Western blotting of glycyrrhetinic acid glucuronides using anti-glycyrrhizin monoclonal antibody

A western blotting method for determining glycyrrhizin by using anti-glycyrrhizin monoclonal antibody was investigated. In this method, we found a new way to separate the glycyrrhizin molecule into two functional parts by using a simple and well-known chemical reaction. The sugar parts are oxidized to give dialdehydes, which react with amino groups on the protein that ties to the adsorbent membrane. The monoclonal antibody binds to the aglycone part of glycyrrhizin molecule. The immunocytolocalization of glycyrrhizin in Glycyrrhiza organs was investigated to indicate that the phloem contains a higher concentration of glycyrrhizin than that of the xylem. This result was confirmed by quantifying glycyrrhizin in the corresponding organs by an enzyme-linked immunosorbent assay using anti-glycyrrhizin monoclonal antibody.