Resumen
Um método utilizando coluna de imunoafinidade para limpeza e cromatografia em camada delgada, foi otimizado em nosso laboratório para determinar Aflatoxina M1 em baixas concentrações no leite. Para a otimização do método, os parâmetros avaliados foram: recuperação, repetitividade, limite de detecção e limite de quantificação. Baseado em adição de padrão nas amostras, os valores das recuperações variaram de 85,83 e 73,86% em níveis de 0,01–0,5 μg/L, respectivamente, e o desvio padrão relativo para repetitividade variou de 7,73-2,08%. O limite de quantificação estabelecido neste método foi de 0,02 μg/L. Os resultados das amostras analisadas por este método tiveram boa correlação quando comparadas com a Cromatografia Líquida de Alta Eficiência. Adicionalmente, o método AOAC também foi otimizado, resultando em um limite de detecção e quantificação de 0,2 e 0,3 μg/L, respectivamente, indicando baixa sensibilidade desse método. A utilização de coluna de imunoafinidade forneceu excelentes resultados na recuperação, sensibilidade e fácil operacionalidade.
Citas
1. Allcroft, L.et al. Metabolism of aflatoxin in sheep: Excretion of the ‘milk toxin’. Nature, 209: 154-5, 1966.
2. AOAC - Association of Official Analytical Chemists. Official Methods of Analysis of AOAC. 12th edition, Washington DC, 1975, chapter 26.
3. CE - European Commitee - Comission Directive 98/53/EC. Laying down the sampling methods and the methods of analysis for the official control of the levels for certain contaminants in foodstuffs, annex II, JOL 201, 17, july, 1998, p.93-101.
4. Chu, F.S. Immunoassay for analisys of mycotoxins. J. Food Prot.,47:562-9, 1984.
5. Cullen, J.M.; Ruebner, B.H.; Hsieh, D.S.P. Comparative hepatocarcinogenicity of aflatoxins B1 and M1 in the rat. Food Chem. Toxicol., 22:1027-8, 1985.
6. Cullen, J.M. et al. Carcinogenicity of dietary aflatoxin M1 in male Fischer rats compared to aflatoxin B1. Cancer Res., 47:1913-17, 1987.
7. De Iongh, H.; Vles, R.O.; van Pelt, J.G. Milk of mammals fed an aflatoxin-containing diet. Nature, 202:466, 1964.
8. Dragacci, S.; Grosso, F.; Gilbert, J. Immunoaffinity Column Cleanup with Liquid Chromatography for Determination of Aflatoxin M1 in Liquid Milk: Collaborative Study. J. Assoc. Off. Anal. Chem., 84:2, 2001.
9. Hansen, T.J. Affinity Column Cleanup and Direct Fluorescence Measurement of Aflatoxin M1 in Raw Milk. J. Food Protec.,53(1):75-7, 1990.
10. IDF/IUPAC/IAEA (International Dairy Federation/International Union of Pure Applied Chemistry/International Atomic Energy Agency). Determination of the content of aflatoxin M1 in milk by immunoafinity clean-up followed by thin layer chromatography. Validation of an IAC-TLC method for AFM1, may, 2001.(não publicado)
11. Mortimer, D.N.; Gilbert, J.; Shepherd, M.J. Rapid and highly sensitive analysis of aflatoxin M1 in liquid and powdered milks using an affinity column cleaup. J. Chromatogr., 407:393-7, 1987.
12. Navas, S.A. Possível exposição de crianças as aflatoxinas M1 e ocratoxina A através do leite materno, na cidade de São Paulo, 2003. 79p. [Dissertação de Mestrado – Faculdade de Engenharia de Alimentos – Universidade de Campinas.]
13. Pong, R.S.; Wogan, G.N. Toxicity and biochemical and fine structural effects of synthetic aflatoxin M1 and B1 in rat liver. J. Natl. Cancer Inst., 47:585-90, 1971.
14. Scott, P.M. Methods for determination of AFM1 In milk and milk products and milk products – a review of performance characteristics. Food Addit. Contam., 6:283-305, 1989.
15. Scott, P.M. Natural Toxins. In Official Methods of Analysis of Association of Official Analytical Chemists. 16th edition, Maryland, 1997, v. II.
16. Scott, P.M.; Trucksess, M.W. Application of Immunoaffinity Columns to Mycotoxin Analysis. J. Assoc. Off. Anal. Chem., 80(5):941-9, 1997.
17. Shepherd, M.J.; Holmes, M.; Gilbert, J. Comparison and critical evaluation of six published extration and clean-up procedures for Aflatoxin M1 J. Chromatogr., 354:305-15, 1986.
18. Stubblefield, R.D.; Shotwell, O.D.; - Reverse phase analytical and preparative high liquid chromatography aflatoxins. J. Assoc. Off. Anal. Chem., 60:784-90, 1977.
19. Sylos, C.M.; Rodriguez-Amaya, D.B.; Carvalho, P.R.N. Occurrence of aflatoxin M1 in milk and dairy products commercialized in Campinas, Brazil. Food Addit. Contam., 13(2):169-72, 1996.
20. Trucksess, M.W.; Rapid analysis (Thin Layer Chromatographic and Immunochemical Methods) for Mycotoxins in Foods and Feeds. In: De Koe, W.J.; Samson, R.A.; van Egmond, H.P.; Gilbert, J.; Sabino, M. ed. Mycotoxins and Phycotoxins in Perspective at the Turn of the Millenium, Wageningen, The Netherlands, 2001, p.29-40.
21. Tuinstra, L.G.M.; Roos, A.H.; van Trijp, J.M.P. Liquid Chromatographic Determination of Aflatoxin M1 in Milk Powder Using Immunoaffinity Columns for Cleanup: Interlaboratory Study. J. Assoc. Off. Anal. Chem., 76(6):1248-54, 1993.
22. Tuinstra, L.M.G.; Brongeest, J.M. Determination of aflatoxin M1 in the milk at the parts per trillion level. J. Chromatogr.,111:448-51, 1975.
23. Wogan, G.N.; Paglialunga, S. Carcinogenicity of synthetic aflatoxin M1 in rats. Food. Cosmet. Toxicol.,Oxford, 12:38-4, 1974.
![Creative Commons License](http://i.creativecommons.org/l/by/4.0/88x31.png)
Esta obra está bajo una licencia internacional Creative Commons Atribución 4.0.
Derechos de autor 2004 Luzia Shundo, Valter Ruvieri, Sandra A. Navas, Myrna Sabino
Deprecated: Non-static method PKPApplication::getRequest() should not be called statically in /var/www/html/plugins/reports/ReviewersReport/ReviewersReport.inc.php on line 22
Deprecated: Non-static method PKPApplication::getRequest() should not be called statically in /var/www/html/plugins/reports/ReviewersReport/ReviewersReport.inc.php on line 22
Deprecated: Non-static method PKPApplication::getRequest() should not be called statically in /var/www/html/plugins/reports/ReviewersReport/ReviewersReport.inc.php on line 22