Determination of nicarbazin and clopidol in poultry feeds by liquid chromatography

A rapid and very effective analytical procedure for the simultaneous liquid chromatographic determination of two coccidiostats, clopidol (CLOP) and nicarbazin (NICA), in poultry feeds was developed and tested. The ground feed samples were extracted using aqueous dimethylformamide after moistening with water. Co-extracted feed constituents were removed with a solid-phase extraction alumina-basic column and the eluates were directly analyzed on an ODS column (250×4.6 mm, 5 μm) with acetonitrile–0.01 M acetate buffer (pH 4.6) as eluent. UV detection of CLOP and the 4,4′-dinitrocarbanilide portion of NICA was carried out at 265 and 345 nm, respectively. The mean recovery from NICA spiked samples was 95% with a RSD of 4% in a concentration range of 2–150 mg/kg while for CLOP it was 98% with a RSD of 5% in a concentration range of 5–150 mg/kg. The limits of detection of NICA and CLOP in feed, based on a detector signal-to-noise ratio of 3, were estimated to be 1 mg/kg and 2.5 mg/kg, respectively, and the lowest levels tested in feeds by this procedure were 2 mg/kg and 5 mg/kg, respectively.