Electrospray 'soft' ionisation (ESI) and multistep mass spectrometry (MS(n)) techniques enable characterisation of a bioactive polymer, poly[(R,S)-3-hydroxybutanoic acid] (a-PHB), containing covalently bonded benzylpenicillin. The chemical structures of individual mass-selected bioactive macromolecules have been determined, and their fragmentation mechanisms have been compared with those of pure penicillin G. Copyright 2000 John Wiley & Sons, Ltd.