CN103808850A - Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis - Google Patents
Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis Download PDFInfo
- Publication number
- CN103808850A CN103808850A CN201210460334.8A CN201210460334A CN103808850A CN 103808850 A CN103808850 A CN 103808850A CN 201210460334 A CN201210460334 A CN 201210460334A CN 103808850 A CN103808850 A CN 103808850A
- Authority
- CN
- China
- Prior art keywords
- described method
- solution
- alcohol
- organic solvent
- thin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention relates to a thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis and relates to a detection method. The method comprises the following steps of (1) preparing a 10-deacetyl baccatin III reference solution; (2) preparing a sample solution; and (3) executing 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 10-deacetylate Bakating III 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.
10-deacetylate Bakating III can be taxol by chemosynthesis, is important so detect 10-deacetylate Bakating III.
Conventional 10-deacetylate Bakating III 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 10-deacetylate Bakating III 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 10-deacetylate Bakating III in Chinese yew, described method comprises the steps:
(1) take 10-deacetylate Bakating III 0.1～1mg, use 1ml organic solvent dissolution, obtain 10-deacetylate Bakating III 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 DMF, N, and it is high that N-METHYLFORMAMIDE dissolves 10-deacetylate Bakating III 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, N, and N-METHYLFORMAMIDE dissolving 10-deacetylate Bakating III 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) 10-deacetylate Bakating III 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 10-deacetylate Bakating III in Chinese yew, described method comprises the steps:
(1) take 10-deacetylate Bakating III standard items 0.5mg, with 1ml methyl alcohol ultrasonic dissolution, obtain 10-deacetylate Bakating III 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 10-deacetylate Bakating III in Chinese yew, described method comprises the steps:
(1) take 10-deacetylate Bakating III 0.1～1mg, use 1ml organic solvent dissolution, obtain 10-deacetylate Bakating III 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 N, N-METHYLFORMAMIDE.
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 N, N-METHYLFORMAMIDE.
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 |
|---|---|---|---|
| CN201210460334.8A CN103808850A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210460334.8A CN103808850A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103808850A true CN103808850A (en) | 2014-05-21 |
Family
ID=50705926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210460334.8A Pending CN103808850A (en) | 2012-11-15 | 2012-11-15 | Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103808850A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105372370A (en) * | 2015-11-23 | 2016-03-02 | 江苏红豆杉药业有限公司 | Method for detecting 10-deacetylbaccatin III in Chinese yew branches and leaves |
| CN107421943A (en) * | 2017-09-04 | 2017-12-01 | 江苏红豆杉生物科技股份有限公司 | 10 DAB rapid semi-quantitative measuring methods in Chinese yew |
| 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 |
|---|---|---|---|---|
| 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 |
| 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 |
-
2012
- 2012-11-15 CN CN201210460334.8A patent/CN103808850A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
Non-Patent Citations (4)
| Title |
|---|
| KANG WANG 等: "Microbial hydrolysis of 7-xylosyl-10-deacetyltaxol to 10-deacetyltaxol", 《JOURNAL OF MOLECULAR CATALYSIS B: ENZYMATIC》 * |
| 张宗勤 等: "南方红豆杉组织培养及紫杉醇的产生", 《西北植物学报》 * |
| 李效贤 等: "南方红豆杉的鉴别研究", 《中药材》 * |
| 赵俊宏 等: "新乡红豆杉中多种紫杉烷类化合物的反相高效液相色谱分析", 《化学研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105372370A (en) * | 2015-11-23 | 2016-03-02 | 江苏红豆杉药业有限公司 | Method for detecting 10-deacetylbaccatin III in Chinese yew branches and leaves |
| CN107421943A (en) * | 2017-09-04 | 2017-12-01 | 江苏红豆杉生物科技股份有限公司 | 10 DAB rapid semi-quantitative measuring methods in Chinese yew |
| 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 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105203657A (en) | Spina date seed reference extract and preparation method and application thereof | |
| CN103822984B (en) | Method for synchronously separating and measuring endogenous abscisic acid, gibberellins and auxin in turfgrass | |
| Ma et al. | Separation of four flavonoids from Rhodiola rosea by on-line combination of sample preparation and counter-current chromatography | |
| Wang et al. | Isolation and purification of six iridoid glycosides from gardenia jasminoides fruit by medium‐pressure liquid chromatography combined with macroporous resin chromatography | |
| CN102293789B (en) | Method for extracting triterpenoids from ganoderma lucidum sporocarp | |
| CN106198832B (en) | A kind of quality of production control method of intensified loquet distillate | |
| Oettl et al. | Dereplication of depsides from the lichen Pseudevernia furfuracea by centrifugal partition chromatography combined to 13C nuclear magnetic resonance pattern recognition | |
| CN103808850A (en) | Thin-layer chromatography detection method of 10-deacetyl baccatin III in taxus chinensis | |
| CN104922196A (en) | General ohwia flavone extract preparation method and general ohwia flavone extract quality detecting method | |
| CN102175790B (en) | HPLC (high performance liquid chromatography) method for synchronously detecting five polyphenols in apples and distinguishing varieties | |
| CN103808851A (en) | Thin-layer chromatography detection method of xylosyl-10-deacetyltaxol in taxus chinensis | |
| CN103784480B (en) | The preparation method of Armillaria luteo-virens antioxidant activity component and application thereof | |
| CN103808854A (en) | Thin-layer chromatography detection method of baccatin III in taxus chinensis | |
| CN103808853A (en) | Thin-layer chromatography detection method of 7-xylosyl-10-deacetyltaxol in taxus chinensis | |
| CN103808852A (en) | Thin-layer chromatography detection method of taxol in taxus chinensis | |
| CN106008445A (en) | Flavone and lignin compound and extracting method thereof | |
| CN103822888B (en) | Catch up with the quality determining method of yellow grass | |
| CN104101661A (en) | Quick detecting method for sudan red in food | |
| Zhou et al. | Evaluation of microwave‐assisted extraction for aristolochic acid from Aristolochiae Fructus by chromatographic analysis coupled with nephrotoxicity studies | |
| CN103940926B (en) | A kind of method of antioxidant content in quick detection Radix Et Rhizoma Rhei seed | |
| CN106596759B (en) | A kind of HPLC analysis method of Ramulus et folium taxi cuspidatae extract and its preparation | |
| CN103197005B (en) | Detection method of betulinic acid, betulin and oleanolic acid in white birch bark | |
| CN106908542A (en) | The quality determining method of antipyretic and itching-stopping lotion | |
| CN105758986A (en) | Thin-layer chromatography identification method of medicinal material fructus podophylli and application of thin-layer chromatography identification method | |
| CN102539574A (en) | Method for detecting polygonaceous plant sourced medicinal materials |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140521 |