CN102453062B - Partricin A separation method - Google Patents
Partricin A separation method Download PDFInfo
- Publication number
- CN102453062B CN102453062B CN201010514332.3A CN201010514332A CN102453062B CN 102453062 B CN102453062 B CN 102453062B CN 201010514332 A CN201010514332 A CN 201010514332A CN 102453062 B CN102453062 B CN 102453062B
- Authority
- CN
- China
- Prior art keywords
- methanol solution
- partricin
- wash
- volume percent
- impurities
- 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.)
- Expired - Fee Related
Links
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a partricin A separation method, which comprises the following steps of: adjusting pH of a partricin-containing fermentation broth, standing and filtering, immersing the filter cake by a polarity organic solvent, filtering and taking a filtrate; adjusting pH of the filtrate, condensing at the temperature of 25-35 DEG C until the water content is 30-35%, standing and filtering to obtain a partricin crude product; dissolving the crude product in a methanol solution containing 25-30% of water, loading the solution into a styrene macroporous anti-phase adsorption resin column, carrying out elution and removing impurities by using a methanol solution containing 18-23% of water, carrying out elution on partricin A by using a methanol solution containing 13-17% of water, collecting a partricin A eluate with the purity being greater than or equal to 80%, condensing the collected partricin A eluate to a methanol solution containing 25-30% of water, loading to the column, carrying out elution and removing impurities by using a methanol solution containing 17-19% of water, carrying out elution by using a methanol solution containing 10-14% of water, and collecting the eluate, wherein the percentage is volume percentage. The method provided by the invention has advantages of simple operation, low cost, little solvent and stable product quality, and is suitable for large-scale industrial production. The purity of the separated product partricin A reaches more than 93%.
Description
Technical field
The present invention relates to the separation method of a kind of partricin A.
Background technology
The separation and purification from pedotheque such as Tiberio Bruzzese in 1971 obtains the raw gold chain mould (Streptomyces aureofaciens) that a streptomycete belongs to, and in the meta-bolites of this bacterium, isolated a kind of Heptaene macrolide class microbiotic, called after partricin (partricin).Partricin is a kind of antimycotic and antiprotozoal efficient medicine, realizes bacteriostatic action, especially have high bacteriostatic activity to Candida albicans by the synthesis of Antifungi cell film.Partricin mainly comprises two kinds of components, partricin A and partricin B.
In raw gold chain mold fermentation product, except product partricin, also generate microbiotic and the impurity of other kinds such as a large amount of tsiklomitsins and duomycin, composition is very complicated, and the technique therefore extracting partricin A from tunning is loaded down with trivial details simultaneously.At present, in prior art, mainly the methods such as traditional organic solvent extraction and charcoal absorption are still adopted to the separation and purification of partricin A.
United States Patent (USP) 3,773, has set forth the separation and purification process of partricin A in 925.Main utilization be the difference of partricin A solubleness in different solvents, after the extraction repeatedly of multi-step, enriching and purifying is carried out in recycling charcoal absorption.The shortcoming of the method have employed a large amount of solvents, very large pressure is caused to environment, and inevitably need in step to adopt gac, and the absorption property of gac is not very stable, even if the gac that same manufacturer produces, the difference of production batch, thus can cause the quality of the finished product unstable.
Summary of the invention
Technical problem to be solved by this invention overcomes existing separation partricin A method to use a large amount of solvent to cause very large pressure to environment, inevitably need in step to adopt gac, and the absorption property instability of gac can cause the defect of the quality instability of the finished product, provide a kind of simple to operate, cost is lower, multi-solvent can not be produced and caused environmental stress, constant product quality, be applicable to the separation method of the partricin A of large-scale industrial production.
The separation method of partricin A of the present invention comprises the steps:
(1) pH will be adjusted to be 3 ~ 5 containing partricin fermented liquid, and leave standstill and filter, filter cake polar organic solvent is soaked, cross leaching filtrate afterwards;
(2) filtrate adjusted pH to be 9 ~ 10, at 25 DEG C ~ 35 DEG C, be concentrated into solution comprises water volume percent 30% ~ 35% afterwards, leave standstill, filter to obtain partricin crude product;
(3) by the methanol solution of partricin dissolving crude product in moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column afterwards, first with the methanol solution wash-out removal of impurities of moisture volume percent 18% ~ 23%, and then with the methanol solution wash-out partricin A of moisture volume percent 13% ~ 17%, collect the partricin A elutriant of purity >=80%;
(4) the partricin A elutriant of collection is concentrated into the methanol solution that solution is moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column again, with the methanol solution wash-out removal of impurities of moisture volume percent 17% ~ 19%, again with the methanol solution wash-out partricin A of moisture volume percent 10% ~ 14%, collect elutriant.
