Determination and pharmacokinetics of 6,7-dimethoxycoumarin in rat plasma after intragastric administration of different decoctions of Yinchenhao Tang

Abstract

A simple and sensitive reversed-phase high-performance liquid chromatography method with UV detection is developed and validated for the determination of 6,7-dimethoxycoumarin in rat plasma and comparative analysis of its pharmacokinetics after intragastric administration of 6,7-dimethoxycoumarin and three different decoctions of Yinchenhao Tang. The plasma samples are deproteinated with acetonitrile. The components are separated on a Kromasil C18 column (250 x 4.6 mm, 5 μm,) with methanol-1% acetic acid solution-tetrahydrofuran (30:63:7, v/v/v) as the mobile phase, and the UV detector is set at 340 nm. Coumarin is used as an internal standard. The linear calibration curve is obtained in the concentration range of 25-2500 ng/mL. The lower limit of quantitation of the method is 25 ng/mL. The intra- and inter-day precision are less than 12%, and the accuracy determined with relative error ranges from -2.9% to 1.7%. The data obtained from rat plasma are analyzed with Topfit 2.0 Pharmacokinetic Software. With pharmacokinetic analysis, the main parameters after intragastric administration of 6,7-dimethoxycoumarin, Herba Artemisiae Scopariae decoction, Artemisiae Scopariae decoction plus Radix et Rhizoma Rhei and Fructus Gardeniae decoction, Yinchenhao Tang are as follows: T1/2 is 0.29, 1.30, 1.07, and 1.75 h, AUC→t is 919.1, 1215.0, 2035.3, and 2537.9 ng·h/mL, AUC0→ is 928.5, 1325.9, 2094.4, and 2612.6 ng·h/mL, respectively.