Adult epicuticular hydrocarbon variation of 14 geographically isolated populations of cactophilic Drosophila mojavensis was assessed to further investigate mechanisms of sexual isolation. Hydrocarbon transfer experiments demonstrated that these compounds are part of the mate recognition system in this species. Sixteen of the 23 epicuticular hydrocarbon components studied differed in amounts between males and females, and 13 differed in quantity between the geographic regions encompassing Baja California and mainland Mexico (Sonora and Sinaloa). Eight hydrocarbon components, seven of which differed in quantity between sexes, showed significant sex-by-region interactions, indicating region-specific sex reversals in hydrocarbon quantities. Such regional variation in epicuticular hydrocarbon profiles suggests that these hydrocarbon differences have also evolved in D. mojavensis since this species invaded mainland Sonora and Sinaloa from Baja California by switching host plants, in addition to a number of key genetic, behavioral, and life-history characters.