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A rapid and sensitive bioassay based on liquid chromatography tandem mass spectrometry (LC–MS/MS) has been developed and validated for the simultaneous determination of eight coumarins in rat plasma. The liquid–liquid extraction method with ethyl acetate was used to prepare the plasma samples after addition of warfarin as an internal standard (IS). Chromatographic separation was performed on an Eclipse plus C18 column (100mm×4.6mm, 1.8μm) using gradient elution when 1mM ammonium acetate aqueous solution – acetonitrile was used as the mobile phase. The lower limit of quantitation (LLOQ) of each coumarin was lower than 2.16ngmL −1 .Intra-day and inter-day precisions were less than 15%. The accuracies were in the range of 88.9–117%. The mean recoveries of coumarins and IS were higher than 84%. The method was successfully applied to a pharmacokinetic study of eight coumarins in rats after oral administration of radix angelicae pubescentis.
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