Abstract: ObjectiveTo develop a sensitive,specific and rapid chemilum inescent immunoassay(CLIA)method for detection of zearalenone(ZEN)in food.MethodsThe optimal conditions of CLIA for ZEN detection including the concentrations of ZEN-bovine serum albumin(BSA)conjugate,the volume of ZEN-BSA solution for immobilization,the working concentrations of both antibody against ZEN and a horse radish peroxidase-labeled anti-antibody were determined by a chessboard titration.Additionally,the linear range and the limit of detection(LOD)were studied as well.Corn and wheat samples were analyzed with the developed CLIA method and there sults were in comparison with those obtained by LCMS/MS.ResultsThe optimized CLIA conditions for ZEN detection were as follows:0.1μg/mL coating concentration of ZEN-BSA,100 L/well of coating volume,1:1 600 of working titer of antibody aginst Z N.The linear range was between 0.01ng/ml and 50ng/mL with 13.04ng/mL of 50% inhibitory concentration,0.15μg/kg of LOD for ZEN in food samples,and the recovery ranged from 66% to 115% with the spiked level bewteen 20μg/kg and 400μg/kg.Twenty three food samples were analyzed by both CLIA and LCMS/MS.A good correlation between the results obtained from both methods was observed(r²=0.98).ConclusionThe developed CLIA method is simple,fast,sensitive and can be employed in the screening of ZEN in food.