Rapid, on-site quantitative determination of higenamine in functional food using a time-resolved fluorescence microsphere test strip

Higenamine (HIG), a benzyltetrahydroisoquinoline alkaloid found naturally in plants, is classified as an S3 Prohibited Substance in the 2020 World Anti-Doping Agency (WADA) report. To avoid problems such as doping violations in competitive events, it is necessary to develop rapid and sensitive detection methods. In this study, a highly-sensitive anti-HIG monoclonal antibody (mAb) was prepared and a time-resolved fluorescent microsphere immunochromatographic test strip (TRFM-ICTS) was established for the rapid quantitative detection of HIG in functional foods. Under optimized conditions, the TRFM-ICTS was compared with colloidal gold immunochromatographic test strip (CG-ICTS), and the half-maximal inhibitory concentration (IC50) of TRFM-ICTS was 1.37 ng/mL. The spiked recoveries ranged from 86.4% to 105.3%, which was consistent with the results of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the detection of real functional food. Therefore, TRFM-ICTS can be a candidate method for doping monitoring in functional foods and a powerful tool for HIG quantification.