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多组分免疫亲和柱净化-高效液相色谱 串联质谱法同步测定植物油中7种真菌毒素
朱建国，李培武，张奇1，张文，张兆威，杨青青，王秀嫔,王素君
中国油料作物学报 ›› 2016, Vol. 38 ›› Issue (5) : 658.
PDF(1317 KB)
PDF(1317 KB)
多组分免疫亲和柱净化-高效液相色谱 串联质谱法同步测定植物油中7种真菌毒素
Simultaneous determination of 7 major mycotoxins in vegetable oil by HPLC-MS/MS with multi-component immunoaffinity column
为提高对植物油中真菌毒素的检测效率，研制免疫亲和柱，建立检测方法。以纯化的抗黄曲霉毒素（AFT，包括AFB1、AFB2、AFG1、AFG2）、抗玉米赤霉烯酮毒素（ZEA）、抗T-2毒素、抗伏马B1毒素（FB1）的四种单克隆抗体为特异性抓捕元件，通过研究比较3种载体材料和2种抗体偶联方法，成功制备出以CNBr-activated Crystarose 4B作为固相载体材料的真菌毒素多组分免疫亲和柱。同时研究比较了提取溶剂、提取方式和上样缓冲液等条件，并结合高效液相色谱串联质谱技术（HPLC-MS/MS），建立了植物油中7种真菌毒素的同步免疫亲和净化—HPLC-MS/MS检测方法。结果表明，该方法对7种真菌毒素的检测线性范围宽，线性回归方程相关系数（R2）均不低于0.992；对AFB1、AFB2、AFG1、AFG2四种真菌毒素的检出限为0.02～0.08 μg/kg，对ZEA、T-2、FB1三种真菌毒素的检出限为0.10～0.50 μg/kg；添加回收实验中，低、中、高三个不同加标水平下的平均回收率为80.22%～106.10%，相对标准偏差为1.05%～6.56%。该方法操作简便、净化彻底、回收率高、有机溶剂消耗量少，为食用植物油中真菌毒素多组分同步确证性检测提供了一种高效、准确、灵敏的技术方法。
A multi-component immunoaffinity column (MIAC) were prepared by covalently coupling 4 high specific monoclonal antibodies, such as anti-aflatoxin (AFT, including AFB1, AFB2, AFG1, AFG2), anti-zearalennone toxin (ZEA), anti-T-2 toxin and anti-fumonisin B1 toxin (FB1), with CNBr-activated Crystarose 4B, after the investigation of 3 carriers and 2 coupling methods. In addition, with the optimization of extraction solvent, extraction method and loading condition, a method of high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) combined with self-made MIAC was established for simultaneous and quantitative determination of these 7 major mycotoxins in vegetable oil. The linear ranges of regression equation were greater and the correlation coefficients were higher than 0.992. The limits of detection to AFB1, AFB2, AFG1 and AFG2 were 0.02-0.08μg/kg and ZEA, T-2 and FB1 were 0.10-0.50μg/kg respectively. The recoveries of these 7 mycotoxins in spiked vegetable oil samples were in the range of 80.22%-106.10% with the relative standard deviation of 1.05%-6.56%. The method had the advantages of simple operation, effective purification, high recovery rate and less organic solvent consumption, which provided an efficient, accurate and sensitive technique of multi-component determination for mycotoxins in edible vegetable oil.
真菌毒素 / 多组分免疫亲和柱 / 高效液相色谱串联质谱 / 同步检测 / 植物油 {{custom_keyword}} /
Mycotoxins / Multicomponent immunoaffinity column (MIAC) / HPLC-MS/MS / Simultaneous detection / Vegetable oil {{custom_keyword}} /
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国家农产品质量安全风险评估重大项目(GJFP2014006)，湖北省科技支撑重点项目 (2014BBB017)，农业行业科技专项 (201203037)
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