CN102180922A - Improved method for purifying erythromycin A - Google Patents
Improved method for purifying erythromycin A Download PDFInfo
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- CN102180922A CN102180922A CN2011100656642A CN201110065664A CN102180922A CN 102180922 A CN102180922 A CN 102180922A CN 2011100656642 A CN2011100656642 A CN 2011100656642A CN 201110065664 A CN201110065664 A CN 201110065664A CN 102180922 A CN102180922 A CN 102180922A
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Abstract
The invention relates to method for purifying erythromycin A from fermented liquid by adopting a resin adsorption method, which includes the following steps: (1) adsorption, (2) washing of impurities, (3) desorption, (4) salifying of eluent and the like. The method overcomes the defects of lower elution rate of 1BV erythromycin, more impurity contents in products and the like in the existing resin adsorption and separation technology. In addition, the invention also provides a resin regeneration method which includes the following main steps: mixed solution consisting of 0.1 mol/L-1.0 mol/L NaOH aqueous solution and a water soluble or partially water soluble organic solvent is adopted for washing resin to be regenerated, so as to solve the problem of poor resin regeneration effect in the prior art.
Description
Technical field
The present invention relates to a kind of method of purification of Erythromycin A, specifically, relate to a kind of method that adopts macroreticular resin absorbing method purification of erythromycin A from fermented liquid.
Background technology
Erythromycin (Erythromycin is called for short EM) is a kind of Macrolide Broad spectrum antibiotics that gets from the streptomyces erythareus fermentation, is one of at present main antibiotic product.Have multiple different isomer such as Erythromycin A, B and C in the erythromycin, its chemical property is all very similar to structure, but wherein the anti-microbial activity of Erythromycin A is stronger, and toxicity is lower, is the main anti-microbial activity composition of medical erythromycin.
The extraction of Erythromycin A refining aspect, resin adsorption method is compared with traditional solvent extration, has advantages such as the solvent loss is little, easy and simple to handle, low in the pollution of the environment, comes into one's own day by day.
Existing relating to, adopt the document of resin adsorption method method of purification of erythromycin A from fermented liquid to have: Chinese microbiotic magazine 2007,32 (5) .284-286,315 and patent documentation CN 101367855, by technical scheme that it disclosed as can be known, the defective that prior art mainly exists is: (1) 1BV erythromycin eluting rate is lower, (2) do not relate to the removal method (so can influence the quality of the finished product) of impurity such as pigment and albumen in the fermented liquid, (3) to the regeneration of adsorption column, do not consider the removal effect of impurity in the fermented liquid, regeneration effect is not good.
Given this, the invention provides a kind of method that adopts resin adsorption method purification of erythromycin A from fermented liquid, overcome the defective that exists in the prior art.
Summary of the invention
Of the present invention from fermented liquid the method for purification of erythromycin A, comprise the steps:
(1) resin absorption
With the erythromycin fermentation liquid is raw material, after adjusting its pH value to 9.0～10.0, gained filtrate behind the membrane filtration carries out the absorption of fixed bed nonpolar macroporous adsorption resin, control erythromycin adsorptive capacity is that 60%～70% of nonpolar macroporous adsorption resin saturated extent of adsorption (descends as if filtrate pH in adsorption process to some extent, then adjust immediately, make it be not less than 9.0 all the time);
(2) resin washing
With the pH value be 8.0～11.0 contain 0.05mol/L (B
4O
7)
2-Borax-NaOH buffered soln or itself and can dissolve each other with water or the organic solvent of partial miscibility [as (but being not limited only to) alcohols, ketone, ethers or ester class etc., being preferably acetone] mixing solutions formed is (in described mixing solutions, organic solvent content is preferably 1v/v%～5v/v%) as washing composition, under 40 ℃ of conditions, nonpolar macroporous adsorption resin in the step (1) is washed, when the pH value of the pH value of the instantaneous effluent liquid in fixed bed exit and washing composition is identical, stop washing;
(3) resin elution
To nonpolar macroporous adsorption resin through step (2) washing, cross post (washing composition in the bed is washed on the top off) with 1BV (BV is the fixed bed volume) deionized water earlier, carry out wash-out with butylacetate as eluent then;
(4) elutriant salify
To by after the Erythromycin A crystallization in first 1BV elutriant of step (3) gained, the drying target product;
Described nonpolar macroporous adsorption resin comprises: product A mberlite XAD16 of U.S. Rohm-hass company or Amberlite XAD 1600 or the product SEPABEADS SP207 of MIT or SEPABEADS SP825 and kin homemade polymeric adsorbent etc.
