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Development of highly specific fluorescence immunoassay and enzyme-linked immunosorbent assay for detection of dimethyl phthalate in water samples

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A direct competitive fluorescence immunoassay (dc-FIA) and a direct competitive enzyme-linked immunosorbent assay (dc-ELISA) for the screening of dimethyl phthalate (DMP) in water samples were developed. The immunoassays utilise polyclonal antibodies against DMP raised in rabbits. The anti-DMP antibodies were linked to horseradish peroxidase (HRP) and fluorescein isothiocyanate (FITC). Under the optimal experimental conditions, the dc-ELISA has a linear working range of 0.1–2000 ng/ml (R 2=0.993) with a limit of detection of 0.09 ng/ml. In the dc-FIA, the linear working range was 0.05–30 ng/ml (R 2=0.996), and the limit of detection was 0.02 ng/ml, which is approximately four-fold more sensitive than the dc-ELISA using the same antibody and coating antigen. The results show low cross-reactivity with other structurally related compounds. The proposed methods are successfully applied to determine the DMP contaminants with a simple extraction procedure, and good recoveries were obtained.
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Keywords: ELISA; contamination; dimethyl phthalate; fluoroimmunoassay; foods; immunoassay; polyclonal antibody; water samples

Document Type: Research Article

Affiliations: Key Laboratory of Chemo-Biosensing, College of Chemistry and Materials Science,Anhui Normal University, WuhuAnhui Province241000, China

Publication date: December 1, 2011

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