Fast and sensitive aflatoxin B1 and total aflatoxins ELISAs for analysis of peanuts, maize and feed ingredients

• Seven highly sensitive monoclonal antibodies against aflatoxins were produced.
• Fast aflatoxin B1 and total aflatoxin ELISAs were developed and validated in peanuts.
• The detection capabilities for aflatoxin B1 were very low: 0.4 μg/kg and 0.3 μg/kg.
• 32 peanut samples were screened and two low contaminated samples were identified.
• High accuracy of the developed assay was confirmed in proficiency tests.

Aflatoxins are a group of carcinogenic compounds produced by Aspergillus fungi that can grow on different agricultural crops. Both acute and chronic exposure to these mycotoxins can cause serious illness. Due to the high occurrence of aflatoxins in crops worldwide fast and cost-effective analytical methods are required for the identification of contaminated agricultural commodities before they are processed into final products and placed on the market. In order to provide new tools for aflatoxin screening two prototype fast ELISA methods: one for the detection of aflatoxin B1 and the other for total aflatoxins were developed. Seven monoclonal antibodies with unique high sensitivity and at the same time good cross-reactivity profiles were produced. The monoclonal antibodies were characterized and two antibodies showing IC50 of 0.037 ng/mL and 0.031 ng/mL for aflatoxin B1 were applied in simple and fast direct competitive ELISA tests. The methods were validated for peanut matrix as this crop is one of the most affected by aflatoxin contamination. The detection capabilities of aflatoxin B1 and total aflatoxins ELISAs were 0.4 μg/kg and 0.3 μg/kg for aflatoxin B1, respectively, which are one of the lowest reported values. Total aflatoxins ELISA was also validated for the detection of aflatoxins B2, G1 and G2. The application of the developed tests was demonstrated by screening 32 peanut samples collected from the UK retailers. Total aflatoxins ELISA was further applied to analyse naturally contaminated maize porridge and distiller's dried grain with solubles samples and the results were correlated with these obtained by UHPLC-MS/MS method.