免疫磁珠亲和纯化-超高效液相色谱串联质谱法快速检测小麦中多种真菌毒素  

Rapid determination of mycotoxins in wheat by immune magnetic beads affinity purification with ultra performance liquid chromatography-tandem mass spectrometry

作　　者：雷方[1] 李承龙[2] 张新[1] 周爽[2] 王丹[2] 吴永宁[1,2] 

机构地区：[1]武汉轻工大学食品科学与工程学院,农产品加工湖北省协同创新中心,武汉430000 [2]国家食品安全风险评估中心,卫生部食品安全风险评估重点实验室,北京100021

出　　处：《食品安全质量检测学报》2016年第3期1252-1260,共9页Journal of Food Safety and Quality

基　　金：国家自然科学基金项目(31471671)~~

摘　　要：目的建立免疫磁珠亲和纯化-超高效液相色谱串联质谱法用于小麦中真菌毒素多组分检测。方法用NaIO_4将抗体Fc段的糖残基氧化生成醛基,与磁珠上的氨基共价偶联,实现抗体在磁珠上的定向固定化。以该方法同时偶联多种抗体,获得定向固定多种真菌毒素抗体的免疫磁珠。进而对其亲和性能及使用条件进行详细表征和优化,建立免疫磁珠亲和纯化—超高效液相色谱串联质谱检测方法,用于小麦中真菌毒素多组分同时检测。结果本研究制备的免疫磁珠对8种真菌毒素有较好的保留效果,建立的免疫磁珠亲和纯化-超高效液相色谱串联质谱方法能够同时测定小麦中8种真菌毒素,对于各毒素的检出限在0.1~2μg/kg之间,方法回收率为78%~113%。结论该方法灵敏度高、特异性强、操作简单、有机试剂使用较少,适用于小麦样品的检测。Objective To establish a method for the determination of mycotoxins in wheat by immune magnetic beads affinity purification with ultra performance liquid chromatography-tandem mass spectrometry.Methods The sugar residue of antibody Fc segment with NaIO_4 oxidant was oxidized to aldehyde,the aldehyde covalent coupled with the amino on the magnetic beads,to realize oriented immobilization of antibody on the magnetic beads.Immune magnetic beads with directional fixed a variety of mycotoxins antibody were gotten using this method to couple various antibodies.The characterization and optimization with affinity purification and using condition of the immune magnetic beads were detailed and a method of immune magnetic beads affinity purification with ultra performance liquid chromatography-tandem mass spectrometry for simultaneous determination of mycotoxins in wheat was established.Results The prepared immune magnetic beads in this study had a good retention on 8 kinds of mycotoxins,and the established method of immune magnetic beads affinity purification with ultra high performance liquid chromatography tandem mass spectrometry could simultaneously determinate 8 kinds of mycotoxins in wheat,the detection limits ranged between 0.1 μg/kg and 2 μg/kg,the recoveries ranged from 78%to 113%.Conclusion The method is of high sensitivity,specificity,simple operation,organic reagents used less,suitable for determination of wheat.

关 键 词：免疫磁珠 小麦 真菌毒素 超高效液相色谱-串联质谱法 

分 类 号：O657.63[理学—分析化学] TS210.7[理学—化学]