A sensitive and reproducible liquid chromatographic method with UV detection was described for the determination of helicidum in beagle dog plasma. Sample preparation was accomplished through protein precipitation with 15% perchloric acid and chromatographic separation was performed on a reversed phase C18 column at 30 °C. Mobile phase consisted of a mixture of acetonitrile–water at a flow rate of 1.0 mL min−1. Wavelength was set at 270 nm. The method was applied to a pharmacokinetic study of two formulations of helicidum. No statistical difference in the t 1/2 (AUC0–5, AUC0–∞) between the two formulations were observed. The t max of helicidum ODTs is significantly shorter than that of conventional tablets (0.84 h vs. 1.33 h, P< 0.05).