CN100549026C - A kind of vancomycin column chromatography purification method - Google Patents
A kind of vancomycin column chromatography purification method Download PDFInfo
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- CN100549026C CN100549026C CNB200610023150XA CN200610023150A CN100549026C CN 100549026 C CN100549026 C CN 100549026C CN B200610023150X A CNB200610023150X A CN B200610023150XA CN 200610023150 A CN200610023150 A CN 200610023150A CN 100549026 C CN100549026 C CN 100549026C
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- vancomycin
- column chromatography
- purification method
- ammonium salt
- chromatography purification
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Abstract
The invention discloses a kind of vancomycin column chromatography purification method, it adopts with Sephadex C series is that the post of filler carries out column chromatography, with certain density salts solution gradient elution, it is characterized in that this salts solution selects C for use
1-3The ammonium salt solution of organic carboxyl acid.The inventive method has overcome and had had the blistered phenomenon of carbonate decomposition in the post process now, thereby has improved stage number, and resolving power improves, and applied sample amount increases; And the content of vancomycin improves, and impurity level reduces.
Description
Technical field
The present invention relates to a kind of extracting and purifying method of antibiotic vancomycin, particularly a kind of vancomycin column chromatography purification method.
Background technology
Vancomycin is a kind of microbiotic of glycopeptide class, is used for the treatment of by catching that gram-positive microorganism causes.Especially the resistance streptococcus aureus and the difficulty that are caused by β-Nei Xiananleikangshengsu are distinguished that clostridium (Clostridium difficile) has curative effect preferably.The applicant's patent application (publication number CN1616651A) discloses with hydrochloric acid and has regulated the fermented liquid pH that fermentation produces, filter the back and cross the D1300 macroporous resin, with acidic ethanol aqueous solution wash-out, elutriant concentrates, employing is that the post of filler carries out column chromatography with Sephadex C series, with certain density salts solution gradient elution, collects significant part, after the activated carbon decolorizing filtering with microporous membrane was removed thermal source, lyophilize obtained the aseptic raw material dry powder of vancomycin hydrochloride.Wherein having a step to adopt with Sephadex C series plastics is that the post of filler carries out column chromatography, with the NH of 0.01-1M
4HCO
3, (NH
4)
2CO
3The method purifying of aqueous solution wash-out, carbonate can decompose foaming in the post process excessively, thereby has influenced separation efficiency; The content that makes vancomycin in the dry powder is 94.4%, and foreign matter content is 1%.
Summary of the invention
The technical problem to be solved in the present invention is and overcomes above-mentioned defective of the prior art, provides a kind of and can improve stage number, resolving power and/or applied sample amount, and can guarantee the vancomycin column chromatography purification method of purifying quality.
For addressing the above problem, the inventor intends adopting and can not decompose blistered salts solution come instead of ammonium carbonate or ammonium bicarbonate soln in crossing the post process.But in R﹠D process, find, many salts solutions commonly used, soluble inorganic salt (as its an alkali metal salt, ammonium salt etc.) as formation such as sulfuric acid, hydrochloric acid, phosphoric acid, such as the sodium/potassium/ammonium salt of acid such as sulfuric acid, hydrochloric acid, phosphoric acid etc., though in crossing the post process, can not decompose foaming, may bring new problem, as oxidized easily, can produce new impurity such as pigment, thereby influence the purifying quality of vancomycin.
The inventor filters out the salts solution that meets requirement of the present invention by attempting different salts solutions, draws the following technical scheme that can solve the technology of the present invention problem thus.
A kind of vancomycin column chromatography purification method, it adopts with Sephadex C series is that the post of filler carries out column chromatography, with certain density salts solution gradient elution, it is characterized in that this salts solution selects C for use
1-3The ammonium salt solution of organic carboxyl acid.
Wherein, this C
1-3The preferred C of the ammonium salt of organic carboxyl acid
1-3Monocarboxylic ammonium salt is more preferably from ammonium formiate, ammonium acetate, propionic acid ammonium and DL-Lactic acid ammonium salt.
And other steps, condition in the vancomycin column chromatography purification method of the present invention, (0.01～1M) etc. all with the disclosed content of above-mentioned CN1616651A patent application as column packing (Sephadex C-15/C-25/C-50), eluent salt strength of solution.
