Determination of brodifacoum in rat plasma by HPLC

OBJECTIVE: A determination method of brodifacoum in rat plasma with bromadiolone as an internal standard was developed. METHODS: A volume of 10 microl internal standard (bromadiolone) was added into rat plasma, and then extracted by 0.5 ml of acetonitrile by shaking for 2 min. The residue was dissolved with 200 microl of mobile phase after centrifugation for 10 min, and evaporation to dryness by Nitrogen blowing. A C18 column and PDA detector were used for separating and detecting. The wavelength was 254 nm, the flow rate was 1.0 ml/min, and the injection volume was 20 microl. RESULTS: The liner range was 1.0-20 microg/ml, and the correlation coefficient was 0.9992. The detection limit was 0.3 microg/ml in plasma (S/N=3). The intra-assay and inter-assay coefficients of variation were 1.89%-2.45% and 2.51%-3.61% respectively. The recoveries in plasma at levels of low, middle and high concentrations were (80.8 +/- 3.1)%, (81.8 +/- 2.7)% and (87.9 +/- 3.6)% (n=6), respectively. The accuracies were 84.1%-91.5% and 86.7%-93.2%, respectively. CONCLUSION: This method is simple, fast and accurate for the determination of brodifacoum in rat plasma.