胶体金免疫层析试纸快速检测甘草酸方法的建立  

Rapid detection of glycyrrhizic acid with colloidal gold immunochromatographic strip

作　　者：孙培培[1] 张洁[2] 郑江伟[2] 贾斌[1] 石峰[2] SUN Pei-pei ZHANG Jie ZHENG Jiang-wei JIA Bin SHI Feng(College of Animal Science and Technology, Shihezi University, Shihezi 832003, China College of Life and Science, Shihezi University, Shihezi 832003, China)

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]石河子大学生命科学学院,石河子832003

出　　处：《药物分析杂志》2017年第10期1769-1775,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金项目(31360226);高层次人才科研启动资金专项(CZX201138)

摘　　要：目的:针对目前甘草酸的检测方法存在成本高、效率低,不能快速检测出甘草酸含量的问题,研制能够快速且高效率的检测甘草酸的方法。方法:首先制备出高效价高灵敏度的抗甘草酸单克隆抗体,然后采用柠檬酸钠还原法制备出30 nm的胶体金颗粒,用来标记抗甘草酸单克隆抗体;接着优化结合垫喷涂量和NC膜上甘草酸包被原的包被量,建立一种基于间接竞争法的胶体金免疫层析试纸快速甘草酸检测技术,并对5个地区的甘草根分别进行甘草酸含量测定。结果:免疫层析试纸快速检测甘草酸的最小检测极限为25 ng·mL^(-1),交叉试验证明试纸条不与非甘草酸类反应,特异性高;试纸条检测结果与HPLC方法测定的数据能达到100%的符合率,具有很好的一致性。结论:该方法具有简单快速,效率高,成本低的特点,适合大规模快速筛查甘草酸的含量测定。Objective：Current test methods for glycyrrhizic acid were costly,inefficient and time-consuming.To solve the above problem,rapid detecting method was developed.Methods：First,glycyrrhizic acid resistant monoclonal antibody with high titer and high sensitivity was prepared.Then colloidal gold of 30 nanoparticles was made to label the antibody,using sodium citrate as the reduction.After the spraying amount of conjugate pad and the amount of the coating antigen were optimized,an indirect competitive ELISA method was established for rapid detection of glycyrrhizic acid.And the method was applied in determination of glycyrrhizic acid in Glycyrrhizae Radix et Rhizoma obtained from 5 provinces.Results：Detection limit of the immunochromatographic strip was 25 ng·mL-1.Results of the cross reaction test indicated that the strip had no cross-reactivity with other constituents.Determination result obtained from the strip was 100% in line with obtained from the HPLC method.Conclusion：The established method was rapid,efficient and low-cost,thus could be applied in high-throughput screening of glycyrrhizic acid.

关 键 词：甘草 甘草酸 甘草甜素 单克隆抗体 试纸条 胶体金 

分 类 号：R917[医药卫生—药物分析学]