CN103788107A - Method for preparing oridonin - Google Patents
Method for preparing oridonin Download PDFInfo
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- CN103788107A CN103788107A CN201210423058.8A CN201210423058A CN103788107A CN 103788107 A CN103788107 A CN 103788107A CN 201210423058 A CN201210423058 A CN 201210423058A CN 103788107 A CN103788107 A CN 103788107A
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- rubescensine
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- water
- moving phase
- methyl alcohol
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/08—Bridged systems
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Extraction Or Liquid Replacement (AREA)
Abstract
The invention relates to a purification method for preparing oridonin, and specifically relates to a method and process for preparing high-pure oridonin. Oridonin is separated and purified by using reverse-phase silica gel or a reverse-phase polymer resin as a filler for a DAC preparation column and a mixed solvent of methanol and water as a mobile phase. A separation process is simple; extraction rate is high; the purity of a product is high; no pollution is caused; and a foundation is provided for further large-scale separation and purification of oridonin by using the preparative liquid chromatography.
Description
Technical field
What the present invention relates to is the method for dynamic axial compression column separation and purification rubescensine A, specifically adopts heating and refluxing extraction, dynamic axial compression preparative column to carry out the method for separation and purification rubescensine A.
Background technology
Rabdosia rubescens is the crack rice dry aerial parts of fork [Rabdosia rubescens (Hamst) C.Y.Wu.et Hsuan] of labiate, be mainly distributed in China Huanghe valley and on the south each department, main product is in various places, southern foot, Taihang Mountain, Henan.Rabdosia rubescens tool relieving superficies by cooling anti-inflammatory analgetic stomach invigorating effect such as invigorate blood circulation, has stronger activity at anti-tumor aspect.Rubescensine A is the effective active composition of Rabdosia rubescens, and it has the larger multiplication capacity that kills and wounds, suppresses to various transplanted tumor cells, is mainly used to clinically treat the disease such as cancer of the stomach, esophagus cancer.Rubescensine A at present mainly from Rabdosia rubescens plant separation and purification obtain, extracting method is mainly ether extraction, ethanol extraction method and middle compression leg chromatography etc., but these extraction process operational cycles are long, and condition is wayward, and the content of the rubescensine A obtaining is low.
Along with developing rapidly of the industry such as pharmacy, biochemical industry, Preparative Liquid Chromatography obtains development and application more and more widely, has become the important method of separation and purifying complex mixture, is particularly useful for the profit of returning of natural product.Dynamic axial compression (DAC) method has many-sided superiority, thereby has obtained more deep research and development.The core technology of dynamic axial compression column be by piston move up and down fill post, maintain post and press and unload post, simplified the process of unloading of filling out of major diameter chromatographic column, the column performance of filling is stable, efficiency is high.DAC post post effect is high, reproducible, loads the time used short, can adopt the filler that particle diameter is less, reduces column length, increases post footpath, thereby reduces wall effect, obtains almost approaching the post effect of analytical column.
The present invention adopt alcohol heating reflux extract, the separation and purification of anti-phase dynamic axial compression column preparative chromatography rubescensine A, separating technology is simple, extraction yield is high, products obtained therefrom purity is high, pollution-free, for further preparation of industrialization type liquid chromatography separation and purification rubescensine A provides the foundation.
Summary of the invention
The object of this invention is to provide a kind of production method of extracting and using dynamic axial compression column separation and purification rubescensine A from Rabdosia rubescens, belong to the extraction separation and purification field of natural product, comprise that ethanol-extracted, dynamic axial compression column separate, the purity detecting of concentrated and finished product.
The present invention adopts following steps for achieving the above object:
A, extraction: powder is crossed to 60 mesh sieves and be placed in extractor, 95% ethanol is solvent, 80 ℃ of water-bath backflow 30min, extract 2-3 time, united extraction liquid, concentrating under reduced pressure becomes thick paste shape;
The dissolving of b, sample: the enriched material of a step dissolves with the mixed solvent of methyl alcohol and water, filters, for subsequent use; The preparation of c, moving phase: moving phase adopt step 1) methyl alcohol and the mixed solvent of water, the concentration of methyl alcohol is between 40-70%;
D, DAC post separate: with sampling pump, to having loaded the sample solution that pumps into above-mentioned dissolving in the dynamic axial compression column of filler, then the moving phase wash-out preparing with step c, collects target main peak, samples HPLC detection;
E, concentrated: the target components of collecting in d step is carried out to concentration, obtain rubescensine A sterling;
The purity detecting of f, finished product: pump: NewstyleNP7000 type pump; Chromatographic column (ODS filler 4.6*250mm, 5 μ are m); Detector: the UV-detector NewstyleNU3000 of Chinese nation, detects wavelength: 239nm, detected temperatures: room temperature; Moving phase: methyl alcohol: water=60:40, flow velocity: 1mL/min.
Embodiment
Embodiment one
1, extract: powder is crossed and taken 100g after 60 mesh sieves and be placed in extractor, use 600mL95% extraction using alcohol, 80 ℃ of water-bath backflow 30min, extract 2 times, united extraction liquid, concentrating under reduced pressure becomes cream;
2, the dissolving of sample: enriched material methyl alcohol: water=55:45 dissolves, and concentration is 10g/mL, filters, for subsequent use;
3, the preparation of moving phase: moving phase is methyl alcohol: water=55:45;
4, DAC post separates: pump into sample solution to having loaded in the dynamic axial compression column that particle diameter is 20um filler (50*500mm) with sampling pump, sample introduction flow velocity is 60mL/min, uses moving phase wash-out, and flow velocity is 100mL/min, collect target component, sampling HPLC detects;
5, concentrated: the target components of collecting is carried out to concentration, obtain rubescensine A sterling, detecting purity through HPLC is 99.2%.
