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In Association with
Development of immunoaffinity column and HPLC analysis method for simultaneous detection of aflatoxin B1, B2, G1, G2 and M1
11th World Congress on Biotechnology and Biotech Industries Meet
Serife Seyda Pirincci, Esin Akcael, Tugrul Kaymak and Selma Ozturk
TUBITAK Marmara Research Center, Turkey Kocaeli University, Turkey Ankara Food Control Laboratory Directorate, Turkey
Posters & Accepted Abstracts: J Biotechnol Biomater
Abstract
Aflatoxins are toxic and carcinogenic secondary metabolites produced by Aspergillus species. Due to their effects on health, its limits in food and feed are regulated by authorities. In determination of Aflatoxin (AFL) levels in food and feed, HPLC analysis upon clean-up with immunoaffinity column (IAC) is the mostly used method. IACs are used to concentrate and purify toxin content of the food or feed sample by the use of antibodies. In this study, 8G8 monoclonal antibody that recognizes AFL B1, B2, G1, G2 and M1 with high affinity was developed, produced in cell culture and immobilized on CNBr activated Sepharose resin without purification of antibody for cost effective and labor saving production of IACs. Antibody immobilized column matrix was filled within column and AFL spiked samples were applied to the columns. Aflatoxin content of eluates was analyzed with HPLC. Recovery % and relative standard deviation of developed IACs is calculated as 99.17% and 2.4 % relatively. These results show developed columns can be used for high performance clean-up of AFL B1, B2, G1, G2 and M1 containing food and feed samples. In addition, we developed an HPLC analysis method to discriminate AFL B1, B2, G1, G2 and M1 peaks for simultaneous detection of these aflatoxin types. By the use of IAC and HPLC analysis method developed in this study, easier and quicker analysis of the samples that have the risk of having multiple aflatoxin types can be achieved.Biography
Email: seyda.pirincci@gmail.com
