism for hydrolyzing 7-xylose group and 13-side chain of taxane |
| CN101477102A (en) * | 2008-12-10 | 2009-07-08 | 上海康桥中药饮片有限公司 | Quality standard of south yew decoction pieces and its detection method |
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| KANG WANG 等: "Microbial hydrolysis of 7-xylosyl-10-deacetyltaxol to 10-deacetyltaxol", 《JOURNAL OF MOLECULAR CATALYSIS B: ENZYMATIC》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109942515A (en) * | 2019-04-28 | 2019-06-28 | 云南汉德生物技术有限公司 | A method of extracting 10- deacetylate taxol |
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Application publication date: 20140521 |