Dual-functional probe-based multi-signal Immunosensor platform for tropane alkaloids: Verification and evaluation

This study aims to realise rapid detecting of tropane alkaloids (TAs) in food. For this purpose, a broad-spectrum single-chain fragment variable was fused with horseradish peroxidase to create an antibody–enzyme complex (AEC) with antigen recognition and catalytic activity. A multi-signal immunosensor platform based on AEC in the direct competitive reaction mode was constructed using 3,3′,5,5′-tetramethylbenzidine and 10-acetyl-3,7-dihydroxyphenoxazine as substrates. The sensitivity of TAs in the immunosensor platform ranged from 0.25 μg/kg to 7912.46 μg/kg. Honey was selected as a representative food sample, and the limit of detection of TAs in honey ranged from 0.02 μg/kg to 409.11 μg/kg, with a recovery rate of 65.7 %–117.1 % and a coefficient of variation less than 21.4 %. Results showed that the immunosensor platform possesses satisfactory accuracy and precision, which highlights its potential for practical applications and its suitability as an ideal tool for rapid screening of TAs in food.