Ultrasound-assisted emulsification microextraction followed by gas chromatography-flame ionization detector (USAEME-GC-FID) was used for determination of thymol in natural honey and mouthwash solution. The effects of various parameters affecting the extraction efficiency, including volume of extraction solvent, salt concentration, emulsification time, and temperature were investigated simultaneously by a full factorial design. The results showed that all factors were significant. In the next step, the optimal conditions were obtained by using a small-central composite design as 73.0mL for volume of extraction solvent, 1.50 (w/v%) for salt concentration, 45oC for emulsification temperature, and 9 min for emulsification time. The limit of detection (LOD) was 0.001 mg ml-1. The linear dynamic range (LDR) was 0.01-50mg ml-1 with determination coefficient (R2) of 0.9994. The relative standard deviation (RSD %) was 1.22 %. Comparing with other previously reported studies, the proposed method is an efficient, fast, simple, low cost, and good alternative for determination of thymol.