Determination of amprolium in chicken meat and egg by high performance liquid chromatography

High performance liquid chromatography (HPLC) with a fluorescence detector was examined for the determination of amprolium (APL) in chicken meat and egg. The methanol extract of samples was evaporated to dryness and hydrochloric acid was added. The suspension from chicken meat was filtered and that from egg was heated on a water bath and then filtered. APL in the filtrate was converted to fluorophores by oxidation with potassium ferricyanide and determined by HPLC on a column of Hitachi Gel 3011 with a mobile phase of methanol-water (8:2). A linear calibration curve was obtained in the range from 0.25 to 1.5μg of APL in the 25ml sample solution. Two fluorophores formed by the oxidation were concluded to be 2-propyl-pyrichromine and 2-propyl-7-methylpyrichromine on the basis of a comparison of their absorption and mass spectra with those of 2-methyl-pyrichromine.