Adopt above-mentioned organic carboxyl acid ammonium salt to replace carbonate as elutriant, overcome and crossed the blistered phenomenon of carbonate decomposition in the post process, thereby improved stage number, resolving power improves, and applied sample amount increases; And guarantee the purifying quality, the content of vancomycin is improved, impurity level reduces.
Embodiment
Embodiment 1
Select for use through the D1300 macroporous resin, with the 80% acidic ethanol aqueous solution wash-out of pH2.0, and the spissated elutriant that contains vancomycin, adopting with Sephadex C-15 series plastics is that the post of filler carries out column chromatography, with the ammonium formiate (HCOONH of 0.05M concentration
4) aqueous solution gradient elution, collect the HPLC area of detection than the part more than 95%, activated carbon decolorizing, after the 0.22um filtering with microporous membrane was removed thermal source, lyophilize obtained the aseptic raw material dry powder of vancomycin hydrochloride.Last liquid measure is 8%; Vancomycin content is 95.5% in the finished product, and single impurity is 0.6%.
Embodiment 2
Employing is that the post of filler carries out column chromatography with Sephadex C-25 series plastics, with the ammonium acetate (CH of 0.1M concentration
3COONH
4) as the column chromatography elutriant, surplus with embodiment 1, make the aseptic raw material of vancomycin hydrochloride in powder.Last liquid measure is 8%; Vancomycin content is 95.3% in the finished product, and single impurity is 0.7.
Embodiment 3
Employing is that the post of filler carries out column chromatography with Sephadex C-50 series plastics, with the propionic acid ammonium (CH of 1M concentration
3CH
2COONH
4) aqueous solution gradient elution, surplus with embodiment 1, make the aseptic raw material dry powder of vancomycin hydrochloride.Last liquid measure is 8%; Vancomycin content is 95.1% in the finished product, and single impurity is 0.8%.
Embodiment 4
Use the DL-Lactic acid ammonium salt [CH of 1M concentration instead
3CH (OH) COONH
4] aqueous solution gradient elution, surplus with embodiment 1, make the aseptic raw material dry powder of vancomycin hydrochloride.Last liquid measure is 8%; Vancomycin content is 95.0% in the finished product, and single impurity is 0.8%.
Claims (3)
1, a kind of vancomycin column chromatography purification method, it adopts with Sephadex C series is that the post of filler carries out column chromatography, with the salts solution gradient elution of 0.01～1M, it is characterized in that this salts solution selects C for use
1-3The ammonium salt solution of organic carboxyl acid.
2, vancomycin column chromatography purification method as claimed in claim 1 is characterized in that this C
1-3The ammonium salt of organic carboxyl acid is C
1-3Monocarboxylic ammonium salt.
3, vancomycin column chromatography purification method as claimed in claim 2 is characterized in that this C
1-3Monocarboxylic ammonium salt is selected from ammonium formiate, ammonium acetate, propionic acid ammonium and DL-Lactic acid ammonium salt.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200610023150XA CN100549026C (en) | 2006-01-06 | 2006-01-06 | A kind of vancomycin column chromatography purification method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200610023150XA CN100549026C (en) | 2006-01-06 | 2006-01-06 | A kind of vancomycin column chromatography purification method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1858061A CN1858061A (en) | 2006-11-08 |
| CN100549026C true CN100549026C (en) | 2009-10-14 |
Family
ID=37297021
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200610023150XA Expired - Fee Related CN100549026C (en) | 2006-01-06 | 2006-01-06 | A kind of vancomycin column chromatography purification method |
Country Status (1)
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|---|---|
| CN (1) | CN100549026C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101613396B (en) * | 2008-06-26 | 2016-08-17 | 浙江医药股份有限公司新昌制药厂 | Non-crystalline form Lyphocin (Fujisawa) and its production and use and its pharmaceutical composition |
| CN101659693B (en) * | 2008-08-27 | 2012-11-21 | 上海医药工业研究院 | Method for preparing pneumocandin B0 |
| CN102050866B (en) * | 2009-11-04 | 2015-02-18 | 上海来益生物药物研究开发中心有限责任公司 | Method for purifying glycopeptide compound |
-
2006
- 2006-01-06 CN CNB200610023150XA patent/CN100549026C/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| N-去甲基万古霉素单组份的分离与精制. 周玉.抗生素，第11卷第5期. 1986 |
| N-去甲基万古霉素单组份的分离与精制. 周玉.抗生素，第11卷第5期. 1986 * |
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|---|---|
| CN1858061A (en) | 2006-11-08 |
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