A sensitive and selective liquid chromatography-mass spectrometry (LC–MS) method for determination of dezocine in rabbit plasma was developed and validated. After addition of diazepam as internal standard (IS), liquid–liquid extraction (LLE) was used for sample preparation, and chromatography involved Agilent SB-C18 column (2.1 mmx50 mm, 3.5 um) using 0.1 % formic acid in water and acetonitrile as a mobile phase with gradient elution. Detection involved positive ion mode electrospray ionization (ESI), and selective ion monitoring (SIM) mode was used for quantification of target fragment ions m/z 245.8 for dezocine and m/z 284.8 for diazepam (internal standard, IS). The assay was linear over the range of 5–500 ng/mL for dezocine, with a lower limit of quantitation (LLOQ) of 5 ng/mL for dezocine. Intra- and inter-day precisions were less than 13 % and the accuracies were in the range of 93.1-105.2 % for dezocine. This developed method was successfully applied for the determination of dezocine in rabbit plasma for pharmacokinetic study.