石墨烯-壳聚糖修饰电极构建石杉碱甲免疫传感器  

Construction of Huperzine A Immunosensor with Graphene-Chitosan Modified Electrode

作　　者：陈媛媛[1] 高璇[1] 刘媛媛 尤金坤 张红艳[1] 余宇燕[1] CHEN Yuan-yuan;GAO Xuan;LIU Yuan-yuan;YOU Jin-kun;ZHANG Hong-yan;YU Yu-yan(Pharmacy College of Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《分析测试学报》2018年第9期1014-1019,共6页Journal of Instrumental Analysis

基　　金：国家自然基金项目(81202914;81703695);福建省科技厅引导性项目(2016Y0056);福建省教育厅科研项目(JAT160238)

摘　　要：通过将石杉碱甲抗体(anti-HupA)固定到由还原态氧化石墨烯/壳聚糖(r GO/CS)复合膜修饰的玻碳电极表面,制备高灵敏度的电化学免疫传感器,利用石杉碱甲抗原抗体特异性反应可阻断[Fe(CN)6]3-/4-氧化还原反应体系电子转移的特点,建立了检测石杉碱甲(Hup A)的电化学免疫传感器分析方法。采用循环伏安法(CV)、交流阻抗法(EIS)及差示脉冲伏安法(DPV)研究Hup A在[Fe(CN)6]3-/4-氧化还原体系的电化学行为,利用DPV法测定Hup A浓度。在优化条件下,15 mmol/L K3[Fe(CN)6]+0. 1 mol/L KCl+0. 1 mol/L PBS(pH 6. 5)测试体系中,Hup A浓度的对数与峰电流差值在1. 0×10-13~1. 0×10-10mol/L范围内呈良好的线性关系,相关系数(r)为1. 000 0,检出限为3. 0×10-14mol/L,回收率为99. 4%~102%。该方法具有特异性强、灵敏度高、操作简单、绿色环保等特点,可用于中药材中石杉碱甲浓度的快速检测。A high-sensitive electrochemical immunosensor was prepared by immobilizing anti-HupA monoclonal antibody onto the glassy carbon electrode modified with a reduced graphene oxide/chitosan composite membrane.And an electrochemical immunosensor for the detection of huperzine A(HupA)was established as the antigen-anibody reaction could block the electron transfer of[Fe(CN)6]3-/4-redox system.The electrochemical behavior of HupA in[Fe(CN)6]3-/4-redox system was investigated by cyclic voltammetry(CV),electrochemical impedance spectroscopy(EIS)and differential pulse voltammetry(DPV).Under the optimized conditions,there existed a good linearity between HupA concentration logarithm and the difference of peak current in the assay system of 15 mmol/L K 3Fe(CN)6+0.1 mol/L KCl+0.1 mol/L PBS(pH 6.5)in range of 1.0×10-13-1.0×10-10 mol/L with a correlation coefficient(r)of 1.000 0.The detection limit was 3.0×10-14 mol/L,and the recoveries were in the range of 99.4%-102%.With the characteristics of strong specificity,high sensitivity,simple operation and environmental protection,this method could be used for the rapid detection of huperzine A in natural herbs.

关 键 词：石墨烯 壳聚糖 石杉碱甲 电化学免疫传感器 差示脉冲伏安法 

分 类 号：O657.1[理学—分析化学] TQ460.72[理学—化学]