Preparation of hapten and monoclonal antibody of diosmin and establishment of enzyme-linked immunosorbent assay

Diosmin, as one of the research hotspots among flavonoids, has been used medicinally in many countries and as a dietary supplement in the United States. It has been confirmed to be effective in the treatment of hemorrhoids, varicose veins, protecting blood vessels, and managing chronic venous diseases. In this study, an artificial antigen of diosmin was prepared using diosmetin, the aglycone of diosmin. Through screening, specific antibodies against diosmin were obtained. The half maximal inhibitory concentration (IC 50 ) of an indirect competitive enzyme-linked immunosorbent assay (icELISA) was 2.62 ng/mL, with a detection range of 0.52 to 26.39 ng/mL. When applied to the detection of diosmin in limons and tablets, the results obtained by icELISA were consistent with those of Ultra Performance Liquid Chromatography - Tandem Mass Spectrometry (UPLC-MS/MS), demonstrating the good accuracy and reproducibility of this method.