The noncovalent interactions in host-guest complexes of alpha-cyclodextrin (alpha-CD) with a series of benzoic acid derivatives (RBA) were investigated by electrospray ionization tandem mass spectrometry and density functional theory (DFT) calculations. The 1:1 stoichiometry of the anionic host-guest complexes was unequivocally confirmed by their mass-to-charge ratios (m/z) and isotope patterns. Collision-induced dissociation experiments revealed exclusive fragmentation into [a-CD] and neutral RBA and afforded the gas-phase kinetic stability trend [alpha-CD center dot 3,5-diMeBA](-) < [alpha-CD center dot 3-MeBA](-) < [alpha-CD center dot BA](-) < [alpha-CD center dot 3-O-HBA](-) < [alpha-CD center dot 3,5-diOHBA](-). This trend follows that of the gas-phase basicities of the guest anions used, indicating that host-guest pairs with more comparable basicities form more stable complexes. DFT calculations at the M06-L/6-31+G(d,p) level of theory provided detailed structural assignments and further elucidated the experimental observations, suggesting that the anionic [alpha-CD center dot RBA](-) inclusion complexes are favored over the nonspecific complexes in the gas phase and that hydrogen bonding constitutes the primary host-guest interaction. Additionally, the results provide an estimated gas-phase basicity Delta G(0) = 325-327 kcal mol(-1) for [alpha-CD](-).