High-performance capillary electrophoresis of a fermentation-derived cyclic peptide analog, animal growth promoter.
We have developed HPCE methods for the analysis of sulfomycin (trivial name) and related compounds (code name, crude material = U82127 = I), which is an animal growth promoter derived from a fermentation beer. The crude material, I, isolated from the fermentation consisted of more than 40 components which were not completely separated by conventional HPLC. Thus, as a complementary analysis method, we have optimized HPCE conditions for I using various capillaries including uncoated, coated, and packed using various buffers. The optimized HPCE conditions were obtained with an uncoated fused-silica capillary and a buffer that consisted of 30 mM Tris-tricine, 10 mM SDS, 10 mM NaCl and 20% MeOH, pH 8.0. Using these HPCE conditions, we were able to separate the one main component collected from the HPLC effluent into two or three components. In order to identify the main components of the fermentation product, an off-line HPLC-HPCE-MS analysis for I was performed. From the MALDI-TOF-MS results, the separated components collected from HPCE had different molecular masses. Four lots of I samples having different characteristics were also analyzed by HPCE to investigate lot-to-lot differences in peak profiles. The four lots of I were found to have very similar peak profiles. In this paper, I refers to the crude fermentation product and sulfomycin to the purified, major HPLC component of I.
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Northeastern University, Department of Chemistry, Boston, MA 02115, USA.
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D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D001515:Beer; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D005285:Fermentation; D006133:Growth Substances / Q000032:analysis*; D010455:Peptides; D010456:Peptides, Cyclic / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000900:Anti-Bacterial Agents; D002021:Buffers; D006133:Growth Substances; D010455:Peptides; D010456:Peptides, Cyclic; C049270:sulfomycin; C102859:Alexomycin