利用表面等离子共振技术检测牛奶中庆大霉素残留方法的建立及验证

Establishment and verification of a method for detecting gentamicin residues in milk based on surface plasmon resonance technology

目的建立一种基于表面等离子共振(SPR)技术快速测定牛奶中庆大霉素残留的方法。方法采用表面SPR技术并利用抗体抗原反应的高特异性特点,通过将庆大霉素-BSA偶联蛋白固定于S系列CM5芯片表面,利用竞争性免疫原理,建立了牛奶中庆大霉素残留的方法并对该方法的准确度、重复性、专属性、检测范围等参数进行考察。结果本实验室建立的牛奶中庆大霉素残留定量方法准确度高(平均回收率为102.60%,RSD值为9.58%)、重复性好(庆大霉素对照品3.2、12.8、51.2ng/mL高中低三个浓度分析重复性所得RSD分别为4.56%、4.83%、3.84%),中间精密度高(RSD值为4.65%),庆大霉素在0.8~102.4ng/mL的范围内,仪器响应值(Ru)与庆大霉素浓度呈现良好的回归关系。结论本研究建立的方法可用于牛奶中庆大霉素残留的检测。

Objective To establish a method for rapid determination of gentamicin residues in milk based on Surface Plasmon Resonance(SPR)technology.Methods Using surface SPR technique and taking advantage of the high specificity of antibody-antigen reaction,the gentamicin-BSA coupled protein was immobilized on the surface of S series CM5 chip.Based on the principle of competitive immunity,a method for gentamicin residue in milk was established,and the accuracy,repeatability,specificity,detection range and other parameters of the method were investigated.Results The method for the determination of gentamicin residues in milk established in our laboratory has high accuracy(average recovery rate was 102.60%,RSD value was 9.58%),excellent repeatability(For high,middle and low concentrations(3.2,12.8 and 51.2ng/mL)of Gentamicin reference substance,the RSD of the analysis repeatability were 4.56%,4.83%and 3.84%,respectively),and high intermediate precision(RSD value was 4.65%).For gentamicin in the concentration range of 0.8-102.4ng/mL,the response value of instrument(Ru)showed a good regression relationship with gentamicin concentration.Conclusion The method established in this study can be used for the detection of gentamicin residues in milk.