Abstract
A method development and its validation are described for determining 31 selected Aspergillus, Fusarium, Penicillium, and Claviceps mycotoxins in small grain cereals (wheat, barley, oats). The method comprises an automated solvent extraction step followed by filtering, concentrating, and the analysis of the crude extract with liquid chromatographic-tandem mass spectrometric determination. The analytes included trichothecenes, zearalenone, fumonisins, moniliformin, enniatins, beauvericin, antibiotic Y, aflatoxins, ochratoxin A, mycophenolic acid, penicillic acid, and ergot alkaloids. These were separated in two consecutive chromatographic runs, both involving negative and positive electrospray ionization modes of mass spectrometry. The validation showed that the method performance was good for (semi-)quantitative work, limits of quantification varying between 1 and 1,250 μg kg−1, recoveries mostly between 51% and 122%, and repeatability being from 2% to 26% within day and from 14% to 28% between days (as relative standard deviation). A distinctive suppressive matrix effect, which depended on the analyte and the matrix, was observed but could be compensated for by using matrix-assisted standards. The developed multi-mycotoxin method permits simultaneous, simple, and rapid determination of several co-existing toxins and is ideal for, e.g., screening-type work or at concentration levels relevant in animal welfare.
Similar content being viewed by others
References
Berthiller F, Schuhmacher R, Buttinger G, Krska R (2005) Rapid simultaneous determination of major type A- and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 1062:209 doi:10.1016/j.chroma.2004.11.011
Biselli S, Hummert C (2005) Development of a multicomponent method for Fusarium toxins using LC-MS/MS and its application during a survey for the content of T-2 toxin and deoxynivalenol in various feed and food samples. Food Addit Contam 22:752 doi:10.1080/02652030500158617
Carabias-Martínez R, Rodríguez-Gonzalo E, Revilla-Ruiz P, Hernández-Méndez J (2005) Pressurized liquid extraction in the analysis of food and biological samples. J Chromatogr A 1089:1 doi:10.1016/j.chroma.2005.06.072
Commission Recommendation 2006/576/EC, Brussels, (2006)
Commission Regulation 1881/2006/EC, Brussels, (2006a)
Commission Regulation 401/2006/EC, Brussels, (2006b)
Commission Regulation 1126/2007/EC, Brussels, (2007)
Directive 2002/32/EC, Brussels, (2002)
Gonzalés Osnaya L, Soriano del Castillo JM, Moltó Cortés JC, Mañes Vinuesa J (2006) Extraction and analysis of ochratoxin A in bread using pressurised liquid extraction and liquid chromatography. J Chromatogr A 1113:32 doi:10.1016/j.chroma.2006.01.130
Juan C, González L, Soriano JM, Moltó JC, Mañes J (2005) Accelerated solvent extraction of ochratoxin A from rice samples. J Agric Food Chem 53:9348 doi:10.1021/jf051560q
Klötzel M, Lauber U, Humpf H-U (2006) A new solid phase extraction clean-up method for the determination of 12 type A and B trichothecenes in cereals and cereal-based food by LC-MS/MS. Mol Nutr Food Res 50:261 doi:10.1002/mnfr.200500234
Klug C, Baltes W, Krönert W, Weber R (1988) A method for the determination of ergot alkaloids in food. Z Lebensm Unters Forsch 186:108 doi:10.1007/BF01042702
Kokkonen M, Jestoi M, Rizzo A (2005) Determination of selected mycotoxins in mould cheeses with liquid chromatography coupled to tandem with mass spectrometry. Food Addit Contam 22:449 doi:10.1080/02652030500089861
Krska R, Schubert-Ullrich P, Molinelli A, Sulyok M, MacDonald S, Crews C (2008) Mycotoxin analysis: an update. Food Addit Contam 25:152 doi:10.1080/02652030701765723
Lantazzio VMT, Solfrizzo M, Powers S, Visconti A (2007) Simultaneous determination of aflatoxins, ochratoxin A and Fusarium toxins in maize by liquid chromatography/ tandem mass spectrometry. Rapid Commun Mass Spectrom 21:3253 doi:10.1002/rcm.3210
Lawrence JF, Niedzwiadek B, Scott PM (2000) Effect of temperature and solvent composition on extraction of fumonisins B1 and B2 from corn products. J AOAC Int 83:604
Lehner AF, Craig M, Fannin N, Bush L, Tobin T (2005) Electrospray[+] tandem quadrupole mass spectrometry in the elucidation of ergot alkaloids chromatographed by HPLC: screening of grass or forage samples for novel toxic compounds. J Mass Spectrom 40:1484 doi:10.1002/jms.933
