Validation of High Performance Liquid Chromatography with Fluorescence Detector Methods for Determination of Aflatoxins in Different Food and Animal Feed Samples — Oak Academic Publishing
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Validation of High Performance Liquid Chromatography with Fluorescence Detector Methods for Determination of Aflatoxins in Different Food and Animal Feed Samples
Department of Chemistry, Faculty of Science, University of Khartoum, Khartoum, Sudan
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National Chemical Laboratories, National Public Health Laboratory, Federal Ministry of Health, Khartoum, Sudan
1 Department of Chemistry, Faculty of Science, University of Khartoum, Khartoum, Sudan
2 National Chemical Laboratories, National Public Health Laboratory, Federal Ministry of Health, Khartoum, Sudan
Method validation for quantitative analysis of aflatoxins, AFB1, AFB2, AFG1 and AFG2 in sorghum, peanut butter, groundnut and animals feed is presented. Aflatoxins were extracted with a mixture of methanol: acetonitrile: water (60:30:10) and cleaned with Alfa test IAC chromatography before analysis by High Performance Liquid Chromatography coupled with fluorescence detection (HPLC-FLD) by adopting an isocratic chromatographic system using a mobile phase consisting acetic acid: acetonitrile: methanol (59:14:27), the separation of the four aflatoxins was achieved in less than 15 minutes. Calibration curves were linear over the concentration range from 2 - 18 ng/mL for AFB1 and AFG1, 0.4 - 3.6 ng/mL for AFB2 and AFG2, respectively. The LOD and LOQ in spiked samples were found to be 0.02 and 0.05 μg/kg for both AFB1 and AFG1, 0.01 and 0.03 μg/kg for both AFB2 and AFG2. The mean recovery values were in range from 84.2 % to 96.9% was obtained. Five samples were found to be contaminated with aflatoxins and the total aflatoxins ranged from 0.02 to 3.26 μg/kg were obtained. Nineteen different samples were found to be contaminated with aflatoxins; the total aflatoxins ranged from 0.27 to 10.48 μg/kg were obtained. The highest total aflatoxins value was obtained in animal feeds.
KeywordsAflatoxinsHPLC-FLDPeanutPeanut ButterSorghum and Animal Feeds
Guan, D., Li, P., Zhang, Q., Zhang, W., Zhang, D. and Jiang, J. (2011) An Ultra-Sensitive Monoclonal Antibody-Based Competitive Enzyme Immunoassay for Aflatoxin M1 in Milk and Infant Milk Products. Food Chemistry, 125, 1359-1364. https://doi.org/10.1016/j.foodchem.2010.10.006
Food and Agriculture Organization and World Health Organization (1997) Evaluation of Certain Food Additives and Contaminants: Forty-Sixth Report of the Joint FAO/WHO Expert Committee on Food Additives.
International Agency for Research on Cancer (1993) Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins. In: Monographs on the Evaluation of Carcinogenic Risks to Humans, Vol., 56, IARC Press, Lyon, 245-395.
Kan, C.A. and Meijer, G.A.L. (2007) The Risk of Contamination of Food with Toxic Substances Present in Animal Feed. Animal Feed Science and Technology, 133, 84-108. https://doi.org/10.1016/j.anifeedsci.2006.08.005
Misra, J.B. (2004) A Mathematical Approach to Comprehensive Evaluation of Quality in Groundnut. Journal of Food Composition and Analysis, 17, 69-79. https://doi.org/10.1016/S0889-1575(03)00102-9
Food and Agriculture Organization (2006) Commission Regulation (EC) No. 401/2006 Laying down the Methods of Sampling and Analysis for the Official Control of the Levels of Mycotoxins in Foodstuffs. Official Journal of the European Union L 70, 12-34.
European Union (2010) Commission Regulation (EC) No. 165/2010 Setting Maximum Levels for Certain Contaminants in Foodstuffs. Official Journal of the European Union L50, 8-12.
Lee, K.Y., Poole, C.F. and Zlatkis, A. (1980) Simultaneous Multi-Mycotoxin Determination by High Performance Thin-Layer Chromatography. Analytical Chemistry, 52, 837-842. https://doi.org/10.1021/ac50056a016
Wei, R.W., Yang, X.L., Qiu, F., Yang, M.H. and Qin, J.P. (2011) Safety Issues and New Rapid Detection Methods in Traditional Chinese Medicinal Materials. China Journal of Chinese Materia Medica, 36, 2342-2346.
