建立了免疫亲和柱净化-柱前化学衍生-高效液相色谱荧光检测器同时检测谷物中T-2毒素和HT-2毒素的方法。样品经溶剂[V(甲醇)∶V(水)=80∶20]提取,通过免疫亲和柱净化(IAC),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苯基柱为分离柱,乙腈-水为流动相进行高效液相色谱分离和检测。在0.005~0.5μg/g内呈良好线性,检出限为0.005μg/g,添加回收率为82.0%~108.0%,RSD<15.5%。

A method was developed for the determination of T-2&HT-2 toxin in grains by liquid chromatography with fluorescence detection after immunoaffinity column clean-up and pre-cloumn chemical derivatization. Samples were extracted with methanol-water( 80: 20,v /v),and purified by immunoaffinity columns containing antibodies specific for T-2&HT-2 toxin. Then derived with 1-anthroylnitrile( 1-AN) and 4-dimethylaminopyridine( DMAP). ZORBAX Eclipse XDB-phenyl column and mobile phase of acetonitrile-water were used for analysis. Recoveries from the different cereals spiked with T-2&HT-2 toxin at levels ranging from 0. 005 to 0. 5μg /g were from 82. 0% to 108. 0%,the relative standard deviations were less than 15. 5% and the limit of detection was 0. 005μg /g. This method has a higher clean-up degree,sensitive,stable,low cost and can be applied widely.