A general mass spectrometric method is described for the absolute assignment of α- or β-acyl naphthalenes, via which the gaseous α- and β-naphthoyl cations of m/z 155 are used as structurally diagnostic fragment ions (SDFI). These stable acylium ions are common and normally abundant fragment ions of acylnaphthalenes in general. Using a pentaquadrupole mass spectrometer, CID experiments with argon and ion/molecule reactions with 2-methyl-1,3-dioxolane, isoprene, acetonitrile and propionitrile were performed but failed to distinguish the two SDFI. Reactions with ethyl vinyl ether and several homologues as well as ethyl vinyl thioether were, however, successful. In reactions with ethyl vinyl ether, the α-SDFI form a pair of diagnostic product ions of m/z 165 and m/z 181, which are absent in the corresponding spectrum of the β-SDFI. Methyl 4-(1-naphthyl)-2,4-dioxobutanoate was used as a test molecule for this class of constitutional isomers and absolute structural assignment as an α-acyl naphthalene was correctly performed via the characterization of its α-SDFI.