Rapid and sensitive determination of amprolium in chicken plasma by high-performance liquid chromatography with post-column reaction |
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| Authors: | K Hamamoto R Koike A Shirakura N Sasaki Y Machida |
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| Affiliation: | aNational Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Tokura 1-15-1, Kokubunji-shi, Tokyo 185, Japan;bDepartment of Clinical Pharmacy, Hoshi University, Ebara 2-4-41, Shinagawa-ku, Tokyo 142, Japan |
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| 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. |
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| Keywords: | Amprolium |
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