食品中玉米赤霉烯酮化学发光酶免疫方法建立

目的建立检测食品中玉米赤霉烯酮(ZEN)快速灵敏化学发光酶免疫方法。方法采用棋盘滴定法确定化学发光检测ZEN最佳工作条件(抗原包被浓度、包被量、一抗和二抗的工作效价、线性范围和检出限等),建立化学发光检测ZEN的方法并对实际样品进行检测,同时将检测结果与液相色谱-串联质谱法(LC-MS/MS)结果进行比较。结果经多次实验,化学发光检测ZEN的工作条件为:玉米赤霉烯酮-牛血清蛋白(ZEN-BSA)最佳包被浓度为0.1μg/mL,包被量为100μL/孔,抗ZEN单克隆抗体的最佳工作效价为1:1600,标准曲线线性范围0.01~50 ng/mL,50%抑制浓度为13.04 ng/mL,ZEN的最低检出浓度为0.007 ng/mL,样品中ZEN的最低检出量为0.15μg/kg,在20~400μg/kg添加水平的回收率范围为66%~115%;用所建方法对23个食品样品进行分析,检测结果与LC-MS/MS结果高度相关(r²=0.98)。结论所建方法准确、灵敏,适用于食品中ZEN污染水平的批量筛选。

Development of a chemiluminescent immunoassay for detection of zearalenone in foods

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.