Development of Highly Sensitive Chemiluminescence Enzyme Immunostaining Assay to Determine Glycyrrhizin Content Using Anti-glycyrrhizin Monoclonal Antibody

Abstract

Licorice, the root of Glycyrrhiza spp., is used in a large number of herbal medicines, such as traditional Chinese medicines, Japanese Kampo medicines, and therapeutic drugs. Since glycyrrhizin (GL) is among the main components in licorice and exhibits numerous beneficial pharmacological activities, the content of GL directly affects biological activity. The quality control based on GL content is an important factor in ensuring biological activity; however, the content of GL in licorice varies depending on plant cultivation environment, genetic factors, and species type. Previously, we prepared an anti-GL monoclonal antibody (anti-GL mAb) and employed it in various immunochemical assays for quality control of licorice and licorice-based products. In this study, we employed the anti-GL mAb in chemiluminescence enzyme immunostaining (CLEIS) to develop a very simple, rapid, specific, and sensitive quality control assay for licorice products, with a limit of detection of 3.9 ng. Furthermore, the CLEIS assay enabled semiquantitative analysis of GL in Kampo medicines. Our results showed that multiple samples can be simultaneously analyzed using CLEIS, and it is a useful tool for determining GL content, as well as ensuring chemical quality control of licoricecontaining products and herbal medicines.

Determination of Glycyrrhizic Acid by Methyl Violet Dual-wavelength Superimposed Visible Spectrophotometry and its Analytical Applications

“…Not only that, but GZA also has a variety of pharmacological effects (Stecanella et al 2021; Wang and Nixon 2001) such as detoxi cation, anti-ulcer, anti-in ammatory, anti-viral, antiatherosclerosis, and anti-cancer, as well as being suitable for patients with diabetes (Zhang et al 2020), asthma (Chen et al 2022), and obesity (Yamamoto et al 2021). Currently, there are many methods for the determination of GZA, such as spectrophotometry (Yu et al 2018), high performance liquid chromatography (Xie et al 2011), chemiluminescence enzyme immunostaining assay (Fujii et al 2022), near infrared spectroscopy (Zhu et al 2018), ultra-high performance liquid chromatography-tandem mass spectrometry (Guo et al 2022), etc. Most of these methods require expensive instruments, timeconsuming, and cumbersome operations, but photometry is a widely used analytical technique in drug analysis because of its simplicity of operation, inexpensive instrumentation, and rapid determination.…”