Synthesis of Novel Haptens and Development of an Enzyme-Linked Immunosorbent Assay for Quantification of Histamine in Foods

• DOI: 10.1021/jf504689x
• Publication Date: 2014-11-24T15:44:06Z
• Authors: Lin Luo; Zhen-Lin Xu; Jin-Yi Yang; Zhi-Li Xiao; Yong-Jun Li; Ross C. Beier; Yuan-Ming Sun; Hong-Tao Lei; Hong Wang; Yu-Dong Shen
• Keywords: Fishes; Enzyme-Linked Immunosorbent Assay; Wine; Yogurt; 01 natural sciences; 0104 chemical sciences; Fish Products; Food, Fortified; Animals; Rabbits; Haptens; Histamine

Novel haptens were designed and synthesized to prepare antibodies against free histamine, but none resulted in producing suitable antibodies for developing an enzyme-linked immunosorbent assay (ELISA). However, an antiserum was obtained having high specificity and affinity to p-nitrobenzoylated histamine (NPHA), which can be easily formed from reaction between histamine and p-nitrobenzoic acid N-hydroxysuccinimide ester (PNBA-OSu) under mild conditions. Based on rabbit polyclonal antibodies, a competitive indirect ELISA (ciELISA) for histamine determination in foods was developed. After ciELISA and derivatization optimization, the assay showed good sensitivity, with limits of detection of 1.8 mg/kg, 93.6 μg/L, and 93.6 μg/kg in fish, red wine, and yoghurt, respectively, with negligible cross-reactivity with related biogenic amines and amino acids. Average recovery of histamine in fortified food samples ranged from 80.9% to 110.1% with coefficients of variation below 16.3%. Good correlation between the ciELISA and liquid chromatography-tandem mass spectrometry was obtained for spiked food samples.