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Issue 11, 2011
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Development and validation of rapid disequilibrium enzyme-linked immunosorbent assays for the detection of Methyl Yellow and Rhodamine Bdyes in foods

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Abstract

Sensitive and specific enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of two illegal synthetic dyes: Methyl Yellow (MY) and Rhodamine B (RB) in food. Polyclonal antibodies were raised against synthesised immunogens and employed in unique direct disequilibrium ELISAs. The time of the assays was only twenty minutes (five minutes for each incubation step with sample and enzyme conjugate and ten minutes with enzyme substrate). The IC50 for MY was in the range 1.4–4.2 ng mL−1 and for RB 0.1–0.5 ng mL−1. A simple sample preparation method was developed for the analysis of a range of sauces. In the case of spices a dispersive solid phase extraction was applied to purify the extracts. The testing of twenty samples took approximately one and a half hours (including sample preparation and analysis). Both assays were validated according to the Commission Decision 2002/657/EC criteria for use in sauces and spices. The detection capability for MY in sauces and spices was determined to be less than 15 ng g−1 and 50 ng g−1, respectively and for RB, 10 ng g−1 for both types of food samples. The precision of the developed assays was determined in a repeatability study. The intra- and inter-assay coefficients of variation were less than 25% for both tests and matrix types. The simplicity and performance of both assays indicate that they will be very reliable screening methods for the detection of the illegal dyes MY and RB in a range of food products.

Graphical abstract: Development and validation of rapid disequilibrium enzyme-linked immunosorbent assays for the detection of Methyl Yellow and Rhodamine B dyes in foods

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Article information


Submitted
23 Nov 2010
Accepted
22 Mar 2011
First published
20 Apr 2011

Analyst, 2011,136, 2403-2410
Article type
Paper

Development and validation of rapid disequilibrium enzyme-linked immunosorbent assays for the detection of Methyl Yellow and Rhodamine B dyes in foods

M. Oplatowska and C. T. Elliott, Analyst, 2011, 136, 2403
DOI: 10.1039/C0AN00934B

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