A systematic study on pharmacokinetics and tissue distribution of danshensu (one of the major active components from Salvia miltiorrhizae) is conducted using a rapid and sensitive high-performance liquid chromatographic (HPLC) method. Before HPLC analysis, biological samples are pretreated with a liquid-liquid extraction. Separation of danshensu and internal standard is achieved on an Agilent Zorbax C18 column with a mobile phase made up of acetonitrile and 0.05% trifluoracetic acid at a flow rate of 0.8 mL/min. The calibration curves in plasma and tissues are linear in the given concentration ranges, with r2 no less than 0.99. The intra-day and inter-day relative standard deviations in the measurement of quality control samples are less than 15%, and the accuracies are in the range of 86-115%. The recoveries of danshensu in plasma and tissues are among 80% to 118%. Meanwhile, the multi-peaks in pharmacokinetic profiles are observed. The method is successfully applied to the pharmacokinetics and tissue distribution study of danshensu after a single oral administration of 50.0 mg/kg sodium danshensu to rats.