Below, further the separation method of partricin A of the present invention is described in detail:
(1) pH will be adjusted to be 3 ~ 5 containing partricin fermented liquid, and leave standstill and filter, filter cake polar organic solvent is soaked, cross leaching filtrate afterwards.
Wherein, described is the microbial bacteria fermented liquid that this area routine prepares partricin containing partricin fermented liquid, is generally obtained through this area ordinary method fermentation culture by raw gold chain mould Streptomyces aureofaciens.The described concrete bacterial classification of raw gold chain mould is not particularly limited, and the bacterial classification that routine can produce partricin all can use, as raw gold chain mould Streptomyces aureofaciens NRRL 3878 etc.Wherein, described raw gold chain mould through cultivation and fermentation obtain containing the concrete bacterial classification of partricin fermented liquid not circumscribed be because partricin is the secondary metabolite of raw gold chain mould Streptomyces aureofaciens, although different concrete bacterial strains is different in the ratio of the output of partricin, impurity, but the main component of fermented liquid is relatively fixing, the fermented liquid that therefore prepared by different concrete bacterial classification raw gold chain mould Streptomyces aureofaciens is all suitable for the inventive method.
Wherein, described tune pH reagent is the acid that this area routine adjusts pH, and be preferably oxalic acid and/or hydrochloric acid, better is oxalic acid.
Wherein, the described viable bacteria act as in abundant removing fermented liquid left standstill, the described time left standstill is preferably >=2 hours, and better is 2 ~ 4 hours.
Wherein, described polar organic solvent is the conventional described polar organic solvent in this area, and be preferably one or more in methyl alcohol, ethanol and acetone, better is methyl alcohol.
Wherein, the consumption of described polar organic solvent is preferably 2 ~ 4 times of filter cake volumes.
Wherein, the effect of described immersion is the partricin fully extracted in filter cake.The time of described immersion is preferably >=2 hours, and better is 2 ~ 4 hours.
Wherein, described immersion and filter operation preferably can also repetitive operations 1 ~ 2 time, merging filtrate afterwards.
(2) filtrate adjusted pH to be 9 ~ 10, at 25 DEG C ~ 35 DEG C, be concentrated into solution comprises water volume percent 30% ~ 35% afterwards, leave standstill, filter to obtain partricin crude product.
Wherein, described tune pH reagent is the alkali that this area routine adjusts pH, is preferably sodium hydroxide and/or potassium hydroxide.
Wherein, because the filter cake in step (1) is moisture, cause water content in the filtrate in step (2) also comparatively large, thus need concentrated further.Described concentrated temperature is preferably 30 DEG C.Described concentrated terminal is preferably for the water in filtrate contains volume percent 33%.
Wherein, described standing acting as makes partricin fully separate out.The described temperature left standstill is preferably 0 DEG C ~ 5 DEG C, and better is 4 DEG C.The described time left standstill is preferably >=24 hours, and better is 24 hours.
(3) by the methanol solution of partricin dissolving crude product in moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column afterwards, first with the methanol solution wash-out removal of impurities of moisture volume percent 18% ~ 23%, and then with the methanol solution wash-out partricin A of moisture volume percent 13% ~ 17%, collect the partricin A elutriant of purity >=80% (HPLC purity).
Wherein, the concentration of described dissolving partricin crude methanol solution is preferably the methanol solution of moisture volume percent 25%.
Wherein, described partricin crude product and the amount ratio of methanol solution are preferably 7mg/mL ~ 9mg/mL, and that better is 8mg/mL.
Wherein, the model of described vinylbenzene macropore reverse phase absorption resin column is preferably NM100, HZ-803 or microballoon 1#.
Wherein, the applied sample amount of described partricin crude product is preferably≤10mg/mL resin.
Wherein, the loading of described partricin crude product to the adsorption flow rate of vinylbenzene macropore reverse phase absorption resin column is preferably 0.2CV/h ~ 0.3CV/h, wherein said CV is this area routine metering term, and full name is resin column volume, English name column volume.
