Monoclonal antibody-based enzyme-linked immunosorbent assay for glycyrrhizin and its aglycon, glycyrrhetic acid.

Monoclonal antibodies (MoAbs) specific for glycyrrhetic acid (GA) were prepared and characterized. Obtained MoAbs (AGA-1, AGA-3, and AGA-6) reacted with GA dose-dependently, but not with glycyrrhizin (GL), carbenoxolone, and steroids. Next, an enzyme-linked immunosorbent assay (ELISA) system using AGA-1 was established. The standard curve showed good linearity between 0.01 and 1000 ng/ml of GA, and the detection limit was 5 pg/ml. Recovery, and intra- and interassay variations of this assay system was satisfactory. GL was also measurable quantitatively after acid-hydrolysis of the samples. The developed ELISA system would be useful to determine GL and GA in biological samples or drugs as an alternative method to high performance liquid chromatography (HPLC).