Abstract: Two kinds of sulfonamide antibiotics(sulfadimidine and trimethoprim) are used as template molecules to prepare a double template molecularly imprinted polymer (MIP) by sol-gel method. And MIP is used as molecular recognition element to construct a molecularly imprinted electrochemical sensor for the determination of sulfadimidine and trimethoprim. The electrochemical performance of the sensor is studied by voltammetry and electrochemical impedance spectra. The parameters of MIP synthesis and electrochemical detection are optimized. Under the optimal conditions, the sensor shows good selectivity, which could realize the simultaneous determination of sulfadimidine and trimethoprim in linear ranges of 0.8~50.0and 1.0~50.0 μmol/L, respectively. The detection limits are 0.14 and 0.26 μmol/L (R_SN=3) for sulfadimidine and trimethoprim, respectively. Finally, the sensor is applied to the simultaneous detection of sulfadimidine and trimethoprim in real samples with recovery of standard addition ranging from 91.20%~103.44%.