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Rapid and sensitive determination of amprolium in chicken plasma by high-performance liquid chromatography with
K Hamamoto1, R Koike, A Shirakura
1National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Kokubunji-shi, Tokyo, Japan.
Abstract:
A rapid, sensitive and reproducible reversed-phase HPLC assay was developed for the determination of amprolium (APL) in chicken plasma. Protein in plasma sample was precipitated with 0.33 M perchloric acid and supernatant solution was injected into the HPLC system. Following the chromatographic separation of APL and the beclotiamine (I.S.) on a C18 column, the derivatives of APL and I.S. were formed by post-column reaction and detected by fluorescence detection (excitation at 400 nm, emission at 460 nm). The method showed excellent precision, accuracy and speed with a detection limit of 2 ng/ml. The intra- and inter-assay variance of this method were less than 11.2%. This method has been successfully applied to plasma determinations after oral administration of APL to chicken.