Development of a Homologous Fluorescence Polarization Immunoassay for Diisobutyl Phthalate in Romaine Lettuce
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Development of a Homologous Fluorescence Polarization Immunoassay for Diisobutyl Phthalate in Romaine Lettuce Yingshan Chen 1 & Xiping Cui 1 & Panpan Wu 1 & Zhengyun Jiang 1 & Luoying Jiao 1 & Qingqing Hu 1 & Sergei A. Eremin 2 & Suqing Zhao 1
Received: 5 April 2016 / Accepted: 29 June 2016 # Springer Science+Business Media New York 2016
Abstract 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 crossreactivity 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.
Keywords Fluorescence polarization immunoassay . Phthalate acid esters . Diisobutyl phthalate . Romaine lettuce
Yingshan Chen and Xiping Cui contributed equally to this work and should be considered co-first authors. * Suqing Zhao [email protected]
Qingqing Hu [email protected]
Yingshan Chen [email protected]
Sergei A. Eremin [email protected]
Xiping Cui [email protected] 1
Department of Pharmaceutical Engineering, Faculty of Chemical Engineering and Light Industry/Guangdong Provincial Key Laboratory of Functional Soft Condensed Matter, Guangdong University of Technology, Guangzhou 510006, People’s Republic of China
2
Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov State University, Moscow 119992, Russia
Panpan Wu [email protected] Zhengyun Jiang [email protected] Luoying Jiao [email protected]
Food Anal. Methods
Introduction Phthalate acid esters (PAEs), which have been widely used as plasticizer in resins and polymers, such as polyvinyl choloride (PVC) products to improve the material’s flexibility and softness, are ubiquitous industrial chemicals (Talsness et al. 2009). In our daily life, we are quite often encompassed with the products containing PAEs, such as medica
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