In monogyne (single queen) ant colonies, worker aggression against intruders, including newly-mated sister queens (gynes) seeking readoption, prevents the development of multi-queen colonies. Some ant species, however, produce multiple queen colonies (polygyny) via the adoption of new gynes in response to ecological, genetic and social parameters. Cuticular chemistry mediates nestmate recognition as well as plays a role in intra-colonial regulation of reproduction. Although aggression against intruders is diminished in some polygyne species, apparently due to depauperate allelic diversity at loci that code for recognition cues or a loss of receptor sites, the mechanism underlying the development of polygyny is largely unknown. Here, we examine the cuticular chemical profiles of new gynes at several stages post-emergence, males and workers from monogyne and polygyne colonies in the facultatively polygyne ant Formica truncorum. Individuals from the two colony types were chemically distinguishable, as were gynes according to developmental stage post-emergence (callow versus mature) and with respect to wing presence and mating status. Overall profile complexity did not differ between populations. These results suggest an association of cuticular hydrocarbon profiles with dispersal tendencies and that the disparate tendency of workers from monogyne and polygyne colonies to adopt new gynes is due to a complex interaction between gyne cuticular chemistry and differential worker sensitivity to those cues.