In this study, the gas phase chemistry of the protonated benzyl esters of proline has been investigated by electrospray ionization mass spectrometry and theoretical calculation. Upon collisional activation, the protonated molecules undergo fragmentation reactions via three primary channels: (1) direct decomposition to the benzyl cation (m/z 91), (2) formation of an ion-neutral complex of [benzyl cation + proline]+, followed by a hydride transfer to generate the protonated 4,5-dihydro-3H-pyrrole-2-carboxylic acid (m/z 114), and (3) electrophilic attack at the amino by the transferring benzyl cation, and the subsequent migration of the activated amino proton leading to the simultaneous loss of (H2O + CO). Interestingly, no hydrogen/deuterium exchange for the fragment ion m/z 114 occurs in the d-labeling experiments, indicating that the transferring hydride in path-b comes from the methenyl hydrogen rather than the amino hydrogen. For para-substituted benzyl esters, the presence of electron-donating substituents significantly promotes the direct decomposition (path-a), whereas the presence of electron-withdrawing ones distinctively inhibits that channel. For the competing channels of path-b and path-c, the presence of electron-donating substituents favors path-b rather than path-c, whereas the presence of electron-withdrawing ones favors path-c rather than path-b. Copyright © 2013 John Wiley & Sons, Ltd.