RATIONALE Polyethylene glycols (PEGs) are soluble molecules utilized in a wide range of applications. Mass spectrometry and fragmentation patterns of positively charged PEG oligomers are well‐known, but decomposition mechanisms of the deprotonated ions have not been studied. METHODS Deprotonated PEGs were generated by electrospray ionization of PEG in water/acetonitrile. Collision‐induced dissociation (CID) experiments were carried out in a tandem mass spectrometer. The anions were studied using a tandem mass spectrometer to carry out CID experiments. A series of small PEG oligomers, with 1 to 8 monomer units, were studied in order to monitor size‐dependent effects on fragmentation reactions. RESULTS Because deprotonated PEG ions have a unique charge site, their dissociation pathways can easily be monitored. The ions fragment by loss of C 2 H 4 O monomer units, with an alternating intensity pattern that suggests the loss of an even number of monomer units is favored. Smaller oligomers and oligomer fragments also yielded fragments corresponding to H 2 elimination and H 2 O loss. H 2 elimination occurs by the generation of a hydride ion which deprotonates an alcohol upon leaving, while dehydration appears to be a charge‐remote process. CONCLUSIONS The fragmentation of deprotonated PEG is dominated by intramolecular S N 2 reactions involving the terminal oxide anion. Copyright © 2013 John Wiley & Sons, Ltd.