Separation of tautomeric forms of [2-nitrophloroglucinol-H]- by an in-electrospray ionization source hydrogen/deuterium exchange approach.

Here we report the observation that, depending on the solvent used for the electrospray, 2-nitrophloroglucinol undergoes a deprotona- tion from different sites forming two tautomeric gas phase ions. Those ions differ bythe collision-induced dissociation [CID] spectra and by the gas phase hydrogen/deuterium (H/D) exchange kinetic. We performed H/D exchange in the electrospray ionization (ESI) source by saturation ESI region with vapors of deuterated solvent (D20). It was observed that [2-nitrophloroglucinol-H]- exchanges two -OH hydrogens when MeOD is used as the spray solvent but when the spray solvent is 50:50 MeOD/DO20 we observed an additional two H/D exchanges at the aromatic ring. We propose that the reaction occurs via a keto-enolt tautomerization mechanism which was found to be energetically favorable.