Fluorescence and electronic action spectroscopy of mass-selected gas-phase fluorescein, 2',7'-dichlorofluorescein, and 2',7'-difluorofluorescein ions.

2',7'-Dichloro- and 2',7'-difluorofluoresceins are superior alternatives to underivatized fluorescein. Although several studies characterizing their condensed-phase photophysical properties have been reported, little is known about their intrinsic characteristics. Here, the gas-phase properties of three charge states of each fluorescein are characterized using a quadrupole ion trap mass spectrometer which has been modified for spectroscopy. Electronic action spectra, constructed by monitoring the extent of photodissociation as a function of excitation wavelength, indicate that the gaseous dianions and cations resemble their solution-phase counterparts. In contrast, a large shift in the electronic action spectra of the monoanions indicates the presence of a different tautomer in the gas phase than that present in solution. The gaseous monoanion is deprotonated on the xanthene ring, rather than being deprotonated on the pendant group as found in soluion. The dianions and cations do not emit detectable fluorescence in the gas phase. In contrast, the monoanions do fluoresce, but the emission intensity is low and the spectra are broad. This work illustrates the effect of halogenation on the intrinsic properties of the dyes and provides useful fundamental understanding that promises to aid the development more robust fluorescent dyes.