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A liquid chromatographic method for the determination of closantel residues in milk and tissues is developed and validated. An acetonitrile-acetone solution (80:20, v/v) is used for the extraction of closantel residues from milk and animal tissues, and the extract is purified by solid-phase extraction with Oasis MAX cartridges and a mixture of formic acid-acetonitrile (5:95, v/v) as the elution solution. A C 18  bonded silica column is used for chromatographic separation. The mobile phase consists of acetonitrile-water (85:15, v/v) containing 0.05% triethylamine at pH 2.5, adjusted with phosphoric acid with the flow-rate set at 1.0 mL/min. Using the fluorescence emission of closantel at λ ex  = 335 nm and λ ex  = 510 nm, the calibration curve is linear, with a correlation coefficient of 0.9999 over the concentration range of 10-5000 µg/kg for the tissue sample and 10-5000 µg/L for the milk sample. The detection limit (s/n = 3) is 3 µg/kg for tissue sample and 3 µg/L for milk sample. The intra- and inter-day repeatabilities are between 3.35–7.66% and 4.04–8.67%, respectively. The proposed method enables the quantitative determination of closantel residues at levels as low as 10 µg/kg in animal tissue samples and 10 µg/L in milk samples.

Determination of Closantel Residues in Milk and Animal Tissues by HPLC with Fluorescence Detection and SPE with Oasis MAX Cartridges