Preparation of a Broad-Spectrum Heterocyclic Aromatic Amines (HAAs) Antibody and Its Application in Detection of Eight HAAs in Heat Processed Meat.

Heterocyclic aromatic amines (HAAs) are potential human mutagens and carcinogens mainly generated in heat-treated meat. In this work, a broad-spectrum HAAs antibody was prepared and used to develop an indirect competitive ELISA (ic-ELISA) for simultaneous determination of eight HAAs, including 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f] quinoline (MeIQ), 2-amino-3-methylimidazo[4,5-f]quinoxaline (IQx), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx), 2-amino-3,4,7,8-tetramethylimidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in grilled and fried meat samples. The limit of detection (LOD, calculated as IC10) and 50% inhibition concentration (IC50) of ic-ELISA were 5.29 μg/L and 99.08 μg/L, respectively. The detection results of this ic-ELISA were in good agreement with the detection results of UPLC-MS/MS in real samples, which indicated that this ic-ELISA can be applied to detect the total content of eight HAAs in heat processed meat. Use of a broad-spectrum antibody is an efficient strategy in developing immunoassay for simultaneous measuring food risk factors with similar structure.