A sensitive, specific and accurate method for determination of arbidol in human plasma was developed. Arbidol and internal standard were extracted from plasma samples by liquid-liquid extraction with diethyl ether. The chromatographic separation was accomplished on a Shiseido C18 3 microm analytical column (100 mm x 2.0 mm i.d.) at a flow rate of 0.3 mL/min isocratically. Detection was performed on a single quadrupole mass spectrometer by selected ion monitoring (SIM) mode via electrospray ionization (ESI) source. The method had a chromatographic run time of 6 min and a good linear relationship over the range 1-1000 ng/mL. The limit of quantitation for arbidol in plasma was 1 ng/mL. The intra-day and inter-day precision (R.S.D.%) was lower than 7% and accuracy ranged from 95 to 105%. The proposed method enables unambiguous identification and quantification of arbidol in vivo and has been successfully applied to study the pharmacokinetics of arbidol in healthy male Chinese volunteers.