Determination of T-2 Mycotoxin in Fusarium strains by HPLC with fluorescence detector

Document Type : Original Article


1 Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

2 Department of Medical Parasitology and Mycology, Faculty of Public Health, Tehran University of Medial Sciences, Iran

3 Department of Medical Mycology, Faculty of Medicine Science, Tarbiat Modares University, Tehran, Iran

4 Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

5 Department of Biochemistry, Faculty of Medicine Science, Tarbiat Modares University, Tehran, Iran

6 Department of Biology, Basic Science Research, Imam Hossein University, Tehran, Iran

7 Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

8 Department of Medical Parasitology and Mycology, Faculty of Medicine, Baqiyatallah University of Medial Sciences, Iran


T-2 toxin is the most poisonous trichothecene produced by Fusarium species especially F.sporotrichioides. T-2 toxin is a biological contaminant in a number of agricultural commodities that can cause severe diseases among humans and animals and even lead to death. The aim of the current study is the analysis of T-2 mycotoxin in Fusarium species by high-performance liquid chromatography (HPLC) combined with florescence detection and derivatization with 1-antroylnitrile (1-AN). Totally, 11Fusariumisolates and reference strains were studied. The isolates were tested for the T-2 toxin production after growing on rice substrate followed by using specific “Multisep 225 Trich Clean up columns” purification. In this study, T-2 toxin production was ranged from 197.05ug/kg to 8503.07ug/kg. This is the first study of T-2 toxin analysis by HPLC-F in Iran.


