Development of a Homologous Fluorescence Polarization Immunoassay for Diisobutyl Phthalate in Romaine Lettuce

In this work, three fluorescein-labeled DiBP derivatives (tracers) with different chemical structures and spacer bridges were synthesized and purified by thin-layer chromatography (TLC). Compared with the heterologous tracer, the homologous tracer exhibited more affinity to the antibody. What is mor...

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Bibliographic Details
Published in:Food analytical methods 2017-02, Vol.10 (2), p.449-458
Main Authors: Chen, Yingshan, Cui, Xiping, Wu, Panpan, Jiang, Zhengyun, Jiao, Luoying, Hu, Qingqing, Eremin, Sergei A., Zhao, Suqing
Format: Article
Language:English
Subjects:
Analytical Chemistry
Antibodies
Chemical synthesis
Chemistry
Chemistry and Materials Science
Chemistry/Food Science
Contamination
Dilution
Fluorescein
Fluorescence
Food contamination
Food safety
Food Science
High performance liquid chromatography
Homology
Immunoassay
Microbiology
Organic chemistry
Polarization
Reliability analysis
Sensitivity analysis
Thin layer chromatography
Tracers
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Summary:In this work, three fluorescein-labeled DiBP derivatives (tracers) with different chemical structures and spacer bridges were synthesized and purified by thin-layer chromatography (TLC). Compared with the heterologous tracer, the homologous tracer exhibited more affinity to the antibody. What is more, the tracer concentration and the antibody dilution were further evaluated to improve the sensitivity of fluorescence polarization immunoassay (FPIA). On the basis of sensitive antibody and tracer, a rapid and specific FPIA has been established for the detection of DiBP contamination in the romaine lettuce, which has rarely been reported before. Under the optimal conditions, the developed FPIA showed a good detection range from 8.82 to 2152.84 ng/mL, with a detection limit of 1.77 ng/mL. In addition, the cross-reactivity to several compounds structurally related to DiBP was less than 7.37 %. Therefore, DiBP contamination in spiked romaine lettuce samples was detected by FPIA, with the recovery from 88.28 to 119.11 %. Moreover, when compared with high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA), the results analyzed by the developed FPIA shows a strong reliability and a high correlation value of 0.978 and 0.980, respectively. Thus, this data, combined with rapidity and simplicity of the assay, demonstrates that the established FPIA is a suitable method for high throughput screening of DiBP contamination in the romaine lettuce.
ISSN:1936-9751
1936-976X
DOI:10.1007/s12161-016-0596-1