In this thesis, differentsample preparation methods such as packed-fiber solid-phase extraction (PFSPE), molecular imprinted solid phase extraction (MISPE) and Hollow fiber-based liquid–liquid–liquid microextraction(HF-LLLME) followed by corona discharge ion mobility spectrometry (CD-IMS)were used for the determination of biological and pharmaceutical compounds. In the first part, Analysis of testosterone in urine sample was accomplished using CD-IMS.Molecular imprinted polymer was used for the extraction and pre-concentration oftestosterone. Under the optimized conditions, the limit ofdetection was calculated to be 0.9 ng/mL. The results from obtained the method were also compared with the standardmethod for analysis of testosterone using SPE-HPLC technique. The results demonstratethe accuracy of the method. In the second work,HF-LLLME combined with CD-IMS was developed for the analysis of amantadine in human urine and plasma samples. All variables affecting the extraction of analyte were optimized. The limits of detection were calculated to be 7.2 and 1.6 ng/mL for plasma and urine samples, respectively. In the third part, the capability of CD-IMS for separation and quantification of sarcosine and L -alanine isomers has been evaluated for the first time. Variables including carrier gas flow rate, injection and cell temperatures were optimized. At the optimized conditions, the detection limit of sarcosine and L -alanine were 0.7 and 0.9 µg/mL, respectively. Furthermore, in this work, since ion peak of sarcosine overlaps with methanol peak, a solvent-free sample introduction system was developed. In the next part, HF-LLLMEfollowed by corona discharge ion mobility spectrometry was presented for analysis of dextromethorphan and pseudoephedrine in urine and plasma samples. The effects of various parameters on HF-LLLME were optimized. Under the optimized conditions, the limits of detection are 0.6 and 0.3 ng/mL for dextromethorphan,and 8.6 and 4.2 ng/mL for pseudoephedrine in plasma and urine samples, respectively.In the next section, electrospunnanofibers from poly(methylmethacrylate) (PMMA) and polystyrene (PS) blend were used as a sorbent to extract tramadol from urine and plasma samples and then determined by CD-IMS. The scanning electron microscopy (SEM) images of the PMMA/PS nanofibers showed a diameter range of 130-600 nm with a smooth morphology. Parameters affecting extraction efficiency were optimized and under the optimized conditions, the detection limits of tramadol were found to be 9.4 and 1.6 ng/mL in plasma and urine samples, respectively. In the last part, a second-order calibration method, Tucker 3, is proposed on CD-IMS data. The results of Tucker 3 were also compared with the PLS, N-PLS and U-PLS models, indicating superiority of the Tucker 3 model over the others.