ASSAY OF BROPIRIMINE IN RAT PLASMA BY MEANS OF ROBOTIC SOLID-PHASE EXTRACTION AND HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A rapid and simple high-performance liquid chromatography (HPLC) metho d incorporating automated solid-phase extraction (SPE) is described fo r the determination of bropirimine, 2-amino-5-bromo-6-phenyl-4(3H)-pyr imidinone, in plasma samples from various species. Using an automated sample processor, plasma samples were loaded onto C-18 SPE columns and the drug eluted with ethanol-methylene chloride (10:90, v/v). The ext racts were analyzed by reversed-phase HPLC using a C, column with a mo bile phase consisting of acetonitrile-water-trifluoroacetic acid (20:8 0:0.1, v/v/v). The UV absorbance of the column effluent was monitored at a wavelength of 292 nm. Linear calibration curves were obtained in the range of 0.01 to 22 mu g/ml. Precision (less than or equal to 4% r elative standard deviation) and accuracy (less than or equal to 5% err or) were acceptable at the concentrations evaluated. Application of th is method to the quantitation of bropirimine in rat plasma for a toxic okinetic/bioavailability study is reported.