Synthesis and application of a T-2 toxin imprinted polymer.

The synthesis of a T-2 toxin imprinted polymer and its application in food analysis are reported for the first time. A molecularly imprinted polymer (MIP) for the selective recognition of T-2 toxin (T-2) was synthesized by bulk polymerization. Methacrylamide and ethyleneglycol dimethacrylate were applied as functional monomer and cross-linker, respectively. Molecularly imprinted solid-phase extraction (MISPE) procedures were optimized for further application in the analysis of T-2. Scatchard plot analysis revealed that two classes of imprinted binding sites were formed in the imprinted polymer. The dissociation constant (KD) of the higher affinity binding sites was 7.0 micromol/l, while the KD of the lower affinity binding sites was 54.7 micromol/l. The performance of the MIP throughout the clean-up of spiked maize, barley and oat sample extracts was compared with the results obtained when using non-imprinted polymer, OASIS HLB and immunoaffinity columns (IAC). Depending on the food matrix and the spiked concentration, recoveries after MISPE and non-imprinted solid-phase extraction varied respectively from 60% to 73% and from 21% to 57%. Recoveries obtained after clean-up using OASIS HLB and IAC were in the range of 74-104% and 60-85%, respectively. Although highest recoveries were obtained with OASIS HLB sorbents, the designed MIP and the IAC were superior regarding selectivity, cross-reactivity, matrix effect, limits of detection (LOD) and limits of quantification (LOQ). Depending on the matrix, LOD after MISPE ranged from 0.4 microg/kg to 0.6 microg/kg and LOQ from 1.4 microg/kg to 1.9 microg/kg. LOD and LOQ after OASIS HLB clean-up varied from 0.9 microg/kg to 3.5 microg/kg and from 3.1 microg/kg to 11.7 microg/kg, respectively. The LOD and LOQ values obtained with IAC were in the range of 0.3-2.3 microg/kg and 1.0-7.7 microg/kg, respectively. Analysis of 39 naturally contaminated samples (maize, barley and oat) by liquid chromatography tandem mass spectrometry revealed that the MIP could be an excellent alternative for clean-up of contaminated food samples.

[1]  C. Maragos,et al.  Synthesis and evaluation of molecularly imprinted polymers as sorbents of moniliformin , 2007, Food additives and contaminants.

[2]  M. Moreno-Bondi,et al.  Molecularly imprinted polymers applied to the clean-up of zearalenone and α-zearalenol from cereal and swine feed sample extracts , 2006, Analytical and bioanalytical chemistry.

[3]  A. Dobson,et al.  Strategies to Prevent Mycotoxin Contamination of Food and Animal Feed: A Review , 2006, Critical reviews in food science and nutrition.

[4]  Karsten Haupt,et al.  Affinity Separations on Molecularly Imprinted Polymers with Special Emphasis on Solid‐Phase Extraction , 2006 .

[5]  H. Humpf,et al.  Acetonitrile: the better extractant for the determination of T-2 and HT-2 toxin in cereals using an immunoaffinity-based cleanup? , 2009 .

[6]  T. Rundberget,et al.  Instrumental methods for determination of nonmacrocyclic trichothecenes in cereals, foodstuffs and cultures , 1998 .

[7]  S. Edwards,et al.  Emerging issues of HT-2 and T-2 toxins in European cereal production , 2009 .

[8]  Sarah De Saeger,et al.  Development of a multi-mycotoxin liquid chromatography/tandem mass spectrometry method for sweet pepper analysis. , 2009, Rapid communications in mass spectrometry : RCM.

[9]  G. Engelhardt,et al.  Occurrence of type A trichothecenes in conventionally and organically produced oats and oat products. , 2007, Molecular nutrition & food research.

[10]  N. Foroud,et al.  Trichothecenes in Cereal Grains , 2009, International journal of molecular sciences.

[11]  S. Patel,et al.  Occurrence and fate of Fusarium mycotoxins during commercial processing of oats in the UK , 2007, Food additives and contaminants.

[12]  A. Visconti,et al.  Determination of T-2 toxin in cereal grains by liquid chromatography with fluorescence detection after immunoaffinity column clean-up and derivatization with 1-anthroylnitrile. , 2003, Journal of chromatography. A.

[13]  M. Schollenberger,et al.  Natural Occurrence of 16 Fusarium Toxins in Grains and Feedstuffs of Plant Origin from Germany , 2005, Mycopathologia.

[14]  R. Krska,et al.  Improving methods of analysis for mycotoxins: molecularly imprinted polymers for deoxynivalenol and zearalenone , 2003, Food additives and contaminants.

[15]  C. Baggiani,et al.  Solid phase extraction of food contaminants using molecular imprinted polymers. , 2007, Analytica chimica acta.

[16]  A. Logrieco,et al.  Epidemiology of Toxigenic Fungi and their Associated Mycotoxins for Some Mediterranean Crops , 2003 .

[17]  S. Patel,et al.  HT-2 toxin and T-2 toxin in commercial cereal processing in the United Kingdom, 2004-2007 , 2009 .

[18]  Kyung Ho Row,et al.  Characteristic and synthetic approach of molecularly imprinted polymer , 2006 .

[19]  V. Lattanzio,et al.  Determination of trichothecenes in cereals and cereal-based products by liquid chromatography–tandem mass spectrometry , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[20]  M. Moreno-Bondi,et al.  Molecularly imprinted polymers with a streamlined mimic for zearalenone analysis. , 2006, Journal of chromatography. A.

[21]  P. Dubruel,et al.  Molecularly imprinted solid-phase extraction of fumonisin B analogues in bell pepper, rice and corn flakes , 2009, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[22]  R. Schothorst,et al.  Report from SCOOP task 3.2.10 "collection of occurrence data of Fusarium toxins in food and assessment of dietary intake by the population of EU member states". Subtask: trichothecenes. , 2004, Toxicology letters.

[23]  Ian A. Nicholls,et al.  On the thermal and chemical stability of molecularly imprinted polymers , 2001 .

[24]  M. C. Leung,et al.  Mycotoxins and the pet food industry: toxicological evidence and risk assessment. , 2007, International journal of food microbiology.

[25]  A. Logrieco,et al.  Diversity in metabolite production by Fusarium langsethiae, Fusarium poae, and Fusarium sporotrichioides. , 2004, International journal of food microbiology.

[26]  P. Cormack,et al.  Molecularly imprinted polymers: synthesis and characterisation. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[27]  C. Hummert,et al.  Development of a multicomponent method for Fusarium toxins using LC-MS/MS and its application during a survey for the content of T-2 toxin and deoxynivalenol in various feed and food samples , 2005, Food additives and contaminants.

[28]  R. Krska,et al.  Rapid simultaneous determination of major type A- and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry. , 2005, Journal of chromatography. A.

[29]  J. Miller,et al.  Analysis of wheat extracts for ochratoxin A by molecularly imprinted solid-phase extraction and pulsed elution , 2004, Analytical and bioanalytical chemistry.

[30]  A. Dossena,et al.  Simultaneous liquid chromatography-fluorescence analysis of type A and type B trichothecenes as fluorescent derivatives via reaction with coumarin-3-carbonyl chloride. , 2004, Journal of chromatography. A.

[31]  P. Zöllner,et al.  Trace mycotoxin analysis in complex biological and food matrices by liquid chromatography-atmospheric pressure ionisation mass spectrometry. , 2006, Journal of chromatography. A.

[32]  C. Maragos.  Recent advances in the development of novel materials for mycotoxin analysis , 2009, Analytical and bioanalytical chemistry.