Synthetic antibiotics known as sulfonamides suppress the synthesis of tetrahydrofolic acid, which cures respiratory tract infections and protozoal infections by preventing the creation of dihydrofolic acid. Electrochemical sensors based on tetrakis(1,10phenanthroline)-μ-(4,4′-bipyridine) dicopper(II) chloride monohydrate ([P 2 Cu-Bip-CuP 2 ]Cl 4 •H 2 O or simply Cu 2 P 4 BCl 4 ) have been successfully applied for the determination of sulfamethoxazole (SMX) and trimethoprim (TMP) from samples. The experimental conditions and parameters were optimized to achieve the best electrode performances for simultaneous quantification of SMX and TMP. Based on the analysis of the effect of scan rate on the peak parameters, the R 2 for the peak current vs square root of the scan rate was greater than that of the peak current vs scan rate, indicating diffusion-controlled behavior of both species. The current intensities of both SMX and TMP were highly improved due to surface activation of the electrodes by electropolymerization. For SMX, the limit of detection was determined to be 27.94 nM, while for TMP, it was 21.56 nM, and the limit of quantifications was 71.88 nM, and the corresponding relative standard deviation for each was 0.74% and 0.11%. The constructed electrode was stored for varying durations ranging from two h to 2 days, and it was found to be above 97% stable after storing for 15 days. The real applicability of the suggested sensor for the simultaneous determination of SMX and TMP was verified by sensing clinical serum and urine samples and their spike recovery studies.