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Development of a simple gel permeation clean-up procedure coupled to a rapid disequilibrium enzyme-linked immunosorbent assay (ELISA) for the detection of Sudan I dye in spices and sauces

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Abstract

Sudan dyes have been found to be added to chilli and chilli products for illegal colour enhancement purposes. Due to the possible carcinogenic effect, they are not authorized to be used in food in the European Union or the USA. However, over the last few years, many products imported from Asian and African countries have been reported via the Rapid Alert System for Food and Feed in the European Union to be contaminated with these dyes. In order to provide fast screening method for the detection of Sudan I (SI), which is the most widely abused member of Sudan dyes family, a unique (20 min without sample preparation) direct disequilibrium enzyme-linked immunosorbent assay (ELISA) was developed. The assay was based on polyclonal antibodies highly specific to SI. A novel, simple gel permeation chromatography clean-up method was developed to purify extracts from matrices containing high amounts of fat and natural pigments, without the need for a large dilution of the sample. The assay was validated according to the Commission Decision 2002/657/EC criteria. The detection capability was determined to be 15 ng g−1 in sauces and 50 ng g−1 in spices. The recoveries found ranged from 81% to 116% and inter- and intra-assay coefficients of variation from 6% to 20%. The assay was used to screen a range of products (85 samples) collected from different retail sources within and outside the European Union. Three samples were found to contain high amounts (1,649, 722 and 1,461 ng g−1) of SI by ELISA. These results were confirmed by liquid chromatography-tandem mass spectrometry method. The innovative procedure allows for the fast, sensitive and high throughput screening of different foodstuffs for the presence of the illegal colorant SI.

Figure (A) Typical standard curve obtained with a rapid disequilibrium Sudan I ELISA, (B) structure of Sudan I and (C) gel permeation chromatography procedure for the clean-up of spices samples

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Acknowledgements

The authors wish to thank Neogen Corporation, USA for funding this study. They also wish to thank Dr. Sam Mitchell and Mr. Colin McRoberts, AFBI, Belfast for their technical advice and Mr. Stephen Holmes, The Food and Environment Research Agency, York for providing food samples.

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Authors and Affiliations

  1. Institute of Agri-Food and Land Use, School of Biological Sciences, Queen’s University Belfast, David Keir Building, Stranmillis Road, BT95AG, Belfast, UK

    Michalina Oplatowska & Christopher T. Elliott

  2. School of Chemistry and Chemical Engineering, Queen’s University Belfast, David Keir Building, Stranmillis Road, BT95AG, Belfast, UK

    Paul J. Stevenson

  3. Eurofins/WEJ Contaminants GmbH, Neuländer Kamp 1, 21079, Hamburg, Germany

    Claudia Schulz & Lutz Hartig

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  1. Michalina Oplatowska
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  2. Paul J. Stevenson
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  3. Claudia Schulz
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  4. Lutz Hartig
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  5. Christopher T. Elliott
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Correspondence to Michalina Oplatowska or Christopher T. Elliott.

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Oplatowska, M., Stevenson, P.J., Schulz, C. et al. Development of a simple gel permeation clean-up procedure coupled to a rapid disequilibrium enzyme-linked immunosorbent assay (ELISA) for the detection of Sudan I dye in spices and sauces. Anal Bioanal Chem 401, 1411–1422 (2011). https://doi.org/10.1007/s00216-011-5185-y

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