Direct Competitive Enzyme-Linked Immunosorbent Assay for Detection of Acrylamide in Food Samples
10.11895/j.issn.0253-3820.140208
- VernacularTitle:直接竞争酶联免疫法测定食品中的丙烯酰胺含量
- Author:
Jing WU
;
Lin LUO
;
Zhili XIAO
;
Jinyi YANG
;
Yuanming SUN
;
Hongtao LEI
;
Yudong SHEN
;
Hong WANG
;
Zhenlin XU
- Publication Type:Journal Article
- Keywords:
Acrylamide;
Polyclonal antibody;
Direct competitive enzyme-linked immunosorbent assay;
Derivatization
- From:
Chinese Journal of Analytical Chemistry
2014;(8):1150-1155
- CountryChina
- Language:Chinese
-
Abstract:
Due to the low molecular weight and simple structure, the production of specific antibodies against acrylamide is unavailable. In this study, a novel hapten was synthesized through the derivatization of acrylamide and 4-mercaptophenylacetic acid. The hapten was then coupled to carrier protein and used to immunize New Zealand rabbits. Polyclonal antibody which showed specific binding to the acrylamide derivative ( hapten) was obtained. The antibody was labeled with horseradish peroxidase ( HRP) and used to develop a direct competitive enzyme-linked immunosorbent assay ( dc-ELISA) . The dc-ELISA was used to determine the content of acrylamide derivative, and then transferred to the content of acrylamide. The assay showed an IC50 value of 45. 49 μg/L, a limit of detection of 3. 0 μg/L and the linear range of 9. 2-195 μg/L for acrylamide. The recovery of acrylamide from spiked food sample was determined ranging from 83 . 6% to 112 . 7%. Good correlations between the results of dc-ELISA and standard HPLC-MS/MS were obtained. The proposed dc-ELISA is suitable for the determination of acrylamide in food samples.
