胶体金免疫层析法定量检测鱼和虾中苯巴比妥残留

建立了一种快速定量检测水产品鱼和虾中苯巴比妥残留的胶体金免疫层析方法。采用胶体金纳米粒标记苯巴比妥单克隆抗体,研究了胶体金免疫层析条件如标记体系p H值、标记抗体浓度、检测T线包被原浓度、质控C线羊抗鼠Ig G浓度、样品前处理方法等对胶体金灵敏度的影响,并借助胶体金试纸条读数仪进行定量检测。结果表明:方法检出限为0.07 ng/mL,线性范围0.08～0.94ng/m L,裸眼消线值(cut-off值)为10.0 ng/mL。方法特异性良好,与巴比妥、戊巴比妥、异戊巴比妥三种结构类似物交叉反应率小于10%。选择罗非鱼、草鱼、鲈鱼和虾进行添加回收试验,样品回收率在73.50%～114.17%之间,相对标准偏差小于15%,且结果与UPLC-MS/MS法一致。该方法具有准确、灵敏、简便、快速等特点,非常适用于水产品鱼和虾中苯巴比妥残留现场快速筛查。

Development of a Colloidal Gold Immunochromatographic Assay for the Quantitative Determination of Phenobarbital Residues in Fish and Shrimp

In this study, a colloidal gold immunochromatography assay for the quantitative determination of phenobarbital residue in fish and shrimp was developed. Anti-phenobarbital monoclonal was labelled with colloidal gold nanoparticles as tracers. The assay conditions, including pH value of labelling, concentrations of antibody for labeling, concentration of the coating antigen (T line) and the goat-anti-mouse IgG (C line), as well as the sample pretreatment were optimized. By using a colloidal gold strip reader, the assay was able to achieve quantitative determination of phenobarbital. Under the optimal condition, the assay showed a limit of detection and detection range were 0.07 ng/mL and 0.08~0.94 ng/mL, respectively. The cut-off value by naked eyes was 10.0 ng/mL. The cross-reaction rates with barbital, pentobarbital and amobarbital were less than 10%, indicating a good specificity of the proposed assay. Recovery test was done by using tilapia, grass carp, perch and shrimp sample and the recoveries ranged from 73.50% to 114.17% with RSDs below 15%. The results had a good agreement with those of UPLC-MS/MS. In conclusion, the proposed assay was accurate, sensitive, convenient and rapid. It can be suitable for rapid screening of phenobarbital residues in aquatic products of fish and shrimp.