A fast, sensitive, high-performance liquid chromatography-tandem mass spectrometry method was developed for the determination of voriconazole in human plasma. Carbamazepine was used as the internal standard and the sample pretreatment involved one-step protein precipitation. Chromatographic separation was conducted on an Ultimate C18 column with a mobile phase consisting of acetonitrile-water (containing 0.1% formic acid; 40:60, v/v) at a flow rate of 0.3 mL/min. The detection of voriconazole was performed on a triple-quadrupole tandem mass spectrometer by multiple reaction monitoring with an electrospray ionization source in the positive mode. The standard curve was linear (r(2) ≥ 0.99) within the concentration range of 2.49-293 ng/mL. The intra-day and inter-day precision values were below 5.3%, and the accuracy was within -4.3-5.7%. The method was applicable to the clinical study of the pharmacokinetics of voriconazole in healthy volunteers following oral administration.