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A study was made for purification of cephamycin C from fermentation of Streptomyces clavuligerus. Initially, the culture broth was clarified by microfiltration and ultrafiltration, after which the resulting permeates were subjected to nonspecific adsorption and ion-exchange chromatography on resin columns. The antibiotic activity was measured by the biological method at each stage by assaying its activity against the Escherichia coli ESS, super sensitive to β-lactam antibiotic. The purification processes were assessed in relation to the variables affecting each step. The purification efficiency by nonspecific adsorption was monitored by UV spectrophotometry, while the ion-exchange adsorption fractions were assessed by NMR spectroscopy. Some of the fractions obtained during purification were also analyzed by mass spectrometry (LC/MS and LC/MS/MS) to identify the cephamycin C molecule. These preliminary results proved the process feasibility.

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The original version of this article unfortunately contained a mistake. The name of the sixth author is incorrect. The correct name is "Alberto Colli Badino". In the article footnote, "A.C. B. Junior" should read as "A.C. Badino".

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.