Étkezési búzák mikotoxin tartalmának meghatározása prevenciós lehetıségek Téren, J., Gyimes, E., Véha, A. 2009. április 15. PICK KLUB Szeged 1
A magyarországi búzát károsító Fusarium fajok 2
A betakarítás elıtti (preharvest) fertızés következtében keletkezı mikotoxinok Trichotecének Fı termelıi a Fusarium nemzetség fajai, de egyéb gombák is elıállítják A típusú pl. T2, DAS B típusú pl. DON, NIV Zearalenon Fı termelıi a F. graminearum és a F. culmorum. A raktári gombák okozta (postharvest) fertızés HOOC NH O ochratoxin A OH Cl O Különbözı élelmiszerek jelentısége az ochratoxin bevitelben 6% 5% 4% 3%1% 8% 10% 13% 50% O A gabonafélékben a Penicillium verrucosum, a trópusi növényekben (kávé, kakaó) az Aspergillus ochraceus termeli az ochratoxint. Jelentıs mikotoxin szennyezıdést okozhatnak. Gabonafélék Kávé Egyéb Kakaó Húsfélék Bor Főszerek Sör Aszalt gyümölcsök A gabonafélék jelentısége kiemelkedı. 3
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Termék Mikotoxin határértékek Az Európai Bizottság 2008.07.01.ig felülvizsgálja Feldolgozatlan gabonafélék Feldolgozatlan durum búza és zab Feldolgozatlan gabonafélékbıl származó valamennyi termék, beleértve a feldolgozott gabonatermékeket és a közvetlen emberi fogyasztásra szánt gabonaféléket Gabonaırlemények Kenyér, tésztafélék, keksz, gabonaszeletek és reggeli gabonapelyhek Tészta (száraz ) Feldolgozott, gabona alapú, csecsemık és kisgyermekek részére szánt élelmiszer és bébiétel (a 856/2005/EK rendelet alapján) DON ( µg/kg ) 1250* 1750 750 500 750 200 * a durum búza, zab és a kukorica kivételével ; ** a kukorica kivételével ZON ( µg/kg ) 100** 75 50 20 OTA ( µg/kg ) 5 3 0,5 HPLCs készülék 5
HPLCs kromatogram 6
ELISA leolvasó 7
A nagymőszeres és a gyors módszer közötti összefüggés 4,5 4 R 2 = 0,9517 DON ELISA (mg/kg) 3,5 3 2,5 2 1,5 1 0,5 0 0,00 2,00 4,00 6,00 DON HPLC (mg/kg) 8
A malmi ırlésre történı elıkészítések hatása a toxin szennyezettségre Rostálás Fajsúly szerinti osztályozás koncentrátor alkalmazásával Felülettisztítás Satake cég PeriTec eljárással A COMPARATIVE STUDY OF FUSARIUMDAMAGED KERNEL AND DEOXINIVALENOL CONTAMINATION IN NATURALLY INFECTED WHEAT SAMPLES. A. Véha1, J. Téren1, E. Gyimes, Á. Mesterházy2 University of Szeged, Faculty of Engineerig, Mars square 7. 6724 Szeged, Hungary, dekan@mk.uszeged.hu 2 Cereal Research Non Profit Company, Szeged, Hungary Fig.1 Regression between HPLC and ELISA methods 3,00 2,50 y = 0,5494x + 0,3341 R2 = 0,7575 2,00 DONELISA(ppm) Introduction Fusarium head blight (FHB) the most important fungal disease of wheat in Hungary caused by mostly by F. graminearum and in a less extent by other Fusarium spp (1., 2.). The toxin contamination in epidemic years is several time higher than the official EU limits are, therefore it is also a severe economic problem. The predominant toxin is deoxynivalenol (DON), therefore in this study we concentrated on this. Results Naturally infected wheat samples were collected in 2006 following an epidemic. They were evaluated for FDK, DON contamination by HPLC and ELISA analyses. We aimed to demonstrate possible reactions between the traits considered. The 30 samples analyzed showed an R2 = 0.7575 between HPLC and ELISA (Veratox DON 5.5 Quantitative Test) test results (Fig.1). The ELISA was less correct, the HPLC maximum was 3.75 mg/kg, and the ELISA maximum was only 2.40. At 10 samples both methods gave lower values than 1.25 