CN102617665A - Method utilizing sulfonic acid cation exchange resin to separate etimicin - Google Patents
Method utilizing sulfonic acid cation exchange resin to separate etimicin Download PDFInfo
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- CN102617665A CN102617665A CN2012100499896A CN201210049989A CN102617665A CN 102617665 A CN102617665 A CN 102617665A CN 2012100499896 A CN2012100499896 A CN 2012100499896A CN 201210049989 A CN201210049989 A CN 201210049989A CN 102617665 A CN102617665 A CN 102617665A
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- etimicin
- exchange resin
- sulfonic acid
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Abstract
The invention discloses a method utilizing sulfonic acid cation exchange resin to separate etimicin, which comprises the following steps: taking 5mL of etimicin fermentation liquid, controlling the flow velocity of loading to be within a range of 0.8-1.2mL/min after acidizing pretreatment is performed to the etimicin fermentation liquid, loading the acidized liquid having undergone acidizing pretreatment to a chromatographic column separating filler, and stewing for 4-24h; controlling elution flow velocity to be within the range of 1.0-5.0 mL/min, sequentially using water of 2 column volumes, 2 column volumes of salt solution with the concentration to be 0.1-0.4mol/L and ammonia water with the concentration to be 3%-45% to perform elution; and obtaining the required etimicin after rotary evaporation of the obtained eluent.
Description
Technical field
The present invention relates to separate the Etimicin field, specifically relate to a kind of method of utilizing polystyrolsulfon acid type Zeo-karb to separate Etimicin.
Background technology
Etimicin (Etimicin) is a kind of semisynthetic water soluble antibiotics.Show that through pharmacology, toxicity, pharmacodynamic study and clinical study widely Etimicin is that the adopted resistance of low, the anti-friendship of untoward reaction is good; The order ototoxicity is far below other aminosugar type of having microbiotic; Be a kind of anti-infective safely and effectively new drug (Lv Yaping, Chen Jianjun, Luo Jun. the stability study [J] of Etimicin in different solvents. Zhejiang Polytechnical University's journal; 2006,34 (3): 291-292).
Etimicin is by gentamicinC
1a1 ethylating product of amido selectivity; So the complexity of Etimicin separation and purification depends on 1 the ethylating selectivity of amido (Liu Jun, Hu Xiaoling, Shen Yiqun; The model a beautiful gem; Zhao Min. the research [J] of associated products during Etimicin is synthetic. Chinese Journal of Pharmaceuticals, 2000,31 (8): 358-360).Because Etimicin is alkalescence, water soluble antibiotics, stable in properties, the method for separating Etimicin at present are to be separating filler with the macroporous resin mostly, but product purity that this method obtains only 90%; Or be separating filler with the acidulous cation resin, but this method will consume a large amount of organic solvent ethanol, and cost is high, not environmental protection.Seek for this reason and a kind ofly can either practice thrift cost, the method that can improve product purity is again separated Etimicin and is had crucial meaning.
Summary of the invention
The objective of the invention is deficiency,, utilize polystyrolsulfon acid type Zeo-karb to separate Etimicin according to the mechanism of action of IX to prior art.
For realizing the object of the invention, the present invention adopts following technical scheme to realize:
A kind of sulfonic acid ion exchange resin separates the method for Etimicin; Described method may further comprise the steps: get 5mL Etimicin fermented liquid; After the Etimicin fermented liquid carried out the acidifying pre-treatment; The flow velocity of appearance will leave standstill 4～24h in the control through the pretreated acidizing fluid material loading of acidifying to the chromatography column of separating filler in 0.8～1.2mL/min scope; The control elution flow rate is in 1.0～5.0mL/min scope, and using 2 cylinder ponding, concentration successively is that 2 column volume salts solutions, the concentration of 0.1～0.4mol/L is that 3%～45% ammoniacal liquor carries out wash-out; The elutriant of above-mentioned gained is obtained required Etimicin behind rotary evaporation;
Described separating filler is selected polystyrolsulfon acid type Zeo-karb for use;
Described salts solution is selected a kind of in acetate solution or the carbonate solution for use;
Described acidifying pre-treatment; May further comprise the steps: get the Etimicin fermented liquid; Using volumetric concentration is that 10～20% sulphuric acid soln is acidified to pH2.0; After leaving standstill 1 hour, using mass concentration again is that 5% sodium hydroxide solution is neutralized to pH6.0 with the Etimicin fermented liquid, promptly gets required Etimicin acidizing fluid;
Described chromatography column selects for use specification to be: aspect ratio is 12.5: 1～55.5: 1 a chromatography column; The preferred specification of described chromatography column is: the chromatography column of (500～755) * (40～15) mm;
It is 18～60 purpose polystyrene-divinylbenzene multipolymers that described polystyrolsulfon acid type Zeo-karb is selected particle diameter for use;
Described acetate solution is selected a kind of in sodium acetate soln or the Spirit of Mindererus for use;
Described carbonate solution is selected a kind of in sodium carbonate solution or the sodium hydrogen carbonate solution for use;
The Etimicin that obtains in the above-mentioned steps is detected its purity with HPLC (HPLC).
