Determination of closantel residues in plasma and tissues by high-performance liquid chromatography with fluorescence detection

The influence of the pH of the mobile phase with some modifiers on the chromatographic behavior and fluorescence properties of closantel have been investigated. At acidic pH values (2-6), the benzamide moiety of the closantel forms a six-membered ring by hydrogen bonding and possesses a native fluorescence. Using the fluorescence emission of closantel at lambda(ex) = 335 nm, lambda(em) = 510 nm, and pH 2.5 of the mobile phase, a linear calibration curve was estimated over a concentration range of about two orders of magnitude with a correlation coefficient larger than 0.992. The limit of the fluorescence detection was 10 microg/kg. This value was at least 10 times lower than that using UV detection. The method was applied to the determination of closantel in plasma and tissue samples, purified by a solid-phase extraction with C18 cartridges.