Determination of Zearalenone Content in Cereals and Feedstuffs by Immunoaffinity Column Coupled with Liquid Chromatography

Abstract

The zearalenone content of maize, wheat, barley, swine feed, and poultry feed samples was determined by immunoaffinity column cleanup followed by liquid chromatography (IAC–LC). Samples were extracted in methanol–water (8 + 2, v/v) solution. The filtered extract was diluted with distilled water and applied to immunoaffinity columns. Zearalenone was eluted with methanol, dried by evaporation, and dissolved in acetonitrile–water (3 + 7, v/v). Zearalenone was separated by isocratic elution of acetonitrile–water (50 + 50, v/v) on reversed-phase C18 column. The quantitative analysis was performed by fluorescence detector and confirmation was based on the UV spectrum obtained by a diode array detector. The mean recovery rate of zearalenone was 82–97% (RSD, 1.4–4.1%) on the original (single-use) immunoaffinity columns. The limit of detection of zearalenone by fluorescence was 10 ng/g at a signal-to-noise ratio of 10:1 and 30 ng/g by spectral confirmation in UV. A good correlation was found (R2 = 0.89) between the results obtained by IAC–LC and by the official AOAC–LC method. The specificity of the method was increased by using fluorescence detection in parallel with UV detection. This method was applicable to the determination of zearalenone content in cereals and other kinds of feedstuffs. Reusability of immunoaffinity columns was examined by washing with water after sample elution and allowing columns to stand for 24 h at room temperature. The zearalenone recovery rate of the regenerated columns varied between 79 and 95% (RSD, 3.2–6.3%). Columns can be regenerated at least 3 times without altering their performance and without affecting the results of repeated determinations.

Occurrence of Zearalenone-4-β-d-glucopyranoside in Wheat

“…Recent studies about the metabolism of zearalenone by cell suspension cultures of maize (Zea mays), wheat (Triticum aestiVum), and different fungal species (Rhizopus sp., Thamnidium elegans, Mucor bainieri) showed that this metabolite is not only reduced to Rand β-zearalenol, but also conjugated to glucose (2,(6)(7)(8). This zearalenone-4-β-D-glucopyranoside (Figure 1) is not determined by current zearalenone analysis (9)(10)(11). However, gastrointestinal digestion releases the aglucone zearalenone (12), which can contribute to the development of a mycotoxicosis.…”

Investigation Of Mycotoxins In Packed Gluten Free Foods

“…Aflatoxin and ochratoxin analyses were done according to AOAC Official Methods 991.31 [21] and 2000.03 [22], respectively. Zearalenone analysis was done as mentioned in the literature [23]. Deoxynivalenol analysis was done with DONPREP, which was supplied from R-Biopharm Rhöne Ltd., and deoxynivalenol analysis in cereal was done using water extraction.…”

Direct and Competitive Optical Grating Immunosensors for Determination of Fusarium Mycotoxin Zearalenone

“…These immunoassays were further amplified with fluorescent quantum dots [ 34 , 35 , 36 ], magnetic nanoparticles [ 37 ], or helical carbon nanotubes [ 38 ]. Alternatively, antibodies [ 24 , 36 , 39 , 40 , 41 , 42 , 43 , 44 ] or molecularly imprinted polymers [ 45 , 46 , 47 ] could be applied for affinity chromatography or pre-column sample purification prior to chromatographic analyses (HPLC, UPLC). Similarly, nanoparticle-assisted lateral flow immunochromatographic strips [ 48 , 49 ] were devised, occasionally with surface-enhanced Raman scattering detection [ 50 ].…”