Glycyrrhizin (GC) is a triterpenoid saponin isolated from the roots of Glycyrrhiza spp., a medicinal plant that is present in 70% of Kampo prescriptions. Since the GC content in Glycyrrhiza spp. affects its various pharmacological activities, Glycyrrhiza spp. is prescribed to contain at least 2% of GC in the Japanese pharmacopoeia, and its quality control based on GC content is required. In this study, a magnetic particles-based enzyme immunoassay (MPs-EIA) was developed using specific monoclonal antibody against GC (MAb 2H2) for the detection of GC in Glycyrrhiza spp. In this system, the immunoreaction time using primary and secondary antibodies was reduced by taking advantage of the wide surface area of magnetic particles (MPs) conjugated with GC by N,N′‑carbonyldiimidazole (CDI)-mediated method. Optimization of MPs-EIA revealed that total assay time (~2 h) was reduced to over half of that of conventional indirect competitive enzyme-linked immunosorbent assay (ELISA) (~5 h). In addition, the GC concentration was detectable within the range from 97.7 to 781 ng/mL, with a limit of detection of 71.4 ng/mL. A series of further validation analyses support the reliability and accuracy of the developed MPs-EIA for the detection of GC in Glycyrrhiza spp. Since the present MPs-EIA overcomes the disadvantage of ELISA in terms of rapidity, it provides a useful approach for the effective quality control of Glycyrrhiza spp., especially when handling multiple samples.

甘草甜素（GC）是从甘草（Glycyrrhiza spp。）的根中分离出的三萜类皂苷，甘草处方中有70％含有这种植物。由于甘草中的GC含量较高。影响其各种药理活性，甘草。在日本药典中，规定其含量至少为2％的GC，因此需要根据GC含量进行质量控制。在这项研究中，使用针对GC（MAb 2H2）的特异性单克隆抗体开发了一种基于磁性粒子的酶免疫分析（MPs-EIA），用于检测甘草中的GC。spp。在该系统中，通过利用N，N'-羰基二咪唑（CDI）介导的方法，利用与GC结合的磁性颗粒（MP）的宽表面积，减少了使用一抗和二抗的免疫反应时间。MPs-EIA的优化表明，总测定时间（〜2 h）减少到传统间接竞争酶联免疫吸附测定（ELISA）（〜5 h）的一半以上。另外，可检测到的GC浓度在97.7至781 ng / mL的范围内，检出限为71.4 ng / mL。一系列进一步的验证分析支持开发的MPs-EIA在甘草中检测GC的可靠性和准确性spp。由于本发明的MPs-EIA在快速度方面克服了ELISA的缺点，因此，它为甘草的有效质量控制提供了一种有用的方法，尤其是在处理多个样品时。