色谱

色谱

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免疫亲和柱净化/柱前衍生化-高效液相色谱荧光检测法测定粮谷中的T-2毒素

李 军 许烨 卫锋 隋凯 赵守成 王玉萍   

  1. 1.Liaoning Entry & Exit Inspection and Quarantine Bureau, Dalian 116001, China; 2.Shenyang Agriculture University, Shenyang 110161, China
  • 收稿日期:2005-05-19 修回日期:2006-03-16 出版日期:2006-05-30 发布日期:1987-03-25
  • 通讯作者: 卫锋

Determination of T-2 Toxin in Cereal Grains by High Performance Liquid Chromatography with Fluorescence Detection after Immunoaffinity Column Clean-Up and Precolumn Derivatization

LI Jun, XU Ye, SUI Kai, WEI Feng, ZHAO Shoucheng, WANG Yuping   

  1. 1.Liaoning Entry & Exit Inspection and Quarantine Bureau, Dalian 116001, China; 2.Shenyang Agriculture University, Shenyang 110161, China
  • Received:2005-05-19 Revised:2006-03-16 Online:2006-05-30 Published:1987-03-25
  • Contact:

    Wei Feng

PDF

838

被引次数

58

摘要： 建立了免疫亲和柱净化/柱前衍生化-高效液相色谱荧光检测器测定粮谷中T-2毒素含量的方法。样品经甲醇-水(体积比为80∶20)混合溶剂提取,通过免疫亲和柱(IAC)净化,以氰酸蒽(1-AN)为衍生化试剂、4-二甲基氨基吡啶(DMAP)为催化剂进行衍生,以ZORBAX Eclipse XDB-C18 柱为分离柱,乙腈-水(体积比为80∶20)为流动相进行高效液相色谱分离及荧光检测，荧光检测的激发波长为381 nm，发射波长为470 nm。T-2毒素的质量浓度为0.01~1.5 mg/L时与峰高呈良好的线性,相关系数为0.9985。在0.01~1.5 μg/g添加水平下，回收率为79.7%~94.5%,相对标准偏差小于7%；检出限（S/N＝3）为0.01 μg/g。该方法净化效果好,灵敏度高,操作简便快速。

关键词: T-2毒素, 高效液相色谱法, 粮谷 , 免疫亲和柱, 荧光检测, 柱前衍生化

Abstract: A method has been developed for the determination of T-2 toxin in cereal grains by high performance liquid chromatography with fluorescence detection after immunoaffinity column clean-up and precolumn derivatization.The derivatization reaction was used to develop a sensitive, reproducible and accurate method for the determination of T-2 toxin in wheat, corn, barley and rice. T-2 toxin was extracted with methanol-water (80∶20, v/v), purified by immunoaffinity columns containing antibodies specific for T-2 toxin, and quantified by reversed-phase high performance liquid chromatography with fluorescence detection (excitation wavelength, 381 nm; emission wavelength, 470 nm) after derivatization with 1-anthroylnitrile （1-AN） and 4-dimethylaminopyridine (DMAP). ZORBAX Eclipse XDB-C18 column and mobile phase of acetonitrile-water (80∶20, v/v) were used for the analysis. Recoveries from the different cereals spiked with T-2 toxin at levels ranging from 0.01 to 1.5 μg/g were from 79.7% to 94.5%, the relative standard deviations were lower than 7% and the limit of detection was 0.01 μg/g based on a signal-to-noise ratio of 3∶1.

Key words: cereal grains , fluorescence detection, high performance liquid chromatography （HPLC）, precolumn derivatization, T-2 toxin, immunoaffinity column