Technique scheme is based on the mechanism of action of IX; Utilize polystyrolsulfon acid type Zeo-karb to separate Etimicin for separating filler; Because polystyrolsulfon acid type Zeo-karb has good reticulated structure and higher specific surface area and specific surface property; And ionic group can adsorb Etimicin and Gentamicin C1a selectively, makes separating effect reach optimum; Simultaneously because polystyrolsulfon acid type Zeo-karb is reusable, so cost reduces greatly, and therefore having forgone needs with a large amount of organic solvent ethanol, so reduce cost, helps environmental protection.
Beneficial effect of the present invention is:
1, the purity of gained medicine is more than 98%, and the recovery is more than 80%.
2, separation method is simple, and used isolating reagent is cheap.
3, used carrier of separating good stability, the life-span is long, is beneficial to recycling, and is easy to regeneration.
4, the ethanol with an organic solvent of having forgone has reduced cost, helps environmental protection.
Embodiment
Through concrete embodiment, the present invention is done detailed description below.
Embodiment 1
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 60 orders;
Chromatography column: 500 * 40mm;
Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH) with 20% sulphuric acid soln, with appearance on the flow velocity of 1.2mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 12h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volume V carries out wash-out successively, and 2 column volume concentration are the sodium hydrogencarbonate of 0.4mol/L, 30% ammoniacal liquor wash-out; The flow velocity of wash-out is 5.0mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 82.8%, and the purity of Etimicin is 98.1%.
Embodiment 2
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 25 orders;
Chromatography column: 400 * 20mm;
Get the Etimicin fermented liquid of 5mL, with 10% sulphuric acid soln be acidified to pH 2.0 (the extensive test paper of pH) with appearance on the flow velocity of 1.0mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 4h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the sodium-acetate of 0.1mol/L, 45% ammoniacal liquor wash-out; The flow velocity of wash-out is 3.0mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 83.1%, and the purity of Etimicin is 98.3%.
Embodiment 3
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 18 orders;
Chromatography column: 755 * 15mm; Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH), to chromatography column, left standstill 1 hour with appearance on the flow velocity of 0.8mL/min with 15% sulphuric acid soln, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 24h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the sodium-acetate of 0.3mol/L, 3% ammoniacal liquor wash-out; The flow velocity of wash-out is 1.5mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 87.1%, and the purity of Etimicin is 98.5%.
Embodiment 4
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 68 orders;
Chromatography column: 500 * 40mm; Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH) with 20% sulphuric acid soln, with appearance on the flow velocity of 1.2mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 8h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the sodium hydrogencarbonate of 0.2mol/L, 20% ammoniacal liquor wash-out; The flow velocity of wash-out is 2.5mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 81.3%, and the purity of Etimicin is 99.2%.
Embodiment 5
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 18 orders;
Chromatography column: 500 * 40mm; Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH) with 10% sulphuric acid soln, with appearance on the flow velocity of 1.2mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 11h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the ammonium acetate of 0.4mol/L, 5% ammoniacal liquor wash-out; The flow velocity of wash-out is 2.0mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 80.5%, and the purity of Etimicin is 98.7%.
Embodiment 6
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 25 orders;
Chromatography column: 400 * 20mm; Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH) with 15% sulphuric acid soln, with appearance on the flow velocity of 0.8mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 17h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the yellow soda ash of 0.3mol/L, 15% ammoniacal liquor wash-out; The flow velocity of wash-out is 3.0mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 82.0%, and the purity of Etimicin is 98.4%.
Embodiment 7
Separation condition: the particle diameter of carrier of separating polystyrolsulfon acid type Zeo-karb is 18 orders;
Chromatography column: 755 * 15mm;
Get the Etimicin fermented liquid of 5mL, be acidified to pH 2.0 (the extensive test paper of pH) with 20% sulphuric acid soln, with appearance on the flow velocity of 1.0mL/min to chromatography column, after left standstill 1 hour, then with 5% sodium hydroxide solution with the fermented liquid pH 6.0 that neutralizes; The fermented liquid that pre-treatment is good to ion exchange column, after the standing adsorption 8h, begins wash-out with appearance on the flow velocity of 1.0mL/min; During wash-out, the water with 2 column volumes carries out wash-out successively, and 2 column volume concentration are the ammonium acetate of 0.2mol/L, 10% ammoniacal liquor wash-out; The flow velocity of wash-out is 1mL/min; The elutriant of collecting gets the Etimicin purified product through rotary evaporation, and the recovery is 82.2%, and the purity of Etimicin is 98.2%.
