Sulfamethazine (SMZ) is effective in the treatment of bacterial infections in food producing animals but its use is prohibited in dairy cows. Nevertheless, a 1988 survey of milk in ten cities conducted by the Food and Drug Administration revealed the presence of SMZ. Therefore, it was apparent that there was a need for rapid screening methods for SMZ. We evaluated commercial immunochemical test kits for SMZ with detectabilities of 1-10 parts per billion (ppb). Manipulations are suggested to effectively optimize immunochemical detection of SMZ in raw and processed fluid milk. The performances of the enzyme immunochemical test kits were evaluated by studying the effects of sample preparation, sample matrix, calibration and detection range of the kits using raw and processed milk samples. Immunochemical results were compared to quantitative high performance thin layer chromatography and high performance liquid chromatography with electrochemical detection. Both chromatographic methods had detectabilities in the low parts per billion range.