Using monoclonal antibodies labeled with Eu 3+ chelates, timeresolved fluoroimmunoassay (TRFIA) methods were developed for the determination of trace sulfamethazine (SMZ), sulfamethoxazole (SMX), and sulfadiazine (SDZ) in environmental waters. Under the optimized conditions, the developed methods offered (i) low detection limits (9.8 ng/L SMZ, 6.1 ng/L SMX, and 5.4 ng/L SDZ, based on 90% inhibition) which were about 1 order of magnitude lower than that of the enzyme-linked immunosorbent assay (ELISA), (ii) high selectivity with no crossreactivity (<0.05%) to similarly structured sulfonamides; (iii) high tolerance to variation of the sample pH (6.0-9.0) and salinity (0-100 mM), as well as the presence of humic acid (0-100 mg/L DOC) and heavy metals (0-1 mg/L concentration each of Cu 2+ , Cd 2+ , Hg 2+ , Pb 2+ , and As(V)) in the samples, and (iv) direct determination with low cost, high sample throughput, and low sample consumption (50-100 µL). The proposed TRFIA procedures were applied to determine sulfonamides in a variety of surface water and wastewater samples without sample pretreatment other than filtration. The satisfactory recoveries (64-127%) and reproducibilities (CV ) 0.2-16%) achieved, as well as the good agreement with those given by liquid chromatography-tandem mass spectroscopy and ELISA methods, demonstrated the applicability of the proposed TRFIA methods for routine screening/quantification of sulfonamides in environmental waters.