Micro high-performance liquid chromatography for the determination of nicarbazin in chicken tissues, eggs, poultry feed and litter

A micro high-performance liquid chromatography (HPLC) method has been developed for the determination of the anti-coccidial drug nicarbazin in chicken tissues, eggs, poultry feeds and litter. The 4,4′-dinitrocarbanilide (DNC) component of nicarbazin was extracted from foods, feeds and litter with acetonitrile. The extracts were purified by liquid-liquid partitioning, evaporated to dryness and taken up in methanol-acetonitrile-water (50:30:20, v/v). Micro HPLC of the 4,4′-dinitrocarbanilide (DNC) portion of nicarbazin is performed isocratically using a small bore column (1 mm I.D.) packed with reversed-phase and a UV detector set at 340 nm. The average recoveries of nicarbazin added to muscle, liver and egg were 92.8, 84.3 and 85.2%, respectively, 95.9% in poultry feed and 76.8–95.9% in different litters. The limit of detection was 25 pg, based on a detector signal-to-noise ratio of 3. These results were achieved with a simplified step extraction without the solid-phase extraction used by various researchers. This method offers a sensitive, selective, rapid, and less expensive alternative to conventional HPLC for such evaluations.