Pallaroni L, von Holst C (2003) Determination of zearalenone from wheat and corn by pressurized liquid extraction and liquid chromatography-electrospray mass spectrometry. J Chromatogr A 993:39 doi:10.1016/S0021-9673(03)00381-9
Pallaroni L, von Holst C (2004) Development of an extraction method for the determination of zearalenone in corn using less organic solvents. J Chromatogr A 1055:247 doi:10.1016/j.chroma.2004.08.147
Razzazi-Fazeli E, Rabus B, Cecon B, Böhm J (2002) Simultaneous quantification of Atrichothecene mycotoxins in grains using liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry. J Chromatogr A 968:129 doi:10.1016/S0021-9673(02)00957-3
Razzazi-Fazeli E, Böhm J, Jarukamjorn K, Zentek J (2003) Simultaneous determination of major Btrichothecenes and the de-epoxy-metabolite of deoxynivalenol in pig urine and maize using high performance liquid chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 796:21 doi:10.1016/S1570-0232(03)00604-4
Ren Y, Zhang Y, Shao S, Cai Z, Feng L, Pan H et al (2007) Simultaneous determination of multicomponent mycotoxin contaminants in foods and feeds by ultra-performance liquid chromatography tandem mass spectrometry. J Chromatogr A 1143:48 doi:10.1016/j.chroma.2006.12.064
Riley RT, Norred WP (1996) Mechanisms of mycotoxicity. In: Howard DH, Miller JD (eds) The mycota, vol. VI. Springer, Berlin, Germany, p 194
Royer D, Humpf H-U, Guy PA (2004) Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry. Food Addit Contam 21:678 doi:10.1080/02652030410001711304
Rundberget T, Wilkins AL (2002) Determination of Penicillium mycotoxins in foods and feeds using liquid chromatography-mass spectrometry. J Chromatogr A 964:189 doi:10.1016/S0021-9673(02)00698-2
Rutschmann J, Stadler PA (1978) Chemical background. In: Berde B, Schild HO (eds) Ergot alkaloids and related compounds. Springer, Berlin, p 29
Spanjer M, Rensen PM, Holten JM (2008) LC-MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs. Food Addit Contam 25:472 doi:10.1080/02652030701552964
Sulyok M, Berthiller F, Krska R, Schuhmacher R (2006) Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize. Rapid Commun Mass Spectrom 20:2649 doi:10.1002/rcm.2640
Sulyok M, Krska R, Schuhmacher R (2007a) Application of a liquid chromatography-tandem mass spectrometric method to multi-mycotoxin determination in raw cereals and evaluation of matrix effects. Food Addit Contam 24:1184 doi:10.1080/02652030701510004
Sulyok M, Krska R, Schuhmacher R (2007b) A liquid chromatography/tandem mass spectrometric multi-mycotoxin method for the quantification of 87 analytes and its application to semiquantitative screening of mouldy food samples. Anal Bioanal Chem 389:1505 doi:10.1007/s00216-007-1542-2
Urraca JL, Marazuela MD, Moreno-Bondi MC (2004) Analysis of zearalenone and α-zearalenol in cereals and swine feed using accelerated solvent extraction and liquid chromatography with fluorescence detection. Anal Chim Acta 524:175 doi:10.1016/j.aca.2004.03.093
van Egmond HP, Schothorst RC, Jonker MA (2007) Regulations relating to mycotoxins in food. Perspectives in a global and European context. Anal Bioanal Chem 389:147 doi:10.1007/s00216-007-1317-9
Van Trijp JMP, Roos AH (1991) Model for the Calculation of Calibration Curves. RIKILT report 91.02., Wageningen
Ware GM, Carman AS, Francis OJ, Kuan SS (1986) Liquid chromatographic determination of ergot alkaloids in wheat. J Assoc Off Anal Chem 69:697
Acknowledgments
The authors wish to thank Laura Ojala and Armi Asola for their excellent technical assistance. Dr. Kristian Fog Nielsen is acknowledged for providing the purified antibiotic Y. This study was partly financed by the Finnish Ministry of Agriculture and Forestry (Dnro 4759/501/2004 and 4775/501/2005).
Author information
Authors and Affiliations
Corresponding author
Additional information
An erratum to this article can be found at http://dx.doi.org/10.1007/s12161-009-9078-z
Rights and permissions
About this article
Cite this article
Kokkonen, M.K., Jestoi, M.N. A Multi-compound LC-MS/MS Method for the Screening of Mycotoxins in Grains. Food Anal. Methods 2, 128–140 (2009). https://doi.org/10.1007/s12161-008-9051-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-008-9051-2