Quantitation of sulfamethazine in pork tissue by thin-layer chromatography

Our earlier method to detect and quantitate sulfamethazine (SMZ) in milk at the 10 ppb level was modified to quantitate SMZ in pork tissue. Sulfabromomethazine (SBZ) is added to the tissue as an internal standard. SMZ and SBZ are extracted from the tissue into water as the supernatant of a centrifuged, aqueous homogenate and are cleaned up and concentrated by a series of solid-phase extractions. The sulfonamide-containing eluate is then separated on a silica gel thin-layer chromatographic plate. SBZ and SMZ are derivatized with fluorescamine, and their fluorescence is quantitated with a scanning densitometer. The limit of detection was estimated at 0.25 ppb (signal-to-noise ratio, 3:1). The average accuracy over the analysis range (0.54-21.8 ppb [micrograms/kg]) was 95.6% (standard deviation = 29.4%, n = 54).