Determination of Aristolochic Acid in Rabbit Plasma by High-Performance Liquid Chromatography with Photodiode-Array Ultraviolet Detection and Its Pharmacokinetics Application

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Abstract
A simple and sensitive high-performance liquid chromatographic method for the determination of aristolochic acid (AA) in rabbit plasma has been developed. Up to 0.1 ml of plasma containing AA was deproteinized by acetonitrile, which contained an internal standard (indomethacin). The supernatant was injected onto a COSMOSIL 5C18-AR column (5 μm) using acetonitrile-0.1 % phosphoric acid (60:40, v/v, pH 2.5–2.8) as the mobile phase. UV detection at 227 nm was followed by ultraviolet spectrum identification (among 200 and 380 nm) with a photodiode-array detector. The method was rapid, easily reproduced, selective and sensitive. It was applied to pharmacokinetic studies of AA in rabbit, after a 5 mg/kg intravenous administration. A biphasic phenomenon with a rapid distribution followed by a slower elimination phase was observed from the plasma concentration-time curve. Compartmental analyses yielded a two-compartment model.

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