Wherein, the concentration of the methanol solution of described wash-out removal of impurities is preferably the methanol solution of moisture volume percent 20%.
Wherein, the consumption of the methanol solution of described wash-out removal of impurities is preferably 2.5CV ~ 4CV, and that better is 3CV.
Wherein, the concentration of the methanol solution of described wash-out partricin A is preferably the methanol solution of moisture volume percent 15%.
Wherein, the detection of the partricin A elutriant of described purity >=80% can be determined conventionally by high performance liquid chromatography detection by this area.Described high performance liquid chromatography detects can by this area ordinary method.
(4) the partricin A elutriant of collection is concentrated into the methanol solution that solution is moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column again, with the methanol solution wash-out removal of impurities of moisture volume percent 17% ~ 19%, again with the methanol solution wash-out partricin A of moisture volume percent 10% ~ 14%, collect elutriant.
Wherein, described concentrated temperature is preferably 25 DEG C ~ 35 DEG C, and better is 30 DEG C.
Wherein, described concentrated after the concentration of methanol solution be preferably the methanol solution of moisture volume percent 25%.
Wherein, described vinylbenzene macropore reverse phase absorption resin column applied sample amount and adsorption flow rate are as previously mentioned.
Wherein, the concentration of the methanol solution of described wash-out removal of impurities is preferably the methanol solution of moisture volume percent 18%.
Wherein, the consumption of the methanol solution of described wash-out removal of impurities is preferably 1.5CV ~ 3CV, and that better is 2CV.
Wherein, the concentration of the methanol solution of described wash-out partricin A is preferably the methanol solution of moisture volume percent 12%.
Wherein, described collection elutriant is preferably partricin A purity >=90%, and detection can be determined conventionally by high performance liquid chromatography detection by this area.Described high performance liquid chromatography detects can by this area ordinary method.
In the present invention, the elutriant that described step (4) is collected, further except desolventizing, namely obtains dry finished product.
In the present invention, relate to the equal recoverable of methanol solution of use.
Agents useful for same of the present invention and raw material are all commercially.
On the basis meeting this area general knowledge, each technical characteristic above-mentioned in the present invention preferably arbitrary combination can obtain preferred embodiments of the present invention.
Positive progressive effect of the present invention is: the separation method of partricin A of the present invention is simple to operate, cost is lower, can not produce multi-solvent and cause environmental stress, constant product quality, be applicable to large-scale industrial production, the partricin A purity being separated the product obtained reaches more than 93%.
Embodiment
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.
In following embodiment, the high performance liquid chromatography testing conditions detecting partricin A is: chromatographic column Agilent C18 5um 250 × 4.6mm, moving phase is acetonitrile: ammonium acetate aqueous solution (0.05mol/L, pH5.5) volume ratio is 40: 60, temperature 35 DEG C, flow velocity 0.8mL/min, wavelength 378nm.
In following embodiment, vinylbenzene macropore reverse phase absorption resin source is:
NM100: Suzhou Nano-micro Technology Co., Ltd.
HZ803: Shanghai Hua Zhen Resins Corporation
Microballoon 1#: Shanghai Hua Zhen Resins Corporation
Per-cent in following embodiment, except specified otherwise, is volume percent.
Preparation embodiment
Prepare partricin fermented liquid, the bacterial classification of use is made a living gold chain mould Streptomyces aureofaciensNRRL 3878 (source american agriculture research DSMZ).
Concrete fermentation culture conditions is:
1, slant culture: cultivate 7 days for 28 DEG C;
Substratum (g/L): Zulkovsky starch 10.0, MgSO
47H
2o 1.0, NaCl 1.0, (NH4)
2sO
42.0, FeSO
47H
2o 0.001, KzHPO
41.0, CaCO
32.0, MnCl
27H
2o 0.001
2, seed culture: 28 DEG C, 28h, shaking speed: 240rpm;
Substratum (g/L): glucose 10.0, peptone 10.0, yeast powder 5.0
3, fermentation culture: 121 DEG C of sterilizing 20min, inoculum size 10%, shaking flask loading amount 100mL/750mL, is placed in 28 DEG C of constant temperature rotary shaker 240rpm and shakes fermentation and obtain final fermented liquid in 7 days;
Substratum (g/L): glucose 40.0, industrial starch 20.0, cold analysis for soybean powder 20.0, dried silkworm chrysalis meal 10.0, yeast powder 2.0, peptone 2.0, NaCl 5.0.
