酶联免疫法快速检测大闸蟹中二噁英污染物的研究

Rapid detection of dioxins in hairy crabs by enzyme-linked immunosorbent assay

目的建立大闸蟹中二噁英的酶联免疫(enzyme linked immunosorbent assay,ELISA)检测方法。方法优化加速溶剂萃取条件,净化过程及前处理方法。样品取可食部分冷冻干燥,经加速溶剂萃取方法提取,酸性硅胶处理,过复合硅胶柱、活性碳小柱配合加压设备加速净化,酶联免疫试剂盒检测。结果二噁英标准品在3.2、25.0、50.0、100.0 pg/5 g干重毒性当量(World health organisation-toxic equivalent,WHO-TEQ)值添加水平的回收率为68.2%~123.1%,相对标准偏差小于30%(n=6),线性相关性(r2)一般大于0.98。样品检测值均低于1.5 WHO-TEQ pg/g湿重的欧盟标准。结论该方法方便、快速、成本低,适用于大闸蟹中二噁英基础定量检测。

Objective To establish an enzyme linked immunosorbent assay(ELISA)method for the determination of dioxin in hairy crabs.Methods Accelerated solvent extraction conditions,purification process and pretreatment method were optimized.The edible part of the sample was freeze-dried,extracted by accelerated solvent extraction method,treated with acidic silica gel,accelerated purified by composite silica gel column,activated carbon column and pressurization equipment,and detected by enzyme-linked immunosorbent assay kit.Results The recoveries of dioxin standard at level of 3.2,25.0,50.0,100.0 pg/5 g of World health organisation-toxic equivalent(WHO-TEQ)dry weight were 68.2%-123.1%,the relative standard deviation was less than 30%(n=6),and the linear correlation(r2)was generally greater than 0.98.The values of samples were lower than the EU standard of 1.5 WHO-TEQ pg/g wet weight.Conclusion The proposed method is cconvenient,fast and low cost,which is suitable for quantitative detection of dioxins in hairy crabs.