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An isocratic high-performance liquid chromatographic method for determination of triptolide and triptonide in human plasma is described. Plasma samples were extracted with Oasis((R))HLB solid-phase extraction (SPE) cartridges. After pretreatment, they were separated on a SymmetryShield(TM)RP18 column with a mobile phase of acetonitrile-water (40:60,v/v) at 40degreesC. The effluent was monitored at UV 217 nm. Linearity (0.010-1.0 mg/L) was good, and the lower limit of detection was 3 ng/mL for triptolide and 4.5 ng/mL for triptonide (S/N = 3). The relative standard deviations of intra- and inter-day assay were less than were better than 80%. The developed method was applied to the determination of triptolide and triptonide concentration in a patient’s plasma after taking the medicament containing Tripterygium wilfordii Hook. F. Copyright (C) 2004 John Wiley Sons, Ltd.