Detection of Acrylamide in Foodstuffs by Nanobody-Based Immunoassays
Yifan Liang, Yuyao Zeng, Lin Luo, Zhenlin Xu, Yu‐Dong Shen, Hong Wang, Bruce D. Hammock
Abstract
Acrylamide is toxic aliphatic amide formed via the Maillard reaction between asparagine and reducing sugars during thermal processing of food. Herein, a specific nanobody termed Nb-7E against the acrylamide derivative xanthyl acrylamide (XAA) was isolated from an immunized phage display library and confirmed to be able to detect acrylamide. First, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established for acrylamide with a limit of detection (LOD) of 0.089 μg/mL and working range from 0.23 to 5.6 μg/mL. Furthermore, an enhanced electrochemical immunoassay (ECIA) was developed based on the optimized reaction conditions. The LOD was as low as 0.033 μg/mL, threefold improved compared to that of ic-ELISA, and a wider linear detection range from 0.39 to 50.0 μg/mL was achieved. The average recoveries ranged from 88.29 to 111.76% in spiked baked biscuits and potato crisps. Finally, the analytical performance of the ECIA was validated by standard ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS).