基于磁性纳米探针的渔用麻醉剂三卡因的快速检测方法

Study on rapid detection of anesthetic tricaine for fish by magnetic immunochromatographic test strips

该研究以超顺磁性纳米材料和三卡因的特异性单抗制备的磁性纳米探针作为标记物,以三卡因结构类似物3-氨基苯甲酸和牛血清蛋白的偶联复合物作为检测线(T线),以羊抗鼠IgG作为质控线(C线)构建层析试纸条,建立了一种渔用麻醉剂三卡因的快速检测方法。结果显示,该试纸条可通过肉眼观察,在5~10 min内检测鱼肉基质(2.5μg/g)、PBS缓冲液(5μg/mL)、水体(5μg/mL)中的三卡因。此外,利用磁性纳米探针的准确定量特性,实现了鱼肉基质和水体中三卡因的快速(20~25 min)定量检测,其检出限为0.150μg/mL(相当于0.075μg/g)、0.141μg/mL,仅为美国水产品中三卡因最大残留限量(1μg/mL)的1/6。该试纸条对苯佐卡因、利多卡因、丁香酚、苯氧乙醇、丙泊酚等常见渔用麻醉剂无交叉反应,特异性高。相较于步骤繁琐、耗时长(2 h)的高效液相色谱,该研究建立的三卡因检测方法具有准确性高(加标回收率81.56%~113.59%)、稳定性好(25℃下可保存半年)且操作简单等特点,可为今后水产品流通过程渔用麻醉剂残留的监测和防控提供技术支撑。

In order to develop a rapid detection method for anesthetic tricaine in aquatic products and water,superparamagnetic nanomaterials and monoclonal antibodies of tricaine were used to prepare magnetic nanoprobe as a detection marker in this study.The conjugate complex of 3-aminobenzoic acid(the structural analogue of tricaine)and bovine serum albumin was used as the detection line(T line),and the goat anti-mouse IgG secondary antibody was used as the control line(C line)to construct the immunochromatographic test strip for rapid detection of the anesthetic tricaine for fish.The results showed that the test strip could be observed by naked eye within 5-10 minutes,and the LODs of 2.5μg/g in fish matrix,5μg/mL in PBS buffer and water were obtained.In addition,a rapid(20-25 min)and highly sensitive quantitative detection method for tricaine in fish meat matrix and water was established with the accurate quantitative characteristic of magnetic nanoprobes,where quantitative LODs were 0.150μg/mL(equivalent to 0.075μg/g),0.141μg/mL in fish meat matrix and water which was 6 times lower than that of American maximum residue threshold(1μg/mL)of tricaine in aquatic products.The developed test strip had no cross-reactivity to common anesthetics for fish such as benzocaine,lidocaine,eugenol,phenoxyethanol and propofol,which also had high specificity.Compared with the cumbersome and time-consuming HPLC detection method(2 h),the detection method established in this study had high accuracy(with spiked recovery rate of 81.56%-113.59%),good stability(could be stored for half a year at 25℃)and simple operation.It could provide technical support for the monitoring and prevention of anesthetic residues for fish during the circulation of aquatic products in the future.