The cuticular hydrocarbons of laboratory-bred and wild-caught females of Phlebotomus martini Parrot and P. celiae Minter, originating from southern Ethiopia, were used to distinguish females of the two sympatric and isomorphic species. Field-caught sandflies were dissected for parasites prior to the analysis. A high degree of separation, was achieved between the two species in both laboratory-bred (91.6%) and wild-caught (91.3%) specimens. However, the discriminating hydrocarbon peaks in laboratory reared specimens were different from those of wild-caught specimens, resulting in a high number of misclassiflcations in the pooled analysis of wild and laboratory specimens in each species. High degrees of hydrocarbon differentiation (84.7%–96.7%) between laboratory-bred and wild caught flies within each species were also obtained.