mg/kg, however, three samples gave at ELISA lower level than this limit and three samples were considered by ELISA as acceptable, but three showed significantly higher values with HPLC than the ELISA test. Three samples with up to 2.50 mg/kg in HPLC were evaluated at 1 mg/kg by ELISA method. Data show that ELISA results higher than 0.6 mg/kg should be controlled by exact analytical methods as the limit values cannot be granted. Samples that contained by ELISA higher than 1.25 mg/kg DON were also higher than this values also by HPLC. The DON (HPLC) and FDK value presented an R2= 0.5183 determination coefficient (Fig.2). Except six samples all were for FDK between 0.00050 %, and DON contamination was between 0.23.2 mg/kg. This supports other results showing that at this low level of FDK the DON contamination cannot be predicted except in the artificially infected wheat samples (Fig.3). 1,50 1,00 0,50 0,00 0,00 0,50 1,00 1,50 2,00 2,50 3,00 3,50 4,00 DON HPLC (ppm) Fig.2 Regression between FDK % and DON concentration 3,00 2,50 2,00 y = 0,5388x 0,4478 R 2 = 0,5183 1,50 FDK% 1 1,00 0,50 Conclusions 0,00 0,00 0,50 1,00 1,50 2,00 2,50 3,00 3,50 4,00 0,50 DON HPLC (ppm) Fig. 3. Regression of FDK and DON contamination of Variety Office materials (3rd year), 2006 90 80 70 DONContent ppm Our conclusion is that all samples should be controlled for DON first with a rapid test, like ELISA. In the second step, however, all samples between 0.50.6 and 1.25 should be controlled by traditional analytical instruments. There are further problems in sampling and labs have at least1015 % differences at ring tests that influence decisions makers in trade and grain industry. The conclusion is that ELISA is suitable for preliminary testing, but a significant part should be measured by traditional way as ELISA is not correct enough. Although the FDK shows significant correlation with DON contamination, the FDK is unsuitable to predict DON values to decide whether the commodity has higher or lower DON than the limit value prescribes. Acknowledgements The authors are indebted to NKTHKPI project 2006ALAP301435/2006 for financial support. References 1. Lamper, Cs., Téren, T., Komoróczy, R., Mesterházy, Á., and Sági, F.: Predicting DON contamination Fusariuminfected wheat grains via determination of the ergosterol content. Cereal Res. Commun. 28. 337344. 2000. 2. Mesterházy, Á., Bartók, T., Mirocha, C.G. and Komoróczy, R.: Nature of wheat resistance to Fusarium head blight and the role of deoxinivalenol for breeding. Plant Breeding 118. 97110. 1999. 3. Mesterházy, Á. and Tóth, B.: Progress of plant resource GH Fusarium project in the CRC Szeged. German Hungarian Seminar,Gaterslaben, 12.12. 2006. y = 1,6936x 4,2653 2 R = 0,7415 60 50 40 30 20 10 0 10 0 5 10 15 20 25 30 35 40 whe at sample infe ction % 3rd International Symposium on Fusarium Head Blight, Szeged 2008. 9
Mycotoxin project Josef Teren, teren@mk.uszeged.hu Fusarium head blight the most important fungal disease of wheat in Hungary. Deoxinivalenol (DON) is a predominant preharvest toxin. Present activities study visual and measuring kernel infection test for cleaning grains by physical means measure the DON contamination with HPLC and various ELISA methods selection of the best ELISA kit ergosterol determination by HPLC investigation of Fus graminearum populations with molecular genetical and traditional methods Köszönjük a figyelmüket! 10