AbstractA simple and sensitive HPLC method to detect tanshinone I, tanshinone IIA, and cryptotanshinone in human urine was developed and validated. Chromatographic separation was achieved on a C18 column (4.6 mm × 250 mm, 5 µm) using a mixture of methanol/water (78:22 v/v, containing 0.5% acetic acid) as the mobile phase at a flow rate of 0.5 mL/min with UV detector at 254 nm. The calibration curve of tanshinones was linear over a range of 1.0 × 10−4 – 1.0 mg/mL. The limit of quantification (LOQ) was found to be 57.0, 76.0, and 45.0 ng/mL, respectively. This method can estimate the pharmacokinetic parameters of tanshinones in human urine after the intravenous and intraperitoneal administration of stanchions.