The Colorado potato beetle, Leptinotarsa decemlineata, reared under diapause-inducing conditions will emerge from the soil as an adult and enter the diapause initiation phase, a period where metabolic reserves are stockpiled before the beetles enter the nonfeeding diapause maintenance phase. Internal and external lipids were characterized during the diapause initiation phase (IP) and compared to the lipid profiles of nondiapausing adults. The primary internal lipids of both diapause IP and nondiapausing adults are triacylglycerols. Only trace amounts of internal lipids were detected in day 1 diapause IP adults. A dramatic increase in internal lipids was observed between day 7 and day 15 post-emergence in the diapause IP adults. The majority of the triacylglycerol isomers were identified as C50, C52 and C54 chain lengths by GC-MS. There were no observed differences in the isomeric distribution of the major internal lipids between diapause IP and nondiapausing adults. External lipids were mainly methyl-branched alkanes containing a 25 to 53 carbon backbone. The quantity of external lipids increased from day 1 to day 7 post-emergence in both the diapause IP and nondiapausing adults, with the bulk of the increase occurring in the longer chain-length methylalkanes.