A Sensitive High-Performance Liquid Chromatographic Method for Detecting Sulfonamide Residues in Swine Serum and Tissues After Fluorescamine Derivatization

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Abstract
A highly sensitive and rapid high-performance liquid chromatographic method for determining sulfonamides (sulfadiazine, sulfamethazine, sulfamonomethoxine, sulfamethoxazole and sulfadimethoxine) in swine serum and tissues is described. The sulfonamides were extracted from the samples, derivatized with fluorescamine, chromatographed on a Nova-Pak C18 (column using acetonitrile-10 mM potassium phosphate (30:70, v/v) as the mobile phase and detected spectrofluororimetrically (excitation 390 nm, emission 475 nm). The retention times were 7.1 to 18.2 min and there was no interference from any co-extractives. The detection limit for each standard sulfonamide solution was 0.1 ng/ml and their calibration curves were linear between 1 and 100 ng/ml. In the presence of sulfadiazine as an internal standard, sulfonamide recovery from spiked serum, muscle, liver and kidney samples (10 ng/ml) was 94.0 ± 4.7 to 97.3 ± 5.9%, 58.5 ± 3.1 to 73.9 ± 5.7%, 65.9 ± 7.1 to 86.9 ± 10.6% and 86.2 ± 4.0 to 92.8 ± 6.4% respectively.