The invention relates to a construction method and application of an electrochemical cell sensor for acetamiprid and imidacloprid joint toxicity evaluation, and belongs to the technical field of celldetection. A size-controllable Ag/His-GQD/rGO three-dimensional composite material is prepared, after a glassy carbon electrode is modified with the composite material, L-cysteine is combined to the composite material through sulfydryl, activated folic acid is combined with L-cysteine through amido bonds, cells are fixed, determination is performed through an electrochemical impedance technology,and the electrochemical cell sensor is prepared. Through combination with a three-dimensional composite material, more binding sites can be provided, and the sensitivity of the sensor is improved. Combined with an electrochemical impedance technology, the constructed sensor can determine the Hep G2 cells quickly, and is relatively low in cost. The Hep G2 cell sensor can be used for evaluating thepesticide toxicity, and has a good application prospect in the aspect of pesticide joint toxicity evaluation.