In this work, the combination of dispersive liquid-liquid microextraction (DLLME) method with secondary electrospray ionization-ion mobility spectrometry (SESI-IMS) was used for determination of imipramine (as a test compound). Some effective parameters on the extraction efficiency were optimized, such as type and volume of extraction and disperser solvents, Centrifugation time and sample pH. Chloroform and ethanol were selected as the extraction and disperser solvents, respectively. The volume of the extraction and disperser solvent were 60 µL and 0.5 mL, Also the sample pH and centrifugation time were 9 and 1 min, respectively. Under the optimized conditions, the limit of detection (LOD) was calculated 9 µg.L -1 , the linearity range (R 2 =0.9976) was found to be 30-800 µg.L -1 . The relative standard deviations (RSDs) of intra- (in a day) and inter-day (in three successive day) were obtained 4% and 6%, respectively. According to the analytical parameters, the high sensitity,speed and repeatability of the purpose method, can be used for determination of the trace amount of imipramine drug in urine and plasma samples.