Embodiment 1 ~ 15 is made a living the obtained partricin fermented liquid of gold chain mould Streptomyces aureofaciens NRRL 3878 fermentation culture.
Embodiment 1
The fermented liquid oxalic acid containing partricin A 543 μ g/mL fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 83.4%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 94.2%.
Embodiment 2
The fermented liquid oxalic acid containing partricin A 528 μ g/ml fermentation obtained adjusts pH to 3.0, and filter cake 2 times of vol acetone are soaked 2h, cross leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.3%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.6%.
Embodiment 3
The fermented liquid oxalic acid containing partricin A 501 μ g/mL fermentation obtained adjusts pH to 3.0, and suction filtration after standing 2h, by filter cake 2 times of volume soaked in absolute ethyl alcohol 2h, is crossed leaching filtrate after soaking, and by filter cake repetitive operation once, merged twice filtrate.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.9%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.9%.
Embodiment 4
The fermented liquid oxalic acid containing partricin A 513 μ g/ml fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 30% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.8%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.9%.
Embodiment 5
The fermented liquid oxalic acid containing partricin A 528 μ g/ml fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 35% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 81.4%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.1%.
Embodiment 6
The fermented liquid oxalic acid containing partricin A 567 μ g/mL fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by macropore reverse phase absorption resin HZ-803, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 81.6%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by macropore reverse phase absorption resin HZ-803, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.3%.
Embodiment 7
The fermented liquid oxalic acid containing partricin A 532 μ g/mL fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by macropore reverse phase absorption resin microsphere 1#, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.5%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by macropore reverse phase absorption resin microsphere 1#, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.9%.
Embodiment 8
The fermented liquid oxalic acid containing partricin A 532 μ g/mL fermentation obtained adjusts pH to 4.0, and filter cake 3 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.8 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 27% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.3%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.8%.
Embodiment 9
The fermented liquid oxalic acid containing partricin A 493 μ g/mL fermentation obtained adjusts pH to 4.0, and filter cake 3 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.8 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 30% water will be used after precipitate collected by suction, and will a small amount of precipitation be there is, filter out, filtrate is by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 81.6%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.1%.
Embodiment 10
The fermented liquid oxalic acid containing partricin A 511 μ g/mL fermentation obtained adjusts pH to 5.0, and filter cake 3 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.4 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 13% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 83.1%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.9%
Embodiment 11
The fermented liquid oxalic acid containing partricin A 521 μ g/mL fermentation obtained adjusts pH to 5.0, and filter cake 3 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.4 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 17% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.3%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.4%.
Embodiment 12
The fermented liquid oxalic acid containing partricin A 507 μ g/mL fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 83.3%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 10% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.7%.
Embodiment 13
The fermented liquid oxalic acid containing partricin A 548 μ g/mL fermentation obtained adjusts pH to 3.0, and filter cake 2 times of volumes methanol are soaked 2h, crossed leaching filtrate, and by filter cake repetitive operation once, merge twice filtrate after soaking by suction filtration after standing 2h.
Adjust pH to 9.0 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.8%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 14% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.4%.
Embodiment 14
Adjust pH to 3.5 with oxalic acid the fermented liquid containing partricin A 514 μ g/mL, suction filtration after standing 2h, filter cake 2.5 times of volumes methanol are soaked 2h, crosses leaching filtrate after soaking, and by filter cake repetitive operation once, merge twice filtrate.
Adjust pH to 9.3 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.6%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 12% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.7%.
Embodiment 15
Adjust pH to 3.5 with oxalic acid the fermented liquid containing partricin A573 μ g/mL, suction filtration after standing 2h, filter cake 2 times of volumes methanol are soaked 2h, crosses leaching filtrate after soaking, and by filter cake repetitive operation once, merge twice filtrate.
Adjust pH to 9.5 filtrate, moisture 33% is slowly concentrated at 30 DEG C, a certain amount of solid is now had to separate out, then be placed in 4 DEG C of refrigerators to leave standstill 24h partricin is fully separated out from mother liquor, the dissolve with methanol solution containing 25% water will be used after precipitate collected by suction, and by NM100 resin, first with the methanol solution prewashing removal of impurities of 3CV containing 20% water after absorption, again with the methanol solution wash-out partricin A containing 15% water, collect the partricin A (HPLC detection) that purity is greater than 80%, after mixing, purity is 82.7%.
