Measuring the Level of Main Mycotoxins in Wheat Samples Collected from Flour Factories Silos in Alborz and Tehran Provinces Using LC-MS/MS

Document Type : Original Research

Authors
G. Heidari 1
S. J. Hashemi Hazaveh 2
M. Bayat 3
B. Daraei 3
1 Department of pathobiology, Science and Research Branch, Islamic Azad University,Tehran,Iran
2 Department of Medical Parasitology and Mycology, School of Public Health, Food Microbiology Research Center, Tehran
3 Departement of Toxicology and Pharmacology, School of pharmacy, Shahid Beheshti University of Medical Sciences,Tehran,Iran
10.29252/iem.6.2.95
Abstract
Aim: Cereals and cereal-based products are prone to be infected by mycotoxin-producing fungi. The aim of this study was to investigate the level of contamination caused by 11 major mycotoxins in wheat samples collected from wheat silos in Tehran and Alborz provinces using UHPLC-MS/MS device.

Materials & Methods: Samples preparation was performed based on the extraction and purification procedures using acetonitrile/water/acetic acid solvents and Myco6in1 immunoaffinity columns, respectively. Selected mycotoxins were detected simultaneously using reversed phase ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) with electrospray ionization technique in positive-ion mode in a 15-minute run in the MRM program. Spiked samples calibration curve was used to overcome the matrix effects and to determine the residual mycotoxins.

Findings: Quantification and detection limits for AFB1 and OTA mycotoxins were 2 and 0.7 ppb; for DON, FB1, and FB2 were 100 and 33.3 ppb; for ZER were 50 and 16.6 ppb: for AFB2, AFG1, AFG2, and T-2 were 5 and 1.6 ppb; and for HT-2 were 20 and 6.6 ppb, respectively. Good precision and linearity was observed for mycotoxins. The average recovery rate of mycotoxins was in the range of 72-123 %, and the relative standard deviation (RSDr), indicating the method accuracy, was between 0.6-24.2 %. The validated method for analyzing the 30 wheat samples was used to evaluate the residual mycotoxins. OTA, T-2, and HT-2 mycotoxins were found in wheat samples. Only in one sample, the level of residual OTA exceeded the allowable limit set by the Iranian National Standards Organization.

Conclusion: The present study results highlighted the need for monitoring wheat and wheat-based products and the implementation of control and preventive measures in wheat fields, storage warehouses, and flour factories.

