HPLC Determination of Brodifacoum in Urine with Fluorophotometric Detection

A method of HPLC determination of brodifacoum (BDFC) in urine with FPD was proposed. Urine sample was acidified with H3PO4 to pH 2.0 and treated by SPE on LC-18 micro-column. Brodifacoum was eluted from the column with eluant containing methanol and methyl tert. butyl ether mixed in the ratio of 1 to 9. The eluate obtained was blown to dryness with nitrogen gas,and the residue was dissolved with 1.0 mL of methanol,from which 10.0 μL was aliquoted and introduced into the instrument for HPLC determination. Chromatographic column of XDB-C18 (150 mm×2.1 mm,5 μm) was used as the stationary phase,and a mixed solution of methanol and dil. acetic acid solution (2+998),mixed in the ratio of 88 to 12,was used as the mobile phase. Fluorophotometric detection at the wavelengths of (λex) 270 nm and (λem) 376 nm was adopted. Linearity range of the method was found in the range of 0.05-10.0 mg·L-1 of BDFC. Determination limit (S/N=10) of the method was found to be 0.07 mg·L-1. Values of recovery and RSD (n=6) found were in the ranges of 79.8%-97.0% and 4.0%-4.9% respectively.