A sensitive, simple and highly reliable high-performance liquid chromatographic method using fluorescence detection is reported for the determination of pindolol in plasma. This method involves a single extraction of pindolol from alkalinized plasma into methyl tert.-butyl ether followed by a back-extraction into dilute hydrochloric acid. Injection of the dilute acid phase directly onto an octyl (LC-8) bonded-phase column provides the final separation, and detection of pindolol is achieved by monitoring the intrinsic fluorescence of pindolol at 315 nm following excitation at 255 nm. The method is sensitive enough to measure with confidence pindolol plasma concentrations of 2 ng/ml using a 2-ml sample. No internal standard is required. This method has been applied to the analysis of 1500 human plasma samples by two different laboratories.