Development and application of  molecularly imprinted polymers (MIP’s) in mycotoxin diagnostics

coordinators: Nicolas Gryson & Peter Maene
Co promoters: Sarah De Saeger & Peter Dubruel
Co-workers: Frederic Dumoulin

Period: 01/01/2009 – 31/12/2011
Grant: IWT-TETRA 080109

Fumonisin (FUM) is a mycotoxin that is mainly present in corn and derived products. The presence of fumonisin in food and feed products may pose a risk to human and animal health.

Current detection methods for fumonisins are based on the use of immuno-affinity columns. Due to several disadvantages related to the development (use of laboratory animals, long development stage) and the use of these columns (high cost, single use), there is an increased demand for alternative user-friendly detection methods.
The goal of this research project is to develop a quick test for the detection of fumonisin in corn products with the aid of molecularly imprinted polymers (MIPs).

A first step involves the development and characterisation of the specific MIP. Main variables in this process are the choice of the functional monomer molecule, the cross-linker molecule and the porogeneous solvent used during polymerisation. The characterisation basically implies the determination of the binding capacity.
In a second step, the procedure for fumonisin extraction from different corn products will be optimised, using a solid-phase extraction method. The optimised method should guarantee a recovery of 70 to 110 %. Based on those two steps, a quick test for fumonisin detection will be developed. This is a semi-quantitative test based on fluorescence detection.
The developed test will be an adequate tool for food and feed companies for the auto control of samples, in order to comply with the current Belgian and European legislation on food and feed safety. The test will be fast, simple, cheap and user-friendly. It will therefore be a convenient tool for all stakeholders in the corn supply chain in the prevention of fumonisin contamination.

Investigation of new analytical challenges for mycotoxin determination: Masked mycotoxins

Promoter: Carlos Van Peteghem
Copromoters: Sarah De Saeger, Dieter Deforce, Geert Haesaert, Mia Eeckhout
Collaborators: Marthe De Boevre, José Diana Di Mavungu, Peter Maene

Period: 01/08/2009 – 31/07/2013
Project number: BOF 05V00709, Research association project

Conjugated mycotoxins, in which the toxin is usually bound to a more polar substance such as glucose, are referred to as masked mycotoxins, as these substances escape established anlatytical techniques but can release their toxic precursors after hydrolysis. Little is known about the occurrence of these compounds in foodstuffs.

High-performance liquid chromatography (HPLC) combined with tandem mass spectrometry (MS/MS) offers a powerful tool for identification and characterization of mycotoxin conjugates. Derivatives can be identified due to their molecular mass and their collision induced dissociation (CID) fragmentation behavior, and unknown mycotoxin conjugates can also be identified with different MS techniques, namely quadrupole time-of-flight (Q-ToF) MS and Fourier transform (FT) MS.

The aim of this project is to evaluate the applicability of LC-MS/MS en high resolution MS tackle the problem of masked mycotoxins. Through this research project, an answer to the following questions will be formulated:

• Which mycotoxin conjugates are found in cereal-based food and feed?

• Is the occurrence of masked mycotoxins dependent on the cereal variety and genotype?

• When and how are conjugated mycotoxins formed?

• What is the share of masked mycotoxins to the total mycotoxin content?

• Do food processing technologies play a role in the occurrence of masked mycotoxins?

• Do mycotoxin conjugates contribute to overestimations of mycotoxin levels with commercial ELISA kits as compared to the traditional LC methods?

Assessment of the chemical risk associated with masked Fusarium toxin-containing

Coordinator: Sarah De Saeger
Copromoters: Carlos Van Peteghem, Mia Eeckhout, Geert Haesaert, Kris Audenaert
Collaborators: José Diana Di Mavungu, Marthe De Boevre, Peter Maene

Period: 01/03/2010 – 31/02/2012
Project number: RT 09/3 MYCOMASK, FOD Volksgezondheit, Veiligheid van de Voedselketen en Leefmilieu

Conjugated or masked mycotoxins first came to attention in the mid-1980s, when animals fed with apparently low mycotoxin contaminated feed showed high severity of mycotoxicosis. The unexpected high toxicity was attributed to the presence of conjugated forms of mycotoxins, possibly generated by the plant metabolism, which were undetected by commonly used analytical methods. These derivatives could be hydrolysed to the precursor toxins in the digestive tracts of the animals or could exert toxic effects comparable to those imputable to free mycotoxins. Little is known about the occurrence, bioavailability and metabolism of these compounds so that a comprehensive study is needed.

This project aims at the following objectives:

1. Develop reliable extraction, purification and quantification methods based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of free and masked Fusarium toxins in cereals and cereal-based foods and feeds;

2. Provide quantitative data concerning the occurrence of masked Fusarium toxins and the circumstances in which they are formed, mainly the glycosylated metabolites of deoxynivalenol (DON), zearalenone (ZEN) and T-2 toxin (T-2) in cereals and cereal-based foods and feeds;

3. Assess the extent in which animals and humans are exposed to masked mycotoxins and carry out a preliminary risk evaluation.