Soleimany, F., Jinap, S., Faridah, A. and Khatib, A. (2012) A UPLC–MS/MS for Simultaneous Determination of Aflatoxins, Ochratoxin A, Zearalenone, DON, Fumonisins, T-2 Toxin and HT-2 Toxin, in Cereals. Food Control, 25, 647-653. https://doi.org/10.1016/j.foodcont.2011.11.012
Mak, A.C., Osterfeld, S.J., Yu, H., Wang, S.H., Davis, R.J., Jejelowo, O.A. and Pourmand, N. (2010) Sensitive Giant Magnetoresistive-Based Immunoassay for Multiplex Mycotoxin Detection. Biosensors and Bioelectronics, 25, 1635-1639. https://doi.org/10.1016/j.bios.2009.11.028
Soleimany, F., Jinap, S. and Abas, F. (2012) Determination of Mycotoxins in Cereals by Liquid Chromatography Tandem Mass Spectrometry. Food Chemistry, 130, 1055-1060. https://doi.org/10.1016/j.foodchem.2011.07.131
Wang, Y.L., Chai, T.J., Lu, G.Z., Quan, C.S., Duan, H.Y., Yao, M.L., Zucker, B.A. and Schlenker, G. (2008) Simultaneous Detection of Airborne Aflatoxin, Ochratoxin and Zearalenone in a Poultry House by Immunoaffinity Clean-Up and High-Performance Liquid Chromatography. Environmental Research, 107, 139-144. https://doi.org/10.1016/j.envres.2008.01.008
Chan, D., MacDonald, S.J., Boughtflower, V. and Brereton, P. (2004) Simultaneous Determination of Aflatoxins and Ochratoxin A in Food Using a Fully Automated Immunoaffinity Column Clean-Up and Liquid Chromatography-Fluorescence Detection. Journal of Chromatography A, 1059, 13-16. https://doi.org/10.1016/j.chroma.2004.09.096
Trucksess, M., Weaver, C., Oles, C., et al. (2006) Determination of Aflatoxins and Ochratoxin A in Ginseng and Other Botanical Roots by Immunoaffinity Column Cleanup and Liquid Chromatography with Fluorescence Detection. Journal of AOAC international, 89, 624-630. https://doi.org/10.1093/jaoac/89.3.624
Trucksess, M.W., Weaver, C.M., Oles, C.J., Rump, L.V., White, K.D., Betz, J.M. and Rader, J.I. (2008) Determination of Aflatoxins B1, B2, G1, and G2 and Ochratoxin A in Ginseng and Ginger by Multitoxin Immunoaffinity Column Cleanup and Liquid Chromatographic Quantitation: Collaborative Study. Journal of AOAC International, 91, 511-523. https://doi.org/10.1093/jaoac/91.3.511
Tarín, A., Rosell, M.G. and Guardino, X. (2004) Use of High-Performance Liquid Chromatography to Assess Airborne Mycotoxins: Aflatoxins and Ochratoxin A. Journal of Chromatography A, 1047, 235-240. https://doi.org/10.1016/S0021-9673(04)01062-3
Erbs, M., Hartmann, N. and Bucheli, T.D. (2007) Determination of the Cross-Re-activities for α-Zearalenol, β-Zearalenol, Zearalanone, α-Zearalanol, and β-Zearalanol on Three Commercial Immunoaffinity Columns Targeting Zearalenone. Journal of AOAC International, 90, 1197-1202. https://doi.org/10.1093/jaoac/90.4.1197
Ofitserova, M., Nerkar, S., Pickering, M., Torma, L. and Thiex, N. (2009) Multiresidue Mycotoxin Analysis in Corn Grain by Column High-Performance Liquid Chromatography with Postcolumn Photochemical and Chemical Derivatization: Single-Laboratory Validation. Journal of AOAC International, 92, 15-25. https://doi.org/10.1093/jaoac/92.1.15
Wen, J., Kong, W.J., Wang, J. and Yang, M.H. (2013) Simultaneous Determination of Four Aflatoxins and Ochratoxin A in Ginger and Related Products by HPLC with Fluorescence Detection after Immunoaffinity Column Clean-Up and Postcolumn Photochemical Derivatization. Journal of Separation Science, 36, 3709-3716. https://doi.org/10.1002/jssc.201300885
