In the study, a fast and selective method based on magnetic separation has been developed for the extraction of nicotine from human plasma using magnetic strong cation exchange (MSCX) resins as adsorbent. MSCX resins were prepared using hydrophobic Fe(3)O(4) magnetite as magnetically susceptible component, styrene and acrylic acid as polymeric matrix components, and acetyl sulfonate as the sulfonation agent. The extraction procedure was carried out in a single step by stirring the mixture of diluted plasma sample and MSCX resins in the vortex for 5 min. Then, the resins with adsorbed nicotine were separated from the sample matrix by applying an appropriate magnetic field. Main factors affecting the extraction of nicotine such as the amount of MSCX resins, pH value of the extraction solvent, extraction time, and washing and eluting conditions were optimized. The nicotine eluted from the resins was determined by liquid chromatography-tandem mass spectrometry. The calibration curve obtained by analyzing matrix-matched standards shows excellent linear relationship (r (2) = 0.9998) in the concentration range of 10-2,500 ng mL(-1). The limit of detection and quantification obtained are 2.9 and 9.7 ng mL(-1), respectively. The relative standard deviations of intra- and inter-day obtained are in the range of 1.9-6.9% and 2.5-7.8% with the recoveries ranging from 78.7% to 99.1%. The proposed method was successfully applied to determine nicotine in human plasma phlebotomized from ten male smokers. Nicotine was detectable with the contents ranging from 44.4 to 221.9 ng mL(-1) in five samples.