Abstract: A simple, accurate and sensitive method has been developed for the simultaneous determination of warfarin, coumatetralyl, bromadiolone, flocoumafen and brodifacoum in whole blood by high performance liquid chromatography (HPLC) with fluorescence detection. The five 4-hydroxycoumarin rodenticides in whole blood were extracted by ethyl acetate, separated on XDB C18 column(150 mm×2.1 mm, 5 μm) by using the mobile phase consisting of methanol-0.2% acetic acid aqueous solution (88∶12, v/v) at a flow rate of 0.5 mL/min and detected with a variational time program for fluorescence wavelength. Each analyte was qualitatively determined with its fluorescence excitation spectrum, fluorescence emission spectrum and retention time being compared with those of the reference standard, and quantified with external calibration method. The linear range was 0.01-10.00 mg/L and the limit of quantification was 0.01 mg/L except warfarin of which the corresponding results were 0.05-10.00 mg/L and 0.05 mg/L. The recoveries were between 81% and 98% and the relative standard deviations (RSDs) were between 3.8% and 8.5%. This method can be used in the diagnosis of the clinical poisoned patients.

Key words: brodifacoum, bromadiolone, coumatetralyl, flocoumafen, fluorescence detection, warfarin, whole blood , high performance liquid chromatography (HPLC)