A high-performance liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS(n)) method has been developed to determine ginsenoside Rd in human plasma and to identify its metabolites in rat urine. The plasma and urine samples were pretreated by solid phase extraction (SPE) prior to analyses. In this work, gentiopicroside was used as the internal standard. The lower limit of quantification (LLOQ) for Rd in human plasma was 3 ng/ml. The average half-life time in plasma was detected as 19.29 h, when 10 mg of ginsenoside Rd was administrated intravenously to the volunteers. Seven metabolites including three oxygenated, two combined and two hydrolyzed components were identified in rat urine samples by using LC-MS and MS-MS, when ginsenoside Rd administered either orally or intravenously.