title: Enzyme-linked immunosorbent assay in analysis of deoxynivalenol: investigation of the impact of sample matrix on results accuracy.
abstract: Enzyme-linked immunosorbent assay (ELISA) represents a bioanalytical strategy frequently used for rapid screening of mycotoxin deoxynivalenol (DON) in cereals and derived products. Due to a considerable affinity of some anti-DON antibodies to structurally similar DON metabolites, such as DON-3-glucoside (DON-3-Glc) and 3-acetyl-DON (3-ADON), a significant overestimation of DON concentrations may occur. A validation study of six commercial DON-dedicated ELISA kits, namely Ridascreen DON, Ridascreen FAST, DON, DON EIA, AgraQuant DON Assay, Veratox 5/5, and Veratox HS was carried out on wheat, barley, and malt matrices. Performance characteristics of all tested ELISAs were determined using aqueous solutions of DON, DON-3-Glc, and 3-ADON analytical standards, further with extracts of artificially spiked blank cereals, and finally with matrix-matched standards of all three compounds. In the final phase, the accuracy of data was assessed through a comparison of DON concentrations determined by particular ELISAs and reference ultra-high-performance liquid chromatography-tandem mass spectrometry method. For this purpose, both quality control materials and a comprehensive set of naturally and artificially contaminated samples of wheat, barley, and malt were analyzed. High cross-reactivities were proved for both DON-3-Glc and 3-ADON in the majority of examined assays, and moreover, a considerable contribution of some matrix components to overestimation of DON results was confirmed.
authors.affiliation: ICT Prague, Technicka 3, 166-28, Prague, Czech Republic.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Dzuman
authors.forename: Marta
authors.initials: M
authors.lastname: Vaclavikova
authors.forename: Ivana
authors.initials: I
authors.lastname: Polisenska
authors.forename: Zdenka
authors.initials: Z
authors.lastname: Veprikova
authors.forename: Marie
authors.initials: M
authors.lastname: Fenclova
authors.forename: Milena
authors.initials: M
authors.lastname: Zachariasova
authors.forename: Jana
authors.initials: J
authors.lastname: Hajslova
mesh_terms: D000906:Antibodies / Q000737:chemistry; D000918:Antibody Specificity; D016477:Artifacts; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards*; D005960:Glucosides / Q000737:chemistry; D001467:Hordeum / Q000737:chemistry / Q000382:microbiology; D009183:Mycotoxins / Q000032:analysis*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D053719:Tandem Mass Spectrometry; D014255:Trichothecenes / Q000032:analysis* / Q000737:chemistry; D014908:Triticum / Q000737:chemistry / Q000382:microbiology
chemical_list: D000906:Antibodies; D005960:Glucosides; D009183:Mycotoxins; D011933:Reagent Kits, Diagnostic; D014255:Trichothecenes; C542050:deoxynivalenol-3-glucoside; C043247:3-acetyldeoxynivalenol; C007262:deoxynivalenol