Kong, W.J., Li, J.Y., Qiu, F., Zheng, Y.G., Yang, M.H., et al. (2013) Development of a Sensitive and Reliable High Performance Liquid Chromatography Method with Fluorescence Detection for High-Throughput Analysis of Multi-Class Mycotoxins in Coix Seed. Analytica Chimica Acta, 799, 68-76. https://doi.org/10.1016/j.aca.2013.08.042
Afsah-Hejri, L., Jinap, S., Arzandeh, S. and Mirhosseini, H. (2011) Optimization Of HPLC Conditions for Quantitative Analysis Of Aflatoxins In Contaminated Peanut. Food Control, 22, 381-388. https://doi.org/10.1016/j.foodcont.2010.09.007
Campone, L., Piccinelli, A.L., Celano, R. and Rastrelli, L. (2011) Application of Dispersive Liquid—Liquid Microextraction for the Determination of Aflatoxins B1, B2, G1 and G2 in Cereal Products. Journal of Chromatography A, 1218, 7648-7654. https://doi.org/10.1016/j.chroma.2011.05.028
Khayoon, W.S., Saad, B., Yan, C.B., Salleh, M.I., Salleh, M.I. and Salleh, B. (2010) Determination Of Aflatoxins In Animal Feeds By HPLC With Multifunctional Column Clean-Up. Food Chemistry, 118, 882-886. https://doi.org/10.1016/j.foodchem.2009.05.082
Rahmani, A., Jinap, S. and Soleimany, F. (2009) Qualitative and Quantitative Analysis of Mycotoxins. Comprehensive Reviews in Food Science and Food Safety, 8, 202-251. https://doi.org/10.1111/j.1541-4337.2009.00079.x
European Union (2002) Commission Regulation (EC) No. 657/2002 Implementing Council Directive 96/23/EC concerning the Performance of Analytical Methods and the Interpretation of Results. Official Journal of the European Union L221, 8-36.
Gobel, R. and Lusky, K. (2004) Simultaneous Determination of Aflatoxins, Ochratoxin A and Zearalenone in Grains by New Immunoaffinity Column/Liquid Chromatography. Journal of AOAC International, 87, 411-416. https://doi.org/10.1093/jaoac/87.2.411
Rahmani, A., Jinap, S. and Soleimany, F. (2010) Validation of the Procedure for the Simultaneous determInation of Aflatoxins Ochratoxin A and Zearalenone in Cereals Using HPLC-FLD. Food Additives & Contaminants: Part A, 27, 1683-1693. https://doi.org/10.1080/19440049.2010.514951
Aycicek, H., Aksoy, A. and Saygi, S. (2005) Determination of Aflatoxin Levels in Some Dairy and Food Products Which Consumed in Ankara, Turkey. Food Control, 16, 263-266. https://doi.org/10.1016/j.foodcont.2004.03.004
Kabbashi, E.B.M. and Abass Ali, S.E. (2014) Aflatoxins in Groundnut Paste in Khartoum State, Sudan. US Open Food Science & Technology Journal, 1, 1-8.
Ali, S.E.A., Mohamed, A.A. and Kabbashi, E.E.B.M. (2012) Aflatoxins in Some Foods and Animal Feed in Khartoum State, Sudan. ACT—Biotechnology Research Communications, 22, 113-118.
Ali, S.E.A. and Elbashir, A.A. (2019) Investigation of Groundnut Cake Contamination by Aflatoxins, Ochratoxins and Zearlenone from Three States in Sudan. Journal of Veterinary Science and Animal Welfare, 3, 38-43.
AhmedI, S.E.A.A. and Elbashir, A.A. (2016) Determination of Aflatoxins in Groundnut and Groundnut Products in Sudan Using AflaTest® y HPLC. Memorias del Instituto de Investigaciones en Ciencias de la Salud, 14, 35-39. https://doi.org/10.18004/Mem.iics/1812-9528/2016.014(02)35-039
Kabbashi, E.B.M. and Abass Ali, S.E. (2014) Quantifying Aflatoxins in Grain and Flour from the Five Largest Storage Facilities in Central Sudan Using AfalTest® HPLC Method. US Open Food Science & Technology Journal, 1, 1-8.
Elbashira, A.A. and Ali, S.E.A. (2014) Aflatoxins, Ochratoxins and Zearalenone in Sorghum and Sorghum Products in Sudan. Food Additives & Contaminants: Part B, 7, 135-140. https://doi.org/10.1080/19393210.2013.859741