Partricin A is collected liquid and be concentrated into the methanol solution of strength of solution containing 25% water, afterwards again by NM100 resin, first with the methanol solution prewashing removal of impurities of 2CV containing 18% water, again with the methanol solution wash-out partricin A containing 14% water, collect the partricin A effluent liquid (HPLC detection) that purity is greater than 90%, after mixing, obtain the partricin A that purity is 93.8%.
Claims (59)
1. a separation method of partricin A, is characterized in that: it comprises the steps:
(1) pH will be adjusted to be 3 ~ 5 containing partricin fermented liquid, and leave standstill and filter, filter cake polar organic solvent is soaked, cross leaching filtrate afterwards; Described polar organic solvent is one or more in methyl alcohol, ethanol and acetone;
(2) filtrate adjusted pH to be 9 ~ 10, at 25 DEG C ~ 35 DEG C, be concentrated into solution comprises water volume percent 30% ~ 35% afterwards, leave standstill, filter to obtain partricin crude product;
(3) by the methanol solution of partricin dissolving crude product in moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column afterwards, first with the methanol solution wash-out removal of impurities of moisture volume percent 18% ~ 23%, and then with the methanol solution wash-out partricin A of moisture volume percent 13% ~ 17%, collect the partricin A elutriant of purity >=80%;
(4) the partricin A elutriant of collection is concentrated into the methanol solution that solution is moisture volume percent 25% ~ 30%, loading vinylbenzene macropore reverse phase absorption resin column again, with the methanol solution wash-out removal of impurities of moisture volume percent 17% ~ 19%, again with the methanol solution wash-out partricin A of moisture volume percent 10% ~ 14%, collect elutriant.
2. separation method as claimed in claim 1, is characterized in that: described obtains through this area ordinary method fermentation culture by giving birth to gold chain mould Streptomyces aureofaciens containing partricin fermented liquid.
3. separation method as claimed in claim 2, is characterized in that: described raw gold chain mould is made a living gold chain mould Streptomyces aureofaciens NRRL 3878.
4. the separation method as described in any one of claims 1 to 3, is characterized in that: in described step (1), and described tune pH reagent is oxalic acid and/or hydrochloric acid; And/or the described time left standstill is >=2 hours.
5. separation method as claimed in claim 4, is characterized in that: in described step (1), and the described time left standstill is 2 ~ 4 hours.
6. the separation method as described in claims 1 to 3,5 any one, is characterized in that: in described step (1), and the consumption of described polar organic solvent is 2 ~ 4 times of filter cake volumes; And/or the time of described immersion is >=2 hours; And/or described immersion and filter operation also repetitive operation 1 ~ 2 time, merging filtrate afterwards.
7. separation method as claimed in claim 6, it is characterized in that: in described step (1), the time of described immersion is 2 ~ 4 hours.
8. separation method as claimed in claim 4, it is characterized in that: in described step (1), the consumption of described polar organic solvent is 2 ~ 4 times of filter cake volumes; And/or the time of described immersion is >=2 hours; And/or described immersion and filter operation also repetitive operation 1 ~ 2 time, merging filtrate afterwards.
9. separation method as claimed in claim 8, it is characterized in that: in described step (1), the time of described immersion is 2 ~ 4 hours.
10. as claims 1 to 3,5, separation method as described in 7 ~ 9 any one, it is characterized in that: in described step (2), described tune pH reagent is sodium hydroxide and/or potassium hydroxide; And/or described concentrated temperature is 30 DEG C; And/or described concentrated terminal be in filtrate water containing volume percent 33%; And/or the described temperature left standstill is 0 DEG C ~ 5 DEG C; And/or the described time left standstill is >=24 hours.
11. separation methods as claimed in claim 10, is characterized in that: in described step (2), and the described temperature left standstill is 4 DEG C; And/or the described time left standstill is 24 hours.
12. separation methods as claimed in claim 4, is characterized in that: in described step (2), described tune pH reagent is sodium hydroxide and/or potassium hydroxide; And/or described concentrated temperature is 30 DEG C; And/or described concentrated terminal be in filtrate water containing volume percent 33%; And/or the described temperature left standstill is 0 DEG C ~ 5 DEG C; And/or the described time left standstill is >=24 hours.
