A simple, sensitive and reproducible high performance liquid chromatographic method was developed for determining valnemulin in feeds. Feed samples were extracted with ethyl acetate under alkaline condition, cleaned up by molecularly imprinted solid-phase extraction, and analyzed by high performance liquid chromatography with ultraviolet detection. The characteristics of the synthesized polymer were evaluated and the loading capacity of the polymer was about 1000 μg analyte/g imprinted polymer. The new procedure for the feed sample cleanup using the prepared polymer cartridge gave higher recoveries and fewer matrix interferences. The assay exhibited a linear dynamic range of 5.0-200 mg kg(-1) with the correlation coefficient above 0.9993. Recoveries of valnemulin from feed samples spiked at 5.0, 20 and 50 mg kg(-1) ranged between 76.0% and 94.4% with relative standard deviations of less than 9%. The limit of detection for valnemulin in feeds was 1 mg kg(-1).
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