Chlorfluazuron (CFZ) is a benzoylphenylurea insecticide that is commonly used in baits for management of subterranean termites. In this study, a new method using GC-MS for the determination and quantification of CFZ in termites was developed and validated. Since a small volume of organic solvent (250 µL) was used in the sample preparation procedure, the extract was analyzed without any evaporation. The assay was simple and rapid, with a short GC run time (10.0 min). The calibration curve was linear over the range of 0.1-2.5 µg/g, and the correlation coefficient was >0.998. This method was sensitive, as demonstrated by the detection and quantification limits of 0.003 µg/g and 0.1 µg/g, respectively. The mean recovery of CFZ from spiked samples was 95.6%. The within-day and between-day precision and accuracy of the assays ranged between 1.19 and 6.43%. This method was used to screen for CFZ transfer between nestmates of a treated Macrotermes gilvus mound. The highest amount of CFZ was detected in workers from the royal chamber, followed by workers in the peripheral zone, workers in the nursery zone, larvae in the nursery zone, and larvae in the peripheral zone.