Abstract A simple, rapid and sensitive LC–MS/MS method for determination of trace amount of new antifungal drug iodiconazole in human plasma was developed (1-(1H-1,2,4-triazole)-2-(2,4-diflurophenyl)-3-[ N-methyl- N-(3-chlor-benzyl)amino]-2-propanol), was used as internal standard (IS). The analytes were extracted by liquid–liquid extraction with n-hexane after internal standard spiked. The separation was performed by a ZORBAX SB-C 18 column (3.5 μm, 2.1 mm × 100 mm) with an isocratic mobile phase consisting of methanol–water–formic acid (50:50:0.05, v/v/v) at a flow rate of 0.3 mL/min. The lower limit of quantification (LLOQ) was 0.10 ng/mL. This method was successfully used to determine the concentration of iodiconazole in human plasma following dermal administration.