CN101024661B - Three single-component separation methods for Ramoplanin - Google Patents
Three single-component separation methods for Ramoplanin Download PDFInfo
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- CN101024661B CN101024661B CN200610024152A CN200610024152A CN101024661B CN 101024661 B CN101024661 B CN 101024661B CN 200610024152 A CN200610024152 A CN 200610024152A CN 200610024152 A CN200610024152 A CN 200610024152A CN 101024661 B CN101024661 B CN 101024661B
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- ramoplanin
- aqueous solution
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- methanol aqueous
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Abstract
The invention discloses the separating method for the three constituents of Ramoplanin. It includes three constituents of Ramoplanin A1, A2 and A3 that are dissolved by methanol, and taking sampling on the upper end of C18 reverse silica gel column, and using methanol solution as flow passing column to take column chromatography separation. The method has the advantages of low cost and good separation effect.
Description
Technical field
The present invention relates to three kinds of single component A1 of a kind of microbiotic ramoplanin, A2, the separation method of A3.
Background technology
(Ramoplanin, A/16686) traditional method of Ti Chuning is high performance liquid chromatography (reversed-phase HPLC, the post: μ-BondapackC with the preparation type described in the United States Patent (USP) (USP4427656) to ramoplanin
18, diameter 7.8mm * 300mm, moving phase: ammonium formiate and acetonitrile mixed solution) separate its A1, A2, three single components of A3, this method devices needed costliness, the single sample size is little, and the preparation liquid phase is owing to need fixed to prepare pillar, usually these pillars all are that producer's direct production is good, so size is fixing, cannot change.
Summary of the invention
The technical problem to be solved in the present invention is to overcome above-mentioned defective of the prior art, and the separation method that a kind of single sample size is big, need not expensive device, three kinds of single components of ramoplanin that separating effect is suitable is provided.
The inventor screens many separator columns that can increase sample size for this, and discovery can solve the problems referred to above by following technical proposal.The separation method of three kinds of single components of ramoplanin of the present invention comprises and will contain three kinds of component A1 of ramoplanin, A2, and the mixture of A3 dissolves with methanol aqueous solution, in C
18Sample on the upper end wet method of reverse phase silica gel post or the dry method is crossed post with methanol aqueous solution as moving phase, carries out column chromatography for separation.
Wherein, the present invention's said " mixture that contains three kinds of components of ramoplanin " can be according to prior art for preparing, for example can be with reference to above-mentioned american documentation literature disclosed method, and particularly the step of example 1～example 3 prepares.
Preferably, dissolving this methanol aqueous solution concentration that contains the mixture of three kinds of components of ramoplanin is 10%～40% (v/v), add methyl alcohol and be in order to increase solvability, the methanol concentration in too high can the influence follow-up wash-out moving phase of methanol concentration, thus cause purification efficiency to descend.
And should be as preferred 50～70% (v/v) of the methanol aqueous solution concentration of moving phase because methanol concentration is too low can't wash-out under ramoplanin, perhaps elution speed is slow excessively, the too high isolation resolution that then influences of concentration.
Sample is preferably the solute drying after earlier the described mixture that contains three kinds of components of ramoplanin being dissolved with methanol aqueous solution on the said dry method of the present invention, and then goes up sample.
The present invention adopts above-mentioned C
18Preferred 0.5～the 2ml/min of the flow velocity of reversed-phase silica gel column chromatography collects under the monitoring of conventional H PLC method and contains A1, A2, the effluent liquid of each component of A3 more than 90%, separates three kinds of single components that obtain ramoplanin.
Separation method of the present invention is compared with existing preparation HPLC separation method, and the single sample size is big when separating three components of equivalent, low price, and separating effect is suitable; And, use C
18Reversed-phase silica gel column chromatography can be used C as required voluntarily
18Reverse phase silica gel filler dress pillar, the volume of filler and pillar, high footpath can be adjusted voluntarily.
Embodiment
Further specify the present invention with embodiment below, but the present invention is not limited.
The mixture that contains ramoplanin three kinds of component A1, A2, A3 among the following embodiment is that the step of example 1～example 3 in the above-mentioned american documentation literature of reference prepares.
Embodiment 1
The mixture that will contain ramoplanin three kinds of component A1, A2, A3 is with 10% methanol aqueous solution dissolving, in C
18(sample on the upper end of post height * diameter: 20cm * 1cm) is crossed post with 50% methanol aqueous solution as moving phase, flow velocity 0.5ml/min to the reverse phase silica gel post.Conventional H PLC monitoring is collected down and is contained the effluent liquid of each component more than 90%.Yield is 83%.
