免疫亲和柱净化-HPLC法同时测定玉米制品中的赭曲霉毒素A和玉米赤霉烯酮

建立了同时测定玉米制品中赭曲霉毒素A和玉米赤霉烯酮的免疫亲和柱净化高效液相色谱方法。试样经乙腈-水溶液(体积比80?20)提取,复合免疫亲和柱富集和净化后,采用Syncronis C18色谱柱(250 mm×4.6 mm,5μm)固定相,以0.5%冰乙酸-乙腈-甲醇(体积比10?45?45)为流动相,等度洗脱,荧光检测器检测(Ex:333 nm,Em:470 nm)。结果表明:赭曲霉毒素A和玉米赤霉烯酮检出限分别为5.65×10-2μg/kg和4.53×10-1-μg/kg;标准曲线的线性范围分别为1~50 ng/mL(r=0.9999)和10~500 ng/mL(r=0.9999);3个不同水平的加标平均回收率为95.81%~98.31%,RSD值为1.24%~1.65%。20批试样中赭曲霉毒素A和玉米赤霉烯酮测定值分别为0.59~0.69μg/kg和3.84~5.22μg/kg,均无过量残留。该方法具有操作简便、灵敏度高、重现性佳等优点,适用于同时检测玉米制品中赭曲霉毒素A和玉米赤霉烯酮。

Simultaneous Determination of Ochratoxin A and Zearalenone in Corn Products by High Performance Liquid Chromatography with Immunoaffinity Column Purification

A method was developed for simultaneous determination of ochratoxin A and zearalenone in corn products by HPLC with immunoaffinity column purification.In the experiments,the samples were extracted by acetonitrile-water(80?20 v/v),then the extract was purified by immunoaffinity column,and the two toxins were separated by reversed-phase HPLC,and then quantified with fluorescence detection(Ex:333 nm,Em:470 nm),and the separation was performed on a Syncronis C18 Column(250 mm×4.6 mm,5μm)with isocratic elution of acetonitrile-methanol-0.5%acetic acid(45?45?10 v/v).The detection limits were 5.65×10-2μg/kg for OTA,4.53×10-1μg/kg for ZEN.The calibration curves of the two toxins were achieved in the concentration range of 1~50 ng/mL(r=0.9999)for OTA,10~500 ng/mL(r=0.9999)for ZEN.The recoveries of OTA and ZEN spiked to corn products was within 95.81%~98.31%and the relative standard deviations were between 1.24%to 1.65%.20 samples measured value is 0.59~0.69μg/kg for OTA,3.84~5.22μg/kg for ZEN,and all samples have no excessive additives.The method is simple and convenient,good reproducibility,suitable for OTA and ZAN simultaneous determination in corn products.