Novel monoclonal antibody-based sensitive enzyme-linked immunosorbent assay and rapid immunochromatographic strip for sensitive detecting aristolochic acid I in herbal remedies

Monoclonal antibodies (mAbs) specific to aristolochic acid I (AAI) were produced from a hybridoma cell line, 10D9E10, and used to establish a rapid and sensitive indirect enzyme-linked immunosorbent assay (ciELISA) and immunochromatographic strip (immunostrip) for detecting AAI in herbal remedies. In the ciELISA, the concentration that caused 50% inhibition of the binding of antibodies to the solid-phase AAI-ovalbumin (OVA) by free AAI (IC50) and the detection limit (IC10) were calculated to be 0.4 and 0.03 ng/mL, respectively. The rapid on-site detection of immunostrip had a detection limit of 1 ng/mL for AAI. Analysis of 24 herbal remedy samples using the ciELISA showed AAI contamination levels ranging from 61 ng/g to 319 µg/g. High-performance liquid chromatography (HPLC) analysis confirmed results obtained from ciELISA and immunostrip, supporting rapid on-site AAI detection in herbal remedies.