Abstract: SPE-LC was applied to the determination of residual amounts of osthol in tobaccos. The tobacco sample was extracted with 50% (φ) methanol solution, followed by purification with liquid-liquid extraction and solid phase extraction, and quantification with RRLC-DAD and identification with Q-TOF. Linear relationship between values of peak area and mass concentration of osthol was kept in the range of 0.25-50.0 mg·L-1, with detection limit (3S/N) of 0.016 mg·kg-1. On the base of blank sample, test for recovery was made by standard addition method; values of recovery found were in the range of 91.5%-95.5%, with intraday RSD′s (n=5) in the range of 3.1%-4.3%.