CN104370896A - Pseudomonas elodea rhzomorph purifying method - Google Patents
Pseudomonas elodea rhzomorph purifying method Download PDFInfo
- Publication number
- CN104370896A CN104370896A CN201310619968.8A CN201310619968A CN104370896A CN 104370896 A CN104370896 A CN 104370896A CN 201310619968 A CN201310619968 A CN 201310619968A CN 104370896 A CN104370896 A CN 104370896A
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- CN
- China
- Prior art keywords
- pseudomycin
- rhzomorph
- crude product
- separation
- pseudomonas elodea
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- 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.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses a pseudomonas elodea rhzomorph purifying method, pseudomonas elodea rhzomorph fermentation liquor is used as a raw material, appropriate column separation conditions can be obtained by optimization of conditions, and the appropriate column separation conditions are further enlarged onto a dynamic axial compression industrial preparative chromatography system for separation for obtaining of high purity pseudomonas elodea rhzomorph. The advantages of the method are that, good effect of separation and purification can be achieved by direct use of the dynamic axial compression industrial preparative chromatography for separation of an extracted crude product, the method has the advantages of simple operation, short period and high efficiency, and the purity can reach more than 98.5%.
Description
Technical field
The invention belongs to biomedicine technical field, be specifically related to a kind of purification process of Pseudomycin.
Technical background
Pseudomycin, medication name mupirocin, it is the inhibitor of aminoacyl-tRNA synthetase (AaRS), be adapted to the infection of the skin soft tissue that gram-positive cocci causes, as dermopathic secondary infections such as the primary infections such as impetigo herpetiformis, furuncle, folliculitis and eczema, dermatitis, ulcer, wounds.Conventional chromatography purifying Pseudomycin product, Technology length consuming time, purity is not high simultaneously, the present invention is directed to these problems and adopts high pressure preparative chromatography, reach satisfied separating effect, therefore this method can become the effective means of natural product, synthesis medical research.
Summary of the invention
The object of the invention is to for the problem that the current time existing in the process of purifying Pseudomycin is long and purity is not high, provide a kind of novel method adopting preparative liquid chromatography system purification of high-purity Pseudomycin, the method comprises the following steps:
A. filter after water-soluble for Pseudomycin crude product, generally Pseudomycin crude product is mixed with saturated solution, when specifically implementing, can adjust according to actual solute effect;
B. get the pseudomonas cellulose solution prepared, be separated with the dynamic axial compression column of populated fixed phase stuffing, collect the cut of the corresponding bands of a spectrum of Pseudomycin;
C. by the solvent evaporate to dryness in the corresponding bands of a spectrum elutriant collected in step b, the Pseudomycin monomer that chromatographically pure is 98% can be obtained.
The invention has the beneficial effects as follows prepared by the Pseudomycin crude product after can directly adopting fermentation, do not need a large amount of pretreatment process, simple, easy to control, Technology simplifies, and is applicable to extensive preparation.In high-performance liquid chromatogram determination, purity, higher than 98%, can be used for the exploitation of medicine completely.
Embodiment
Embodiment 1
1. after getting 1g Pseudomycin fermentation liquor treatment, crude product is water-soluble, is mixed with saturated solution, is placed in filtration unit, solids removed by filtration particle;
2. Pseudomycin solution is pumped into dynamic axial compression column preparing chromatography system, column packed is of a size of Φ 50 × 250mm, and filler is 18 alkyl silica gel, and particle diameter is 10 ~ 30um, applied sample amount 1g.The volume ratio of moving phase is 30:70, and after Pseudomycin wash-out, moving phase switches to acetonitrile at high proportion, is gone out by rear end impurity, and a separation cycle terminates.The determined wavelength of the UV, visible light luminosity detector adopted is 240nm, collects the cut of retention time at 35 ~ 49min, through HPLC purity assay >=98.8%.
Embodiment 2
1. after getting 1.5g Pseudomycin fermentation liquor treatment, crude product is water-soluble, is mixed with saturated solution, is placed in filtration unit, solids removed by filtration particle;
2. Pseudomycin solution is pumped into dynamic axial compression column preparing chromatography system, column packed is of a size of Φ 50 × 250mm, and filler is 18 alkyl silica gel, and particle diameter is 10 ~ 15um, applied sample amount 1.5g.The volume ratio of moving phase is 26:74, and after Pseudomycin wash-out, moving phase switches to acetonitrile at high proportion, is gone out by rear end impurity, and a separation cycle terminates.The determined wavelength of the UV, visible light luminosity detector adopted is 240nm, collects the cut of retention time at 31 ~ 47min, through HPLC purity assay >=98.5%.
