In this research, a suitable molecularly imprinted polymer (MIP) as a solid phase extraction (SPE) sorbent has been synthesized for the selective separation and cleaning up of salicylic acid from the biological samples,plasma and urine. in negative mode electrospray ionization ion mobility spectrometry (ESI-IMS) was used for the first time as a detection system. In this study salicylic acid, an acidic drug which can form intramolecular hydrogen-bond between its two functional groups -COOH and –OH. Although the process of recognition of MIP is similar to that of macrobiomoleculessuch as enzymes and antibodies, MIP have considerable advantages over biomoleculesas recognition systems, because of their chemical and physical stability Ion mobility spectrometry with low cost, inexpensive maintenance and very fast analysis makes an attractive technique for determination of salicylic acid. The imprinted polymer was prepared by non-covalent method using salicylic acid as the template 4-vinyle pyridin (4-vpy) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker agent, and 2,2'-azobis (2-isobutyronitrile) (AIBN) as initiator. In this work, Several factors affecting the performance of MIP such as water sample pH, washing volume raito, and its speed were studied. Sample pH was adjusted at pH=5. MIP, sorbent was conditioned first with methanol and then with purified water (2.0 mL each solvent) before being used for the first time. The water sample (2.0 mL in each time) was passed through the colmun with speed of 0.04 mL/min. The solid phase was then washed with 2.0 mL mixer water and methanol (95:5) to remove interference The analytes were then eluted with 20 mL methanol at speed of 0.04 mL/min. then 200.0 ?L of elution solvent was dried by a gentle stream of nitrogen gas.. The eluent was injected to ESI-IMS injector. In addition, these experiments were compared with the non-imprinted polymer (NIP) and the results indicated that NIP had no specific affinity to adsorb salicylic acid The retention capacity of the prepared MIP sorbent was 2.6 mg/g. Under optimum conditions, the proposed method with respect to other reported methods shows high sensitivity and selectivity, with detection limit of 0.008µg/mL. The linear dynamic range of about 2 orders of magnitude (from 0.02 to 2.00 µg/mL ) was obtained for this compound. and the relative standard deviations were under 6% (good precision). Also, the proposed method was used to analyze various real samples so, Once, the drug spiked human plasma and urine, Once again, Someone volunteers to take medication and his urine and plasma were analyzed and the obtained results confirmed the capability of ESI-IMS for detection of salicylic acid. Recoveries were obtained as higher as 84 to 99%.