Direct competitive enzyme-linked immunosorbent assay for sulfamethazine

A direct competitive enzyme-linked immunosorbent assay (ELISA) for screening sulfamethazine (SMZ) in pork tissues was developed. The assay was made with the affinity-purified polyclonal antibody-coated microtiter plate. A cross reactivity of IgG was observed at 3.5 microg/g of sulfamerazine among nine kinds of sulfonamide tested. Pork tissues fortified with SMZ was mixed with octadecyl silica (C18), and extracted with dichloromethane. The extracted SMZ was measured by homemade ELISA, commercial ELISA, and HPLC. The results were correlated (r=0.993, p<0.01). The homemade ELISA was sensitive to determine SMZ at the maximum residue level (MRL) as commercial one. During stability test of the IgG coated microtiter plate performed at 40 degrees C for 14 days, no difference in sensitivity was observed. We developed homemade ELISA with a detection limit of 10 ng of SMZ per g of pork tissues, and it could be used to screen SMZ in pork tissues.