Abstract
A sensitive and reliable analytical method based on immunoaffinity chromatography cleanup followed by HPLC separation and fluorimetric detection is described for the quantitative determination of aflatoxin M1 in milk. The chromatographic separation is accomplished by using a C18 column and a gradient elution with methanol, acetonitrile, and water. No extraction solvent process is required and a minimal milk sample cleanup is performed by a direct loading of the immunoaffinity columns and elution with methanol. The method has been successfully validated according to Decision EC No 657/2002 by using the conventional validation approach. The results of the validation process demonstrate the agreement of the method with the provisions of Regulation EC No 401/2006.
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References
Peraica M, Radic B, Lucic A, Pavlovic M (1999) Toxic effects of mycotoxins in human. B World Health Organ 77:754–766
Eaton DL, Groopman JD (1994) The toxicology of aflatoxins. Academic Press, New York
Sweeney MJ, Dobson ADW (1998) Mycotoxin production by Aspergillus, Fusarium and Penicillium species. Int J Food Microbiol 43:141–158
van Egmond HP (1989) Mycotoxins in dairy products. Elsevier Applied Science, London, NY, pp 1–54
International Agency for Research on Cancer (IARC). (2002) IARC Monographs on the Evaluation of Carcinogenic Risks to Humans 56:171–176
Magliulo M, Mirasoli M, Simoni P, Lelli R, Portanti O, Roda A (2005) Development and validation of an ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M1 in milk. J Agric Food Chem 53:3300–3305
Badea M, Micheli L, Messia MC, Cardigliota T, Marconi E, Mottram T, Valesco-Garcia M, Moscone D, Palleschi G (2004) Aflatoxin M1 determination in raw milk using a flow-injection immunoassay system. Anal Chim Acta 520:141–148
Gilbert J, Anklam E (2002) Validation of analytical methods for determining mycotoxins in foodstuffs. TrAC – Trend Anal Chem 21:468–486
Cavaliere C, Foglia P, Pastorini E, Samperi R, Laganà A (2006) Aflatoxin M1 determination in cheese by liquid chromatography-tandem mass spectrometry. J Chromatogr A 1101:69–78
Muscarella M, Lo MS, Palermo C, Centonze D (2007) Validation according to European Commission Decision 2002/657/EC of a confirmatory method for aflatoxin M1 in milk based on immunoaffinity columns and high performance liquid chromatography with fluorescence detection. Anal Chim Acta 594:257–264
European Commission. Regulation (EC) No 401/2006. Off J Eur Union L 70: 12–34
European Commission. Regulation (EC) No 1881/2006. Off J Eur Union L 364:5–24
European Commission. Regulation (EC) No 882/2004. Off J Eur Union L 165:1–141
Acknowledgments
The authors gratefully acknowledge P. D’Antini (Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata) for the technical assistance. This work was supported by Ministero della Salute (Rome, Italy).
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Muscarella, M., Magro, S.L., Palermo, C., Centonze, D. (2011). A Confirmatory Method for Aflatoxin M1 Determination in Milk Based on Immunoaffinity Cleanup and High-Performance Liquid Chromatography with Fluorometric Detection. In: Holst, O. (eds) Microbial Toxins. Methods in Molecular Biology, vol 739. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-102-4_17
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DOI: https://doi.org/10.1007/978-1-61779-102-4_17
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Publisher Name: Humana Press, Totowa, NJ
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