13. separation methods as claimed in claim 12, is characterized in that: in described step (2), and the described temperature left standstill is 4 DEG C; And/or the described time left standstill is 24 hours.
14. separation methods as claimed in claim 6, is characterized in that: in described step (2), described tune pH reagent is sodium hydroxide and/or potassium hydroxide; And/or described concentrated temperature is 30 DEG C; And/or described concentrated terminal be in filtrate water containing volume percent 33%; And/or the described temperature left standstill is 0 DEG C ~ 5 DEG C; And/or the described time left standstill is >=24 hours.
15. separation methods as claimed in claim 14, is characterized in that: in described step (2), and the described temperature left standstill is 4 DEG C; And/or the described time left standstill is 24 hours.
16. as claims 1 to 3,5,7 ~ 9, separation method as described in 11 ~ 15 any one, it is characterized in that: in described step (3), the concentration of described dissolving partricin crude methanol solution is the methanol solution of moisture volume percent 25%; And/or the amount ratio of described partricin crude product and methanol solution is 7mg/mL ~ 9mg/mL; And/or the model of described vinylbenzene macropore reverse phase absorption resin column is NM100, HZ-803 or microballoon 1#; And/or the applied sample amount of described partricin crude product is≤10mg/mL resin; And/or the loading of described partricin crude product to the adsorption flow rate of vinylbenzene macropore reverse phase absorption resin column is 0.2CV/h ~ 0.3CV/h.
17. separation methods as claimed in claim 16, is characterized in that: in described step (3), described partricin crude product and the amount ratio of methanol solution are 8mg/mL.
18. separation methods as claimed in claim 4, is characterized in that: in described step (3), and the concentration of described dissolving partricin crude methanol solution is the methanol solution of moisture volume percent 25%; And/or the amount ratio of described partricin crude product and methanol solution is 7mg/mL ~ 9mg/mL; And/or the model of described vinylbenzene macropore reverse phase absorption resin column is NM100, HZ-803 or microballoon 1#; And/or the applied sample amount of described partricin crude product is≤10mg/mL resin; And/or the loading of described partricin crude product to the adsorption flow rate of vinylbenzene macropore reverse phase absorption resin column is 0.2CV/h ~ 0.3CV/h.
19. separation methods as claimed in claim 18, is characterized in that: in described step (3), described partricin crude product and the amount ratio of methanol solution are 8mg/mL.
20. separation methods as claimed in claim 6, is characterized in that: in described step (3), and the concentration of described dissolving partricin crude methanol solution is the methanol solution of moisture volume percent 25%; And/or the amount ratio of described partricin crude product and methanol solution is 7mg/mL ~ 9mg/mL; And/or the model of described vinylbenzene macropore reverse phase absorption resin column is NM100, HZ-803 or microballoon 1#; And/or the applied sample amount of described partricin crude product is≤10mg/mL resin; And/or the loading of described partricin crude product to the adsorption flow rate of vinylbenzene macropore reverse phase absorption resin column is 0.2CV/h ~ 0.3CV/h.
21. separation methods as claimed in claim 20, is characterized in that: in described step (3), described partricin crude product and the amount ratio of methanol solution are 8mg/mL.
22. separation methods as claimed in claim 10, is characterized in that: in described step (3), and the concentration of described dissolving partricin crude methanol solution is the methanol solution of moisture volume percent 25%; And/or the amount ratio of described partricin crude product and methanol solution is 7mg/mL ~ 9mg/mL; And/or the model of described vinylbenzene macropore reverse phase absorption resin column is NM100, HZ-803 or microballoon 1#; And/or the applied sample amount of described partricin crude product is≤10mg/mL resin; And/or the loading of described partricin crude product to the adsorption flow rate of vinylbenzene macropore reverse phase absorption resin column is 0.2CV/h ~ 0.3CV/h.
23. separation methods as claimed in claim 22, is characterized in that: in described step (3), described partricin crude product and the amount ratio of methanol solution are 8mg/mL.
24. as claims 1 to 3,5,7 ~ 9,11 ~ 15, separation method as described in 17 ~ 23 any one, it is characterized in that: in described step (3), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 20%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 2.5CV ~ 4CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 15%; And/or the detection of the partricin A elutriant of described purity >=80% is determined by high performance liquid chromatography detection.
25. separation methods as claimed in claim 24, is characterized in that: the consumption of the methanol solution of the wash-out removal of impurities described in described step (3) is 3CV.
