title: Synthesis of novel haptens and development of an enzyme-linked immunosorbent assay for quantification of histamine in foods.
abstract: 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.
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Luo
authors.forename: Zhen-Lin
authors.initials: ZL
authors.lastname: Xu
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Yang
authors.forename: Zhi-Li
authors.initials: ZL
authors.lastname: Xiao
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Li
authors.forename: Ross C
authors.initials: RC
authors.lastname: Beier
authors.forename: Yuan-Ming
authors.initials: YM
authors.lastname: Sun
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Lei
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Shen
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D005396:Fish Products / Q000032:analysis*; D005399:Fishes; D005527:Food, Fortified / Q000032:analysis; D006241:Haptens / Q000737:chemistry*; D006632:Histamine / Q000032:analysis*; D011817:Rabbits; D014920:Wine / Q000032:analysis*; D015014:Yogurt / Q000032:analysis*
chemical_list: D006241:Haptens; D006632:Histamine
keywords: ELISA; haptens; histamine