鱼肉中LMG的分子印迹仿生ELISA检测方法-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Detection of Leuco Malachite Green in Fish by Biomimetic ELISA Method

Author(s):: PENG Aihong; WANG Xiya; LIN Zhengzhong; CHEN Xiaomei; HUANG Zhiyong; （College of Food and Biological Engineering,Jimei University,Xiamen 361021,China）

Keywords:: leuco malachite green; ELISA; molecularly imprinted biomimetic antibody; fish

摘要:: 针对水产品中隐性孔雀石绿（leuco malachite green,LMG）残留的问题，建立了鱼肉中LMG的分子印迹仿生酶联免疫吸附法（enzyme linked immunosorbent assay,ELISA)检测方法。以LMG为模板分子，在pH=8.5的Tris-HCl介质中，多巴胺（dopamine,DA）经氧化自聚合4 h即可在微孔板表面形成聚合物膜，经V(甲醇)∶V(乙酸)=9∶1混合溶液洗脱模板分子后，形成LMG分子印迹聚合物膜（membrane of molecularly imprinted polymer,MIP）。以LMG-MIP膜为仿生抗体，建立了鱼肉中LMG残留的仿生ELISA检测方法。该方法灵敏度为 5.18 μg/L，检出限达0.03 μg/L。利用隐性结晶紫和孔雀石绿这２种LMG的结构类似物进行方法的特异性试验，交叉反应率为21.3%。实际样品加标回收实验表明，方法的回收率为71.8%~73.5%，RSD≤4.2%。结果表明，该检测方法可用于鱼肉中LMG残留的快速筛查。

Abstract:: For the detection of leuco malachite green (LMG) in aquatic products,a biomimetic ELISA method was established based on the biomimetic antibody membrane of molecularly imprinted polymer (MIP).Using LMG as template molecules,the LMG-MIP membrane could be formed on the surface of microplates through the dopamine (DA) self-polymerization for 4 h in Tris-HCl (pH=8.5).After elution of LMG by methanol-acetic acid at the volume ratio of 9∶1,the LMG-MIP membrane on microplates could be used as biomimetic antibody of LMG in ELISA.The sensitivity estimated with IC50 was 5.18 μg/L and the detection limit was 0.03 μg/L.The cross-reactivities for two structural analogues of LMG were 21.3% for malachite green.The recovery tested by LMG standard spiking in fish samples ranged from 71.8% to 73.5%,and the average relative standard deviation was less than 4.2%.The results showed that the method could be used for the rapid detection of LMG residues in fish.