26. separation methods as claimed in claim 4, is characterized in that: in described step (3), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 20%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 2.5CV ~ 4CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 15%; And/or the detection of the partricin A elutriant of described purity >=80% is determined by high performance liquid chromatography detection.
27. separation methods as claimed in claim 26, is characterized in that: the consumption of the methanol solution of the wash-out removal of impurities described in described step (3) is 3CV.
28. separation methods as claimed in claim 6, is characterized in that: in described step (3), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 20%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 2.5CV ~ 4CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 15%; And/or the detection of the partricin A elutriant of described purity >=80% is determined by high performance liquid chromatography detection.
29. separation methods as claimed in claim 28, is characterized in that: the consumption of the methanol solution of the wash-out removal of impurities described in described step (3) is 3CV.
30. separation methods as claimed in claim 10, is characterized in that: in described step (3), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 20%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 2.5CV ~ 4CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 15%; And/or the detection of the partricin A elutriant of described purity >=80% is determined by high performance liquid chromatography detection.
31. separation methods as claimed in claim 30, is characterized in that: the consumption of the methanol solution of the wash-out removal of impurities described in described step (3) is 3CV.
32. separation methods as claimed in claim 16, is characterized in that: in described step (3), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 20%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 2.5CV ~ 4CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 15%; And/or the detection of the partricin A elutriant of described purity >=80% is determined by high performance liquid chromatography detection.
33. separation methods as claimed in claim 32, is characterized in that: the consumption of the methanol solution of the wash-out removal of impurities described in described step (3) is 3CV.
34. as claims 1 to 3,5,7 ~ 9,11 ~ 15,17 ~ 23, separation method as described in 25 ~ 33 any one, it is characterized in that: in described step (4), described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
35. separation methods as claimed in claim 34, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
36. separation methods as claimed in claim 4, is characterized in that: in described step (4), and described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
37. separation methods as claimed in claim 36, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
38. separation methods as claimed in claim 6, is characterized in that: in described step (4), and described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
39. separation methods as claimed in claim 38, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
40. separation methods as claimed in claim 10, is characterized in that: in described step (4), and described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
41. separation methods as claimed in claim 40, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
42. separation methods as claimed in claim 16, is characterized in that: in described step (4), and described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
43. separation methods as claimed in claim 42, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
44. separation methods as claimed in claim 24, is characterized in that: in described step (4), and described concentrated temperature is 25 DEG C ~ 35 DEG C; And/or described concentrated after the concentration of methanol solution be the methanol solution of moisture volume percent 25%; And/or described applied sample amount is≤10mg/mL resin; And/or the adsorption flow rate of described loading is 0.2CV/h ~ 0.3CV/h.
45. separation methods as claimed in claim 44, is characterized in that: in described step (4), and described concentrated temperature is 30 DEG C.
46. as claims 1 to 3,5,7 ~ 9,11 ~ 15,17 ~ 23,25 ~ 33, separation method as described in 35 ~ 45 any one, it is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
47. separation methods as claimed in claim 46, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
48. separation methods as claimed in claim 4, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
49. separation methods as claimed in claim 48, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
50. separation methods as claimed in claim 6, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
51. separation methods as claimed in claim 50, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
52. separation methods as claimed in claim 10, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
53. separation methods as claimed in claim 52, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
54. separation methods as claimed in claim 16, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
55. separation methods as claimed in claim 54, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
56. separation methods as claimed in claim 24, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
57. separation methods as claimed in claim 56, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
58. separation methods as claimed in claim 34, is characterized in that: in described step (4), the concentration of the methanol solution of described wash-out removal of impurities is the methanol solution of moisture volume percent 18%; And/or the consumption of the methanol solution of described wash-out removal of impurities is 1.5CV ~ 3CV; And/or the concentration of the methanol solution of described wash-out partricin A is the methanol solution of moisture volume percent 12%; And/or partricin A purity >=90% in described collection elutriant.