Embodiment 3
1. after getting 3g Pseudomycin fermentation liquor treatment, crude product is water-soluble, is mixed with saturated solution, is placed in filtration unit, solids removed by filtration particle;
2. Pseudomycin solution is pumped into dynamic axial compression column preparing chromatography system, column packed is of a size of Φ 150 × 250mm, and filler is 18 alkyl silica gel, and particle diameter is 10um, applied sample amount 1g.The volume ratio of moving phase is 22:78, and after Pseudomycin wash-out, moving phase switches to acetonitrile at high proportion, is gone out by rear end impurity, and a separation cycle terminates.The determined wavelength of the UV, visible light luminosity detector adopted is 240nm, collects the cut of retention time at 42 ~ 53min, through HPLC purity assay >=98.1%.
Claims (4)
1. a purification process for Pseudomycin, is characterized in that comprising the following steps:
(1) by the pre-treatment of Pseudomycin fermented liquid, concentrated, dry as being separated crude product;
(2) preparing chromatography system is injected after dissolving crude product by sampling pump or sampling valve;
(3) adopt acetonitrile/damping fluid to do moving phase, wash-out obtains Pseudomycin monomer.
2. method according to claim 1, is characterized in that: described Pseudomycin crude product is fermentation gained.
3. method according to claim 2, is characterized in that: described chromatographic system adopts dynamic axial compression column, and to fill stationary phase be 18 alkyl silica gel filler.
4. method according to claim 3, is characterized in that: in moving phase, volume ratio (V/V) scope of acetonitrile and damping fluid is 20:80 ~ 35:65.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310619968.8A CN104370896A (en) | 2013-11-29 | 2013-11-29 | Pseudomonas elodea rhzomorph purifying method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310619968.8A CN104370896A (en) | 2013-11-29 | 2013-11-29 | Pseudomonas elodea rhzomorph purifying method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104370896A true CN104370896A (en) | 2015-02-25 |
Family
ID=52550131
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310619968.8A Pending CN104370896A (en) | 2013-11-29 | 2013-11-29 | Pseudomonas elodea rhzomorph purifying method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104370896A (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4071536A (en) * | 1971-06-12 | 1978-01-31 | Beecham Group Limited | Antibiotics |
| US4222942A (en) * | 1977-09-30 | 1980-09-16 | Beecham Group Limited | Isolation of organic acids |
| CN1345377A (en) * | 1999-02-03 | 2002-04-17 | 拜奥盖尔药厂有限公司 | Method for separating pseudomonic acid A from culture solution containing pseudomonic acid complex |
| CN1670217A (en) * | 1999-02-03 | 2005-09-21 | 特瓦药厂有限公司 | Process for the preparation of pseudomonic acid A antibiotic by microbiological method |
| CN101124221A (en) * | 2005-02-21 | 2008-02-13 | 阿尔法马股份公司 | Purification of mupirocin |
| CN101511826A (en) * | 2006-08-30 | 2009-08-19 | 阿克塞里尔制药公司 | Preparation and purification of mupirocin calcium |
| CN101591333A (en) * | 2009-07-02 | 2009-12-02 | 北京博尔莱生物技术有限公司 | A kind of method of purifying pseudomonas acid A |
| CN102863433A (en) * | 2012-09-26 | 2013-01-09 | 北京仁峰科技有限公司 | Mupirocin purification method |
-
2013
- 2013-11-29 CN CN201310619968.8A patent/CN104370896A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4071536A (en) * | 1971-06-12 | 1978-01-31 | Beecham Group Limited | Antibiotics |
| US4222942A (en) * | 1977-09-30 | 1980-09-16 | Beecham Group Limited | Isolation of organic acids |
| CN1345377A (en) * | 1999-02-03 | 2002-04-17 | 拜奥盖尔药厂有限公司 | Method for separating pseudomonic acid A from culture solution containing pseudomonic acid complex |
| CN1670217A (en) * | 1999-02-03 | 2005-09-21 | 特瓦药厂有限公司 | Process for the preparation of pseudomonic acid A antibiotic by microbiological method |
| CN101124221A (en) * | 2005-02-21 | 2008-02-13 | 阿尔法马股份公司 | Purification of mupirocin |
| CN101511826A (en) * | 2006-08-30 | 2009-08-19 | 阿克塞里尔制药公司 | Preparation and purification of mupirocin calcium |
| CN101591333A (en) * | 2009-07-02 | 2009-12-02 | 北京博尔莱生物技术有限公司 | A kind of method of purifying pseudomonas acid A |
| CN102863433A (en) * | 2012-09-26 | 2013-01-09 | 北京仁峰科技有限公司 | Mupirocin purification method |
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150225 |
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| WD01 | Invention patent application deemed withdrawn after publication |