Abstract: Objective To establish a reversed phase high-performance liquid chromatography (HPLC) system for determining plasma concentration of olanzapine and analyze the pharmacokinetics of olanzapine in healthy Chinese volunteers. Methods Ten healthy male subjects received a single oral dose of 20 mg olanzapine tablets. The plasma concentrations of olanzapine were determined by HPLC, and the data were analyzed using 3P97 program. Results The plasma concentration curve of olanzapine following a single oral dose conformed to the two compartment open model. The main pharmacokinetics parameters of olanzapine were as follow: Cmax was 113.7±33.1 μg/L, Tmax 5.07±0.65 h, t1/2 35.44±4.21 h, AUC0-144 2 235±257 μg·h·L-1, and AUC0-inf 2 516±301 μg·h·L-1. Conclusion The system established in this study allows for highly sensitive, selective and accurate determination of plasma concentration of olanzapine, and provides valuable information for clinical trials.