Purification of pneumocandin B1 and B5 impurities based on preparative high performance liquid chromatography
Objective This research was carried out on the enrichment and purification of components with low content.The purification of impurities B1 and B5 from Glarea lozoyensis fermentation broth(i.e.,the crude product of pneumocandin B0)was prepared,following their structures identification.Method A two-step purification method based on preparative high performance liquid chromatography(prep-HPLC)was established.The HILIC mode stationary phase was used in the first step in order to enrich the targeted impurities,and the RPLC mode stationary phase was used in the second step in order to purify the impurities.The two impurities were identified by mass spectrometry(MS)and nuclear magnetic resonance(NMR).Results The two impurities were B1 and B5,with a relative molecular mass of 1049 Da and chromatographic purity of 97.83％and 98.13％,respectively.By NMR identification,B1 is a homo tyrosine analogue of B0 and B5 is an ornithine analogue of B0.Conclusion The two-step purification method based on prep-HPLC developed in this study provides a reference for the isolation of components of low content.The first-step separation was to enrich the target components.The second-step separation that used an orthogonal column to improve the separation selectivity.As a result,impurities B1 and B5 were successfully isolated.It is helpful in establishing the impurity profile of B0 and furthering its quality control.
PneumocandinImpurityPreparative high performance liquid chromatographyPurificationStructure identification
万方数据
维普
