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Synthesis of Novel Haptens and Development of an Enzyme-Linked Immunosorbent Assay for Quantification of Histamine in Foods
journal contribution
posted on 2014-12-24, 00:00 authored by 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 ShenNovel
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.