title: Development of an indirect ELISA for the determination of ethyl carbamate in Chinese rice wine.
abstract: The widespread occurrence of ethyl carbamate (EC, 89.09 Da), a group 2A carcinogen, in fermented foods and alcoholic beverages has raised worldwide public health concern. Immunoassay for EC is unavailable due to the simple and small structure of EC. In this work, an initial attempt to produce antibody specific for EC, by using 4-((ethoxycarbonyl)amino)butanoic acid as hapten, was made but failed. However, since EC can easily react with 9-xanthydrol to form xanthyl ethyl carbamate (XEC), two haptens based on XEC structure were designed and synthesized. Polyclonal antibody against XEC, instead of EC was obtained and then used to develop a competitive indirect ELISA for EC via a pre-analysis derivatization. After optimization, the ciELISA was applied in analyzing Chinese rice wine with detection limit of 166 μg/L, and negligible cross-reactivity with EC analogs. Recoveries of EC in fortified samples were from 84.4% to 100.9%, with coefficients of variation below 10%. Results for analysis of real samples by the ci-ELISA correlated well with that by reference method GC-MS, suggesting the good accuracy and reproducibility of the proposed method. This is the first report of an immunoassay capable of detecting EC, which is suitable for monitoring EC in a large amount of 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.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Lei
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Yang
authors.affiliation: College of Light Industry and Food Science, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China.
authors.forename: Gong-Liang
authors.initials: GL
authors.lastname: Liu
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China. Electronic address: gzsyming@163.com.
authors.forename: Yuan-Ming
authors.initials: YM
authors.lastname: Sun
authors.affiliation: College of Light Industry and Food Science, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China.
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Bai
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Shen
authors.affiliation: Guangdong Meiweixian Flavoring Foods Co.,Ltd., China.
authors.forename: Sui
authors.initials: S
authors.lastname: Chen
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China. Electronic address: jallent@163.com.
authors.forename: Zhen-Lin
authors.initials: ZL
authors.lastname: Xu
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000737:chemistry; D057230:Limit of Detection; D012275:Oryza; D015203:Reproducibility of Results; D014520:Urethane / Q000032:analysis*; D014920:Wine / Q000032:analysis*
chemical_list: D006241:Haptens; D014520:Urethane
keywords: Chinese rice wine; Ethyl carbamate; Hapten; Immunoassay; Polyclonal antibody