Embodiment two
1, extract: powder is crossed and taken 150g after 60 mesh sieves and be placed in extractor, use 1200mL95% extraction using alcohol, 80 ℃ of water-bath backflow 30min, extract 3 times, united extraction liquid, concentrating under reduced pressure becomes cream;
2, the dissolving of sample: enriched material methyl alcohol: water=55:45 dissolves, and concentration is 15g/mL, filters, for subsequent use;
3, the preparation of moving phase: moving phase is methyl alcohol: water=55:45;
4, DAC post separates: pump into sample solution to having loaded in the dynamic axial compression column that particle diameter is 20um filler (50*700mm) with sampling pump, sample introduction flow velocity is 80mL/min, uses moving phase wash-out, and flow velocity is 120mL/min, collect target component, sampling HPLC detects;
5, concentrated: the target components of collecting is carried out to concentration, obtain rubescensine A sterling, detecting purity through HPLC is 99.03%.
Embodiment three
1, extract: powder is crossed and taken 200g after 60 mesh sieves and be placed in extractor, use 1500mL95% extraction using alcohol, 80 ℃ of water-bath backflow 30min, extract 3 times, united extraction liquid, concentrating under reduced pressure becomes cream;
2, the dissolving of sample: enriched material methyl alcohol: water=60:40 dissolves, and concentration is 20g/mL, filters, for subsequent use;
3, the preparation of moving phase: moving phase is methyl alcohol: water=60:40;
4, DAC post separates: pump into sample solution to having loaded in the dynamic axial compression column that particle diameter is 30um filler (100*500mm) with sampling pump, sample introduction flow velocity is 150mL/min, uses moving phase wash-out, and flow velocity is 200mL/min, collect target component, sampling HPLC detects;
5, concentrated: the target components of collecting is carried out to concentration, obtain rubescensine A sterling, detecting purity through HPLC is 99.35%.
Above-mentioned embodiment is used for the present invention that explains, rather than limits the invention, and in the protection domain of spirit of the present invention and claim, any modification and change that the present invention is made, all fall into protection scope of the present invention.
Claims (7)
1. a method of preparing rubescensine A, comprising:
The preparation of a, rubescensine A crude product: Rabdosia rubescens hay powder is sieved and is placed in extractor, and 95% ethanol is solvent, 80 ℃ of water-bath backflow 30min, extract 2-3 time, united extraction liquid, concentrating under reduced pressure becomes thick paste shape;
The dissolving of b, sample: rubescensine A crude product is dissolved with the mixed solvent of methyl alcohol and water, filter, for subsequent use;
The preparation of c, moving phase: moving phase adopts the methyl alcohol of step a and the mixed solvent of water, and the concentration of methyl alcohol is between 40-70%;
D, upper prop wash-out: pump into installing in the dynamic axial compression preparative column of filler the sample that step a has dissolved with sampling pump, then the moving phase wash-out preparing with step b, collect rubescensine A, sampling detects purity with HPLC;
E, concentrated: the target components of collecting is carried out to concentration, obtain rubescensine A sterling.
2. method according to claim 1, is characterized in that in a step, each consumption that refluxes 95% ethanol used is 5 ~ 10 times of amounts of Rabdosia rubescens hay powder.
3. method according to claim 1, is characterized in that the concentration of b step sample is between 10-30g/mL.
4. method according to claim 1, is characterized in that adopting single needle upper prop to adsorb, resolve and carry out Fractional Collections in c step; Or adopt the spininess continuous sample introduction mode of continuous mode upper prop, parsing in batches, and every batch of desorbed solution is carried out to Fractional Collections.
5. method according to claim 1, is characterized in that the dynamic axial compression column adopting is the preparative column of the each serial different diameter of 50-1000mm, and described filler is C18 and C8, and packing material size is 20-40um.
6. method according to claim 1, the loading flow velocity that it is characterized in that c step sample solution is 0.02BV/min ~ 0.10BV/min, elution flow rate is 0.10 BV/min ~ 0.20BV/min.
7. according to the method described in claim 1 ~ 6 any one, it is characterized in that repeating c step, until gained rubescensine A HPLC detection level reaches more than 99%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210423058.8A CN103788107A (en) | 2012-10-30 | 2012-10-30 | Method for preparing oridonin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210423058.8A CN103788107A (en) | 2012-10-30 | 2012-10-30 | Method for preparing oridonin |
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| Publication Number | Publication Date |
|---|---|
| CN103788107A true CN103788107A (en) | 2014-05-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210423058.8A Pending CN103788107A (en) | 2012-10-30 | 2012-10-30 | Method for preparing oridonin |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105732653A (en) * | 2016-02-03 | 2016-07-06 | 河南中医学院 | Method for preparing oridonin from isodon japonica |
-
2012
- 2012-10-30 CN CN201210423058.8A patent/CN103788107A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105732653A (en) * | 2016-02-03 | 2016-07-06 | 河南中医学院 | Method for preparing oridonin from isodon japonica |
| CN105732653B (en) * | 2016-02-03 | 2017-12-15 | 河南中医药大学 | A kind of method that Oridonin is prepared from Isodon Japonica Hara |
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Application publication date: 20140514 |