This paper describes a simple technique for extraction and a sensitive high-performance liquid chromatographic method for separation and quantitation of ondansetron in human plasma. The procedure involved liquid-liquid extraction of ondansetron from plasma, reversed-phase HPLC separation and ultraviolet detection at 305 nm. The internal standard method was applied for quantitation. The recovery of ondansetron was >85%. Linearity was good throughout the concentration range anticipated in human plasma from investigations in panic disorder (0.5-15 ng/ml, r2 ranging from 0.9953 to 0.9988). This method was applied to the determination of plasma concentrations of ondansetron in humans.