Biological entities, such as DNA bases or proteins, possess numerous tautomers and isomers that lie close in energy, making the experimental characterization of a unique tautomer challenging. We apply VUV synchrotron-based experiments combined with state-of-the-art ab initio methodology to determine the adiabatic ionization energies (AIEs) of specific gas-phase cytosine tautomers produced in a molecular beam. The structures and energetics of neutral and cationic cytosine tautomers were determined using explicitly correlated methods. The experimental spectra correspond to well-resolved bands that are attributable to the specific contributions of five neutral tautomers of cytosine prior to ionization. Their AIEs are experimentally determined for the first time with an accuracy of 0.003 eV. This study also serves as an important showcase for other biological entities presenting a dense pattern of isomeric and tautomeric forms in their spectra that can be investigated to understand the charge redistribution in these species upon ionization.