Development of an Indirect Competitive ELISA for Analysis of Alternariol in Bread and Bran Samples
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Alternariol (AOH) is one of the major mycotoxins produced by various species of Alternaria fungi. Natural occurrences of AOH have been reported in various foods, including fruits; processed fruit products such as apple juice, tomato products; wheat and other grains; oilseeds and products thereof, such as sunflower seeds, oilseed rape meal, and flax seed/linseed; and pecans. In this study, AOH-specific polyclonal antibodies were generated and developed an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for monitoring AOH in bread and bran samples. The assay was very sensitive with a limit of detection (LOD) of 2.4 ± 0.6 ng/g and a half maximal inhibitory concentration (IC50) of 15.2 ± 2.6 ng/g in bread and a LOD of 8.4 ± 1.2 ng/g and IC50 of 52.8 ± 10.8 ng/g in bran extract. The assay was very specific to AOH and showed no cross-reactivity to alternariol monomethyl ether, altertoxin, altenuene, tentoxin, or tenuazonic acid. The effect of organic solvents on the assay was tested. The ELISA system tolerated methanol and acetonitrile as co-solvents at up to 5% content without significant loss of IC50 value. Recoveries in all cases were greater than 75%, and the results using this method were comparable to those obtained from mass spectrometry methods. We conclude that this method is suitable for rapid detection of AOH in bread and bran samples, without expensive analytical equipment or time-consuming sample preparation.
KeywordsMycotoxin Alternariol Polyclonal antibodies Indirect ELISA Food samples
The authors thank Wendy Zhao, FRD Bureau of Chemical Safety, Health Canada, for providing the commercial samples which were analyzed by LC-MS/MS method in our division.
Compliance with Ethical Standards
Conflict of Interest
Gurmit Singh declares that he has no conflict of interest. Ligia Velasquez declares that she has no conflict of interest. Beth Brady declares that she has no conflict of interest. Terry Koerner declares that he has no conflict of interest. Anne-Catherine Huet declares that she has no conflict of interest. Philippe Delahaut declares that he has no conflict of interest.
This article does not contain any studies with human subjects. All animal experiments that described in the present study were performed in CER Groupe, Belgium, following all institutional and national guidelines for the care and use of laboratory animals.
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