Select the reason of borax-NaOH buffered soln to be in the above-mentioned steps (2): (1) experiment finds that the pigment molecule in the erythromycin filtrate is soluble in basic solution, and it is acid that protein molecule wherein is mostly, so best with impurity effects such as the pigment in the alkaline buffer solution flush away polymeric adsorbent, albumen; (2) Erythromycin A higher, that stable p H value helps being adsorbed on the resin further transfers the free alkali that is molecular state at short notice and does not produce alkalescence destruction in the buffered soln, has increased the eluting rate of follow-up elution process; (3) than other buffered soln with organic acid or organic bases configuration, this buffered soln low price saves production cost; (4) contain higher Na in this buffered soln
+Ionic strength, Na
+Water and effect can effectively reduce the solubleness of Erythromycin A in the aqueous solution, reduce the loss of Erythromycin A in the washing process; (5) contain high valence ion (as Ca in the erythromycin filtrate of upper prop absorption
2+, Mg
2+And Fe
2+Deng), can not use the buffered soln that contains carbanion.
In addition, of the present invention from fermented liquid the method for purification of erythromycin A, also comprise specifically comprising the steps: the regeneration step of used nonpolar macroporous adsorption resin
The nonpolar macroporous adsorption resin that wash-out in the step (3) is finished, earlier cross post with 1BV acetone, the butylacetate in the bed is washed on the top off, use then by concentration be 0.1mol/L～1.0mol/L the NaOH aqueous solution with can dissolve each other with water or the organic solvent of partial miscibility [as (but being not limited only to): alcohols, ketone, ethers or ester class etc., being preferably acetone] (in described mixing solutions, organic solvent content is preferably 20v/v%～80v/v%) used nonpolar macroporous adsorption resin is regenerated for the mixing solutions formed.
Positively effect of the present invention is:
(1) be that to carry out resin absorption and pH value be that 8.0～11.0 buffered soln washs the resin that has adsorbed target compound for 9.0～10.0 filtrate with the pH value among the present invention, guaranteed that Erythromycin A all exists with the free alkali of molecular state in the technological process, make 1BV eluting rate in the follow-up elution process greater than 90%, elution peak is more concentrated, thereby improved the yield of technology, saved the consumption of eluent.
(2) the resin washing methods flush away among the present invention be adsorbed on impurity such as partial pigment on the resin and albumen, solved the fermented liquid impurity removal problem in original technology, improved the quality of final Erythromycin A product; The rate of loss that higher ionic strength makes Erythromycin A in the washing process in the washings has guaranteed the preparation of high yield, high quality Erythromycin A product less than 2%.
(3) resin regeneration method among the present invention has solved the resin regeneration problem in original technology.Color of resin after the regeneration uses repeatedly the back that the adsorptive capacity of Erythromycin A in the filtrate is remained on more than 90% the regeneration effect ideal near primary colors repeatedly.
(4) technology of the present invention is succinct, and easy handling is practical, and good economy performance can directly apply to commercial scale production.
Description of drawings
Fig. 1 is the schematic flow sheet of purification of erythromycin A method of the present invention.
Embodiment
For a better understanding of the present invention, the invention will be further described below by embodiment, but for embodiment do not limit protection scope of the present invention.
Embodiment
(1) resin absorption
Erythromycin fermentation liquid puts jar after the 50nm ceramic membrane filter after gained filtrate adjustment pH is 9.30, feeds in the glass chromatography column that macroporous adsorbent resin SEPABEADS SP825 is housed and carries out resin absorption, and the control upper column quantity is 63.7% of a resin saturated extent of adsorption.
(2) resin washing
To the resin that the absorption of last step finishes, feeding pH value is borax-NaOH buffered soln of 9.78, carries out the resin washing under 40 ℃, stops washing when instantaneous effluent liquid pH is 9.78, and the erythromycin rate of loss is 0.92% in the washing process.
(3) resin elution
Washed the resin that finishes the last step, earlier cross post with the 1BV deionized water, feed butylacetate then and carry out resin elution, the erythromycin chemical titer is 70062u/mL in the 1BV elutriant, and eluting rate was 91.6% (eluting rate of the 1BV of prior art is about 50%～70%).
(4) elutriant salify
Erythromycin A in the 1BV elutriant of gained of last step is crystallized into thiocyanate-, get the red product of sulphur after the drying.The product biological value is 869.3u/mg, and the Erythromycin A component concentration is 85.6%, and whole process yield is not less than 85.3%.
(5) resin regeneration
To the resin that wash-out in the step (3) finishes, cross post with 1BV acetone earlier, the mixing solutions that feeds 60% acetone and 40%0.4mol/LNaOH solution then carries out resin regeneration, and is with deionized water that the resin flushing is extremely neutral at last.The saturated extent of adsorption of regeneration back resin is 93.1% of a fresh resin saturated extent of adsorption.