Keywords

Subjects

1. Bhat, R., Rai, R.V., and Karim, A.A. Mycotoxins in food and feed: present status and future concerns. Comprehensive Reviews in Food Science and Food Safety., 2010 ,9: 57-81.
2. Cheng, CT., Perak, M., Mass Poisoning Tale of the 9 Emperor Gods and Rat Tail Noodles, Am J Forensic Med Pathol., 1992, 3: 13-26
3. Bryden, W.L. Mycotoxin contamination of the feed supply chain: Implications for animal productivity and feed security. Anim. Feed Sci. Technol., 2012, 173: 134–158.
4. Marquardt, R.R. Effects of molds and their toxins on livestock performance: A western Canadian perspective. Anim. Feed Sci. Technol., 1996, 58: 77–89.
5. Richard,J. L. Some major mycotoxins and their mycotoxicoses-an overview.Int. J. Food Microbiol., 2007, 119:3-10.
6. Kim D-H, Hong S-Y, Kang JW, Cho SM, Lee KR, An TK, et al. Simultaneous determination of multi-mycotoxins in cereal grains collected from South Korea by LC/MS/MS. Toxins., 2017, 9(3):106.
7. Vaclavikova M, MacMahon S, Zhang K, Begley TH. Application of single immunoaffinity clean-up for simultaneous determination of regulated mycotoxins in cereals and nuts. Talanta., 2013,117:345-51.
8. Park J, Kim D-H, Moon J-Y, An J-A, Kim Y-W, Chung S-H, et al. Distribution analysis of twelve mycotoxins in corn and corn-derived products by LC-MS/MS to evaluate the carry-over ratio during wet-milling. Toxins., 2018,10(8):319.
9. Sarrafian,M.R. Country report:Iran.Fouurth Session of the Technical Committee of APCAEM. Chiang Rai, Thailand., 2009.
10. CS Reddy, Mycotoxin Contamination of Rice. Access on 2013-04-03 www .rkmp.co.in /.../Mycotoxin%20Contamination%20of%20Rice.pdf
11. Zamani-Zadeh, H. R., and Khoursandi, H. Occurrence of Fusarium species and their mycotoxins in wheat in Mazandaran province, Iran.,J. plant Pathol., 1995, 31: 23-37.
12. Sulyok,M., Berthiller,F., Krska,R., and Schuhmacher,R. Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize. Rapid Communications in Mass Spectrometry., 2006,20: 2649-2659.
13. Rasmussen,R.R., Storm,I.M.L.D., Rasmussen,P.H., Smedsgaard,J., and Nielsen,K.F. Multi-mycotoxin analysis of maize silage by LC-MS/MS. Analytical and Bioanalytical Chemistry., 2010, 397: 765-776.
14. Sorensen, L. K., & Elbaek, T. H. Determination of mycotoxins in bovine milk by liquid chromatography tandem mass spectrometry. Journal of Chromatography B., 2005, 820: 183–196.
15. Frenich, AG., Vidal, J.L.M., Romero-Gonzalez, R., Aguilera-Luiz, M.M., Simple and high-throughput method for the multi mycotoxin analysis in cereals and related foods by ultra-highperformance liquid chromatography/tandem mass spectrometry.Food Chem., 2009, 117: 705-712.
16. Razzazi-Fazeli.E. A Review of Recent Trends in Applications of Liquid Chromatography-Mass Spectrometry for Determination of Mycotoxins, Journal of Liquid Chromatography & Related Technologies., 2008, 31(11):1641 – 1686.
17. diMavungu,J.D.,Monbaliu,S.,Scippo,M.L.,MaghuinRogister,G.,Schneider,Y.J., Larondelle,Y.,Callebaut,A., Robbens,J., van Peteghem,C., and de Saeger,S. LC-MS/MS multi-analyte method for mycotoxin determination in food supplements. Food Additives and Contaminants-Part A., 2009, 26: 885-895.
18. Monbaliu, S., Van Poucke, C., van Peteghem, C., Van Poucke, K., Heungens, K., and de Saeger, S. Development of a multi-mycotoxin liquid chromatography/ tandem mass spectrometry method for sweet pepper analysis.Rapid Communications in Mass Spectrometry., 2009, 23: 3-11.
19. Boyd, R.K., Basic, C., and Bethem, R.A. Trace Quantitative Analysis by Mass Spectrometry. John Wiley & Sons Ltd., Chichester., 2008.
20. Ediage E E, Diana Di Mavungu J, Monbaliu S, Van Peteghem C, De Saeger S. A validated multianalyte LC–MS/MS method for quantification of 25 mycotoxins in cassava flour, peanut cake and maize samples. Journal of agricultural and food chemistry., 2011,59(10):5173-80.
21. Ediage EN, Van Poucke C, De Saeger S. A multi-analyte LC–MS/MS method for the analysis of 23 mycotoxins in different sorghum varieties: The forgotten sample matrix. Food chemistry., 2015,177:397-404.
22. Commission Regulation (EC) No 1881/2006, Commission Directive 2006/1881/EC of 19 December 2006. setting maximum levels for certain contaminants in food stuffs. Official Journal of the European Communities (2006) L364, 5–24.
23. F.Soleimany,S.Jinap,F.Abas.,Determination of mycotoxins in cereals by liquid chromatography tandem mass spectrometry. Food Chemistry.,2012,130(4) :1055-1060.
24. Kumar, R., Ansari, K. M., Saxena, N., Dwivedi, P. D., Jain, S. K., & Das, M., Detection of Ochratoxin A in wheat samples in different regions of India. Food Control., 2012, 26(1), 63–67.
25. Zhou, Q., Li, F., Chen, L., & Jiang, D.,Quantitative Analysis of 10 Mycotoxins in Wheat Flour by Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry with a Modified QuEChERS Strategy. Journal of Food Science, 2016, 81(11):2886–2890.
26. Hussein HS, Brasel JM. Toxicity, metabolism, and impact of mycotoxins on humans and animals. Toxicology., 2001,167(2):101-34.
27. Yazdanpanah, H., Miraglia, M., Calfapietra, F. R., & Brera, C.Natural occurrence of aflatoxins and ochratoxin A in corn and barley from Mazandaran and Golestan in north provinces of Iran. Mycotoxin Research., 2001, 17: 21-30.
28. A.Zinedine,C.Brera,S.Elakhdari,C.Catano,.F.Debegnach,S.Angelinib,.De Santis ,M . Faidc,M.Benlemlih,V.Minardi,M.Miraglia,. Natural occurrence of mycotoxins in cereals and spices commercialized in Morocco, Food Control., 2006, 17(11) 868-874.
29. Heidari G., Hashemi Hazaveh j., Daraei B., Bayata M. Validation of an UHPLC-MS/MS Method for Simultaneous Analysis of 11 Mycotoxins in Wheat Flour Using Immunoaffinity Column. Iranian Journal of Pharmaceutical Research (2019), 18 (Special Issue): 182-189
Infection Epidemiology and Microbiology
Volume 6, Issue 2
June 2020
Pages 95-107