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A sensitive and selective direct competitive enzyme‐linked immunosorbent assay for fast detection of Sudan I in food samples
Author(s) -
Wang Yuzhen,
Yang Hong,
Wang Bin,
Deng Anping
Publication year - 2011
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.4392
Subject(s) - enzyme , chemistry , chromatography , food science , biochemistry
BACKGROUND: Sudan I, a synthetic azo dye, is considered to be a genotoxic carcinogen and is prohibited in foodstuffs for any purpose at any level worldwide. In this study, a sensitive and specific direct competitive enzyme‐linked immunosorbent assay (dc‐ELISA) for fast detection of Sudan I in food samples was developed for the first time. The monoclonal antibody against Sudan I was used as capture protein, while horseradish peroxidase labeled Sudan I conjugate prepared by the periodate method via ovalbumin (OVA) as a bridge was used as enzyme tracer. RESULTS: The standard curve of dc‐ELISA for Sudan I was constructed in the range 0.1–100 ng mL −1 and the assay time was within 80 min. Sensitivity was 2.6 ng mL −1 and the limit of detection was 0.08 ng mL −1 . Cross‐reactivity values of the assay with Sudan II, III and IV were 5.78%, 1.72% and 0.64%; no cross‐reactivity was found with six other edible colorants. The assay was tolerated to 30% of methanol and 10% of acetonitrile without significant loss of IC 50 . Recoveries of spiked Sudan I in five different samples including chilli powder, tomato sauce, hotpot seasoning and chilli sauce I and II were within 88.4–113.2% and the intra‐assay relative standard deviation was less than 14%. The dc‐ELISA was confirmed by conventional high‐performance liquid chromatography and the correlation coefficient of the two methods was 0.9902. CONCLUSION: The proposed dc‐ELISA method provides an alternative method for sensitive, specific and fast determination of Sudan I in food samples. Copyright © 2011 Society of Chemical Industry