59. separation methods as claimed in claim 58, is characterized in that: in described step (4), the consumption of the methanol solution of described wash-out removal of impurities is 2CV.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010514332.3A CN102453062B (en) | 2010-10-21 | 2010-10-21 | Partricin A separation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010514332.3A CN102453062B (en) | 2010-10-21 | 2010-10-21 | Partricin A separation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102453062A CN102453062A (en) | 2012-05-16 |
| CN102453062B true CN102453062B (en) | 2015-03-04 |
Family
ID=46036784
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010514332.3A Expired - Fee Related CN102453062B (en) | 2010-10-21 | 2010-10-21 | Partricin A separation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102453062B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3773925A (en) * | 1970-11-03 | 1973-11-20 | Prodotti Antibiotici Spa | Partricin |
| US4014994A (en) * | 1974-08-29 | 1977-03-29 | Spa-Societa Prodotti Antibiotici S.P.A. | Process for the recovery and purification of partricin |
| CN101367855A (en) * | 2008-09-24 | 2009-02-18 | 华东理工大学 | Method for purification of erythromycin A |
-
2010
- 2010-10-21 CN CN201010514332.3A patent/CN102453062B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3773925A (en) * | 1970-11-03 | 1973-11-20 | Prodotti Antibiotici Spa | Partricin |
| US4014994A (en) * | 1974-08-29 | 1977-03-29 | Spa-Societa Prodotti Antibiotici S.P.A. | Process for the recovery and purification of partricin |
| CN101367855A (en) * | 2008-09-24 | 2009-02-18 | 华东理工大学 | Method for purification of erythromycin A |
Non-Patent Citations (6)
| Title |
|---|
| Robert C. Tweit,等.Characterization of The Antifungal and Antiprotozoal Antibiotic Partricin and Structural Studies on Partricins A and B.《The Journal of Antibiotic》.1982,第XXXV卷(第8期),第998页第2段. * |
| 王海燕,等.大孔树脂法分离纯化那他霉素的工艺研究.《中国抗生素杂志》.2010,第35卷(第3期),全文. * |
| 蔡寅,等.大孔吸附树脂在微生物制药分离纯化应用上的最新进展.《离子交换与吸附》.2008,第24卷(第5期),全文. * |
| 顾觉奋,等.大孔网状吸附剂在微生物制药分离纯化上的应用.《离子交换与吸附》.2002,第18卷(第3期),全文. * |
| 顾觉奋.大孔网状吸附剂在微生物制药分离纯化上的应用.《微生物药品研发动态——新技术、新产品及市场信息》.化学工业出版社,2005,(第1版),第122-131页. * |
| 顾觉奋.大孔网状聚合物吸附剂在微生物制药生产上的应用.《分离纯化工艺原理》.中国医药科技出版社,2002,(第1版),第153-155页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102453062A (en) | 2012-05-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102746348B (en) | A kind of separation method of lincomycin | |
| CN102336796B (en) | Preparation method of nemadectin | |
| CN101481715B (en) | Method for purifying tacrolimus by biofermentation | |
| CN101665446B (en) | Extract method of capsaicine and capsanthin | |
| CN102465164B (en) | Preparation method of pristinamycin | |
| EP2123765B1 (en) | Method for production of microbial fermentation product | |
| CN102952179B (en) | A kind of preparation method of high-purity micafungin precursor compound | |
| CN102443012B (en) | A kind of method of purifying rapamycin from fermented liquid | |
| US10316052B2 (en) | Fidaxomicin purification method | |
| CN102086226B (en) | Method for preparing cyclosporine A | |
| CN105585578B (en) | A kind of preparation method of rapamycin | |
| CN102453062B (en) | Partricin A separation method | |
| CN109053638B (en) | Extraction and purification method of fumagillin | |
| CN102453063B (en) | Method for separation of partricin B | |
| CN1931835A (en) | Process of extracting citrulline from water melon | |
| CN100436392C (en) | Method of culturing and extracting veralkaol using polygonum cuspidatum produced verakaol endogenic bacteria | |
| CN108570016B (en) | PF1022A separation and purification method | |
| CN107778357B (en) | Extraction and purification method of pneumocandin B0 | |
| CN112409426B (en) | Preparation method of sisomicin sulfate | |
| CN103224547A (en) | Daptomycin separation and purification method | |
| CN102863433A (en) | Mupirocin purification method | |
| CN102399210A (en) | Method for separating and extracting high-purity brefeldin A from fermentation liquor | |
| CN103087117B (en) | A kind of preparation method of high purity Elaiophylin | |
| CN1306035C (en) | Method for extracting natural abscisic acid | |
| CN104805146A (en) | Method for preparing L-tyrosine from fermentation broth of Chinese prickly ash endogenous brevibacterium frigoritolerans |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150304 Termination date: 20191021 |