In this study, simple and efficient ultrasound-assisted emulsification microextraction (USAEME) combined with gas chromatography-flame ionization detector (GC-FID) was developed for the preconcentration and determination of carbamazepine in biological samples. In this method, the fine droplets of 1-octanol were formed and dispersed in the sample with the help of ultrasonic waves, which accelerated the formation of the fine cloudy solution without using disperser solvents. Several factors influencing the extraction efficiency such as the nature and volume of organic solvent, extraction temperature, ionic strength and centrifugation time were investigated and optimized. The new method (USAEME) provided detection limits of 0.6 µg L
-1 and 1.2 µg L
-1 in urine and plasma samples, respectively. The calibration graphs were linear in the range of 2.5-500 µg L
-1 and 5.0-500 µg L
-1 in urine and plasma, respectively. This proposed method was successfully applied to the analysis of carbamazepine in biological samples.