  1. Boutrif, E., Canet, C., Mycotoxin prevention and control: FAO programs. Rev Med Vet, 1998, vol. 149, pp. 681-94.
  2. Ueno, Y., Trichothecenes Chemical, Biological and Toxicological Aspects. Development in food science. Vol 4, Amsterdam: Elsevier, 1983.
  3. Ler, S.G., Lee, F.K., Gopalakrishnakone, P., Trends in detection of warfare agents Detection methods for ricin,staphylococcal entrotoxin B and T-2 toxin. J Chroma A, 2006, vol. 1133, pp. 1-12.
  4. Li, Y., Wang, Z., Beier, R.C., Shen, J., De,Smet. D. De,Saeger. S. Zhang, S. T-2 toxin, a trichothecenemycotoxin: review of toxicity, metabolism, and analytical methods. J Agric Food Chem , 2011, vol. 59, pp. 3441-53.
  5. Omurtag, G.Z., Yazicioglu, D., Determination of T-2 toxin in grain and grain products by HPLC and TLC. J InvironSci Health B, 2000, vol. 35, pp. 797- 807.
  6. Ueno, Y., Trichothecenes in food. In: Krogh P. Mycotoxins in food. London: Academic Press, 1987, pp. 123–148.
  7. Joffe, A.Z., Fusariumpoae, F., sporotrichioides as principal causal agents of alimentary toxic aleukia. In: Wyllie RD, Morehouse LG. Mycotoxic Fungi, Mycotoxins,MycotoxicosesAnEncylopedic Handbook. New York: Marcel  Dekker,1978, pp.21–86.
  8. Beardall, J.M., Miller, J.D., Diseases in humans with mycotoxins as possible causes. In: Miller JD, Trenholm HL, eds. Mycotoxins in grain: Compounds other than aflatoxin. St.Paul: EaganPress, 1994.
  9. Wang, Z.G., Feng, J.N., Tong, Z. Human toxicosis caused by moldy rice contaminated with Fusarium and T-2 toxin. Biomed Environ Sci, 1993, vol. 6, pp. 65-70.
  10. Ueno, Y., Ishii, K., Kanaeda. S., Tsunoda, H., Tanaka, T., Enomoto, M., Toxicological approaches to the metabolites of Fusaria. IV. Microbial survey on "bean-hulls poisoning of horses" with the isolation of toxic trichothecenes, neosolaniol and T-2 toxin of Fusariumsolani M-1-1. Japan J exp Med, 1972, vol. 42, pp. 187-203.
  11. Yazar, S., Omurtag, G. Z., Fumonisins, Trichothecenes and Zearalenone in Cereals. Int J  MolSci, 2008,vol. 9, pp. 2062-2090.
  12. Joffe, A.Z., Toxic Fusarium species and varieties in nature and in laboratory conditions. In: Fusarium Species: Their Biology and Toxicology, Wiley- Interscience ,NewYork 1986, pp. 9–79.
  13. Wannemacher, R.W., Wiener, S.L., TrichotheceneMycotoxinsIn:Medical Aspects of Chemical and Biological Warfare, 1994, vol. 34, pp. 655-676.
  14. Leslie, J.F., Summerell, B.A., The Fusarium Laboratory Manual. Blackwell Publishing, 2006.
  15. Thrane, U., Adler, A., Clasen, P.E. Diversity in metabolite production by Fusariumlangsethiae , Fusariumpoae and Fusariumsporotrichioides. Int J Food Microbiol, 2004, vol. 95, pp. 257-266.
  16. Omurtag, G.Z., Yazicioglu, D., Occurance of T-2 inprocessed cereals and pulses in turkey determine by HPLC and TLC. Food additcontam, 2001, vol. 18, pp. 844-849.
  17. Pascale, M., Haidukowski, M., Visconti, A.,  Determination of T-2 toxin in cereal grains by liquid chromatography with fluorescence detection after immunoaffinity column clean-up and derivatization with 1-anthroylnitrile. J Chroma A, 2003, vol. 989, pp. 257-264.
  18. Mateo, J.J., Mateo, R., Jimenez, M., Accumulation of type A trichothecene in maize, wheat and rice by Fusariumsporotrichioides isolates under diverse culture conditions. Int J Food Microbiol, 2002, vol. 72, pp. 115-123.
  19. Lincy, S.V., Latha, R., Chandrashekar, A., Manonmani, H.K., Detection of toxigenic fungi and quantification of type A trichothecene levels in some food and feed materials from India. Food Control, 2008, vol. 19, pp. 962–966.
  20. Kachuei, R., Yadegari, M.H., Rezaie, S., Allame, A., Safaie, N., Zaini, F., KhanezadYazdi, F.Investigation of stored wheat mycoflora, reporting the Fusarium cf. langsethiae in three provinces of Iran during 2007. Ann Microbiol, 2009, vol. 59, pp. 383-390.
  21. Lauren, D. R., Agnew, M. P., Multitoxin screening for Fusariummycotoxins grain. J Agric Food Chem, 1991, vol. 39, pp. 502-507.
  22. Marasas, W.F.O., Yagen, B., Sydenham, E.W. Combrinck, S. Thiel PG. Comparative yields of T-2 toxin and related trichothecenes from five toxicologically important strains of Fusariumsporotrichioides. Appl  Environ Microbiol,1987, vol. 53, pp. 693–696.
  23. Milanez, T.V., Valente-Soares, L.M., Gas chromatography- Mass spectrometry determination of trichothecenemycotoxins in commercial corn harvested in the state of sao Paulo, Brazil. J Braz Chem Soc, 2006, vol. 17, pp. 412-416.
  24. Torp, M.Langseth, W. Production of T-2 toxin by a Fusarium resembling Fusariumpoae. Mycopathol, 1999, vol. 147, pp.89– 96.
  25. Cullen, D., Smalley, E.B., Caldwell, R.W. New process for T-2 toxin production. Appl Environ Microbiol, 1982, vol. 44, pp. 371–375.
  26. Marasas, W.F.O., Nelson, P.E., Toussoun, T.A., Toxigenic Fusarium Species: Identity and Mycotoxicology, Pennsylvania State University Press, University Park, 1986.
  27. Visconti, A., Lattanzio, V.M.T., Pascale, M., Haidukowski, M., Analysis of T-2 and HT-2 toxins in cereal grains by immunoaffinity cleanup and liquid chromatography with fluorescence detection. J Chroma A, 2005, vol. 1075, pp. 151-158.
  28. Lippolis, V., Pascale, M., Maragos, C. M., Visconti A. Improvement of detection sensitivity of T-2 and HT-2 toxins using different fluorescent labeling reagents by high-performance liquid chromatography. Talanta, 2008, vol. 74, pp. 1476-1483.
  29. Daraei, B., Yazdanpanah, H., Saberi, H.,  Natural occurrence of Fusariummycotoxins in a wheat-based product from the Iranian market. Toxicolet, 1998, vol. 95, pp.156.
  30. Riazipour, M., Imani-Fooladi, A.A., Razzaghi-Abyaneh, M., Natural occurrence of T2-toxin in domestic and imported rice. Iranian J Public Health, 2009, vol. 38, pp. 111-116.
  31. Torp, M., Nirenberg, H. I., Fusariumlangsethiae sp. Nov. on cereals in Europe. Int J Food Microbiol, 2004, vol. 95, pp. 247-256.
  32. CEN (European Committee for Standardization), TC275-WG5, Biotoxin, Doc. No. 195, CEN Report CR 13505, Brussels, 1999.