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

中文关键词:  免疫磁珠  小麦  真菌毒素  超高效液相色谱-串联质谱法

英文关键词:immunomagnetic beads  wheat  mycotoxin  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      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NaIO4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0.1~2 μg/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。

英文摘要:

      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 NaIO4 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.