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.

Critical evaluation of indirect methods for the determination of deoxynivalenol and its conjugated forms in cereals

“…The results obtained in this study are doubtful for several reasons. First of all, AgraQuant ® was found to be a kit that overestimates the DON content compared to LC-MS/MS results [ 23 – 25 ]. Two comprehensive studies performed on four commercially available DON ELISA kits revealed that DON overestimation is caused by both cross-reactivity with structurally related compounds and by matrix effects.…”

“…Two comprehensive studies performed on four commercially available DON ELISA kits revealed that DON overestimation is caused by both cross-reactivity with structurally related compounds and by matrix effects. Although the kit manufacturer stated that the cross-reactivity with 3ADON was >100 %, two studies reported it to be more than 300 % based on the solvent standard [ 24 , 25 ]. Cross-reactivity with D3G was found to be 113 % in the latter study [ 25 ], while Tran and Smith [ 15 ] based their calculations on a mere 4.8 %, as specified by the manufacturer.…”

“…Although the kit manufacturer stated that the cross-reactivity with 3ADON was >100 %, two studies reported it to be more than 300 % based on the solvent standard [ 24 , 25 ]. Cross-reactivity with D3G was found to be 113 % in the latter study [ 25 ], while Tran and Smith [ 15 ] based their calculations on a mere 4.8 %, as specified by the manufacturer. Furthermore, a matrix contribution to DON overestimation was reported [ 24 ] several years before the published hydrolytic method of Tran and Smith [ 15 ].…”

“…In general, it is recommended that matrix-matched calibration curves should be used in ELISA analysis in order to decrease overestimation. It has been proven that every matrix behaves differently in terms of the cross-reactivity caused by matrix co-extracts [ 24 , 25 ]. Based on our LC-MS/MS validation data, this is also true of nonhydrolyzed and hydrolyzed versions of the same matrix.…”

Biosensors Determination of Aflatoxin B1 in Agricultural Products

“…Паралельно з цим, в останні роки збільшилась увага до більш простих у використанні та високочутливих імуноферментних методів аналізу (ELISA). В літературі є повідомлення про використання ELISA для аналізу дезоксиніваленолу [20,21], афлатоксину B1 [21,22], зераленону та охратоксину [21], одночасного визначення афлатоксину В1 і афлатоксину М1 [23].…”