孔雀绿分子印迹96孔检测板的制备及应用研究

Study on Preparation and Application of Malachite Green-MIP-96-Wells Detecting Plates

为更加快速、灵敏、简单地检测水产品中的孔雀绿（MG）残留,通过原位聚合法,以乙腈/甲苯溶液（V/V=3∶1）为溶剂、甲基丙烯酸（MAA）为功能单体、二甲基丙烯酸乙二醇酯（EGDMA）为交联剂（模板分子与之比例为1∶4∶20）,制备了分子印迹聚合物（MIP）。通过包被MIP到96孔板上研发了以化学发光免疫分析法（CLIA）检测的MG-MIP-96孔检测板,其标准曲线为y=-29197.49x＋1945998.45,R2=0.9416,线性范围为30~66.64 ng/m L,检测限为3.355ng/m L。批内变异系数范围为7.38%~18.07%,批内回收率范围为103.41%~144.45%;批间变异系数范围为12.45%~17.52%,批间回收率范围为100.07%~143.20%。以孔雀石绿结构类似物副品红（PA）和结晶紫（CV）进行特异性检测的交叉率分别为27.97%、30.97%。说明此新型检测方法具有很大的研究价值和实际应用价值。

To detect malachite green（ MG） in the aquatic product easily and sensitively,molecularly imprinted polymer（ MIP） microspheres were prepared via in-situ polymerization method with MG as template,α-methacrylic acid（ MAA） as functional monomer,ethylene glycol dimethacrylate（ EGDMA） as cross-linker and acetonitrile /toluene（ V / V = 3 ∶ 1） solution as solvent. Then the MG-MIP-96-well detecting plates detected by the method of chemilumineseent immunoassay（ CLIA） were developed by coating the MIPs in the 96-well plates. The standard curve of as-developed method is y =- 29197. 49 x ＋ 1945998. 45,R2= 0. 9416,linearity range is 30 ~ 66. 64 ng /m L,the LOD is 3. 355 ng / m L,the intra and inter-assay coefficient of variation are 7. 38% ~ 18. 07% and 12. 45% ~17. 52%,the intra and inter-assay recovery are 103. 41% ~ 144. 45% and 100. 07% ~ 143. 20%,and the crossover rates are 27. 97% and 30. 97% with specifically detecting the MG’s analogue,pararosaniline（ PA） and crystal violet（ CV）. The experiment results showed that the as-developed method has great potential in practical application.