Detection of sulfonamides in animal feeds

The animal feed sample is extracted with ethanol or acetone and the extract is evaporated to dryness. Another portion of the same sample, spiked at the 3 ppm level with 8 sulfonamides, is similarly extracted and the extract is evaporated to dryness. The residue from each solution is dispersed with 5 ml 0.1N NaOH and, following the addition of 1 ml 1N HCl and mixing, the solution is filtered. The filtrate is mixed with Celite, transferred to a column, and eluted with ammoniacal ether. Aliquots of the concentrated sample and control eluates are spotted on a neutral Adsorbosil-1 thin layer chromatographic (TLC) plate. Following development in chloroform-methanol (95+5) and drying, the plate is sprayed with an alcoholic solution of p-dimethylaminobenzaldehyde until the control chromatogram shows 6 yellow spots which are, from top to bottom; sulfadimethoxine (SO), the combined sulfamerazine (SM)-sulfamethazine (SH)-sulfaquinoxoline (SQ) spot, sulfadiazine (SD), sulfapyridine (SP), sulfathiazole (SZ), and sulfaguanidine (SG). A spot on the sample chromatogram can be identified if the Rf is identical to one of the 5 sulfonamides not overlapping with another compound. If the Rf of the sample spot is approximately the same as that of the combined SM-SH-SQ spot, more definite identification can be obtained by using basic Adsorbosil-1 TLC plates, with chloroform-methanol (92+8) as the developing solvent.