Embodiment 2
The mixture that will contain three kinds of components of ramoplanin is with 40% methanol aqueous solution dissolving, in C
18(sample on the upper end of post height * diameter: 20cm * 1cm) is crossed post with 70% methanol aqueous solution as moving phase, flow velocity 2ml/min to the reverse phase silica gel post.Conventional H PLC monitoring is collected down and is contained the effluent liquid of each component more than 90%.Yield is 85%.
Embodiment 3
The mixture that will contain three kinds of components of ramoplanin is with 20% methanol aqueous solution dissolving, with solute vacuum-drying, in C
18(sample on the upper end of post height * diameter: 20cm * 1cm) is crossed post with 60% methanol aqueous solution as moving phase, flow velocity 1ml/min to the reverse phase silica gel post.Conventional H PLC monitoring is collected down and is contained the effluent liquid of each component more than 90%.Yield is 82%.
Used C in the foregoing description
18The reverse phase silica gel column packing is the easy science and technology limited Company 20 μ m-50 μ m ball-type C of the intelligent moral in Beijing
18The reverse phase silica gel column packing; Surplus reagent is conventional commercially available chromatographically pure.
Claims (5)
1. the separation method of three kinds of single components of a ramoplanin, it comprises and will contain three kinds of component A1 of ramoplanin, A2, the mixture of A3 dissolves with methanol aqueous solution, in C
18Sample on the upper end wet method of reverse phase silica gel post or the dry method is crossed post with methanol aqueous solution as moving phase, carries out column chromatography for separation.
2. separation method as claimed in claim 1, it is characterized in that dissolving this methanol aqueous solution concentration that contains the mixture of three kinds of components of ramoplanin is 10%～40%.
3. separation method as claimed in claim 1 is characterized in that this methanol aqueous solution concentration as moving phase is 50～70%.
4. separation method as claimed in claim 1 is characterized in that sample on the said dry method comprises the solute drying of mixture after with the methanol aqueous solution dissolving that will describedly contain three kinds of components of ramoplanin earlier, and then on sample.
5. separation method as claimed in claim 1, the flow velocity that it is characterized in that described column chromatography is 0.5～2ml/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200610024152A CN101024661B (en) | 2006-02-24 | 2006-02-24 | Three single-component separation methods for Ramoplanin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200610024152A CN101024661B (en) | 2006-02-24 | 2006-02-24 | Three single-component separation methods for Ramoplanin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101024661A CN101024661A (en) | 2007-08-29 |
| CN101024661B true CN101024661B (en) | 2010-05-12 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN200610024152A Expired - Fee Related CN101024661B (en) | 2006-02-24 | 2006-02-24 | Three single-component separation methods for Ramoplanin |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103360470A (en) * | 2013-07-26 | 2013-10-23 | 成都摩尔生物医药有限公司 | Purification method for ramoplanin fermentation liquor |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102838662B (en) * | 2011-12-02 | 2014-05-14 | 华北制药集团新药研究开发有限责任公司 | Preparation method of high purity ramoplanin single components |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4427656A (en) * | 1980-08-16 | 1984-01-24 | Gruppo Lepetit S.P.A. | Antibiotic A/16686 factor A2, the process for the preparation thereof and the co-produced antibiotic A/16686 factors A1 and A3 |
| US5925550A (en) * | 1986-09-11 | 1999-07-20 | Biosearch Italia S.P.A. | Method for selectively increasing the ratio of single major components of antibiotic A/16686 complex |
-
2006
- 2006-02-24 CN CN200610024152A patent/CN101024661B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4427656A (en) * | 1980-08-16 | 1984-01-24 | Gruppo Lepetit S.P.A. | Antibiotic A/16686 factor A2, the process for the preparation thereof and the co-produced antibiotic A/16686 factors A1 and A3 |
| US5925550A (en) * | 1986-09-11 | 1999-07-20 | Biosearch Italia S.P.A. | Method for selectively increasing the ratio of single major components of antibiotic A/16686 complex |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103360470A (en) * | 2013-07-26 | 2013-10-23 | 成都摩尔生物医药有限公司 | Purification method for ramoplanin fermentation liquor |
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| CN101024661A (en) | 2007-08-29 |
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