免疫亲和柱–高效液相色谱法测定饲料中的T–2毒素.doc

免疫亲和柱-高效液相色谱荧光检测法测定饲料中的T-2毒素 林淼，赵志辉*，韩薇 （）摘 要: 建立了高效液相色谱测定的方法。色谱柱，。结果表明：的线性范围为~200 μg/L，加标回收率为~92.1%，相对标准偏差(RSD)该方法可用于饲料中的测定。 关键词: 高效液相色谱；饲料； Determination of T-2 Toxin in feed by High Performance Liquid Chromatography with Fluorescence Detection after immunoaffinity Column Clean-Up LIN Miao，ZHAO Zhi-hui*，Han Wei (Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciencesy, Shanghai 201403, China) Abstract：A method has been developed for the determination of T-2 toxin in feed 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 feed. 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 after derivatization with 1-anthroylnitrile ( 1-AN ) 第一作者：林淼（1981－），女，研究实习员，主要从事农产品检测研究。E－mail:linmiao0224@ 通讯联系人：赵志辉（1970－），男，研究员，主要从事农产品检测研究。E－mail: zhao9912@ 基金项目：1、国家标准《饲料中T-2毒素的测定 免疫亲和柱净化-高效液相色谱法》编制说明，项目编号T-469,2、上海市科技兴农重点攻关项目，项目编号：沪农科攻字（2009）第6-1号 and 4-dimethylam inopyridine (DMAP). The results show the linear range of carnitine is 10~200μg/L, the recovery is 76.9~92.1%, RSD is under 6 %. The method has been applied to the determination of T-2 toxin in feed with satisfactory result. Keywords：immunoaffinity precolumn derivatization; high performance liquid chromatography; feed; T-2 toxin T-2毒素（T-2 Toxin)是一种倍半萜烯化合物，学名为4β-1, 5-二乙酰氧基-8α- (3-甲基丁酰氧基) -3α-羧基-12, 13-环氧单端孢霉-9-烯，为白色针状结晶, 室温下稳定, 烹调过程不易将其破坏[1]；熔点为150～151 ℃；难溶于水；易溶于极性溶剂。分子式为C24 H34 O9 , 相对分子质量为466.51 [2]，其结构式如图一。 T-2 毒素是单端孢霉烯族毒素(trichothecenes)中危害最大的毒素之一，T-2 毒素作为一种常见的真菌毒素，在自然界中广泛存在，主要污染小麦、大麦、玉米等粮食作物，人误食污染的谷物后，可引起呕吐、腹泻、发烧等中毒症状。受T-2毒素污染的饲料可引发动物发生食欲不振，精神沉郁，唇、口腔黏膜溃疡和坏死，免疫功能下降，产蛋