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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: To solve the problem that glycyrrhizic acid detection method has the disadvantages of high cost and low efficiency, the glycyrrhizic acid content can not be detected quickly and the method for rapidly and efficiently detecting glycyrrhizin can be developed. Methods: Monoclonal anti-glycyrrhizin antibody with high titer and high sensitivity was prepared and then 30 nm colloidal gold particles were prepared by sodium citrate reduction method to label anti-glycyrrhizin monoclonal antibody. Then, NC coated glycyrrhizin coating on the original volume, the establishment of a method based on indirect competition colloidal gold immunochromatographic test strip rapid glycyrrhizic acid detection techniques, and licorice roots in five regions were measured glycyrrhizic acid content. Results: The minimum limit of detection of glycyrrhizin for immunochromatographic test strips was 25 ng · mL -1. The cross-test showed that the strip did not react with non-glycyrrhizic acid and had high specificity. The test strip test results were compared with HPLC The data measured by the method can reach 100% coincidence rate with good consistency. Conclusion: The method is simple, rapid, high efficiency, low cost, suitable for large-scale rapid screening of glycyrrhizin content determination.