Claims (7)
1. a sulfonic acid ion exchange resin separates the method for Etimicin, and it is characterized in that: described method may further comprise the steps:
Get 5mL Etimicin fermented liquid, the Etimicin fermented liquid is carried out the acidifying pre-treatment after, in the control appearance flow velocity in 0.8～1.2mL/min scope, will through the pretreated acidizing fluid material loading of acidifying to the chromatography column of separating filler, leave standstill 4～24h; The control elution flow rate is in 1.0～5.0mL/min scope, and using 2 cylinder ponding, concentration successively is that 2 column volume salts solutions, the concentration of 0.1～0.4mol/L is that 3%～45% ammoniacal liquor carries out wash-out; The elutriant of above-mentioned gained is obtained required Etimicin behind rotary evaporation;
Described separating filler is selected polystyrolsulfon acid type Zeo-karb for use;
Described salts solution is selected a kind of in acetate solution or the carbonate solution for use.
2. a kind of sulfonic acid ion exchange resin according to claim 1 separates the method for Etimicin, and it is characterized in that: described acidifying pre-treatment may further comprise the steps:
Get the Etimicin fermented liquid, using volumetric concentration is that 10～20% sulphuric acid soln is acidified to pH2.0, leave standstill 1 hour after, using mass concentration again is that 5% sodium hydroxide solution is neutralized to pH6.0 with the Etimicin fermented liquid, promptly gets required Etimicin acidizing fluid.
3. a kind of sulfonic acid ion exchange resin according to claim 1 separates the method for Etimicin, and it is characterized in that: described chromatography column selects for use specification to be: aspect ratio is 12.5: 1～55.5: 1 a chromatography column.
4. a kind of sulfonic acid ion exchange resin according to claim 3 separates the method for Etimicin, and it is characterized in that: described chromatography column selects for use specification to be: the chromatography column of (500～755) * (40～15) mm.
5. a kind of sulfonic acid ion exchange resin according to claim 1 separates the method for Etimicin, and it is characterized in that: it is 18～60 purpose polystyrene-divinylbenzene multipolymers that described polystyrolsulfon acid type Zeo-karb is selected particle diameter for use.
6. a kind of sulfonic acid ion exchange resin according to claim 1 separates the method for Etimicin, it is characterized in that: described acetate solution is selected a kind of in sodium acetate soln or the Spirit of Mindererus for use.
7. a kind of sulfonic acid ion exchange resin according to claim 1 separates the method for Etimicin, it is characterized in that: described carbonate solution is selected a kind of in sodium carbonate solution or the sodium hydrogen carbonate solution for use.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105388046A (en) * | 2014-09-04 | 2016-03-09 | 中国科学院大连化学物理研究所 | Applications of sulfonated porous polystyrene-divinylbenzene particles |
| CN114307252A (en) * | 2021-12-30 | 2022-04-12 | 南京工业大学 | Process for separating sugar and acid by using quasi-two-dimensional chromatography |
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| CN1100467A (en) * | 1993-04-23 | 1995-03-22 | 江苏省微生物研究所 | 1-N-ethyl gentamicin derivative and its preparing method |
| CN101614712A (en) * | 2009-07-27 | 2009-12-30 | 南京工业大学 | A kind of analytical approach of Etimicin |
| CN101928312A (en) * | 2010-03-26 | 2010-12-29 | 常州方圆制药有限公司 | Preparation method of 1-N-ethyl gentamicin C1a sulfate |
| US20110172411A1 (en) * | 2008-06-26 | 2011-07-14 | Danisco A/S | Process for separation of ca- or mg-sulfite spent liquor to yield crystalline xylose |
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2012
- 2012-02-29 CN CN2012100499896A patent/CN102617665A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1100467A (en) * | 1993-04-23 | 1995-03-22 | 江苏省微生物研究所 | 1-N-ethyl gentamicin derivative and its preparing method |
| US20110172411A1 (en) * | 2008-06-26 | 2011-07-14 | Danisco A/S | Process for separation of ca- or mg-sulfite spent liquor to yield crystalline xylose |
| CN101614712A (en) * | 2009-07-27 | 2009-12-30 | 南京工业大学 | A kind of analytical approach of Etimicin |
| CN101928312A (en) * | 2010-03-26 | 2010-12-29 | 常州方圆制药有限公司 | Preparation method of 1-N-ethyl gentamicin C1a sulfate |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105388046A (en) * | 2014-09-04 | 2016-03-09 | 中国科学院大连化学物理研究所 | Applications of sulfonated porous polystyrene-divinylbenzene particles |
| CN105388046B (en) * | 2014-09-04 | 2018-08-14 | 中国科学院大连化学物理研究所 | A kind of application of sulfonated porous polystyrene-divinylbenzene particle |
| CN114307252A (en) * | 2021-12-30 | 2022-04-12 | 南京工业大学 | Process for separating sugar and acid by using quasi-two-dimensional chromatography |
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Application publication date: 20120801 |