Efficient HPLC method for the determination of nicarbazin, as dinitrocarbanilide in broiler liver

A simple, fast and reliable HPLC-UV method has been developed for the determination of dinitrocarbanilide residues in broiler liver. Liver samples (2 g) were extracted with two portions of acetonitrile (10 and 5 ml), defatted with hexane and evaporated to dryness under nitrogen. Extracts were reconstituted in acetonitrile-water (70/30, v/v, 500 μl), loaded onto C18 solid phase (SPE) cartridges and eluted with acetonitrile-water (70/30, v/v, 2.5 ml) into clean test-tubes. Extracts were evaporated to dryness and reconstituted in acetonitrile-water (80/20, v/v, 500 μl). An aliquot of the extract was assayed by high performance liquid chromatography (HPLC) with UV detection at 350 nm. The method was validated according to EU guidelines using liver tissues fortified at levels of 100, 200 and 300 μg/kg, with dinitrocarbanilide. The decision limit (CCα) and the detection capability (CCβ) were calculated from the within laboratory repeatability data to be 228 and 266 μg/kg, respectively. The mean recovery was typically >70% and the limits of quantitation was 12.5 μg/kg (based on the lowest standard on the calibration curve).