Liquid Chromatographic Determination of Mebendazole and its Metabolites, Aminomebendazole and Hydroxymebendazole, in Eel Muscle Tissue

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Abstract
An analytical method is presented for liquid chromatographic (LC) determination of mebendazole (MBZ), hydroxymebendazole (MBZ-OH), and aminomebendazole (MBZ-NH2) in eel muscle tissue. Muscle tissue is extracted with ethyl acetate at pH 7.5. After addition of n-hexane, the extract is cleaned up and concentrated on an aminopropyl solid-phase extraction column. The test solutions are analyzed isocratically on a ChromSpher B LC column with acetonitrile–phosphate buffer, pH 6.2, as mobile phase. Limits of detection and quantitation were 0.7 and 1.1 ¼g/kg, respectively, for MBZOH; 1.4 and 2.3 ¼g/kg, respectively, for MBZ; and 1.5 and 2.1 ¼g/kg, respectively, for MBZ-NH2. Interand intraday coefficients of variation were 3.5 and 3.4%, respectively, for MBZ-OH; 2.5 and 3.1%, respectively, for MBZ; and 5.8 and 4.8%, respectively, for MBZ-NH2. Mean recoveries were 90% for MBZ, 74% for MBZ-NH2, and 92% for MBZ-OH. A linear range of applicability of at least 10–1000 ¼g/kg was found for each analyte. Incurred MBZ-NH2 (181.3 ¼g/kg) was identified in eel muscle tissue apart from MBZ (23.7 ¼g/kg) after 48 h exposure ina treatment bath containing MBZ at 1 mg/L.

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