Claims (5)
1. the method for a purification of erythromycin A from fermented liquid is characterized in that, described method comprises the steps:
(1) be raw material with the erythromycin fermentation liquid of putting jar, carry out fixed-bed resin absorption after gained filtrate behind the membrane filtration is adjusted its pH value to 9.0～10.0, control erythromycin adsorptive capacity is 60%～70% of a resin saturated extent of adsorption;
(2) with the pH value be 8.0～11.0 contain 0.05mol/L (B
4O
7)
2-Borax-NaOH buffered soln or itself and can dissolve each other with water or mixing solutions that the organic solvent of partial miscibility is formed as washing composition, under 40 ℃ of conditions, the nonpolar macroporous adsorption resin that has adsorbed target compound in the step (1) is washed, when the pH value of the pH value of the instantaneous effluent liquid in fixed bed exit and washing composition is identical, stop washing;
(3) to nonpolar macroporous adsorption resin through step (2) washing, cross post with the 1BV deionized water earlier, carry out wash-out with butylacetate as eluent then;
(4) to by obtaining target product after the Erythromycin A crystallization in first 1BV elutriant of step (3) gained, the drying;
Described nonpolar macroporous adsorption resin is: Amberlite XAD16, Amberlite XAD1600, SEPABEADSSP207 or SEPABEADS SP825.
2. in accordance with the method for claim 1, it is characterized in that, described in the step (2) by the pH value be 8.0～11.0 contain 0.05mol/L (B
4O
7)
2-Borax-NaOH buffered soln with can dissolve each other with water or mixing solutions that the organic solvent of partial miscibility is formed in the content of organic solvent be 1v/v%～5v/v%.
3. according to claim 1 or 2 described methods, it is characterized in that described method also comprises the steps:
The nonpolar macroporous adsorption resin that wash-out in the step (3) is finished, earlier cross post with 1BV acetone, the butylacetate in the bed is washed on the top off, and using then by concentration is that the NaOH aqueous solution of 0.1mol/L～1.0mol/L washs used nonpolar macroporous adsorption resin with the mixing solutions that can dissolve each other with water or the organic solvent of partial miscibility is formed.
4. in accordance with the method for claim 3, it is characterized in that the content of organic solvent is 20v/v%～80v/v% in the wherein said mixing solutions.
5. according to claim 2 or 4 described methods, it is characterized in that wherein said organic solvent is an acetone.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100656642A CN102180922A (en) | 2011-03-18 | 2011-03-18 | Improved method for purifying erythromycin A |
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|---|---|---|---|
| CN2011100656642A CN102180922A (en) | 2011-03-18 | 2011-03-18 | Improved method for purifying erythromycin A |
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| CN102180922A true CN102180922A (en) | 2011-09-14 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102408462A (en) * | 2011-12-02 | 2012-04-11 | 伊犁川宁生物技术有限公司 | Preparation method of erythromycin thiocyanate |
| CN103159812A (en) * | 2011-12-08 | 2013-06-19 | 胡先念 | Method of extracting erythromycin from erythromycin aqueous solution |
| CN103694295A (en) * | 2014-01-08 | 2014-04-02 | 华东理工大学 | Method for optimizing available mycin components |
| CN105699566A (en) * | 2016-01-05 | 2016-06-22 | 南京大学 | Passive sampling instrument for measuring macrolide antibiotic in water and application thereof |
| CN108373490A (en) * | 2017-11-24 | 2018-08-07 | 宁夏启元药业有限公司 | A kind of separation method of erythromycin impurity B |
| CN109553650A (en) * | 2017-09-25 | 2019-04-02 | 联邦制药(内蒙古)有限公司 | The aqueous extraction method of erythromycin fermentation liquid |
-
2011
- 2011-03-18 CN CN2011100656642A patent/CN102180922A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 宋应华等: "大孔树脂提纯红霉素的研究", 《中国抗生素杂志》 * |
| 雷引林等: "硼酸络合法脱糖的研究", 《中国抗生素杂志》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102408462A (en) * | 2011-12-02 | 2012-04-11 | 伊犁川宁生物技术有限公司 | Preparation method of erythromycin thiocyanate |
| CN102408462B (en) * | 2011-12-02 | 2014-10-22 | 伊犁川宁生物技术有限公司 | Preparation method of erythromycin thiocyanate |
| CN103159812A (en) * | 2011-12-08 | 2013-06-19 | 胡先念 | Method of extracting erythromycin from erythromycin aqueous solution |
| CN103159812B (en) * | 2011-12-08 | 2016-03-23 | 湖南中创化工股份有限公司 | A kind of method extracting erythromycin from the erythromycin aqueous solution |
| CN103694295A (en) * | 2014-01-08 | 2014-04-02 | 华东理工大学 | Method for optimizing available mycin components |
| CN103694295B (en) * | 2014-01-08 | 2016-01-20 | 华东理工大学 | A kind of method optimizing rokitamycin component |
| CN105699566A (en) * | 2016-01-05 | 2016-06-22 | 南京大学 | Passive sampling instrument for measuring macrolide antibiotic in water and application thereof |
| CN109553650A (en) * | 2017-09-25 | 2019-04-02 | 联邦制药(内蒙古)有限公司 | The aqueous extraction method of erythromycin fermentation liquid |
| CN109553650B (en) * | 2017-09-25 | 2020-11-20 | 联邦制药(内蒙古)有限公司 | Water phase extraction method of erythromycin fermentation liquor |
| CN108373490A (en) * | 2017-11-24 | 2018-08-07 | 宁夏启元药业有限公司 | A kind of separation method of erythromycin impurity B |
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Application publication date: 20110914 |