title: Hapten synthesis and development of a competitive indirect enzyme-linked immunosorbent assay for acrylamide in food samples.
abstract: The high level of acrylamide in widely consumed processed foods poses a potentially significant risk to human health, which has led to an increasing demand for rapid, simple, and selective analytical methods. In the present work, several haptens for acrylamide were designed in an attempt to prepare antibodies with acrylamide affinity, but they failed their purpose. However, a polyclonal antibody was produced against 4-mercaptophenylacetic acid (4-MPA)-derivatized acrylamide, which showed high binding affinity to the derivative. As acrylamide easily reacted with 4-MPA at high derivation yield, a competitive indirect enzyme-linked immunosorbent assay (ciELISA) for acrylamide via a preanalysis derivatization was developed. The derivatization and ELISA conditions were fully optimized to produce a method for acrylamide assay that exhibited an IC50 of 2.86 μg/kg, limit of detection at 0.036 μg/kg, and linear range of 0.25-24.15 μg/kg. The results of preanalysis recovery tests of acrylamide-spiked food samples and screening of blind food samples by both ciELISA and HPLC-MS/MS indicated the proposed ciELISA's good accuracy and reliability. This method was thus deemed suitable for routine acrylamide screening in food samples at low cost.
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Shen
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Lei
authors.forename: Yuan-Ming
authors.initials: YM
authors.lastname: Sun
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Yang
authors.forename: Zhi-Li
authors.initials: ZL
authors.lastname: Xiao
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Zhen-Lin
authors.initials: ZL
authors.lastname: Xu
mesh_terms: D020106:Acrylamide / Q000032:analysis*; D000917:Antibody Formation; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D006241:Haptens / Q000096:biosynthesis*; D053719:Tandem Mass Spectrometry
chemical_list: D006241:Haptens; D020106:Acrylamide