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石墨烯-壳聚糖修饰电极构建石杉碱甲免疫传感器
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作者单位
陈媛媛,高璇,刘媛媛,尤金坤,张红艳,余宇燕 福建中医药大学药学院 
基金项目: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)
中文摘要: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(rGO/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(HupA)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HupA在[Fe(CN)6]3-/4-氧化还原体系的电化学行为,利用DPV法测定HupA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,HupA浓度的对数与峰电流差值在1.0×10-13~1.0×10-10 mol/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3.0×10-14 mol/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。
中文关键词: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
 
Construction of Huperzine A Immunosensor with Graphene-Chitosan Modified Electrode
Abstract: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 K3Fe(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.
Key Words:graphene  chitosan  huperzine A  electrochemical immunosensor  differential pulse voltammetry(DPV)
引用本文:陈媛媛,高璇,刘媛媛,尤金坤,张红艳,余宇燕.石墨烯-壳聚糖修饰电极构建石杉碱甲免疫传感器[J].分析测试学报,2018,37(9):1014-1019.
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