CN103808852A - Thin-layer chromatography detection method of taxol in taxus chinensis - Google Patents
Thin-layer chromatography detection method of taxol in taxus chinensis Download PDFInfo
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- CN103808852A CN103808852A CN201210460859.1A CN201210460859A CN103808852A CN 103808852 A CN103808852 A CN 103808852A CN 201210460859 A CN201210460859 A CN 201210460859A CN 103808852 A CN103808852 A CN 103808852A
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Abstract
The invention relates to a thin-layer chromatography detection method of taxol in taxus chinensis, and relates to a detection method. The method comprises the following steps of (1) preparing a taxol 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 taxol 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.
Conventional taxol 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 taxol 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 taxol in Chinese yew, described method comprises the steps:
(1) take taxol 0.1～1mg, use 1ml organic solvent dissolution, obtain taxol 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 machine solvent having in described method step is preferably methyl alcohol, absolute ethyl alcohol or N, N-METHYLFORMAMIDE, and N, it is high that N-METHYLFORMAMIDE dissolves taxol solubility, 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, N, and it is high that N-METHYLFORMAMIDE dissolves taxol solubility, 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) taxol 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 taxol in Chinese yew, described method comprises the steps:
(1) take taxol standard items 0.5mg, with 1ml methyl alcohol ultrasonic dissolution, obtain taxol 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 taxol in Chinese yew, described method comprises the steps:
(1) take taxol 0.1～1mg, use 1ml organic solvent dissolution, obtain taxol 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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210460859.1A CN103808852A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of taxol in taxus chinensis |
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| CN201210460859.1A CN103808852A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of taxol in taxus chinensis |
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| CN103808852A true CN103808852A (en) | 2014-05-21 |
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| CN201210460859.1A Pending CN103808852A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of taxol in taxus chinensis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110441439A (en) * | 2019-07-01 | 2019-11-12 | 杭州师范大学 | It is a kind of for distinguishing the metabolic markers and its detection method of heavy foliage Chinese yew and taxusyunnanensis |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0940567A (en) * | 1995-07-27 | 1997-02-10 | Taisho Pharmaceut Co Ltd | Production of taxol and taxane terpenoid |
| US20030073617A1 (en) * | 1996-03-12 | 2003-04-17 | Pg-Txl Company, L.P. | Water soluble paclitaxel derivatives |
| EP1818328A1 (en) * | 2006-02-10 | 2007-08-15 | 6570763 Canada Inc. | Chromatographic method for the isolation and purification of taxane derivatives |
| CN101113976A (en) * | 2006-07-27 | 2008-01-30 | 黑龙江大学 | Portable measuring method for measuring content of paclitaxel compounds in fermentation liquor |
-
2012
- 2012-11-15 CN CN201210460859.1A patent/CN103808852A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0940567A (en) * | 1995-07-27 | 1997-02-10 | Taisho Pharmaceut Co Ltd | Production of taxol and taxane terpenoid |
| US20030073617A1 (en) * | 1996-03-12 | 2003-04-17 | Pg-Txl Company, L.P. | Water soluble paclitaxel derivatives |
| EP1818328A1 (en) * | 2006-02-10 | 2007-08-15 | 6570763 Canada Inc. | Chromatographic method for the isolation and purification of taxane derivatives |
| CN101113976A (en) * | 2006-07-27 | 2008-01-30 | 黑龙江大学 | Portable measuring method for measuring content of paclitaxel compounds in fermentation liquor |
Non-Patent Citations (5)
| Title |
|---|
| B.V.SOROCHINSKII 等: "Method of isolating taxol from Taxus baccata", 《CHEMISTRY OF NATURAL COMPOUNDS》 * |
| KOPPAKA V.RAO 等: "Taxol and Related Taxanes. I. Taxanes of Taxus brevifolia Bark", 《PHARMACEUTICAL RESEARCH》 * |
| 符德欢 等: "三种栽培红豆杉的生药学研究", 《云南中医学院学报》 * |
| 谈增毅: "薄层色谱检识云南红豆杉化学成分的研究", 《基层中药杂志》 * |
| 赵凯 等: "抗癌药物紫杉醇的提取与分离纯化技术", 《生物技术通讯》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110441439A (en) * | 2019-07-01 | 2019-11-12 | 杭州师范大学 | It is a kind of for distinguishing the metabolic markers and its detection method of heavy foliage Chinese yew and taxusyunnanensis |
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Application publication date: 20140521 |