3. Fusarium mycotoxins in cereals - Plant Research International ...

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4. Analysis of relevant Fusarium mycotoxins in cereals - the state of the art W.A. van Osenbruggen and H. Pettersson 4.1 Introduction Toxin producing Fusarium species and their toxins are commonly occurring on European cereals. The most common Fusarium toxins found in cereals are the trichothecenes(deoxynivalenol, nivalenol, HT-2 and T-2 toxins), zearalenone and the fumonisins. High concentrations of deoxynivalenol (DON) have also been found in North American and European wheat during recent years. The risks for human and animal health by these fusarium toxins have been addressed by the international organisations WHO/FAO, the European Commission and national authorities in many countries. Toxic evaluation and risk assessments are going on or have been done for many of the toxins. The Nordic Working Group on Food Toxicology and Risk Assessment has presented a Risk Assessment of the most common fusarium toxins in cereals (Eriksen & Alexander, 1998). The European Commissions Scientific Committee on Food has given an opinion about DON, zearalenone, fumonisin B1, nivalenol and T- 2/HT-2 toxin. JECFA (Joint FAO/WHO Expert Committee on Food Additives) has evaluated zearalenone, fumonisins and the trichothecenes (DON, T-2 toxin and HT-2 toxin) (JECFA, 2001). U.S. Food and Drug Administration have recently made an evaluation for fumonisins and published a final paper and recommendation on Internet http://www.cfsan.fda.gov/~dms/fumongui.html. Maximal limits for certain Fusarium toxins are already introduced in some countries or they are under preparation in the European Union and other countries. The procedure for DON in the EU seems to be quickened compared to earlier for other mycotoxin limits. There is a need for good and standardised analytical methods to make surveys and control of these toxins, but in most cases they are still lacking. The methods for many of the toxins are complicated and often leading to high variations both within and between laboratories. Activities supported by the European Commission are going on to improve the analytical methods. Results from those studies and others concerning method development will be presented to give an overview of the analytical methods for Fusarium toxins. Screening methods for the Fusarium toxins are also necessary for rapid detection of contaminated cereals. Some of the ELISA methods can be used for such purposes, but additional investigations are going on making use of techniques like NearInfraRed or 42
Biosensor-technology. Also results on cytotoxicity screening of trichothecenes with colorimetric bioassays will be presented. 4.2 Fusarium mycotoxins - analysis Trichothecenes The working group for Biotoxins of the European Committee for Standardisation (CEN) has expressed the need for a standardised method for DON. There is already an AOAC collaboratively studied gas chromatographic method for DON (Ware et al., 1986). In that study there was, however, a very high variation (31-52%) between laboratories even for samples with relatively high concentration. In the BCR certification of reference materials for DON they obtained much lower variation (1-7%) by mainly using HPLC methods (Gilbert et al., 1992b). Other inter-comparison studies with different methods have also found high between laboratory variations for DON 19-60%, nivalenol 20-33%, HT-2 16-103% and T-2 19-131% (Gilbert, 1992a; Pettersson, 1995, 1998b; Schuhmacher et al., 1997; Joseph & Krska, 1998). The variations were mostly lower when a common calibrant was used. The high variation even for DON is remarkable. Capillary gas chromatographic methods using EC or MS-detection have been preferred for their higher sensitivity and selectivity. In the EU-project Intercomparision of Trichothecene Analysis (the Standard, Measurement and Testing, SMT Program) have those methods been studied for the analysis of DON, nivalenol, HT-2 and T-2 toxins during the last four years. The project has been co-ordinated by Hans Pettersson, Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden. The purity and the amount/concentration of commercial trichothecenes for calibration do vary and is one important cause for between laboratory variation. The project has revealed even more serious GC-method problems (Pettersson, 1998a,b; Pettersson & Langseth, 2002a). The GC-responses of the trichothecenes are higher in presence of matrix than as pure calibrants. This is mainly due to different adsorption of the derivatized trichothecenes in the injector and column in presence and absence of matrix and may lead to an overestimation of the trichothecenes in analysed samples by sometimes more than 100%. This matrix effect was more or less pronounced in all laboratories and with all methods. Regular changing of dirty injector liner and cutting/shorting the column will reduce but not completely eliminate the problem. Using cool on-column injector instead of splitless injector and injection of low sample amount 43
Page 2 and 3: This document has been prepared wit
Page 5: Contents Preface Acknowledgements S
Page 9: Acknowledgements The Concerted Acti
Page 12 and 13: high-yielding resistant varieties e
Page 15 and 16: 1. The Concerted Action Mycotochain
Page 17 and 18: 2. Crop production 2.1 Introduction
Page 19 and 20: ot is unlikely to occur in wheat (J
Page 21 and 22: interact differentially with host c
Page 23 and 24: haploid mapping population with `Su
Page 25 and 26: longer period of time on a great ac
Page 27 and 28: 2.3 Ring test with selected Europea
Page 29 and 30: 1 days after 1 January 2 average of
Page 31 and 32: The correlation between DON and per
Page 33 and 34: anthesis with a mixed conidial susp
Page 35 and 36: At present, the challenge of contro
Page 37 and 38: good host for F. graminearum, its r
Page 40 and 41: 3. Fusarium mycotoxins in cereals:
Page 42 and 43: Fusarium spp. and production of myc
Page 44 and 45: Ca(OH)2) gave 100% reduction of fum
Page 46 and 47: under certain baking and processing
Page 48 and 49: epresent a significant proportion o
Page 50 and 51: Deoxynivalenol was converted to 3-k
Page 54 and 55: can also reduce the overestimation.
Page 56 and 57: The BrdU-method together with Swiss
Page 58 and 59: 81-84% and fumonisin B2 76-82% but
Page 60: Recently a new coating based on sel
Page 63 and 64: immunosuppressive; deoxynivalenol i
Page 65 and 66: • Establish critical limits • E
Page 67 and 68: Principle 3. Establish critical lim
Page 69 and 70: 5.4 Good Agricultural Practice and
Page 71 and 72: the government of EU member states
Page 73 and 74: 5.6 Harmonisation of standards for
Page 76 and 77: 6. The Chain-wide approach: Final c
Page 78: PROCESSING POST- PRE-HARVEST HARVES
Page 81 and 82: Beck, R. & J. Lepschy, 2000. Ergebn
Page 83 and 84: Gang, G., 1996. Genetische, physiol
Page 85 and 86: Mesterházy, A., 1989. Progress in
Page 87 and 88: Parry, D.W., T.R. Pettitt, P. Jenki
Page 89 and 90: Turner, J.A., P. Jennings & P. Nich
Page 91 and 92: Kaminski, E. & E. Wasowicze, 1991.
Page 93 and 94: Cahill, L.M., S.C. Kruger, B.T. McA
Page 95: Chapter 5 FAO, 1979. Recommended pr
Page 98 and 99: • O.C. (Otto) Knottnerus Tel +31-
Page 100 and 101: Partner 8: State Institute for Qual
Page 102 and 103:
Other specialists • C. (Claudio)
Page 104 and 105:
Partner 20: University of Hohenheim
Page 106:
• Novel test kits for mycotoxins
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