Fixed-Bed Column Process as a Strategy for Separation and Purification of Cephamycin C from Fermented Broth

Abstract

Fixed-bed column processes using the anionic resin Q Sepharose XL were evaluated for cephamycin C (CepC) purification from fermentation broth. Breakthrough and desorption curves were obtained for different flow rates (2.5, 5.0, and 7.5 mL/min). The elution method consisted of a stepwise gradient using NaCl solutions (0.1%, 0.3%, and 0.5%), which resulted in the separation of CepC from other antibiotics. The flow rate did not interfere in adsorption during loading of the column, but band broadening was observed during elution as the flow rate was increased. After the ion-exchange process, the fractions containing CepC were subjected to solid-phase extraction using a C18 cartridge to remove salts. Analyses of the broth used to feed the column and of the salt-free fractions by LC–MS showed a reduction in the concentration of some contaminants (possibly penicillin N, deacetylcephalosporin C, and deacetoxycephalosporin C) compared to the concentration of CepC. In conclusion, ion exchange followed by adsorption on a C18 adsorbent was demonstrated to be a selective and efficient procedure for the purification of CepC from fermentation broth.