Method for separation and purification of daptomycin RS-8 impurity
A daptomycin and RS-8 technology, applied in the direction of peptides, etc., can solve the problems of daptomycin resistance, changing the properties of cell plasma membrane, and not specifically involving the separation method of daptomycin impurities
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Embodiment 1
[0025] After the daptomycin fermentation broth is filtered, the filtrate is chromatographed on FPDA13 resin, and the column is washed with 2BV of 0.03N sodium chloride, 0.06N sodium acetate, and acetic acid to adjust the pH to 6.5-7.0. 0.3N sodium chloride, 0.06N acetic acid Sodium, acetic acid adjusted pH6.5-7.0 solution elution, collect the daptomycin RS-8 impurity with chromatographic purity greater than 40%, through preparative chromatography (mobile phase is acetonitrile:0.01% trifluoroacetic acid=25:75, volume ratio) was separated and purified, and was tracked and detected by high performance liquid phase (the detection conditions were the same as the method disclosed in the European patent "PROCESSFOR THE PURIFICATION OF DAPTOMYCIN" (EP 1 586 580 A2)), and the daptomycin RS- 8 impurity preparation solution.
[0026] The preparation solution was vacuum freeze-dried, and the obtained solid daptomycin RS-8 impurity was detected by high performance liquid chromatography (th...
Embodiment 2
[0028] After filtering the daptomycin fermentation broth, apply FPDA13 resin chromatography on the filtrate, wash the column with 0.03N sodium chloride, 0.06N sodium acetate, and acetic acid to adjust the pH6.5-7.0 solution 2BV, 0.3N sodium chloride, 0.06N sodium acetate , acetic acid to regulate pH6.5-7.0 solution elution, collect the daptomycin RS-8 impurity with chromatographic purity greater than 40%, through preparative chromatography (mobile phase is acetonitrile:0.01% trifluoroacetic acid=30:70, volume ratio ) was separated and purified, and tracked and detected by high performance liquid phase (detection conditions were the same as the method disclosed in the European patent "PROCESS FORTHE PURIFICATION OF DAPTOMYCIN" (EP 1 586 580 A2)), and collected daptomycin RS-8 with a chromatographic purity>90% Impurity preparation solution.
[0029] The preparation solution was vacuum freeze-dried, and the obtained solid daptomycin RS-8 impurity was detected by high performance ...
Embodiment 3
[0031] After filtering the daptomycin fermentation broth, apply FPDA13 resin chromatography on the filtrate, wash the column with 0.03N sodium chloride, 0.06N sodium acetate, and acetic acid to adjust the pH6.5-7.0 solution 2BV, 0.3N sodium chloride, 0.06N sodium acetate , acetic acid to regulate pH6.5-7.0 solution elution, collect the daptomycin RS-8 impurity with chromatographic purity greater than 40%, through preparative chromatography (mobile phase is acetonitrile:0.01% trifluoroacetic acid=33:67, volume ratio ) was separated and purified, and tracked and detected by high performance liquid phase (detection conditions were the same as the method disclosed in the European patent "PROCESS FORTHE PURIFICATION OF DAPTOMYCIN" (EP 1 586 580 A2)), and collected daptomycin RS-8 with a chromatographic purity>90% Impurity preparation solution.
[0032] The preparation solution was vacuum freeze-dried, and the obtained solid daptomycin RS-8 impurity was detected by high performance ...
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