传感器类型
电化学生物传感器
检测对象
新霉素(neomycin),样品基质为牛奶(milk samples)
检测原理
该传感器以滤纸为支撑,羧基化单壁碳纳米管(SWNTs)与抗新霉素抗体复合后浸渍干燥形成导电免疫识别层。检测时,纸基传感器作为工作电极,在含新霉素的缓冲液中孵育,新霉素与固定抗体特异性结合形成免疫复合物。结合事件使抗体—抗原复合物远离相邻SWNTs,加宽纳米管间间隙结,降低电荷转移效率并增大接触电阻,从而降低纸基传感器整体导电性。在0.5 V(vs. Ag/AgCl)恒电位下记录计时电流曲线,取稳定平台电流作为输出信号;新霉素浓度越高,电流下降越明显。循环伏安法也显示电流随新霉素浓度增加而降低,验证了阻抗/导电性变化机制。
检测灵敏度
LOD: 0.04 ng mL−1;线性范围: 0.2–125 ng mL−1;校准方程: Y = 1.805 − 0.0483X;相关系数: 0.9907
效应效果
该传感器对同属氨基糖苷类的庆大霉素(gentamicin)无明显电流响应,表明对目标新霉素具有较高特异性。实际牛奶样品经乙酸处理、稀释、离心和过滤后检测,三个加标水平(0.5、5、50 ng/mL)的回收率为93.25%–110.47%,相对标准偏差(CV)低于10%,说明方法重现性和准确性良好。其检出限0.04 ng/mL低于欧盟牛奶中新霉素限量,且制备仅需滤纸和浸渍—干燥循环,成本低、操作简便,适合批量制备和现场快速筛查。作者认为该纸基免疫传感器可用于牛奶中多种抗生素残留检测,但仍需进一步验证其他抗生素及多目标同时检测。
传感器的构成
- 基底/工作电极:滤纸(filtration paper)条,作为纸基支撑和电化学工作电极,提供导电通道。
- 纳米导电修饰层:羧基化单壁碳纳米管(carboxylated SWNTs),分散后浸渍于滤纸,形成高导电网络。
- 分散稳定剂:聚苯乙烯磺酸钠(PSS),用于稳定SWNTs分散并提高蛋白生物相容性。
- 识别元件:抗新霉素兔多克隆抗体(anti-neomycin rabbit polyclonal antibody),与SWNTs复合,特异性结合新霉素。
- 信号转导层:SWNT–抗体复合涂层,新霉素结合后改变纳米管接触电阻和电荷转移,引起电流/阻抗变化。
- 外部电极:Pt对电极和Ag/AgCl参考电极,与纸基工作电极组成三电极体系进行电化学测量。
中文摘要
本文报道一种基于滤纸支撑的电化学免疫传感器,用于牛奶中抗生素残留的快速、低成本检测。研究采用单壁碳纳米管(SWNTs)和抗新霉素抗体,通过简单浸渍—干燥法将二者复合涂覆于滤纸条,制备出具有较高导电性的纸基生物传感器。当新霉素与固定抗体结合后,纸基传感器整体阻抗和电流随新霉素浓度升高而下降,从而实现电化学信号响应。优化条件下,该方法对新霉素的检出限为0.04 ng/mL,线性范围为0.2–125 ng/mL,明显低于欧盟对牛奶中新霉素的限量要求。经简单样品处理后,该传感器可用于实际牛奶样品中新霉素的测定,加标回收率为93.25%–110.47%。作者指出,采用类似制备流程还可用于牛奶中多种抗生素残留的检测。
英文摘要
Paper supports were used to develop a simple, inexpensive, fast and sensitive electrochemical immunosensor for the analysis of antibiotic residues in milk samples, where single-walled carbon nanotubes (SWNTs) and a simple dip-dry coating method were employed to prepare the highly sensitive biosensor. Well-dispersed SWNTs were impregnated with an antibody against neomycin to obtain a composite coating solution, followed by dipping the filtration paper in the solution to fabricate the sensitive biosensor which had high electrical conductivity. Based on the impedance change in the entire paper supported biosensor with increased concentrations of neomycin, the limit detection of the optimized method was 0.04 ng mL(-1) and a linear detection range from 0.2 to 125 ng mL(-1), well below the European Union regulations for neomycin in this matrix. This paper supported biosensor was applied to determine neomycin in milk samples after a simple sample treatment, with spiked recoveries which ranged from 93.25 to 110.47%. A variety of antibiotic residues in milk samples could be determined following similar sensor preparation.