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Quantification of 2,4-dichlorophenoxyacetic acid in oranges and mandarins by chemiluminescent ELISA.

Food chemistry (2013-06-26)
Marina M Vdovenko, Alexandra S Stepanova, Sergei A Eremin, Nguyen Van Cuong, Natalia A Uskova, Ivan Yu Sakharov
ABSTRACT

Direct competitive enzyme-linked immunosorbent assay (ELISA) for 2,4-dichlorophenoxyacetic acid (2,4-D) was developed. Varying the concentrations of monoclonal anti-2,4-D-antibody and the conjugate of soybean peroxidase and 2,4-D the conditions of ELISA performance were optimised. The chemiluminescent method based on peroxidase-catalysed oxidation of luminol was applied to measure the enzyme activity of the conjugate. A mixture of 3-(10'-phenothiazinyl)propane-1-sulfonate and 4-morpholinopyridine was used as potent enhancer of chemiluminescence signal. It was shown that the values of the lower detection limit, IC50 and the working range were 1.5, 64.0, and 6.5-545ng/mL, respectively. The recovery values of CL-ELISA from 10 spiked samples of oranges (n=5) and mandarins (n=5) cultivated in green house without use of 2,4-D and containing different 2,4-D concentrations (10-300ng/mL) were ranged from 92% to 104% that indicated on the absence of matrix effect for the fruit extracts of interest. Determination of 2,4-D in peel of five oranges and five mandarins purchased from stores in Vietnam showed that 2,4-D content in oranges fruits (79-104μg/kg) was significantly higher than that in mandarins (1.66-2.82μg/kg).

MATERIALS
Product Number
Brand
Product Description

Supelco
2,4-D, certified reference material, TraceCERT®
Supelco
2,4-D, PESTANAL®, analytical standard
Supelco
2,4-D, PESTANAL®, analytical standard
Supelco
2,4-D solution, certified reference material, 5000 μg/mL in acetonitrile
Sigma-Aldrich
2,4-Dichlorophenoxyacetic acid, ≥95%, crystalline
Sigma-Aldrich
2,4-Dichlorophenoxyacetic acid, 97%